EURL-FV European Union Reference Laboratory Pesticide Residues in Fruits and Vegetables

NUEVA REGLAMENTACION Y GUIAS DE

CONTROL DE CALIDAD EUROPEAS

Amadeo R. Fernández-Alba

Food

Residue Control

Results

Harmonization

World Trade

EU Regulation

Social Perception

Pesticide

Manufacturers

New/Upgrade analytical

Techniques

1.EU REGULATIONS

2.SOCIAL PERCEPTION

3.PESTICIDE MANUFACTURERS

4.WORLD TRADE

5.RESULTS HARMONIZATION

6. NEW/UPGRADE ANAL. TECH.

I. EU PESTICIDE RESIDUE REGULATION

Regulation

Directive

What is a MRL?

Maximum residue level means the upper legal level of a concentration for a pesticide residue in or on food and feed based on

good agricultural practice and the lowest consumer exposure necessary to protect vulnerable consumers.

What is the main principle in setting a MRL? Maximum residue level should be set at the lowest achievable level

consistent with good agricultural practice for each pesticide with a

view to protecting vulnerable groups such as children and the unborn.

The EU approach and legislation on

pesticides – residues

Community Legislation

• Regulation (EC) No 396/2005 and amendments:

– Regulation (EC) No 299/2008 of the European Parliament and of

the Council of 11 March 2008 amending Regulation (EC) No 396/2005 on maximum residue levels of pesticides in or on food and feed of plant and animal origin, as regards the implementing powers conferred on the Commission

– Regulation (EC) No 396/2005 of the European Parliament and the Council of 23 February 2005 on maximum residue levels of pesticides in or on food and feed of plant and animal origin and amending Council Directive 91/414/EEC.

Available at: http://ec.europa.eu/sanco_pesticides/public/index.cfm

Reg. (EC) No. 1097/2009 Reg. (EC) No. 304/2010

Reg. (EC) No. 256/2009

Reg. (EC) No. 1050/2009 Reg. (EC) No. 750/2010

Reg. (EC) No. 765/2010

Reg. (EC) No. 893/2010

Reg. (EC) No. 178/2006

Reg. (EC) No. 149/2008

Reg. (EC) No. 839/2008

Reg. (EC) No. 822/2009 Reg. (EC) No. 459/2010

Not yet published

Not yet published

Reg. (EC) No. 260/2008

List of Commodities

Existing EU MRLS

EU temporary MRLs

Pesticides with

no MRL (low risk)

Pesticides with

default limit other than

0.01mg/Kg

Conversion

Factor of MRLs

Fumigants with

special derogations

Comments

Food Law, EFSA and Food Safety

Official Controls on imports

Coordinated Multiannual Community Control Programme, planned within 3 years and including

Baby Food (2006/125/EC and 2006/141/EC) and Organic farming samples to be analysed

Annex I

Annex II

Annexes III A III B

Annex

IV

Annex V

Annex

VI

Annex

VII

Placing PPP in the market

Reg. (EC) No.

299/2008

Harmonised Legislation

Legislation

Reg. (EC) No.178/2002

Reg. (EC) No.16/2011

Reg. (EC) No.882/2004

Reg. (EC) No.669/2009

Reg. (EC) No.1213/2008

Reg. (EC) No.901/2009 Reg. (EC) No.915/2010

Reg. (EC) No.

396/2005

Directive 91/414/EEC Regulation (EC) No. 1107/2009

Regulation (EC) No. 1141/2010

EFSA

FVO DG SANCO

Mechanisms of Control specially through the legislation in place

- Pesticide Legislation: Reg. 299/2008

- Coordinated Programs: Reg. 915/2010

- RASFF: Reg. 16/2011

- MRL Database

- Risk Assessment: Reg.178/2002 & Reg.16/2011

- Data Collection of EU Monitoring: Reg. 82/2004

- Inspections: Reg. 882/2004 & Reg. 669/2009

530 pesticides

Before Reg. No 396/2005

250 pesticides

National

MRLs None, except for feed/fish

0,01 mg/kg

EU-MRLs

850 pesticides

Default

MRLs Only in 2 EU Member

States

Harmonisation of pesticide MRLs

The EU approach and legislation on

pesticides – residue

The EU approach and legislation on pesticides – residue

Which products are covered by legislation?

• 315 commodities – 10 main groups

• 190 existing entries from past Directives

• 125 new entries • Mainly minor crops (e.g. cassava, thyme, lupins),

but also cocoa, coffee, sugar plants, honey

• Fish and plants used in animal feed listed, but for the time being exempted from EU-MRL setting

DATABASE on Active substances

http://ec.europa.eu/sanco_pesticides/public/index.cfm

The EU approach and legislation on pesticides – residue

EU MRLs

AZOXYSTROBIN MRL (mg/kg) IPRODIONE MRL (mg/kg)

Lettuce 3.0 Lettuce 10.0

Grapes 2.0 Cucumber 2.0

Tomatos 2.0 Apples 10.0

Brocoli 0.05 Oranges 0.02

Spinach 0.05 Asparagus 0.02

Tomatoe Pepper Cucumber

Abamectine 0.01 0.01 0.01

Benomyl 0.50 0.10 0.50

Cyromazin 0.50 2.00 0.10

Dimetomorf 0.50 0.02 0.50

Ethiofencarb 2.00 2.00 2.00

Flufenoxuron 0.50 0.50 0.20

Hexaflumuron 0.05 0.50 0.05

Hexitiazox 0.05 0.50 0.10

Imidacloprid 0.10 0.50 0.10

Lufenuron 0.02 0.02 0.02

Methomyl 1.00 1.00 0.02

Oxamyl 2.00 2.00 2.00

Teflubenzuron 0.50 0.50 0.20

Tiabendazole 0.10 1.00 1.00

Acrinathrin 0.10 0.20 0.02

Bifenthrin 0.20 0.20 0.01

Buprofecin 0.50 0.50 0.20

Chlorpyriphos-me 0.50 0.50 0.05

Endosulfan 1.00 1.00 1.00

Malathion 3.00 3.00 3.00

Methalaxyl 0.20 0.50 0.05

Methamidophos 0.50 0.01 1.00

Pyrimiphos-methyl 0.50 1.00 0.20

Procymidone 2.00 2.00 1.00

Maximum Residue Levels (MRLs) authorized in EU (mg/kg)

600

30 40

400

900

30

Pesticide US Codex EU Brazil Korea Taiwan

Bentazone 0.05 0.1 0.1 0.02 0.05 0.5

Clomazone 0.02 --- 0.01 0.1 0.1 ---

Deltamethrin 1 2 2 1 1 0.05

Glyphosate 0.1 30 0.1 0.2 0.1 0.1

Comparison of the different MRLs in rice established by the Codex

Alimentarius, Brazil, US, Taiwan, Korea and the European Union for the most

used herbicides in rice crops and the principal pesticides found in rice

samples.

Pesticide Annex I

91/414

Registered

(General)

In Turkey

Pyrithiobac Not listed Yes

Oxolinic Acid Not listed Yes

Imazapic Not listed Yes

Fluthiacet-Methyl Not listed Yes

Cyclosulfamuron Not listed Yes

Chloroneb Not listed Yes

Brodifacoum Not listed Yes

Anilofos Not listed Yes

Pyrimidifen Not listed Yes

Triosulfuron Not listed Yes

EPN Not listed Yes

Acephate Out Yes

Agrobacterium radiobacter Out Yes

Alachlor Out Yes

Aldicarb Out Yes

Pesticide Annex I

91/414

Registered

(General)

In Turkey

Amitraz Out Yes

Atrazine Out Yes

Imazapic Out Yes

Azinphos-methyl Out Yes

Benfuracarb Out Yes

Benomyl Out Yes

Beta-cypermethrin Out Yes

Brodifacoum Out Yes

Bromacil Out Yes

Bromopropylate Out Yes

Bronopol Out Yes

Cadusafos Out Yes

Carbofuran Out Yes

Carbosulfan Out Yes

Chlorfenapyr Out Yes

MONITORING

2,4-D Carbofuran Dimethoate Fluazifop Lufenuron Phosmet Tebufenpyrad

4,4′-Methoxychlor Carbosulfan Dimethoate Fludioxonil Malathion Phoxim Tecnazene

Abamectin Chlordane Omethoate Flufenoxuron Maneb group Pyraclostrobin Teflubenzuron

Acephate Chlorfenapyr Dimethomorph Fluquinconazole Mepanipyrim Pirimicarb Tefluthrin

Acetamiprid Chlorfenvinphos Dinocap Flusilazole Mepiquat Pirimiphos-methyl Tetraconazole

Acrinathrin Chlormequat Diphenylamine Flutriafol Metalaxyl Prochloraz Tetradifon

Aldicarb Chlorobenzilate Endosulfan Folpet Metconazole Procymidone Thiabendazole

Amitraz Chlorothalonil Endrin Formetanate Methamidophos Profenofos Thiamethoxam

Amitrole Chlorpropham Epoxiconazole Fosthiazate Methidathion Propamocarb Thiacloprid

Azinphos-ethyl Chlorpyriphos Ethephon Glyphosate Methiocarb Propargite Thiophanate-methyl

Azinphos-methyl Chlorpyriphos-methyl Ethion Haloxyfop Methomyl Propiconazole Tolcloflos-methyl

Azoxystrobin Clofentezin Etofenprox HCB Methoxyfenozide Propyzamide Tolylfluanid

Benfuracarb Clothianidin Ethoprophos Heptachlor Monocrotophos Prothioconazole Triadimefon

Bifenthrin Cyfluthrin Fenamiphos Alpha-HCH Myclobutanil Pyrazophos Triadimenol

Bitertanol Cypermethrin Fenarimol Beta-HCH Oxadixyl Pyrethrins Triazophos

Boscalid Cyproconazole Fenazaquin Gamma-HCH Oxamyl Pyridaben Trichlorfon

Bromide ion Cyprodinil Fenbutatin oxide Hexaconazole Oxydemeton-methyl Pyrimethanil Trifloxystrobin

Bromopropylate DDT Fenbuconazole Hexythiazox Paclobutrazole Pyriproxyfen Triflumuron

Bromuconazole Deltamethrin Fenhexamid Imazalil Parathion Quinoxyfen Trifluralin

Bupirimate Diazinon Fenitrothion Imidacloprid Parathion-Methyl Quintozene Triticonazole

Buprofezin Dichlofluanid Fenoxycarb Indoxacarb Pencycuron Resmethrin Vinclozolin

Cadusafos Dichlorvos Fenpropathrin Iprodione Penconazole Spinosad Zoxamide

Camphechlor Dicloran Fenpropimorph Iprovalicarb Pendimethalin Spiroxamine

Captan Dicofol Fenthion Kresoxim-methyl Permethrin Taufluvalinate

Carbaryl Dieldrin Fenvalerate/Esfenvalerate Lambda-cyhalothrin Phenthoate Tebuconazole

Carbendazim Difenoconazole Fipronil Linuron Phosalone Tebufenozide

Pesticide monitored in Coordinated Plan 2010

177 compounds governed by

Regulation (EC) No 396/2005 amending

Directive 91/414/EEC

Overall frequency of samples with and without measurable residues in the

2008 EU coordinated program (≤MRL: Samples with measurable residues

below or at the MRL; >MRL: Samples with residues above the MRL). Total

number of samples: 11610.

Origin of samples (EU: EU, Iceland and Norway; Imported: third countries); surveillance and enforcement samples of fruit,

vegetables, cereals, processed commodities and baby food

Number of residues found in individual surveillance

samples from the national and EU coordinated pesticide monitoring programmes 2008

II. Social Perception and

Consumer Trends

Changes in the Attitude towards Food

P r o d u c t i o n

C o n s u m e r

Year 1945 1950 1960 1970 1980 1990 2000

Increase

Agric.production

Increase

Product Quality

Increase Product Safety

Protect the Environment

Concern Hunger Appetite

Is there anything to

eat? What is there

to eat?

How Safe is Food

How was the Production

In A

nle

hnung

an F

lacho

wsky

2002

Motives for Buying Organic-Food

47

30

29

21

16

15

14

0 10 20 30 40 50

Absence of chemicals and additives

Reminds me of former times

Prevent Illnesses

Better Taste

Safe the planet for future generations

Friendliness to Animals

Support organic farmers

%

Source: Frankfurter Rundschau 07/10/2004

From Michelangelo Annastassiades (CRL Stuttgart)

III. Plant Protection

Industry

Diazinon

Pirimifos-metil

Endosulfan

Chlorothalonil

Chlorpyrifos

Endosulfan sulfato

Vinclozolina

Procimidona

Cipermetrina

Pesticides (1st generation)

Pesticides (new generation)

Methamidophos Methiocarb

Azoxystrobin

Spinosad

Chemical structures for some well-known and new pesticides

Acephate Lindane

Acrinathrin Malathion

Bromopropylate Mecarbam

Chlorothalonil Methamidophos

Chlorpyriphos Methidathion

Chlorpyriphos-methyl Methiocarb

Chlozolinate Omethoate

Dichlofluanid Parathion-methyl

Dichlorvos Procymidone

Dicofol Pyrazophos

Dimethoate Pyridaphenthion

Endosulfan Pyrimiphos-methyl

Fenpropathrin Tetradifon

Fenthion Trichlorfon

Flufenoxuron Quinalphos

Imidacloprid Vinclozoline

Pesticides covered in 1998

Number of target pesticides (in Almería area)

•Decrease of the number of authorized substances

•Absence sometimes of and effective authorized

substances

•Residue appearance of misussed or not authorized

pesticides

IV. Results Harmonization

European Community

27 countries with quite different

size, population, culture, economy….

Harmonization is in progress….

Social perception = Reality?

Labs within Network (EU: N=264)

8+2 10

13 21 35

35 36

40

Belgium

Burgaria

Greece

France

Spain

Germany

Poland

Italy

6 9

Romania

Hungary

5 5

Portugal

UK

4 4

Denmark

Slovenia

2 2

3 3 3

3 4

4

Lithuania

Ireland

Austria

Estonia

Finland

Netherlands

Slovakia

Czech Rep.

2 2

Cyprus

Sweden

1 1

Malta

Latvia

264 1

Average EU

Luxemburg

(1.1) 0.8

0.9 3

1.3

2.4 1.1

1.5

3.6 1.1

2.1 12.2

1.4 0.5

Labs COUNTRY Population

per Lab (mio)

1.7 2.2

2.8 0.4 1.8

5.4 1.4

2.6

0.4 4.5

0.4 2.3

1.9 0.5

Labs COUNTRY Population

per Lab (mio)

No of EU Samples done by MRM

0

1000

2000

3000

4000

5000

EU

PT

-FV

10

-001

EU

PT

-FV

10

-002

EU

PT

-FV

10

-004

EU

PT

-FV

10

-007

EU

PT

-FV

10

-011

EU

PT

-FV

10

-015

EU

PT

-FV

10

-017

EU

PT

-FV

10

-018

EU

PT

-FV

10

-020

EU

PT

-FV

10

-022

EU

PT

-FV

10

-024

EU

PT

-FV

10

-026

EU

PT

-FV

10

-028

EU

PT

-FV

10

-033

EU

PT

-FV

10

-053

EU

PT

-FV

10

-058

EU

PT

-FV

10

-062

EU

PT

-FV

10

-064

EU

PT

-FV

10

-067

EU

PT

-FV

10

-069

EU

PT

-FV

10

-070

EU

PT

-FV

10

-071

EU

PT

-FV

10

-072

EU

PT

-FV

10

-074

EU

PT

-FV

10

-078

EU

PT

-FV

10

-079

EU

PT

-FV

10

-082

EU

PT

-FV

10

-087

EU

PT

-FV

10

-088

EU

PT

-FV

10

-091

EU

PT

-FV

10

-092

EU

PT

-FV

10

-098

EU

PT

-FV

10

-099

EU

PT

-FV

10

-102

EU

PT

-FV

10

-106

EU

PT

-FV

10

-111

EU

PT

-FV

10

-112

EU

PT

-FV

10

-127

EU

PT

-FV

10

-128

EU

PT

-FV

10

-132

EU

PT

-FV

10

-003

EU

PT

-FV

10

-005

EU

PT

-FV

10

-006

EU

PT

-FV

10

-008

EU

PT

-FV

10

-009

EU

PT

-FV

10

-010

EU

PT

-FV

10

-012

EU

PT

-FV

10

-013

EU

PT

-FV

10

-014

EU

PT

-FV

10

-016

EU

PT

-FV

10

-019

EU

PT

-FV

10

-021

EU

PT

-FV

10

-023

EU

PT

-FV

10

-025

EU

PT

-FV

10

-027

EU

PT

-FV

10

-029

EU

PT

-FV

10

-030

EU

PT

-FV

10

-031

EU

PT

-FV

10

-032

EU

PT

-FV

10

-034

EU

PT

-FV

10

-035

EU

PT

-FV

10

-036

EU

PT

-FV

10

-037

EU

PT

-FV

10

-038

EU

PT

-FV

10

-039

EU

PT

-FV

10

-040

EU

PT

-FV

10

-041

EU

PT

-FV

10

-042

EU

PT

-FV

10

-043

EU

PT

-FV

10

-044

EU

PT

-FV

10

-045

EU

PT

-FV

10

-046

EU

PT

-FV

10

-047

EU

PT

-FV

10

-048

EU

PT

-FV

10

-049

EU

PT

-FV

10

-050

EU

PT

-FV

10

-051

EU

PT

-FV

10

-052

EU

PT

-FV

10

-054

EU

PT

-FV

10

-055

EU

PT

-FV

10

-056

EU

PT

-FV

10

-057

EU

PT

-FV

10

-059

EU

PT

-FV

10

-060

EU

PT

-FV

10

-061

EU

PT

-FV

10

-063

EU

PT

-FV

10

-065

EU

PT

-FV

10

-066

EU

PT

-FV

10

-068

EU

PT

-FV

10

-073

EU

PT

-FV

10

-075

EU

PT

-FV

10

-076

EU

PT

-FV

10

-077

EU

PT

-FV

10

-080

EU

PT

-FV

10

-081

EU

PT

-FV

10

-083

EU

PT

-FV

10

-084

EU

PT

-FV

10

-085

EU

PT

-FV

10

-086

EU

PT

-FV

10

-089

EU

PT

-FV

10

-090

EU

PT

-FV

10

-093

EU

PT

-FV

10

-094

EU

PT

-FV

10

-095

EU

PT

-FV

10

-096

EU

PT

-FV

10

-097

EU

PT

-FV

10

-100

EU

PT

-FV

10

-101

EU

PT

-FV

10

-103

EU

PT

-FV

10

-104

EU

PT

-FV

10

-105

EU

PT

-FV

10

-107

EU

PT

-FV

10

-108

EU

PT

-FV

10

-109

EU

PT

-FV

10

-110

EU

PT

-FV

10

-113

EU

PT

-FV

10

-114

EU

PT

-FV

10

-115

EU

PT

-FV

10

-116

EU

PT

-FV

10

-117

EU

PT

-FV

10

-118

EU

PT

-FV

10

-119

EU

PT

-FV

10

-120

EU

PT

-FV

10

-121

EU

PT

-FV

10

-122

EU

PT

-FV

10

-123

EU

PT

-FV

10

-124

EU

PT

-FV

10

-125

EU

PT

-FV

10

-126

EU

PT

-FV

10

-129

EU

PT

-FV

10

-130

EU

PT

-FV

10

-131

No of EU Samples done by MRM

0

1000

2000

3000

4000

5000

EU

PT

-FV

10

-001

EU

PT

-FV

10

-002

EU

PT

-FV

10

-004

EU

PT

-FV

10

-007

EU

PT

-FV

10

-011

EU

PT

-FV

10

-015

EU

PT

-FV

10

-017

EU

PT

-FV

10

-018

EU

PT

-FV

10

-020

EU

PT

-FV

10

-022

EU

PT

-FV

10

-024

EU

PT

-FV

10

-026

EU

PT

-FV

10

-028

EU

PT

-FV

10

-033

EU

PT

-FV

10

-053

EU

PT

-FV

10

-058

EU

PT

-FV

10

-062

EU

PT

-FV

10

-064

EU

PT

-FV

10

-067

EU

PT

-FV

10

-069

EU

PT

-FV

10

-070

EU

PT

-FV

10

-071

EU

PT

-FV

10

-072

EU

PT

-FV

10

-074

EU

PT

-FV

10

-078

EU

PT

-FV

10

-079

EU

PT

-FV

10

-082

EU

PT

-FV

10

-087

EU

PT

-FV

10

-088

EU

PT

-FV

10

-091

EU

PT

-FV

10

-092

EU

PT

-FV

10

-098

EU

PT

-FV

10

-099

EU

PT

-FV

10

-102

EU

PT

-FV

10

-106

EU

PT

-FV

10

-111

EU

PT

-FV

10

-112

EU

PT

-FV

10

-127

EU

PT

-FV

10

-128

EU

PT

-FV

10

-132

EU

PT

-FV

10

-003

EU

PT

-FV

10

-005

EU

PT

-FV

10

-006

EU

PT

-FV

10

-008

EU

PT

-FV

10

-009

EU

PT

-FV

10

-010

EU

PT

-FV

10

-012

EU

PT

-FV

10

-013

EU

PT

-FV

10

-014

EU

PT

-FV

10

-016

EU

PT

-FV

10

-019

EU

PT

-FV

10

-021

EU

PT

-FV

10

-023

EU

PT

-FV

10

-025

EU

PT

-FV

10

-027

EU

PT

-FV

10

-029

EU

PT

-FV

10

-030

EU

PT

-FV

10

-031

EU

PT

-FV

10

-032

EU

PT

-FV

10

-034

EU

PT

-FV

10

-035

EU

PT

-FV

10

-036

EU

PT

-FV

10

-037

EU

PT

-FV

10

-038

EU

PT

-FV

10

-039

EU

PT

-FV

10

-040

EU

PT

-FV

10

-041

EU

PT

-FV

10

-042

EU

PT

-FV

10

-043

EU

PT

-FV

10

-044

EU

PT

-FV

10

-045

EU

PT

-FV

10

-046

EU

PT

-FV

10

-047

EU

PT

-FV

10

-048

EU

PT

-FV

10

-049

EU

PT

-FV

10

-050

EU

PT

-FV

10

-051

EU

PT

-FV

10

-052

EU

PT

-FV

10

-054

EU

PT

-FV

10

-055

EU

PT

-FV

10

-056

EU

PT

-FV

10

-057

EU

PT

-FV

10

-059

EU

PT

-FV

10

-060

EU

PT

-FV

10

-061

EU

PT

-FV

10

-063

EU

PT

-FV

10

-065

EU

PT

-FV

10

-066

EU

PT

-FV

10

-068

EU

PT

-FV

10

-073

EU

PT

-FV

10

-075

EU

PT

-FV

10

-076

EU

PT

-FV

10

-077

EU

PT

-FV

10

-080

EU

PT

-FV

10

-081

EU

PT

-FV

10

-083

EU

PT

-FV

10

-084

EU

PT

-FV

10

-085

EU

PT

-FV

10

-086

EU

PT

-FV

10

-089

EU

PT

-FV

10

-090

EU

PT

-FV

10

-093

EU

PT

-FV

10

-094

EU

PT

-FV

10

-095

EU

PT

-FV

10

-096

EU

PT

-FV

10

-097

EU

PT

-FV

10

-100

EU

PT

-FV

10

-101

EU

PT

-FV

10

-103

EU

PT

-FV

10

-104

EU

PT

-FV

10

-105

EU

PT

-FV

10

-107

EU

PT

-FV

10

-108

EU

PT

-FV

10

-109

EU

PT

-FV

10

-110

EU

PT

-FV

10

-113

EU

PT

-FV

10

-114

EU

PT

-FV

10

-115

EU

PT

-FV

10

-116

EU

PT

-FV

10

-117

EU

PT

-FV

10

-118

EU

PT

-FV

10

-119

EU

PT

-FV

10

-120

EU

PT

-FV

10

-121

EU

PT

-FV

10

-122

EU

PT

-FV

10

-123

EU

PT

-FV

10

-124

EU

PT

-FV

10

-125

EU

PT

-FV

10

-126

EU

PT

-FV

10

-129

EU

PT

-FV

10

-130

EU

PT

-FV

10

-131

NRL No NRL

20000 25000 11000

per year

TOTAL: 151631

1213

CRL

CRL

CRL

CRL

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

EURL-Central Homepage: www.eurl-pesticides.eu

many laboratories have recently

invested in new higher mass accuracy MS systems that allow them to greatly increase the scope of their

multiresidue methods

Another reason for conducting this screening method proficiency test is to gain information from the

laboratories on the type of software that they use. Whether they are using

commercial software and databases, or whether they are internally constructed and searched manually.

This information will provide an overall view of the purpose of this type of

test, and if, in a near future, there is the need to develop new software.

The EURL-FV aim is to be able to use mass spectrometry based screening

methods routinely, following validation. This is in line with the new Document SANCO/10684/2009

(“Method validation and quality control procedures for pesticide

residues analysis in food and feed”)

General European Commission Proficiency Test Scheme

Data Analysis

Lab 1 Lab 2 Lab 3 Lab 4 Lab 5

CRL-FV

Scientific Group

Reports

Samples and Protocol

INFORMATION

EUPT Web Page Data Analysis

EURL-FV

Scientific Group

Samples and Protocol

INFORMATION

EUPT Web Page

EUPTs performed

Photo description of the test sample

preparation in different EUPT commodities.

A B C D

E F G H

I J K L

A B C D

E F G H

I J K L

12,753 EUPT z-Score Results over

the 10 tests

10 EUPTs z-Score Results

-5,0

-4,0

-3,0

-2,0

-1,0

0,0

1,0

2,0

3,0

4,0

5,0

0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300

EUPT1 EUPT2 EUPT3 EUPT4 EUPT5 EUPT6 EUPT7 EUPT8 EUPT9 EUPT10

25%

FFP RSD

50%

25%

50%

65%

of data

86%

of data

3%

of data

4%

of data

3%

of data

4%

of data

z-Scores

10 EUPTs z-Score Results

-5,0

-4,0

-3,0

-2,0

-1,0

0,0

1,0

2,0

3,0

4,0

5,0

0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300

EUPT1 EUPT2 EUPT3 EUPT4 EUPT5 EUPT6 EUPT7 EUPT8 EUPT9 EUPT10

10 EUPTs z-Score Results

-5,0

-4,0

-3,0

-2,0

-1,0

0,0

1,0

2,0

3,0

4,0

5,0

0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300

EUPT1 EUPT2 EUPT3 EUPT4 EUPT5 EUPT6 EUPT7 EUPT8 EUPT9 EUPT10

25%

FFP RSD

50%

25%

50%

65%

of data

86%

of data

3%

of data

4%

of data

3%

of data

4%

of data

z-Scores

10 EUPTs z-Score Results

-5,0

-4,0

-3,0

-2,0

-1,0

0,0

1,0

2,0

3,0

4,0

5,0

0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300

EUPT1 EUPT2 EUPT3 EUPT4 EUPT5 EUPT6 EUPT7 EUPT8 EUPT9 EUPT10

Qn RSD for 10 European Commission

Proficiency Tests

0

10

20

30

40

50

60

25% FFP - RSD

EUPT - 1 EUPT - 2 EUPT - 3 EUPT - 4 EUPT - 5 EUPT - 6 EUPT - 7 EUPT - 8 EUPT - 9 EUPT - 10

0

10

20

30

40

50

60

25% FFP - RSD

EUPT - 1 EUPT - 2 EUPT - 3 EUPT - 4 EUPT - 5 EUPT - 6 EUPT - 7 EUPT - 8 EUPT - 9 EUPT - 10

V. WORLD TRADE

¿Violative o non Violative?

§§ MRL =

1,0 mg/kg

1,65 mg/kg

Result = 1,65 0,82 mg/kg (k = 2; 95%)

Lab A Lab B

Con expresión de la

incertidumbre (k = 2; 95%)

Lab A Lab B

Sin evaluación de la

incertidumbre

(solo precisión)

Lab A Lab B

0.8

1.2

1.0

1.4

1.6

mg

kg

-1

Valor

mg k

g-1

Expanded uncertainty 50%

Results with Uncertainty (50%)

(k = 2; 95%)

1. 0

1. 8

1. 4

2. 2

2. 4

Results without Uncertainty

MRL = 1.00 mg/kg

Non violative Violative

1.65 mg/kg

2.00 mg/kg

Document N° SANCO/2007/3131

V. NEW ANALYTICAL/INSTRUMENTAL

APPROACHES/CHALLENGES

SOLVENT EXTRACTION METHOD

LC/GC - MS

High Sensitivity (LOQ)

Multiclass compounds

High identification criteria

Centrifugue 5 min. 3700 r.p.m.

Add 15 AcN with 1% of acetic

acid and 200 µL TPP (25 ppm)

Take the supernant to a

vial for analysis

Weigh 15 g of sample

in a 50 ml Falcon tube

Add 6 g MgSO4 anhidrous

+ 2.5 g NaAc·3 H2O and shake vigorously for 4 minutes

Take a 5 ml aliquot from

supernant and add 750 mg MgSO4

anh. + 250 mg PSA

Centrifugue 5 min. 3700 r.p.m.

Centrifugue 5 min. 3700 r.p.m.

Add 10 EtOAc and 200 µL

TPP (25 ppm). Shake 3 sec.

Take the supernant

to a vial for analysis

Weigh 10 g of sample

in a 50 ml Falcon tube

Add 8 g MgSO4 anhidrous

+ 1.5 g NaCl and shake vigorously for 4 minutes

QuEChERS EtOAc method

•20 min ( analysis time) +

10 min (equilibration) = 30 min/analysis

Around 50 analysis/day

EI/PCI - MS/MS

GC-IT-MS (Internal Ionization)

Dic

lorv

os

M

eta

mid

ofo

s/

10 15 20 25 30 35 40 minutos

0

100

200

300

400

500

600

kCounts

Acefa

to

Eto

pro

fos

Dim

eto

ato

Lin

dano

Pro

piz

am

ida

Dia

zin

on

Pir

imeta

nil

Clo

rpirifos

Metil

Vin

clo

zolin

a

Para

tio

n M

etil

Meta

laxil

Pir

imifos M

etil

Metio

carb

C

lorp

irifos

Fen

tio

n

Die

tofe

nca

rb

Tri

ad

imefo

n

Cip

rodin

il P

en

co

nazo

l M

ecarb

am

Quin

alfos

Pro

cim

idona

Tri

adim

enol

Quin

om

etionato

E

nd

osu

lfan I

Hexaco

nazo

l F

en

am

ifo

s B

up

rofe

zin

Mic

lobuta

nil

Bup

irim

ato

En

do

sulfan II

O

xad

ixil

Ofu

race

Tri

azofo

s

Benala

xil En

do

sulfan S

ulfato

Nuari

mo

l D

ico

fol /

Ipro

dio

na

B

rom

opro

pila

to

Bifen

trin

F

en

pro

patrin

T

ebufe

np

irad

Tetr

ad

ifon

Fo

salo

ne

Fen

ari

mol

Pir

azo

fos

Acri

natr

in

Perm

etr

in

Pir

idaben

Cip

erm

etr

inas

Difen

oco

nazo

l D

eltam

etr

in

Spiked pepper extract

(0.50 mg/kg)

PCI

7.00 7.25 7.50 7.75 8.00

0

5

10

15

20

Dic

lorv

os

Me

tam

ido

fos

GC-QqQ-MS/MS

Ion: 126

75 100 125 150 175 200 m/z

0

25

50

75

100

79

94

109

126

141

METAMIDOFOS

min

75 100 125 150 175 200 m/z

0

25

50

75

100

109 127

145

221

DICLORVOS

Ion: 145

Analyst, (2001)

1

2

3

4

5

6

7

8 9

10 11

12 13

14

15

16

17 18

19

20

21

23

24

25

26

1 Dinocap 2 Thiocyclam 3 Thiabendazole, Parathion

4 Methomyl 5 Fenuron

6 Acetamiprid 7 Thiacloprid 8 Imazalil, Aldicarb

9 Monuron 10 Metolcarb

11 Carbofuran 12 Prometryn 13 Diuron, Ethiofencarb

14 Difenoxuron 15 Dimetomorph

16 Methiocarb 17 Terbuthylazine, Spinosyn A 18 Methidation

19 Promecarb 20 Azoxystrobin, Prochloraz, Spinosyn D

21 Diflubenzuron 22 Chlorfenvinphos, Triflumizole 23 Benalaxyl

24 Parathion, Difenoconazole 25 Buprofezin

26 Spiromesifen

22

5 10 15 20 25 30 35 40 45 50 55

Time, min

0,0

1,0e4

2,0e4

3,0e4

4,0e4

5,0e4

6,0e4

7,0e4

8,0e4

9,0e4

1,0e5

1,1e5

1,2e5

1,3e5

Inte

nsity, c

ps

30,00

36,51 5,16

4,80

21,11 5,78

22,69

29,21 16,82 4,12

19,59 39,80 13,12

7,40 35,25

9,94 15,83 26,40

14,62

25,47 9,65 37,03

6,12 49,04

LC-QqQ-MS

CHALLENGES

Matrix Effects In

Pesticide Residue Analysis:

Is The Battle Over?

Problems Technical

Identification

Quantification

LC-QqQ-MS/MS Skimer

After 20 injections

GC-Q-MS Liner after 40 injections

XIC of +MRM (297 pairs): 226.2/170.0 amu from Sample 1 (MRMs 100) of Data MRM pesticides_02.wiff (Turbo Spray) Max. 1.1e6 cps.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Time, min

0.00

5.00e4

1.00e5

1.50e5

2.00e5

2.50e5

3.00e5

3.50e5

4.00e5

4.50e5

5.00e5

5.50e5

6.00e5

6.50e5

7.00e5

7.50e5

8.00e5

8.50e5

9.00e5

9.50e5

1.00e6

1.05e6

Inte

nsity

, cp

s

11.79

Results

4x10

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

5.5

6

6.5

7

+ TIC MRM 50 ppb std chromatogram 0.1%HCOOH

1 1 2 2 3 3 4 4 5 5

Counts vs. Acquisit ion Time (min)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32

4x10

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

5.5

6

6.5

7

+ TIC MRM 50 ppb std chromatogram 0.1%HCOOH

1 1 2 2 3 3 4 4 5 5

Counts vs. Acquisit ion Time (min)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32

0.1% formic

acid 100 ppb

Distribution of retention times take into account the peak width; HPLC application using

0.1% formic acid as modifier

0 0

4

14

56

32

4 4

7

3

1

9

1413

14

22

16

18

33

16

19

21 21

1312

56

3

0 0 00

5

10

15

20

25

30

35

40

45

50

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33

Retention time (min)

Nu

mb

er

of

co

mp

ou

nd

s

I. II. III. IV. V.

23,420,918,114,10 Start time (min) of

time windows

Time windows

Results

Examples for common transitions:

specific

specific

2x10

0

1

2

3

4

+ MRM (223.00000 -> 86.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

1

2

3

4+ MRM (223.00000 -> 148.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

0.5

1

1.5

+ MRM (223.00000 -> 106.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

0.5

1

1.5

+ MRM (223.00000 -> 166.00000) 50 ppb std chromatogram 0.1%HCOOH

Counts vs. Acquisit ion Time (min)

0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14

223.0/86.0

223.0/148.0

223.0/106.0

223.0/166.0

Aldicarb sulfone=Aldoxycarb

2x10

0

1

2

3

4

+ MRM (223.00000 -> 86.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

1

2

3

4+ MRM (223.00000 -> 148.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

0.5

1

1.5

+ MRM (223.00000 -> 106.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

0.5

1

1.5

+ MRM (223.00000 -> 166.00000) 50 ppb std chromatogram 0.1%HCOOH

Counts vs. Acquisit ion Time (min)

0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14

223.0/86.0

223.0/148.0

223.0/106.0

223.0/166.0

2x10

0

1

2

3

4

+ MRM (223.00000 -> 86.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

1

2

3

4+ MRM (223.00000 -> 148.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

0.5

1

1.5

+ MRM (223.00000 -> 106.00000) 50 ppb std chromatogram 0.1%HCOOH

2x10

0

0.5

1

1.5

+ MRM (223.00000 -> 166.00000) 50 ppb std chromatogram 0.1%HCOOH

Counts vs. Acquisit ion Time (min)

0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14

223.0/86.0

223.0/148.0

223.0/106.0

223.0/166.0

Aldicarb sulfone=Aldoxycarb

butoxycarboxin – aldicarb sulfone

scanned – unscanned transitions are common

Counts vs. Acquisit ion Time (min)

3x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

+ MRM (233.00000 -> 72.00000) 50 ppb std chromatogram0.1%HCOOH

2x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

+ MRM (233.00000 -> 160.00000) 50 ppb std chromatogram 0.1%HCOOH

14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5

Counts vs. Acquisit ion Time (min)

3x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

+ MRM (233.00000 -> 72.00000) 50 ppb std chromatogram0.1%HCOOH

2x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

+ MRM (233.00000 -> 160.00000) 50 ppb std chromatogram 0.1%HCOOH

14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5

3x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

+ MRM (233.00000 -> 72.00000) 50 ppb std chromatogram0.1%HCOOH

2x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

+ MRM (233.00000 -> 160.00000) 50 ppb std chromatogram 0.1%HCOOH

14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5

223.0/72.0

223.0/160.0

Counts vs. Acquisit ion Time (min)

3x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

+ MRM (233.00000 -> 72.00000) 50 ppb std chromatogram0.1%HCOOH

2x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

+ MRM (233.00000 -> 160.00000) 50 ppb std chromatogram 0.1%HCOOH

14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5

Counts vs. Acquisit ion Time (min)

3x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

+ MRM (233.00000 -> 72.00000) 50 ppb std chromatogram0.1%HCOOH

2x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

+ MRM (233.00000 -> 160.00000) 50 ppb std chromatogram 0.1%HCOOH

14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5

3x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

+ MRM (233.00000 -> 72.00000) 50 ppb std chromatogram0.1%HCOOH

2x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

+ MRM (233.00000 -> 160.00000) 50 ppb std chromatogram 0.1%HCOOH

14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5

223.0/72.0

223.0/160.0

fluometuron - diuron

both scanned transitions are common

specific

specific

Without dilution

Tomato

Orange

Leek

Tomato

Orange

Leek

Distribution of mass diferences between ions in database and matrix.

84

38

17

34

83

6

39

9

17

7

76

16

74

38

10

03

60

5

37

6

18

2

91

28

57

62

67

2

38

8

21

0

10

1

45

19

28

79

58

5

31

6

16

8

97

66

27

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

0 - 1 Da 0 - 0.1 Da 0 - 0.04 Da 0 - 0.02 Da 0 - 0.01 Da 0 - 0.005 Da 0 - 0.001 Da

Garlic Coliflower Rice Pepper

R=15000=300/0.02 R=7500=300/0.04

Leek Dil10 Standard 100ppb

Ratio 0.6

Triflumuron? 359.2 > 156.0

Triflumuron? 359.2 > 138.9

Ratio 0.7

Triflumuron 359.2 > 156.0

Triflumuron 359.2 > 138.9

6x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

+ESI Scan (9.994 min) Frag=90.0V EUPT_SC_1PPB_PUERRO.d Subtract

* 361.0288* 360.0336

* 359.0409

359.1971

362.0287

358.3313361.1944359.3360 360.2012

Counts vs. Mass-to-Charge (m/z)

357.8 358 358.2 358.4 358.6 358.8 359 359.2 359.4 359.6 359.8 360 360.2 360.4 360.6 360.8 361 361.2 361.4 361.6 361.8 362 362.2

The fragment and the isotopic profile

of the triflumuron doesn’t appear

TRIFLUMURON in LC-TOF-FULL SCAN

EIC 359,0405

6 x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

+ESI EIC(359.0405) Scan Frag=90.0V EUPT_SC_1PPB_PUERRO.d Smooth

Counts vs. Acquisition Time (min) 8.6 8.8 9 9.2 9.4 9.6 9.8 10 10.2 10.4 10.6 10.8 11 11.2 11.4 11.6 11.8 12 12.2

TRIFLUMURON 11,22 359,0405 [M+H]+ C15H11ClF3N2O3

TRIFLUMURON Cl37 11,22 361,0375 [M+H+2]+ C15H1137ClF3N2O3

TRIFLUMURON F1 11,22 156,0211 C7H7ClNO

TRIFLUMURON F1 Cl37 11,22 158,0181 C7H737ClNO

Problems Quantification

Without Clean-up

With Clean-up

Matrix Matched Calibration

Istopically Labeled Internal Standards

QC PROCEDURES FOR

PEST. RES. ANAL.

Guidelines for Residues

Monitoring Control in the

European Union

Document 7826/VI/97

ISO 17025

Practical design of a batch of samples

STANDARDS

SOLUTIONS

A) 100 Standards x 40 €/Standard = 4000 €

B) 200 Standards x 40 €/Standard = 8000 €

C) 600 Standards x 40 €/Standard = 24000 €

1-2 YEARS

QC Double Check

(every day)

3-6 months

Stability?

How to overcome the problems?

I. Easy and very practical solutions II. Specific strategies III. Most powerful MS technologies

HIGH SENSITIVE LC-MS

QuEChERS

Dilution 2x

No dilution

LC-MS/MS

Dilution 10x

Preparation of the samples

Dilution 5x

Dilution 8x

Dilution 15x

Dilution 50x

Dilution 25x

Dilution 100x

XIC of +MRM (114 pairs): 223.000/56.000 amu Expected RT: 9.6 ID: Acetamiprid2 from Sample 59 (1 ppb zumo mix naranja dil5x) of QuECh... Max. 4.9e4 cps.

8.8 9.0 9.2 9.4 9.6 9.8 10.0 10.2 10.4 10.6 10.8 11.0 11.2 11.4Time, min

0.0

2000.0

4000.0

6000.0

8000.0

1.0e4

1.2e4

1.4e4

1.6e4

1.8e4

2.0e4

2.2e4

2.4e4

2.6e4

2.8e4

3.0e4

3.2e4

3.4e4

3.6e4

3.8e4

4.0e4

4.2e4

4.4e4

4.6e4

4.8e4

Inte

ns

ity

, c

ps

9.70

Acetamiprid

XIC of +MRM (114 pairs): 297.800/255.900 amu Expected RT: 9.9 ID: Imazalil2 from Sample 50 (1 ppb zumo mix pera ) of QuEChERS efecto... Max. 9.8e5 cps.

9.1 9.2 9.3 9.4 9.5 9.6 9.7 9.8 9.9 10.0 10.1 10.2 10.3 10.4 10.5 10.6 10.7Time, min

0.0

5.0e4

1.0e5

1.5e5

2.0e5

2.5e5

3.0e5

3.5e5

4.0e5

4.5e5

5.0e5

5.5e5

6.0e5

6.5e5

7.0e5

7.5e5

8.0e5

8.5e5

9.0e5

9.5e5

9.8e5

Inte

ns

ity

, c

ps

9.83

Imazalil

XIC of +MRM (114 pairs): 222.000/123.100 amu Expected RT: 12.1 ID: Carbofuran2 from Sample 8 (1 ppb puerro dil5x) of .wiff (Turbo Spray... Max. 1.8e5 cps.

11.5 11.6 11.7 11.8 11.9 12.0 12.1 12.2 12.3 12.4 12.5 12.6 12.7 12.8 12.9 13.0Time, min

0.0

1.0e4

2.0e4

3.0e4

4.0e4

5.0e4

6.0e4

7.0e4

8.0e4

9.0e4

1.0e5

1.1e5

1.2e5

1.3e5

1.4e5

1.5e5

1.6e5

1.7e5

1.8e5

Inte

ns

ity

, c

ps

12.30

Carbofuran

Standard 1 μg/l

Matrix Dil.5x

Matrix no dilution

x2

x7

x2

Matrix effect

Acetamiprid

0,0

10,0

20,0

30,0

40,0

50,0

60,0

70,0

0 2 4 6 8

Leek

Orange

Tomato

Matrix effect %

Sig

nal S

upre

ssio

n

No Dil Dil15x Dil10x Dil8x Dil5x Dil2x Dil25x Dil50x

Acetamiprid

0,00E+00

2,00E+06

4,00E+06

6,00E+06

8,00E+06

1,00E+07

1,20E+07

1,40E+07

0 5 10 15 20 25

Solvent

Dil 15x

Dil 5x

No Dil

Matrix effect

Leek

m/z 321.00911 (Matrix)

Thiacloprid (m/z 253.03092)

Leek

No Dil Dil15x Dil10x Dil8x Dil5x Dil2x

% S

ignal S

upre

ssio

n

Dil25x Dil50x Dil100x

Matrix effect

Ethiofencarb

0,0

10,0

20,0

30,0

40,0

50,0

60,0

70,0

80,0

90,0

0 2 4 6 8

Leek

Orange

Tomato

No Dil Dil15x Dil10x Dil8x Dil5x Dil2x Dil25x Dil50x

Ethiofencarb

0,00E+00

1,00E+06

2,00E+06

3,00E+06

4,00E+06

5,00E+06

6,00E+06

7,00E+06

8,00E+06

0 5 10 15 20 25

Solvent

Dil 15x

Dil 5x

Sin dil

Matrix effect

Naranja

Orange

35%

48%

0%

17%No matrix effect

Improves before or with Dil.15x

Improves at high dilutions

(50x-100x)

Doesn´t improve with dilution

Tomato

68%

26%

6%0%

No matrix effect

Improves before or with Dil.15x

Improves at high dilutions

(50x-100x)

Doesn´t improve with dilution

Leek

7%

76%

4%13%

No matrix effect

Improves before or with Dil.15x

Improves at high dilutions

(50x-100x)

Doesn´t improve with dilution

Orange

Tomato

Leek

Orange

35%

48%

0%

17%No matrix effect

Improves before or with Dil.15x

Improves at high dilutions

(50x-100x)

Doesn´t improve with dilution

FULL SCAN High resolution

Mass accuracy

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 Time, min

2.0e6

6.0e6

1.0e7

1.4e7

1.8e7

2.2e7

2.6e7

3.0e7

3.4e7

3.8e7

4.2e7

Inte

nsity

, cp

s

16.0 18.0 20.0 22.0 24.0 26.0 28.0

Time, min

24.0

(a) XIC [ m/z 404.0 0.5]

16.0 18.0 20.0 22.0 24.0 26.0 28.0

Time, min

24.0

22.5

(b) XIC [ m/z 404.1 0.1]

16.0 18.0 20.0 22.0 24.0 26.0 Time, min

24.0

28.0

(c) XIC [ m/z 404.12 0.02]

13.0 15.0 17.0 19.0 21.0 Time, min

16.3

(a) XIC [ m/z 223.0 0.5]

13.0 15.0 17.0 19.0 21.0 Time, min

16.3

(b) XIC [ m/z 223.0 0.1]

13.0 15.0 17.0 19.0 21.0 Time, min

16.3

(c) XIC [ m/z 223.07 0.01]

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 Time, min

Inte

nsity

, cp

s

16.0 18.0 20.0 22.0 24.0 26.0 28.0

Time, min

24.0

(a) XIC [ m/z 404.0 0.5]

16.0 18.0 20.0 22.0 24.0 26.0 28.0

Time, min

24.0

22.5

(b) XIC [ m/z 404.1 0.1]

16.0 18.0 20.0 22.0 24.0 26.0 Time, min

24.0

28.0

(c) XIC [ m/z 404.12 0.02]

13.0 15.0 17.0 19.0 21.0 Time, min

16.3

(a) XIC [ m/z 223.0 0.5]

13.0 15.0 17.0 19.0 21.0 Time, min

16.3

(b) XIC [ m/z 223.0 0.1]

13.0 15.0 17.0 19.0 21.0 Time, min

16.3

(c) XIC [ m/z 223.07 0.01]

Azoxystrobin

Thiacloprid

5 ppm

Sulfadimethoxine

1 GHz 4 GHz

C12H15N4O4S+

Exact Mass: 311.0809

H3N N

N

OCH3

OCH3

S

NH2

O O

N

N

OCH3

OCH3

H2N

Exact Mass: 156.0768

S

O O

H2N

Exact Mass: 156.0114

RESOLUTION

DATABASE SEARCH

10

0.15

PESTICIDES in solvent (20-500ppb)

10

0.15

DATABASE SEARCH

PESTICIDES in solvent (20-500ppb)

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time, min

1.0e4

3.0e4

5.0e4

7.0e4

9.0e4

1.1e5

1.3e5

1.5e5

1.7e5

1.9e5

Inte

ns

ity, c

ps

16.2

XICXIC

218.0 219.0 220.0 221.0 222.0 223.0 224.0 225.0 226.0 227.0 228.0 229.0m/z, amu

4000.0

8000.0

1.2e4

1.6e4

2.0e4

2.4e4

2.8e4

3.2e4

3.6e4

4.0e4

4.4e4

4.8e4

5.2e4

5.6e4

6.0e4

6.4e4

6.8e4

Inte

ns

ity, c

ou

nts

223.0745

225.0715

m/zm/z 223.0745223.0745

Chlorine isotopic profileChlorine isotopic profile

[M+H]+

FIGURE 1

N

N

Cl

N

CH3CH3

N

CC1010

HH1111

NN44ClClCC

1010HH

1111NN

44ClCl

ACETAMIPRID

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time, min

1.0e4

3.0e4

5.0e4

7.0e4

9.0e4

1.1e5

1.3e5

1.5e5

1.7e5

1.9e5

Inte

ns

ity, c

ps

16.2

XICXIC

218.0 219.0 220.0 221.0 222.0 223.0 224.0 225.0 226.0 227.0 228.0 229.0m/z, amu

4000.0

8000.0

1.2e4

1.6e4

2.0e4

2.4e4

2.8e4

3.2e4

3.6e4

4.0e4

4.4e4

4.8e4

5.2e4

5.6e4

6.0e4

6.4e4

6.8e4

Inte

ns

ity, c

ou

nts

223.0745

225.0715

m/zm/z 223.0745223.0745

Chlorine isotopic profileChlorine isotopic profile

[M+H]+

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time, min

1.0e4

3.0e4

5.0e4

7.0e4

9.0e4

1.1e5

1.3e5

1.5e5

1.7e5

1.9e5

Inte

ns

ity, c

ps

16.2

XICXIC

218.0 219.0 220.0 221.0 222.0 223.0 224.0 225.0 226.0 227.0 228.0 229.0m/z, amu

4000.0

8000.0

1.2e4

1.6e4

2.0e4

2.4e4

2.8e4

3.2e4

3.6e4

4.0e4

4.4e4

4.8e4

5.2e4

5.6e4

6.0e4

6.4e4

6.8e4

Inte

ns

ity, c

ou

nts

223.0745

225.0715

m/zm/z 223.0745223.0745

Chlorine isotopic profileChlorine isotopic profile

[M+H]+

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time, min

1.0e4

3.0e4

5.0e4

7.0e4

9.0e4

1.1e5

1.3e5

1.5e5

1.7e5

1.9e5

Inte

ns

ity, c

ps

16.2

XICXIC

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time, min

1.0e4

3.0e4

5.0e4

7.0e4

9.0e4

1.1e5

1.3e5

1.5e5

1.7e5

1.9e5

Inte

ns

ity, c

ps

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time, min

1.0e4

3.0e4

5.0e4

7.0e4

9.0e4

1.1e5

1.3e5

1.5e5

1.7e5

1.9e5

Inte

ns

ity, c

ps

16.2

XICXIC

218.0 219.0 220.0 221.0 222.0 223.0 224.0 225.0 226.0 227.0 228.0 229.0m/z, amu

4000.0

8000.0

1.2e4

1.6e4

2.0e4

2.4e4

2.8e4

3.2e4

3.6e4

4.0e4

4.4e4

4.8e4

5.2e4

5.6e4

6.0e4

6.4e4

6.8e4

Inte

ns

ity, c

ou

nts

223.0745

225.0715

m/zm/z 223.0745223.0745

Chlorine isotopic profileChlorine isotopic profile

[M+H]+

FIGURE 1

N

N

Cl

N

CH3CH3

N N

N

Cl

N

CH3CH3

N

CC1010

HH1111

NN44ClClCC

1010HH

1111NN

44ClClCC

1010HH

1111NN

44ClClCC

1010HH

1111NN

44ClCl

ACETAMIPRID

PROBLEMS

sensitivity

quantification

ident. Param.

RETROSPECTIVE ANALYSIS

EASY TO IMPLEMENT

Additional Advantages

Accurate Mass of Diagnostic Frag. Ions Pesticide Family Compound Diagnostic Ion Accurate Mass of

Diagnostic Ion

Phenylureas Diuron Fenuron

Isoproturon Monuron

72.0444

Organophosphates Chlorpyrifos methyl Dimethoate

Malathion

124.9821

Triazines Deethylatrazine Deethylterbuthylazine

Deisopropylatrazine Atrazine

Terbuthylazine

146.0228

146.0228 + 174.0541

Fluorobenzylureas Diflubenzuron Lufenuron Flufenoxuron Teflubenzuron

Hexaflumuron

158.0412

Neonicotinoids Acetamiprid Thiacloprid

126.0105

Conazole Fungicides Bromuconazole Propiconazole

Imazalil

158.9763

S

P

O

O

N O

N

N

N

Cl

NH3H2N N

N

N

Cl

NH3NH

NH3

O

F

F

NCl

Cl

Cl

NON TARGET Searching for UNKNOWN pesticides in food by LC/TOF-MS

(Using the chlorine isotope signature)

8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 16.0 17.0 18.0 19.0 20.0 21.0 22.0 23.0 24.0 25.0 26.0 27.0 28.0 29.0 30.0 Time [min]

0.0

2.0e6

4.0e6

6.0e6

8.0e6

1.0e7

1.2e7

1.4e7

1.6e7

1.8e7

2.0e7

2.2e7

2.4e7

2.6e7

2.8e7

3.0e7

3.2e7

3.4e7

3.6e7

3.8e7

4.0e7

4.2e7

4.4e7

4.6e7

4.8e7

5.0e7

5.2e7

5.4e7

5.6e7

5.8e7

6.0e7

Inte

ns

ity, c

ps

26.73

26.09

25.71 28.11

18.01

22.98 28.42 21.66 23.24 25.04

28.85 20.70 24.13

29.06 19.39 20.26

230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 m/z, amu

0.0

5000.0

1.0e4

1.5e4

2.0e4

2.5e4

3.0e4

3.5e4

4.0e4

4.5e4

5.0e4

5.5e4

6.0e4

6.5e4

7.0e4

7.5e4

8.0e4

8.5e4

9.0e4

9.5e4

1.0e5

1.1e5

1.1e5

1.2e5

1.2e5

1.3e5

1.3e5

1.4e5

1.4e5

1.5e5

1.5e5

1.6e5

Inte

nsity

, co

un

ts

311.0716

313.0687

297.0556

299.0529

312.0741

315.0662

298.0588 301.0510

316.0684

Imazalil

14.0160 + CH2

Methyl substituted Imazalil

DBE 7.5

DBE 7.5 Cl

Cl

O

N

N

-CH3

Imazalil methyl-derivate

An impurity from commercial

formulations containing imazalil

Ion Trap MS/MS

m/z 311 269

m/z 297 255

Applications

Code Matrix Origin Screening by LC-TOF-MS Confirmation by LC-QQQ-MS

6

Pomegranate

India

Below TOF LOD Acefate (3ppb)

Carbendazim (155 ppb) Carbendazime (315ppb)

Difenoconazole (14.7 ppb) Difenoconazole (41ppb)

Below TOF LOD Methiocarbsulfoxide (4ppb)

Below TOF LOD Omethoate * (1.8ppb)

7

Pear

Sudáfrica

Below TOF LOD Metoxifenozide (0.2ppb)

Thiacloprid (13 ppb) Thiacloprid (52ppb)

8 Pear Chile Iprodione (214 ppb) Iprodione (415ppb)

10

Lime

Brasil

Imazalil (9 ppb) Imazalil (44ppb)

Prochloraz (242 ppb) Prochloraz (490ppb)

11 White Grape

Argentina Myclobutanil* (8 ppb) Myclobutanil * (8ppb)

12 Red Plum Sudáfrica Iprodione (81.69 ppb) Iprodione (116ppb)

13 White Plum Sudáfrica FREE OF PESTICIDES FREE OF PESTICIDES

14 Kiwi Italia Fenhexamid (2399 ppb) Fenhexamide (5884 ppb)

15 Bean Marruecos FREE OF PESTICIDES FREE OF PESTICIDES

16 Bean Marruecos FREE OF PESTICIDES FREE OF PESTICIDES

17

Pineapple

Costa Rica

Triadimefon * ( 54 ppb) Triadimefon * (74ppb)

Triadimenol * (120 ppb) Triadimenol * (219ppb)

18 Mango Perú Thiabendazole (188 ppb) Thiabendazole (210ppb)

*Out of Annex I **Pending ***Never notified N.A. Not analyzed Applications

4x10

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

Cpd 1: 4.096: +ESI EIC(192.0553, 193.0603, 383.1085) Scan Frag=190.0V garlic_2.d

1 1

3x10

1

2

3

4

5

6

7

8

Cpd 1: 4.096: +ESI ECC Scan Frag=190.0V garlic_2.d

4.096Cpd 1: 4.096

Counts vs. Acquisition Time (min)

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

INTERFERENCE OF CARBENDAZIM IN GARLIC

3x10

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

5.5

Cpd 1: 4.096: +ESI MFE Spectrum (4.000-4.172 min) Frag=190.0V garlic_2.d

192.0553(M+H)+

383.1085(2M+H)+

Response vs. Mass-to-Charge (m/ z)

100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600 625 650 675 700 725 750 775 800 825 850 875 900 925 950 975 1000

¿?

5x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

+ESI Scan (3.195 min) Frag=190.0V mix160pest_100ppb_29abr.d Subtract

192.0762

160.0502

Counts vs. Mass-to-Charge (m/ z)

135 140 145 150 155 160 165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240

[M+H]+ [C10H9O]+

[C9H10N3O3]+

MASS SPECTRUM OF

CARBENDAZIM

Cpd1: 4.096

383.0885

192.0453

160.0502

192.0762

MASS SPECTRUM OF

THE INTERFERENCE [M+H]+

[2M+H]+

Bupirimate

LC/Q/TOF

TOF MODE

7 x10

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

1.3

1.4

1.5

1.6

+ESI EIC(317.1642) Scan Frag=90.0V Sens1mix160pestFS20ppbPeppers_1.d Noise (PeakToPeak) = 25410.00; SNR (9.166min) = 1851.6

9.166

8.622

8.490 9.314

9.446

Counts vs. Acquisition Time (min)

8.45 8.5 8.55 8.6 8.65 8.7 8.75 8.8 8.85 8.9 8.95 9 9.05 9.1 9.15 9.2 9.25 9.3 9.35 9.4 9.45 9.5 9.55 9.6 9.65 9.7 9.75 9.8 9.85 9.9

6 x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

+ESI Scan (9.166 min) Frag=90.0V Sens1mix160pestFS20ppbPeppers_1.d

* 317.1659

318.1674

309.1312

319.1629

Counts vs. Mass-to-Charge (m/z)

309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329

[M+H]+

Calculated Mass:

317.1642

FULL SCAN SPECTRUM

C13H24N4O3S

Bupirimate

LC/Q/TOF

Q/TOF MODE

5 x10

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

+ESI EIC Product Ion Frag=90.0V (** -> 88.0757, 108.0117, 166.0972, 210.1588) Sens1mix160pestMSMS20ppbPeppers_1.d Smooth Noise (PeakToPeak) = 115.49; SNR (9.176min) = 13910.1

9.176

Counts vs. Acquisition Time (min) 6.5 6.6 6.7 6.8 6.9 7 7.1 7.2 7.3 7.4 7.5 7.6 7.7 7.8 7.9 8 8.1 8.2 8.3 8.4 8.5 8.6 8.7 8.8 8.9 9 9.1 9.2 9.3 9.4 9.5 9.6 9.7 9.8 9.9 10

4 x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

2.6

2.8

3

3.2

+ESI Product Ion (9.190 min) Frag=90.0V CID@20.9 (317.1642[z=1] -> **) Sens1mix160pestMSMS20ppbPeppers_1.d

166.0936

317.1591

210.1557 108.0079

237.2030 272.1021

150.0985

194.1253

Counts vs. Mass-to-Charge (m/z) 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450

[M+H]+

Calculated Mass:

317.1642

EIC

MS/MS SPECTRUM (P.I)

C13H24N4O3S

Target analysis

SENSITIVITY

Non target analysis

SENSITIVITY + SEARCHING CAP. & SOFTWARE