The quality of extravirgin olive oil obtained by an innovative extraction technology

Zinnai A Venturi F Sanmartin C DrsquoAgata M Taglieri I Andrich GDAFE University of Pisa

44th International Conference on Food Processing amp Technology10-12 August London UK

The product what is the The product what is the extravirgin olive oil extravirgin olive oil

The extravirgin olive oil (EVOO) is obtained only by physical processes without using any treatment different from washing settling centrifugation and filtration

This product has to reach a score ge 65 during official sensorial tasting session (in a range between 0 and 9) and be lacking in defects at the same time its free acidity (expressed like oleic acid grams vs 100 g of oil) has to be lower than 08

(Reg CE 15132001 23th July 2001)

Extraction Extraction technologytechnology

Quality of Quality of extracted extracted

oiloil

Why EVOO represents one of the main elements in the Mediterranean diet

Anti-inflammatory effect

Cardiovascular protection

Digestive health benefits

Anti-cancer activity

Anti-inflammatory effect

Cardiovascular protection

Digestive health benefits

Anti-cancer activity

Its daily intake produces several health benefits

Its daily intake produces several health benefits

Only some references (among many papers)Huang CL Sumpio BE ldquoOlive oil the mediterranean diet and

cardiovascular healthrdquo J Am Coll Surg 207 407ndash416 (2008)V Llorente ndash Cortes R Estruch MP Mena E Ros MA Gonzales M Fitograve RM

Lamuela-Raventograves L Badimon ldquoEffect of Mediterranean diet on the expression of pro-atherogenic genes in a population at high cardiovascular

riskrdquo Atherosclerosis August 2009Reduction in the incidence of the type Diabetes 2

With the Mediterranean diet Salas-Salvado et al Diabetes Care 34 14-19 2011

What was the idea What was the idea Knowing that the addition of Knowing that the addition of carbonic snow to grapes let to produce high quality carbonic snow to grapes let to produce high quality

wines hellipwines hellip

Our research group decided to apply Our research group decided to apply this technology to extravirgin olive oil this technology to extravirgin olive oil

extraction extraction

The method The method An innovative EVOO extraction technology (Patent ndeg IT1405173-B) involving the addition of a cryogen (ex solid CO2) to the olives was developed in order to increase the extraction yield and obtain an oil characterized by an higher concentration of its health compounds (ex phenols tocopherols) and a stronger link with its production area

bull Nontoxic and donrsquot leave residuesNontoxic and donrsquot leave residues

bull Not inflammable Not inflammable

bull Good solvent (non-polar)Good solvent (non-polar)

bull Inert by an organoleptic point of Inert by an organoleptic point of viewview

bull Triple point easy to reach (-Triple point easy to reach (-575degC 51atm)575degC 51atm)

bull Easy to find and useEasy to find and use

bull low costs low costs

bull Anti-microbial and anti-oxidant Anti-microbial and anti-oxidant actionaction

bull Enzymes inhibitor (ex lypoxidase)Enzymes inhibitor (ex lypoxidase)

Among all the Among all the possible possible cryogens cryogens

suitable for food suitable for food technology technology

(N(N2L2L He HeLL Ar ArLL etc) etc)

COCO2s2s (or carbonic snow) (or carbonic snow)shows a lot of advantagesshows a lot of advantages

CO2s (-78degC 1

atm)

Olives (T= 12-16degC)

-Q-Q

+Q+Q

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

The product what is the The product what is the extravirgin olive oil extravirgin olive oil

The extravirgin olive oil (EVOO) is obtained only by physical processes without using any treatment different from washing settling centrifugation and filtration

This product has to reach a score ge 65 during official sensorial tasting session (in a range between 0 and 9) and be lacking in defects at the same time its free acidity (expressed like oleic acid grams vs 100 g of oil) has to be lower than 08

(Reg CE 15132001 23th July 2001)

Extraction Extraction technologytechnology

Quality of Quality of extracted extracted

oiloil

Why EVOO represents one of the main elements in the Mediterranean diet

Anti-inflammatory effect

Cardiovascular protection

Digestive health benefits

Anti-cancer activity

Anti-inflammatory effect

Cardiovascular protection

Digestive health benefits

Anti-cancer activity

Its daily intake produces several health benefits

Its daily intake produces several health benefits

Only some references (among many papers)Huang CL Sumpio BE ldquoOlive oil the mediterranean diet and

cardiovascular healthrdquo J Am Coll Surg 207 407ndash416 (2008)V Llorente ndash Cortes R Estruch MP Mena E Ros MA Gonzales M Fitograve RM

Lamuela-Raventograves L Badimon ldquoEffect of Mediterranean diet on the expression of pro-atherogenic genes in a population at high cardiovascular

riskrdquo Atherosclerosis August 2009Reduction in the incidence of the type Diabetes 2

With the Mediterranean diet Salas-Salvado et al Diabetes Care 34 14-19 2011

What was the idea What was the idea Knowing that the addition of Knowing that the addition of carbonic snow to grapes let to produce high quality carbonic snow to grapes let to produce high quality

wines hellipwines hellip

Our research group decided to apply Our research group decided to apply this technology to extravirgin olive oil this technology to extravirgin olive oil

extraction extraction

The method The method An innovative EVOO extraction technology (Patent ndeg IT1405173-B) involving the addition of a cryogen (ex solid CO2) to the olives was developed in order to increase the extraction yield and obtain an oil characterized by an higher concentration of its health compounds (ex phenols tocopherols) and a stronger link with its production area

bull Nontoxic and donrsquot leave residuesNontoxic and donrsquot leave residues

bull Not inflammable Not inflammable

bull Good solvent (non-polar)Good solvent (non-polar)

bull Inert by an organoleptic point of Inert by an organoleptic point of viewview

bull Triple point easy to reach (-Triple point easy to reach (-575degC 51atm)575degC 51atm)

bull Easy to find and useEasy to find and use

bull low costs low costs

bull Anti-microbial and anti-oxidant Anti-microbial and anti-oxidant actionaction

bull Enzymes inhibitor (ex lypoxidase)Enzymes inhibitor (ex lypoxidase)

Among all the Among all the possible possible cryogens cryogens

suitable for food suitable for food technology technology

(N(N2L2L He HeLL Ar ArLL etc) etc)

COCO2s2s (or carbonic snow) (or carbonic snow)shows a lot of advantagesshows a lot of advantages

CO2s (-78degC 1

atm)

Olives (T= 12-16degC)

-Q-Q

+Q+Q

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Why EVOO represents one of the main elements in the Mediterranean diet

Anti-inflammatory effect

Cardiovascular protection

Digestive health benefits

Anti-cancer activity

Anti-inflammatory effect

Cardiovascular protection

Digestive health benefits

Anti-cancer activity

Its daily intake produces several health benefits

Its daily intake produces several health benefits

Only some references (among many papers)Huang CL Sumpio BE ldquoOlive oil the mediterranean diet and

cardiovascular healthrdquo J Am Coll Surg 207 407ndash416 (2008)V Llorente ndash Cortes R Estruch MP Mena E Ros MA Gonzales M Fitograve RM

Lamuela-Raventograves L Badimon ldquoEffect of Mediterranean diet on the expression of pro-atherogenic genes in a population at high cardiovascular

riskrdquo Atherosclerosis August 2009Reduction in the incidence of the type Diabetes 2

With the Mediterranean diet Salas-Salvado et al Diabetes Care 34 14-19 2011

What was the idea What was the idea Knowing that the addition of Knowing that the addition of carbonic snow to grapes let to produce high quality carbonic snow to grapes let to produce high quality

wines hellipwines hellip

Our research group decided to apply Our research group decided to apply this technology to extravirgin olive oil this technology to extravirgin olive oil

extraction extraction

The method The method An innovative EVOO extraction technology (Patent ndeg IT1405173-B) involving the addition of a cryogen (ex solid CO2) to the olives was developed in order to increase the extraction yield and obtain an oil characterized by an higher concentration of its health compounds (ex phenols tocopherols) and a stronger link with its production area

bull Nontoxic and donrsquot leave residuesNontoxic and donrsquot leave residues

bull Not inflammable Not inflammable

bull Good solvent (non-polar)Good solvent (non-polar)

bull Inert by an organoleptic point of Inert by an organoleptic point of viewview

bull Triple point easy to reach (-Triple point easy to reach (-575degC 51atm)575degC 51atm)

bull Easy to find and useEasy to find and use

bull low costs low costs

bull Anti-microbial and anti-oxidant Anti-microbial and anti-oxidant actionaction

bull Enzymes inhibitor (ex lypoxidase)Enzymes inhibitor (ex lypoxidase)

Among all the Among all the possible possible cryogens cryogens

suitable for food suitable for food technology technology

(N(N2L2L He HeLL Ar ArLL etc) etc)

COCO2s2s (or carbonic snow) (or carbonic snow)shows a lot of advantagesshows a lot of advantages

CO2s (-78degC 1

atm)

Olives (T= 12-16degC)

-Q-Q

+Q+Q

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

What was the idea What was the idea Knowing that the addition of Knowing that the addition of carbonic snow to grapes let to produce high quality carbonic snow to grapes let to produce high quality

wines hellipwines hellip

Our research group decided to apply Our research group decided to apply this technology to extravirgin olive oil this technology to extravirgin olive oil

extraction extraction

The method The method An innovative EVOO extraction technology (Patent ndeg IT1405173-B) involving the addition of a cryogen (ex solid CO2) to the olives was developed in order to increase the extraction yield and obtain an oil characterized by an higher concentration of its health compounds (ex phenols tocopherols) and a stronger link with its production area

bull Nontoxic and donrsquot leave residuesNontoxic and donrsquot leave residues

bull Not inflammable Not inflammable

bull Good solvent (non-polar)Good solvent (non-polar)

bull Inert by an organoleptic point of Inert by an organoleptic point of viewview

bull Triple point easy to reach (-Triple point easy to reach (-575degC 51atm)575degC 51atm)

bull Easy to find and useEasy to find and use

bull low costs low costs

bull Anti-microbial and anti-oxidant Anti-microbial and anti-oxidant actionaction

bull Enzymes inhibitor (ex lypoxidase)Enzymes inhibitor (ex lypoxidase)

Among all the Among all the possible possible cryogens cryogens

suitable for food suitable for food technology technology

(N(N2L2L He HeLL Ar ArLL etc) etc)

COCO2s2s (or carbonic snow) (or carbonic snow)shows a lot of advantagesshows a lot of advantages

CO2s (-78degC 1

atm)

Olives (T= 12-16degC)

-Q-Q

+Q+Q

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

The method The method An innovative EVOO extraction technology (Patent ndeg IT1405173-B) involving the addition of a cryogen (ex solid CO2) to the olives was developed in order to increase the extraction yield and obtain an oil characterized by an higher concentration of its health compounds (ex phenols tocopherols) and a stronger link with its production area

bull Nontoxic and donrsquot leave residuesNontoxic and donrsquot leave residues

bull Not inflammable Not inflammable

bull Good solvent (non-polar)Good solvent (non-polar)

bull Inert by an organoleptic point of Inert by an organoleptic point of viewview

bull Triple point easy to reach (-Triple point easy to reach (-575degC 51atm)575degC 51atm)

bull Easy to find and useEasy to find and use

bull low costs low costs

bull Anti-microbial and anti-oxidant Anti-microbial and anti-oxidant actionaction

bull Enzymes inhibitor (ex lypoxidase)Enzymes inhibitor (ex lypoxidase)

Among all the Among all the possible possible cryogens cryogens

suitable for food suitable for food technology technology

(N(N2L2L He HeLL Ar ArLL etc) etc)

COCO2s2s (or carbonic snow) (or carbonic snow)shows a lot of advantagesshows a lot of advantages

CO2s (-78degC 1

atm)

Olives (T= 12-16degC)

-Q-Q

+Q+Q

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

bull Nontoxic and donrsquot leave residuesNontoxic and donrsquot leave residues

bull Not inflammable Not inflammable

bull Good solvent (non-polar)Good solvent (non-polar)

bull Inert by an organoleptic point of Inert by an organoleptic point of viewview

bull Triple point easy to reach (-Triple point easy to reach (-575degC 51atm)575degC 51atm)

bull Easy to find and useEasy to find and use

bull low costs low costs

bull Anti-microbial and anti-oxidant Anti-microbial and anti-oxidant actionaction

bull Enzymes inhibitor (ex lypoxidase)Enzymes inhibitor (ex lypoxidase)

Among all the Among all the possible possible cryogens cryogens

suitable for food suitable for food technology technology

(N(N2L2L He HeLL Ar ArLL etc) etc)

COCO2s2s (or carbonic snow) (or carbonic snow)shows a lot of advantagesshows a lot of advantages

CO2s (-78degC 1

atm)

Olives (T= 12-16degC)

-Q-Q

+Q+Q

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

CO2s (-78degC 1

atm)

Olives (T= 12-16degC)

-Q-Q

+Q+Q

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Diffusion of cellular components in the Diffusion of cellular components in the extracellular liquid phase extracellular liquid phase

CELLULAR

CRASH

WATER FREEZING

ICE VOLUME gt water volume

ADDITION CO2s

Mechanical action

Mechanical action

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

COCO2s2s + Olivesolive paste + Olivesolive paste

Able to prevent Able to prevent the possible the possible oxidation of oxidation of extracted oilextracted oil

Increase of the Increase of the extraction yieldextraction yield

Wished Wished aromatic aromatic profile profile

Higher Higher antioxidant antioxidant

contentcontent

COCO2g2g

Modulation of Modulation of the working the working

variables related variables related toto thethe

MALAXATION MALAXATION

Inert gaseous layer Inert gaseous layer

((COCO22gg airair = 15 = 15))

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

- they were determined the values for the main parameters (no reference are available) to use for the extraction of oil

What was doneWhat was done

- It was verified the influence of cryogen addition - It was verified the influence of cryogen addition (CO(CO2s2s) to the olives during pre-milling phase on ) to the olives during pre-milling phase on the the yield of the oil yield of the oil obtained and on obtained and on oil quality oil quality in in order to evaluate the suitability of the new method order to evaluate the suitability of the new method for EVOO productionfor EVOO production

Our research group

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

OIL

1

2

3

4

7

20-30 kg of olives

5

The micro oil mill used Oliomio Baby reg (TEM)The micro oil mill used Oliomio Baby reg (TEM)

6

Water

CO2(s)

Receiving hopper(1)

Wet olive residues

Knives crusher (2)

Horizontal continuous

malaxer (3)2-phases decanter

(7)

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

Where did the olives come fromWhere did the olives come from They are They are monovarietal and polyvarietal (monovarietal and polyvarietal (mix of some mix of some

varietiesvarieties) olives collected from Tuscany olive-) olives collected from Tuscany olive-yards in two different years yards in two different years

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

The olives utilized in each experimental runs The olives utilized in each experimental runs (( 60 kg) were preliminary mixed to ensure 60 kg) were preliminary mixed to ensure homogeneous feeds Each sample was split homogeneous feeds Each sample was split

in two fractions (30 kg)in two fractions (30 kg)

Traditional extraction

process

Traditional extraction

process

Addiction of CO2s

Addiction of CO2s

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Origin and cultivar of the raw matter Origin and cultivar of the raw matter utilizedutilized

Sample ID Geographical Cultivar

Traditional

Cryo origin

A 1 Tuscany (GR 2010)

Frantoio

B 2 Tuscany (SI 2010)

mix

C 3 Tuscany (SI 2010)

mix

D 4 Tuscany (SI 2010)

mix

E 5 Tuscany (GR 2014)

Leccino

F 6 Tuscany (GR 2014)

Frantoio

G 7 Tuscany (GR 2014)

Moraiolo

Frantoio Leccino Correggiolo Frantoio Leccino Correggiolo

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

CryoCryo TraditionalTraditional

Cryogenolives (ww)Cryogenolives (ww) 01 divide 02 01 divide 02 00

Temperature of olivesTemperature of olives (degC)(degC) ~-2~-2 115115

Temperature of pasteTemperature of paste (degC)(degC) ~240~240 ~240~240

Time of MalaxationTime of Malaxation (s)(s) 24002400 24002400

Time of ExtractionTime of Extraction (s)(s) 49004900 43004300

Water addedolivesWater addedolives ()() 8585 9393

Main operating variables adopted Main operating variables adopted during the experimental runs during the experimental runs

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method

In order to evaluate the effect induced by the addition of the cryogen on the oil extraction yield it was determined the Extractability

Index Variation (EIV) as the ratio between the difference of the oil extracted using CO2s and

that obtained by a traditional extraction process and the amount of product obtained by

traditional method EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100EIV = (EC-ET)ET EIV = (EC-ET)ET 100 100

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Oil extraction yieldOil extraction yieldOil extraction yieldOil extraction yield

Run EC ET

1A 858 825

2B 855 772

3C 912 827

4D 841 748

5E 808 793

6F 736 689

7G 844 774

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

The direct addition of cryogen to the olives during pre-milling phase could induce a

general increase in the oil extraction yield

[(EC-ET)ET]10040 108 103 124 19 68 94

[(EC-ET)ET]10040 108 103 124 19 68 94

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Whatrsquos about the oil extraction yield

Whatrsquos about the oil extraction yield

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

If itrsquos necessary this method allows to harvest olives early in order to reduce greatly the damages due to the third fly attack of Bactrocera oleae that can induce a significant loss in oil production (up to 60) as well as a decrease in oil quality

Water content

inside the fruit

Water content

inside the fruit

CELLULAR CRASH

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Percentage Increase () in tocopherols content (PITC)

60 06

Percentage Increase in Total phenols (PITPh)

51 106

Effects on oil Effects on oil quality of the new quality of the new extraction methodextraction method

p=005p=005

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Oil general chemical parameters

There arenrsquot reliable differences due to the new extraction method

This evidence is probably due to the high efficience of the

extraction plant (even in absence of cryogen addition)

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

RunFree acidity ( oleic acid)

Peroxide value (meq O2Kg)K232 K270 K

A 03 a 626 a 152 a 009 a 000

1 03 a 675 a 145 a 009 a 000

B 06 a 653 a 144 a 010 a 000

2 04 b 482 b 137 a 007 a 000

C 07 a 652 b 127 a 012 a 000

3 07 a 768 a 126 a 010 a 000

D 05 a 680 a 104 b 011 b 000

4 05 a 714 a 178 a 017 a 000

E 06 a 611 a 148 b 011 b 000

5 06 a 521 b 157 a 015 a 000

F 05 a 1067 b 166 a 013 b 000

6 05 a 1127 a 168 a 014 a 000

G 05 a 972 a 160 a 012 a 000

7 05 a 847 b 162 a 011 a 000

Limit value for EVOO le 08 le 20 le 25 le 022 000

Oil general chemical parameters Oil general chemical parameters

In relation to each couple of runs (A1divideG7) for each parameter values not sharing the same letter have significantly different mean concentration

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

CONCLUSIONSCONCLUSIONS

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

bull The general analytical parametersThe general analytical parameters (Reg 256891) donrsquot show reliable differences due to the new extraction process and prove the efficiency of micro oil mill used (Oliomio Babyreg)

bull The concentration of PHENOLS PHENOLS tends to increase but there is a great variability in experimental data (this variability doenrsquot seem to be related to the amount of water remaining in the oil)

bull The concentration of TOCOPHEROLS TOCOPHEROLS increases using the new extraction method

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

CONCLUSIONSCONCLUSIONS

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

bull The CO2s addition seems to increase the extraction yield (this effect is greater when the olives water content is greater)

bull The extraction yield is greater when itrsquos used an higher ration between CO2s added olives squeezed

bull The possibility of using olives early harvested could limit the losses due to Bactrocera oleae (this can be useful in ldquodifficul yearrdquo)

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

CONCLUSIONSCONCLUSIONS

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

bull The new extraction method could allow to obtain an EVOO with an higher concentration in bioactive compounds

EVOO can increases its nutritional properties and its antioxidant activity

and so its SHELFLIFE

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

- to control the changes in chemical and sensory qualities both for oil

extracted with the new system and the traditional one as a function of storage

time

What will be doneWhat will be done

- To set up a ldquomore complexrdquo innovative plant to - To set up a ldquomore complexrdquo innovative plant to verify the possibility to introduce other verify the possibility to introduce other modifications to the extraction process (ex verify if modifications to the extraction process (ex verify if the cryogen addition can be done even (or) during the cryogen addition can be done even (or) during the malaxation) the malaxation)

This project was selected for the

presentation at Expo Milan 24th August

2015 Italian Pavillon

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

ReferencesReferencesReferencesReferences

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Solid carbon dioxide to promote the extraction of extra virgin olive oil

AZinnai F Venturi C Sanmartin F Favati G AndrichGrasas y Aceitas (in press)

Extra-virgin olive oil production with the addition of solid CO2

AZinnai F Venturi C Sanmartin I Taglieri G Andrich

Agro FOOD Industry Hi Tech - vol 26(3) - MayJune 2015

Preliminary results on the influence of carbonic snow addition during the olive processing oil extraction yield and elemental

profile C Zoani G Zappa F Venturi C Sanmartin G Andrich and

A ZinnaiJournal of Nutrition and Food Sciences 4 277 (2014)

doi 1041722155-96001000277

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Thanks to hellipThanks to hellipbull Research groups of Prof Fabio Favati and

Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

bull Research groups of Prof Fabio Favati and Drssa Susanna Bartolini for their useful collaboration in the analytical phase of this research

bull ldquoSara Arrigonirdquo farm - La Spezia (Italy) that took part in the experimental activity

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Research supported by

SEE YOU IN MILAN TO EXPO 2015

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

Total OIL entrance(kg of crushed fruits x of oil inside the olives)

= Total OIL output

(kg of the olive residues x of oil inside the olive residues)

+EXTRACTED OIL

MASS BALANCE during extraction MASS BALANCE during extraction processprocess

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

0

10

20

30

40

50

60

000 005 010 015 020

CO2s olive (kg kg)

Fen

oli

To

tali

(pp

m a

cid

o g

alli

co)

0

003

006

009

012

015

018

Res

a e

stra

ttiv

a

(oli

o e

stra

tto

o

live

lav

ora

te)

(kg

k

g)

fenoli totali

resa estrattiva

a

ab

b

ab

Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno estratto kg olive lavorate) ottenute aggiungendo o meno

diverse quantitagrave di criogeno alle olive processate diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate) ( 0 010 015 020 kg criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive addizionando diverse quantitagrave di criogeno alle olive

(0 010 015 020 kg di criogeno kg olive lavorate) (0 010 015 020 kg di criogeno kg olive lavorate)

lettere diverse sottolineano lrsquoesistenza di differenze statisticamente significative (p = 005)

0

50

100

150

200

250

300

350

0 10 15 20

criogeno ( kg CO2s kg olive )

Vit

amin

a E

(pp

m a

-to

cofe

rolo

)

b a a a

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

We need 08 kg of CO2s to lower 100 kg of olives temperature by 1degC

Cost of 1 kg of CO2s= euro050

Cost to turn down the average temperature of 100

kg of olives by 10degC(15degC 5degC )

08kgdegC x 10degC x 050 eurokg=4euro

Considering an average oil yield of 15

The addition cost for a kg of extravirgin olive oil is

4 euro100 kg olives x 15100kg oilkg olives

lt 03eurokg oil

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Extraction yieldExtraction yield

01630152

01320146

000

005

010

015

020

A1 A2Extr

acti

on

yie

ld(k

g o

ilkg o

lives)

CCSC

The CO2s addition is more useful if the olives have a greater content in water (fruits less ripe )

EC

ET

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Run CRIO Yield TRAD Yield EIV

A11 122 110 109

A12 130 118 102

A2 163 132 235

A3 155 157 -13

A4 160 142 127

A5 152 146 41

B1 158 152 40

C1 120 128 -62

D1 148 145 22

D2 164 150 93

D3 173 162 68

Ea 88 85 35

Eb 92 85 82

Ec 126 85 482

Ripe index increasing

CO2g addedincreasing

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Determination of oil elemental profile sample pre-treatment

High PressureMicrowave Digestion

Milestone 1200 Mega

05 golive oil

Mineralized oil sample

(final V = 25 ml)

2 ml H2O2 + 6 ml HNO3

Different mixtures testedH2O2 30vv (1 divide 2 ml)+ HNO3 699vv (2 divide

6 ml)

Total dissoluti

on

high-purity water

(gt 18 MΩ)

time Power Pressure

1 min 250 WFree

pressure race

1 min 0 W

5 min 400 W

5 min 650 W

Vent = 5 min

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Circulation of the thermal fluid

Circulation of the thermal fluid

Addiction of cryogen

Addiction of cryogen

Thermal probes Thermal probes

Olives Olives

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

Preliminary results about oil extraction Preliminary results about oil extraction yield yield discussion discussion

The number of the experimental runs carried out until now is quite reduced

The number of the experimental runs carried out until now is quite reduced

BUTBUT

The results appear very encouraging and the new method seem very suitable for olive oil extraction

The results appear very encouraging and the new method seem very suitable for olive oil extraction

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

During the next crop season we will increase the number of experimental runs adopting several combinations of the working parameters (ie amount of cryogenamount of olives fruit ripening stage etc)

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

TQ mineralized

olive oil solution

Varian Vista MPX(axial configuration

simultaneous 112 Mpixel CCD detector)

study of spectral interferencesstudy of spectral interferences

preliminary qualitative analysis72 elements

preliminary qualitative analysis72 elements

quantitative analysisquantitative analysis

Gas flows optimization based on signal-to-background ratio (SB)

Auxiliary flow (lmin)

Nebulizer flow(lmin)

10 15 09 10 11 12

SBMg 3296 11720 13948 17998 19921 18412Mn 0128 0197 0245 0312 0303 0294Na 1827 1940 1827 3187 6121 4371

Zn - 213857 nm more sensitive but interfered by Fe

Zn - 206200 nm - less sensitive but without interferences

Analytical wavelengths (nm)

Al - 396152 nm K - 766491 nm Rb - 780026 nm

As - 188980 nm Li - 670783 nm S - 181972 nm

B - 249772 nm Mg - 279553 nm Sb - 206834 nm

Ba - 455403 nm Mn - 257610 nm Si - 251611 nm

Ca - 396847 nm Na - 589592 nm Sr - 407771 nm

Cr - 267716 nm Ni - 231604 nm Ti - 334941 nm

Cu - 327395 nm P - 213618 nm V - 311837 nm

Fe - 238204 nm Pb - 220353 nm Zn ndash 206200 nm

Instrumental conditions

RF Frequency 40 MHz (free running air-cooled)

RF Power 12 kW

Gas (plasma auxiliary nebulizer) Argon

Plasma flow 150 lmin

Auxiliary flow 15 lmin

Nebulizer flow 110 lmin

Replicates 5

Replicate read time 5 s

Operating parameters for ICP-AES quantitative analysis

ICP-AES Analysis

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39

Bruker Aurora M90 (90 degree ion mirror ion optics Collision Reaction Interface)110 diluted

mineralized olive oil solution

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Tuning with a 5 microgl Be Mg Co In Ba Ce Ti Pb and Th solution for

sensitivity and resolution optimization and mass calibration

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Check of the level of oxide ions by the CeO+Ce+ ratio lt 2

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

Monitoring of double charged ions by the signal 137Ba++137Ba+ lt 3

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

CheckCorrection of isobaric interferences examination of more isotopes of the same element(eg Rb85 Rb 87 Sr88Sr87 Nd142 Nd146 Nd144 Se77 Se78 Sn147 Sm148 Sm152 Gd157 Gd154 Er166 Er168 Yb170 Yb172) application of correction equations with respect to isotopes of other elements potentially interfering (eg Nd142 corrected by analyzing Ce140 Nd144 by Sm147 Sm152 by Gd157 Rb87 by Sr88 ) use of the Collision Reaction Interface ndash CRI (for As and Se)

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

Working rangesbull005 divide 5 microgl - As Cd Ce Er Eu Gd La Mn Nd Pb Pr Rb Sb Sc Se Sm Sr Th V Y Ybbull01 divide 10 microgl - Co Cr Cu Mg Ni

External calibration5 standard solutions + blankCurve Fit LinearWeighted Fit NoCorrelation coefficientsR gt 09999

Normal Sensitivity Mode (without CRI) High Sensitivity Mode As and Se ndash Normal mode with CRI

ICP-MS Analysis

• PowerPoint Presentation
• The product what is the extravirgin olive oil
• Why EVOO represents one of the main elements in the Mediterranean diet
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• What was done
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Effects on oil quality of the new extraction method
• Slide 19
• Slide 20
• CONCLUSIONS
• Slide 22
• Slide 23
• What will be done
• Slide 25
• Thanks to hellip
• Slide 27
• Slide 28
• Slide 29
• Fenoli totali ( ppm acido gallico) e rese estrattive (kg olio estratto kg olive lavorate) ottenute aggiungendo o meno diverse quantitagrave di criogeno alle olive processate ( 0 010 015 020 kg criogeno kg olive lavorate)
• Vitamina E (ppm di α-tocoferolo) presente negli oli estratti ottenuti addizionando diverse quantitagrave di criogeno alle olive (0 010 015 020 kg di criogeno kg olive lavorate)
• Slide 32
• Extraction yield
• Slide 34
• Determination of oil elemental profile sample pre-treatment
• Slide 36
• Slide 37
• Slide 38
• Slide 39