Multiresidue screening and quantitation of pesticides...

Routine multiresidue screening and quantitation of pesticidesin waters by LC-MS/MS

RESIDUE ANALYSIS MEETINGTuesday, 28th March 2006 - Bern

Didier ORTELLI – Patrick EDDER – Emmanuelle [email protected]

Service de Protection de la Consommation22 quai Ernest Ansermet, 1211 Genève-4

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Service de Protection de la Consommation

History

Bureau de la salubrité1892

Institut d'hygiène

2006

Laboratoire Cantonal

Service de protectionde la consommation

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Pesticides residue analysis

Food control ~1800 samples/yearFruits and vegetablesCerealsAnimal foodstuff (meat, fish, honey, …)Processed products (oil, wine, babyfood, …)

Water analysis ~100 samples/yearDrinking water (tap water, mineral water, …)Geneva lake surveyRiver pollution

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Pesticides residue analysis

Three major challengesDetectionIdentificationQuantification

Required performancesHigh sensitivityHigh specificityReproducibilityRobustness

http://www.equistaff.com/images/haystack.gif

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Identified or unknown analytes?Identified analytes

Specific analytical methodDedicated sample preparation

selective SPE (ex : cation exchange)Immuno-affinity cartridgeMIP

LC or GC analysisGC-ECD, GC-NPD, GC-MSLC-DAD

Unknown analytesTheoretical approachAnalytical approach

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Theoretical approach

PropertiesSolubilityStabilitySoil adsorptionToxicity…

UsageAgricultural practiceDomestic usageIndustrial productionUrban usage...

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Analytical approach

Mutiresidue methods are the required tools for risk assessment and monitoring of a great number of chemical compounds.

ObjectivesTo develop a multiresidue method for the determination of as much as possible pesticides in waters

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Analytical approach

LC-MS/MS is the method of choice due to its capacity to detect analytes at ng/L range associated with high selectivity

Sample preparationLow selectivity : solid phase extractionHigh concentration factor

LC-MS/MS analysisEfficient chromatographyMRM detection

Mutiresidue screening: 1 precursor and 1 product ionConfirmation : 1 precursor and at least 2 products ions

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LC-MS/MS

Mass filter Q1 Collision cell Q2 Mass filter Q3

Collision gas(Argon)

Detection

10 µL

V

LC System : Acquity UPLC (Waters)Column : Atlantis dC18 3µm 2.1 x 100 mm (Waters)Mobile phase : Gradient with water and methanol containing 0.1 % of formic acidIonisation source : Electrospray (positive and negative ionisation modes) MSMS system : QuattroMicro QqQ (Waters)Acquisition : mode MRMLC : 25min, 2 injections per extract (herbicides and fungicides/insecticides)

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Optimisation of MS/MS conditionsCyproconazole, 10 µg/ml, 10 µl/min, cone 25 V; Collision 30V

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400m/z0

100

%

0

100

%

CYPROCONAZOLE 1 (0.505) Scan ES+ 3.05e860

56

292

1588570141117

214236

294

CYPROCONAZOLE3 1 (0.505) Daughters of 292ES+ 1.23e770

125

139

N

N

N

ClOH

CH3

CyproconazoleMH+292

Daughter scanof 292

MS scan

12570

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Optimisation of MS/MS conditions

More than 250 compounds have been tested

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Chromatography

Column: Atlantis dC18 3µm 2.1 x 100 mm (Waters)Mobile [A] 0.1% of formic acid in waterphase [B] 0.1% of formic acid in MeOHFlow rate: 300 µl/minInjection: 10 µL

% B

Time [min]0 1 4 15 21 250

40

80

100Gradient

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Acquisition MS/MS pour le dépistage

43 MS/MStransitions

simultanées

Cycle time: ~2.5 sec

Acquisition is carried out in 12 acquisition groups1 MS/MS transition per substancesDwell time of 40ms for each MS/MS transitions

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ChromatographyAb

unda

nce

Abun

danc

e

88Herbicides

146Fungicides

InsecticidesAcaricides

Time 0 105 15 20 25 min.

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Sample preparation

Adapted for a large number of substancesFast, simple, robustConcentration factor : 5000x

Sample EvaporationExtraction ReconstitutionElution

MeOH

Elution

100 µLMeOH/H2O

500 ml

40°C

NITR

OG

EN

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Solid phase extraction

CartridgesOasis HLBChromabond C18-ECVarian BondElut C18Varian BondElut C8MN C18-hydra MN HR-PBaker Florisil

MethodEnrichment of 500 ml of water spiked et 20 ng/L of each pesticides. Elution with 4 ml of MeOH. Evaporation under N2. Reconstitution in 100 µl of MeOH/H2O.

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Solid phase extraction

-

50

100

150

200

250

Nom

bre

de p

estic

ides

C8 C18 C18ec C18hydra HR-P Oasis SiOH

nb 0-20%

20-50%

50-70%

>70%

Results for 234 pesticides

Recovery

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Validation results

Validation scheme: 5 QC level [2 - 200 ng/L], 2 repetitions, 5 days88 herbicides + 146 insecticides, acaricides and fungicidesResults

Recovery %(n=2, k=5, j=5)

Herbicides Insecticides fungicides

50 -120 % 65 9120 - 50 % 7 27

< 20 % 16 28

Fidelity : RSD intra-day 8 -18 %; RSD between-days 11 - 38 %Limit of detection : 1 ng/L for 72%, between 5 and 20 ng/L for 21% and between 20 and 100 ng/L for 7% of compounds

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Matrix effects

Effets de matrice lors de la préconcentration (réduction de la capacité) et à la détection par les phénomènes de suppression d'ionisation dues à des substances co-éluantes.

Test des effets de matrice sur des eaux clairesRéseauNappeLacRivière

La dispersion due aux effets de matrice est similaire à la fidélité inter-jour, sauf en cas de forte turbidité de l'échantillon.

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Applications

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Pesticides residue in commercial mineral waters

Somme des résidus de pesticides trouvés dans 30 eaux minérales

0102030405060708090

100

A B C D E F G H I J K L M N O P Q R S T U V W X Y Z

Rés

idus

de

pest

icid

es [

ng/L

]

Echantillons

LMR substance individuelle 100 ng/LLMR pesticides totaux 500 ng/L

13 (43 %) sans pesticides17 (57 %) avec pesticides

Mise en évidence de 19 pesticides différentsMajoritairement des triazines, des phényl-urées et quelques fongicides

Valeur individuelle max : 25 ng/L

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River pollution

Pollution of the river Allondon occuringafter an important rain in july 2005

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Pesticides monitoring in Geneva lake

Commission International pourla Protection des Eaux du Léman

The main role of the CIPEL is to survey the evolution of the water quality of lake Geneva and its tributaries. It issues recommendations to Swiss and French governments, encouraging them to take measures to fight sources of pollution. It contributes towards the coordination of water policy for the lake Geneva basin and promotes public awareness of the importance of water protection.

Basin of Geneva lake~8'000km2

~1'500'000 inhabitants

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Recherche des pesticides dans les eaux du lac Léman

Point SHL2 – 30 m du 10 octobre 2005

Foramsulfuron (H) 81 ng/LMetalaxyl (F) 52 ng/LAtrazine (H) 27 ng/LAmidosulfuron (H) 19 ng/LAtrazine-deséthyl (H) 21 ng/LMetolachlor (H) 13 ng/LAtrazine-deisopropyl (H) 17 ng/LFenarimol (F) 15 ng/LTerbutylazine (H) 11 ng/LSimazine (H) 17 ng/LPropiconazole 11 ng/LPymetrozine 10 ng/LIodosulfuron-methyl (H) 10 ng/L

+ 23 autres substances < 10 ng/L

Somme : 401 ng/L

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Example of chromatograms

Eau du lac – surfaceSeptembre 200571 ng/L

25 ng/L 19 ng/L

16 ng/L

13 ng/L

13 ng/L

21 ng/L

5 ng/L

9 ng/L

6 ng/L

4 ng/L

2.6 ng/L

0.5 ng/L

1.6 ng/L

1.8 ng/L

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Recherche des sources de pollutionSélection de pesticides

0.000

0.020

0.040

0.060

0.080

0M 1M 5M 7.5M 10M 30M 100M 305M FondProfondeur

Con

cent

ratio

n [µ

g/L] Zone d’insertion

du Rhône

Concentration en foramsulfuronmesurée dans le lac = 165 ansd’usage agricole selon les volumes utilisés actuellement, même en estimant que la totalité se retrouve dans le lac

+

Présence d'industries sur le bassin versant produisant ou conditionnant ces substances

+

La preuve par l'analyse des eaux de sortie des STEP industrielles :Utilisation de la même méthodeRésultats semi-quantitatifsIdentification sans équivoque des produits incriminésAvantage de l'utilisation d'une méthode robuste dont la

sélectivité est basée sur la détection !

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And tap water ?Geneva lake water accounts for 80% of the drinking water resources for the Canton of Geneva and for 600'000 inhabitants around the lake.

GenèveBellevue

CoppetFounex

Nyon

Rolle

St-Sulpice Lutry

Vevey

EvianYvoire

PreoxydationAcidification

FloculationSand filtration

OzonationCharcoal filtration

Steps of waterprocessing

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Efficacité de la potabilisation

0.30

0.22

0.10

0.0020.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

Sum

of p

estic

ides

resi

dues

Tota

l con

cent

ratio

n [µ

g/L]

Tapwater

Service Industrielsde Genève

StationLe Prieuré

PreoxydationAcidificationFloculation

Sand filtrationOzonation

Charcoalfiltration

In collaboration with

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Autres stations de potabilisationRégion Vevey

Teneur totale (somme des pesticides)

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

Brute - Vevey Traitée -Vevey/Gonelles

Traitée - Lutry Traitée - St-Sulpice

[µg/

L]

Région EvianTeneur totale (somme des pesticides

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

Brute - Evian Traitée - Evian Traitée - Yvoire

[µg/

L]

Région NyonTeneur totale (somme des pesticides)

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

Brute - Nyon Traitée -Nyon

Traitée -Rolle

Traitée -Coppet

Traitée -Founex

Traitée -Yvoire

[µg/

L]

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Avantages de la méthodologie développée

Vision globale des problèmes de pollutionIncrémentation aisée de la méthode avec de nouvelles substances d'intérêtCapacité de réaction élevéeBonnes performances quantitatives Identification des substances assurée

Possibilité de procéder à des analyses semi-quantitatives dans des matrices plus complexes

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Acknowledgements

Dr Patrick EDDER

Dr Emmanuelle COGNARD

Gaëlle RAVION

Philippe JAN

CIPEL

Dr Stefan RAMSEIER

www.geneve.ch/consommation

www.cipel.org

www.sig-ge.ch

Multiresidue screening and quantitation of pesticides...

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