130 annual meeting & exposition - nacrw · european reference laboratory - fv 130th annual...

European Reference Laboratory - FV

130th Annual Meeting & Exposition


21st September 2016



OfLs OfLs

Main Objectives of the EURLs



EURL

EURL

EURL

EURL

NRL

NRL

NRL

NRL NRL

NRL

NRL

NRL

NRL

NRL

NRL NRL

NRL

NRL

NRL

NRL

NRL

NRL

NRL

NRL

OFL OFL OFL

OFL OFL

OFL

OFL

OFL

OFL

OFL

OFL

OFL

OFL

OFL

OFL

OFL OFL

OFL OFL

OFL

OFL

OFL OFL

OFL

OFL

OFL

OFL

OFL OFL

OFL

OFL

OFL

OFL

OFL

OFL

OFL OFL

OFL

OFL

OFL

OFL

OFL OFL

OFL OFL

OFL OFL

OFL

OFL

EURL

Network



EURLs = 4

NRLs = 31

OfLs = 134



EURLs = 4

NRLs = 31



EURLs = 4

NRLs = 31

OfLs = 134



Colaboration with – NON EU countries (18)



Organization of

Workshops

Specific trainings

and NRL visits

Technical Assistance

to the COM

Administrative

Duties

New method

Development and

Validation

Networking with NRLs

and Official Labs

Collaboration with Third

Countries’ Labs

Evaluation of new

analytical platforms

www.eurl-pesticides.eu

EURLs DUTIES/ACTIVITIES



Proficiency Tests - History

2004 2005 2006 2007 2008 2009



Proficiency Tests - History

2010 2011 2012 2013 2014

ISO/IEC 17043

2015

http://www.eurl-pesticides.eu/docs/public/tmplt_article.asp?CntID=877&LabID=500&Lang=EN

http://www.eurl-pesticides.eu/docs/public/tmplt_article.asp?CntID=877&LabID=500&Lang=EN

http://www.onlineremedies.in/2014/02/coriander-hara-dhaniya-coriander-remedies-coriander-uses-coriander-benefits.html



Acephate Difenoconazole Kresoxim-methyl Pirimicarb

Acetamiprid Diflubenzuron Lambda-Cyhalothrin Pirimphos-methyl

Acrinathrin Dimethoate Linuron Prochloraz

Aldicarb(sum) Diphenilamine Malathion Procydimone

Amitraz (DMPF&DMF) Dithiocarbamates(CS2) Mecarbam Propoxur

Azinphos-methyl Endosulfan Metalaxyl Propyzamide

Azoxystrobin EPN Metamidophos Prothiofos

Bifenthrin Fenamiphos Methidathion Pyraclostrobin

Boscalid Fenamiphos sulfone Methiocarb sum Pyridaben

Bromopropylate Fenamiphos sulfoxide Methiocarb Pyrimethanil

Bupirimate Ethion Methomyl Pyriproxifen

Buprofezin Fenhexamid Methoxyfenozide Quinoxyfen

Cadusafos Fenitrothion Methyl-Tiophanate Quintozene

Captan Fenpropathrin Mevinphos Spinosad

Carbaryl Fludioxinil Monocrotophos Spirodiclofen

Carbendazim Flufenoxuron Myclobutanil Tebuconazole

Carbofuran sum Fluopicolide Omethoate Tetraconazole

Chlorpropham Flutolanil Ortophenilphenol Thiabendazole

Chlorpyrifos Folpet Oxamyl Thiacloprid

Chlorpyrifos-methyl Fosthiazate Oxydemeton-methyl Tolclofos-methyl

Chlothalonil Hexythiazox Parathion Tolyfluanid

Cypermethrin Imazalil Penconazole Triadimenol

Cyprodinil Imidacloprid Pencycuron Triazophos

Delthamethrin Indoxacarb Pendimethalin Trifloxystrobin

Diazinon Iprodione Permethrin Triflumuron

Dichlofluanid Iprovalicarb Phosalone Vinclozolin

Dicofol Isofenphos-methyl Phosmet

Pesticides Used to Treat the Sample in last ten EUPT-FVs

183 compounds



Summary of Results



Evolution of data dispersion along the EUPT-FV

history (Qn RSD, %)

25



Qualifying Results with

Uncertanity Measurements

U′ = 2 ∙ 𝑢



EUPT-17



Revisions of the SANTE – AQC Document

1. Doc. 7826/VI/97 (1st EU AQC Meeting,1997, Portugal)

2. Doc. SANCO/3103/2000 (2nd EU AQC Meeting ,1999, Greece)

3. Doc. SANCO/10476/2003 (3rd EU AQC Meeting, 2003, UK)

4. Doc. SANCO/10232/2006 (4th EU AQC Meeting, 2005, Sweden)

5. Doc. SANCO/2007/3131 (2007 EURL Joint Workshop, Spain)

6. Doc. SANCO/10684/2009 (2009 EURL Joint Workshop, Denmark)

7. Doc. SANCO/12495/2011 (2011 EURL Joint Workshop, Germany)

8. Doc. SANCO/12571/2013 (2013 EURL Joint Workshop, Spain)

9. Doc. SANTE/11945/2015 (2015 EURL Joint Workshop, Germany)

www.eurl-pesticides.eu

NEXT 2017 !



EU Guidelines on Analytical

Quality Control and

Method Validation

Procedures for Pesticide

Residues Analysis in Food and Feed

Document Nº SANTE/11945/2015

2007 2009 2011

2013

2015



EU Guidelines on

Analytical Quality

Control and Method

Validation Procedures

for Pesticide Residues

Analysis in Food and Feed

Document Nº SANTE/11945/2015



Analytical Quality Control and Validation

Procedures for Pesticide Residues

Analysis in Food and Feed

AQC SANTE Guidance Document

Document SANTE/11945/2015

Revised by the AQC SANTE

Advisory Group - 2015

Evaluated by the Advisory Group and

then discussed and agreed by the

EU-National Reference Laboratories



Revisions of EU-SANTE – AQC Document

1. Doc. 7826/VI/97

9. Doc. SANTE/11945/2015 ----2017

• Harmonization with other international guidelines

• Updating criteria to include new technical developments

• Updating criteria to include the real labs experiences

• Improving the practical application of criteria

• Clarifying doubts and misinterpretations from labs

• Keeping the document as simple, practical and flexible as possible

• To support compliance with, and specific implementation of ISO/IEC

17025 (accreditation standard)

Reasons to justify the revisions



Structure of Document SANTE/11945/2015

A.- Introduction and legal background

B.- Sampling, transport, traceability and storage of samples

C.- Sample Analysis

D.- Identification of analytes and confirmation of results

E.- Reporting results

F.- Pesticide standards, stock and calibration solutions

G.- Analytical method validation and performance criteria

H.- Additional recommendations

Annex A and Appendices

Annex A (Commodity groups and representative matrices)

Appendix A (Method validation procedure: outline and example approaches)

Appendix B (Examples of conversion factors)

Appendix C (Examples for the estimation of measurement uncertainty)

Appendix D (Glossary)

Valid

atio

n

Qu

ality

Co

ntro

l



Initial full validation



- High water content

- High acid content (and high water content)

- High sugar content (and low water content)

- High oil content (and very low water content)

- High oil content (and intermediate water content)

- High starch and/or protein content (low water & fat content)

- “Difficult or unique commodities”

VEGETABLES, FRUITS, and CEREALS (7 Groups)

Validation Commodity Groups for Representative Matrices

(Annex A)

Document N° SANTE/11945/2015



CG, LC-High Resolution

Accurate Mass Spectrometry



Linuron

Standard in solvent

Ion ratio: 1.8

Linuron

Real sample of coriander

Ion ratio: 2.4

FALSE NEGATIVE

LC-TQ-MS/MS



EUPT Sample of coriander analyzed by

LC-HRAMS

Linuron

Matrix interfetence



72 hours

NO Target List



Laboratories analising by HRAMS

in the last three EUPTs-FV-SM


53rd Annual North American Chemical Residue Workshop

HRAMs

No HRAMs Homogeneity: 0,085 mg/kg

+25%

-25%

-50%

50%

EU S-Proficiency Test



SENSITIVITY (10 µg/kg)

LINEARITY (2-3 orders)

+ REPRODUCIBILITY

(20%)

LC, GC-HRAMS performance requirements

Quant.



MASS ACCURACY (5 ppm)

RESOLUTION (¿?)

𝟐𝟎𝟎. 𝟎𝟎𝟏 − 𝟐𝟎𝟎. 𝟎𝟎𝟎

𝟐𝟎𝟎. 𝟎𝟎𝟎=

𝟎. 𝟎𝟎𝟏

𝟐𝟎𝟎. 𝟎𝟎𝟎= 𝟓 ∙ 𝟏𝟎−𝟔

Qual.



Ethirimol 0.1 mg/kg in tomato, MS2 spectrum

ppm or mDa?



0

20

40

60

80

0.1 0.01 0.005 0.001 0.0005

495 49 25 5 2 (Da)

(ppm)

Thiabendazole

C10H7N3S (m/z 202.0433)

C0-25H0-50N0-5O0-5Cl0-5Br0-5S0-5

Nu

mb

er

of

po

ssib

le m

ole

cula

r fo

rmu

las

5 ppm

~ 2 000 ~ 20 000 ~ 40 000 ~ 200 000 ~ 400 000 (FWHM) Resolution

Resolution ~ 70 000 FWHM

9 molecular formulas


- Cl, Br

~ 70 000 ~25 000

Absolute mass error relative to Thiabendazole

Resolution ~ 25 000 FWHM



- Cl, Br

RESOLUTION (¿?)



MS detector/ characteristics

Typical

systems (examples)

Acquisition

Requirements for identification

minimum number of ions other

Unit mass resolution

quadrupole,

ion trap, TOF full scan, limited m/z range, SIM 3 ions

S/N ≥ 3e)

Analyte peaks in

the extracted ion

chromatograms

must fully overlap.

Ion ratio within

±30% (relative)

of average

of calibration

standards from same sequence

MS/MS

triple

quadrupole,

ion trap,

Q-trap,

Q-TOF, Q-Orbitrap

selected or multiple reaction

monitoring (SRM, MRM), mass

resolution for precursor-ion

isolation equal to or better than unit mass resolution

2 product ions

Accurate mass

measurement

High resolution

MS: (Q-)TOF

(Q-)Orbitrap

FT-ICR-MS

sector MS

full scan, limited m/z range, SIM,

fragmentation with or without

precursor-ion selection, or combinations thereof

2 ions with

mass accuracy

≤ 5 ppma,b, c)

combined single stage MS and

MS/MS with mass resolution for

precursor-ion isolation equal to or better than unit mass resolution

2 ions:

1 molecular ion,

(de)protonated molecule

or adduct ion with mass acc. ≤ 5 ppma,c

plus

1 MS/MS product iond)

a) preferably including the molecular ion, (de)protonated molecule or adduct ion

b) including at least one fragment ion

c) < 1 mDa for m/z < 200

d) no specific requirement for mass accuracy

e) in case noise is absent, a signal should be present in at least 5 subsequent scans

Conditions for Identification

By using Target List (with retention time)

•Table 4 in Document Nº SANTE/2015/11945 . Identification criteria for different MS techniques



MS mode MS1 MS/MS MS1 and MS/MS

EIC: signal requirement

(absolute)

A criterion to be set by one of the following methods:

(1) a S/N threshold ≥ 3;

(2) an intensity ratio relative to the comparison standard

equal or above a preset threshold

EIC: retention time,

relative to comparison

standard

A criterion to be set by one of the following methods:

(1) ≤ 0.2 min, or

(2) within ±2.5%, not to exceed 0.5 min, or

(3) within an established error range, not to exceed 0.5 min

MS: number of structurally

significant ions Minimum 2 Minimum 2

Minimum 2

combined

MS: mass accuracy ≤ 5 ppm ≤ 10 ppm MS1: ≤ 5 ppm;

MS/MS: ≤ 10 ppm

Table 1. Summarized Requirements for Confirmation of Identity

FDA Acceptance Criteria for Confirmation

of Identity of Chemical Residues using Exact Mass Data



64%

30%

5% 1%

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

0-0.2 mDa 0.21-0.5 mDa 0.51-1 mDa >1 mDa

Errors in full scan MS (n = 7444)

45% 38%

16%

1% 0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

0-0.2 mDa 0.21-0.5 mDa 0.51-1 mDa >1 mDa

Errors in ddMS2 (n = 30623)

Is it necessary 10 ppm

in MS2?



Full scan MS 20-70k

GC-EI-HRAMS

Real Samples: Fluopyram

Orbitrap GC-EI-MS System

50 µg/kg Std in Tomato Real Sample: Pear

C = 26 µg/kg

Ratio1: 50%

Ratio1: 30%

Ratio1: 35%

Ratio1: 52%

Ratio1: 29%

Ratio1: 34%

The ratios are within ± 4% with

respect to the standard and

mass accuracy below 0.2 mDa

Ion 145.0260: 0.14 mDa

Ion 195.0057: -0.08 mDa

Ion 223.0244: 0.02 mDa

Ion 173.0209: 0.01 mDa



20-70k 20-70 k

TARGET ANALYSIS (DATA DEPENDENT ACQUISITION)

Full scan MS MS2

Time Time

20-70k

NON TARGET MS2 (DATA INDEPENDEN ACQUISITION)

20-70 k

LC-ESI-HRAMS Is it necessary ion ratio within 30%?

Is it achievable when MS and MS2

Is selected?



80%

85%

90%

95%

100%garlic

onion

spinach

green bean

lettuce

appleorange

cucumber

parsley

leek

tomato

0.1 mg/kg 0.01 mg/kg

80%82%84%86%88%90%92%94%96%98%

100%garlic

onion

spinach

green bean

lettuce

appleorange

cucumber

parsley

leek

tomato

0.1 mg/kg 0.01 mg/kg

0.1 mg/kg without ion ratio 0.01 mg/kg without ion ratio

a) b)

Percentage of identified compounds with:

a) dd MS2 [one fragment ion with mass error <5 ppm]

b) AIF [one fragment ion with mass error <5 ppm and variation of ion ration < 30%]

Ion ratio + 30% Ion ratio+ 30%



Fragment ion Abundance in solvent Abundance in leek

114.9615 39% 43%

142.9386 100% 100%

170.9699 82% 74%

199.0013 29% 26%

Ethion

Full scan

384.9949 ± 5 ppm

ddMS2

384.9949 ± 0.5 Da

Solvent Leek

10,66 10,66

114.9615

142.9386

113.0423

170.9699

199.0013

142.9384

170.9696

199,0008

Dd MS2

(two fragment ions with mass error < 5 ppm)



C:\Xcalibur\...\Puerro_10ppb_AIF 09/13/16 14:04:11

RT: 9.06 - 10.21

9.1 9.2 9.3 9.4 9.5 9.6 9.7 9.8 9.9 10.0 10.1 10.2

Time (min)

0

20

40

60

80

100

0

20

40

60

80

100

Re

lative

Ab

un

da

nce

9.50

9.51

9.49

9.529.48

9.539.47 9.549.46

9.569.45 9.589.43

NL:

0

m/z=

152.5253-

152.5268 MS

Puerro_10ppb_

AIF

NL:

1.46E6

Base Peak

m/z=

153.1012-

153.1028 MS

Puerro_10ppb_

AIF

Puerro_10ppb_AIF #1808 RT: 9.52 AV: 1 NL: 6.58E7

T: FTMS + p ESI Full ms2 [email protected] [66.7000-1000.0000]

100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200

m/z

0

1

2

3

4

5

6

7

8

Re

lative

Ab

un

da

nce

107.0860

194.9968

147.0298142.9985

177.1123

136.9549119.0858

149.0234

121.1015163.0247

185.0067123.0807

186.9713169.0795 199.1254138.9706 179.0019153.0346

188.9683159.1170

165.0404 173.1326

128.0623

193.9889184.0733

196.0002

Diazinon (305.1083) MS2 spectrum Retention time 9.48 ± 0.1 min Precursor selection Quadrupole window width 1 Da

Diazinon MS2 spectrum No precursor selection Quadrupole window width 934 Da (66 – 1000 Da)

Puerro_10ppb_EC_opt_012min 09/13/16 00:34:23

RT: 9.05 - 10.19

9.1 9.2 9.3 9.4 9.5 9.6 9.7 9.8 9.9 10.0 10.1

Time (min)

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

9.48 9.50

9.47

9.51

9.52

9.529.45

9.53

9.53

9.559.43

9.429.57 9.57

9.41 9.63 9.68

NL:

3.92E6

Base Peak m/z=

305.1068-

305.1098 MS

Puerro_10ppb_

EC_opt_012min

Puerro_10ppb_EC_opt_012min #2085 RT: 9.53 AV: 1 NL: 3.35E5

T: FTMS + p ESI d Full ms2 [email protected] [50.0000-330.0000]

100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200

m/z

0

1

2

3

4

5

6

7

8

Re

lative

Ab

un

da

nce

105.0700

131.0856107.0858112.0344 119.0855

145.1008121.1012

109.0649 135.0801

157.1009143.0854123.0804

149.0958

129.0696 161.0956

124.9821

101.0426163.1118 171.1171138.0107

114.9614 153.1020

169.0791

114.9617

153.1024

169.0795


RT: 9.05 - 10.19

9.1 9.2 9.3 9.4 9.5 9.6 9.7 9.8 9.9 10.0 10.1

Time (min)

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

9.48 9.50

9.47

9.51

9.52

9.529.45

9.53

9.53

9.559.43

9.429.57 9.57

9.41 9.63 9.68

NL:

3.92E6

Base Peak m/z=

305.1068-

305.1098 MS

Puerro_10ppb_

EC_opt_012min


T: FTMS + p ESI d Full ms2 [email protected] [50.0000-330.0000]

153.00 153.02 153.04 153.06 153.08 153.10 153.12 153.14 153.16 153.18 153.20

m/z

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

153.1020

C:\Xcalibur\...\Puerro_10ppb_AIF 09/13/16 14:04:11

RT: 9.06 - 10.21

9.1 9.2 9.3 9.4 9.5 9.6 9.7 9.8 9.9 10.0 10.1 10.2

Time (min)

0

20

40

60

80

100

0

20

40

60

80

100

Re

lative

Ab

un

da

nce

9.50

9.519.4910.14

9.52 10.13 10.1510.029.489.53 10.0710.009.93 10.169.469.22 9.26 9.299.07 9.929.439.17 9.659.309.13 9.769.33 9.949.899.56 9.989.70 9.779.37 9.659.58 9.79 9.829.71

9.66

9.50

9.51

9.49

9.529.48

9.539.47 9.549.46

9.569.45 9.589.43

NL:

3.52E6

m/z=

168.2896-

169.9762 MS

Puerro_10ppb_

AIF

NL:

1.46E6

Base Peak

m/z=

153.1012-

153.1028 MS

Puerro_10ppb_

AIF

Puerro_10ppb_AIF #1808 RT: 9.52 AV: 1 NL: 1.03E6

T: FTMS + p ESI Full ms2 [email protected] [66.7000-1000.0000]

153.00 153.02 153.04 153.06 153.08 153.10 153.12 153.14 153.16 153.18 153.20

m/z

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

153.0346

153.0548

153.1024

153.0733

153.0912

153.1276

153.0701

Not necessary

80

168.90 168.92 168.94 168.96 168.98 169.00 169.02 169.04 169.06 169.08 169.10 169.12 169.14 169.16 169.18 169.20

m/z

0

10

20

30

40

50

60

70

80

90

100

Rela

tive A

bundance

169.0791


RT: 0.00 - 12.11

0 1 2 3 4 5 6 7 8 9 10 11 12

Time (min)

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

NL: 1.43E6

TIC F: FTMS + p ESI d

Full ms2

[email protected]

[50.0000-330.0000] MS

Puerro_10ppb_EC_opt_0

12min


F: FTMS + p ESI d Full ms2 [email protected] [50.0000-330.0000]

114.70 114.75 114.80 114.85 114.90 114.95 115.00 115.05 115.10 115.15 115.20

m/z

0

10

20

30

40

50

60

70

80

90

100

Re

lative

Ab

un

da

nce

114.9614

115.0379

114.9614

Not necessary

Not necessary



CONCLUSIONS



INFORMATION + INTERACTION + HARMONIZATION



INFORMATION + INTERACTION + HARMONIZATION

PT

AQC


130th Annual Meeting & Exposition http://www.eurl-pesticides.eu

130 annual meeting & exposition - nacrw · european reference laboratory - fv 130th annual...

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