Validation of a Simple and Rapid Multiresidue Method (QuEChERS) and its Implementation in Routine Pesticide Analysis
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1 Validation of a Simple and Rapid Multiresidue Method (QuEChERS) and its Implementation in Routine Pesticide Analysis Michelangelo Anastassiades, Ellen Scherbaum and Dorothea Bertsch Chemisches und Veterinäruntersuchungsamt Stuttgart, Schaflandstrasse 3/2, Fellbach Poster presented at the MGPR Symposium (May 2003, Aix en Provence, France) CONTENTS Introduction Mini-Multiresidue-Method (QuEChERS) Validation studies Solvent consumption Conclusions Reference INTRODUCTION Many pesticide multiresidue methods (MRMs) used are complicated, laborious, time consuming, require high amounts of solvents and are therefore expensive. What makes traditional multiresidue methods inefficient? Main Drawbacks Large Sample-Sizes Macro-Approach Too Many, Analytical Steps Complicated Tasks Consequences Wasteful: High Solvent & Material Consumption Time-Consuming and Troublesome Procedures End Result Environmentally Critical Expensive Error-Prone Considering that the time spent for instrumental analysis is also continuously growing due to the introduction of new analytes and instrument techniques, laboratories are not able to analyse the number of samples they would like to. In addition, some important analytes can not be satisfactorily covered by many common MRMs (e.g. basic, acidic and very polar compounds). In order to cover such analytes, laboratories have to additionally perform laborious single analyte methods, which is often not possible. This results in a large grey area of pesticides which are not routinely monitored by most laboratories. In the last decade there has been a general trend to develop faster analytical methods. The automated instrument based extraction procedures SFE and ASE,
2 which were introduced in the mid 1990s to speed up extraction, did not succeed to replace traditional multiresidue approaches. Ideally, a multiresidue method should be fast and easy to perform, require a minimum amount of chemicals, provide a certain degree of selectivity to avoid complicated cleanup procedures and still cover a sufficiently broad spectrum of analytes. Analysts accustomed to performing complex traditional analytical procedures often hesitate to switch to simpler ones assuming that a simpler and faster analytical procedure can not be accurate enough and should, if at all, only be used for screening procedures. In reality, however, the more analytical steps a procedure entails and the more complicated it is the more likely is the introduction of systematic and random errors. MINI-MULTIRESIDUE-METHOD (QUECHERS) During the development of the method the principal aim was to make it as streamlined as possible by avoiding complicated and time consuming analytical steps. Time Consuming, Complicated or Error-Prone Steps Sample Processing/Homogenization Blending (e.g. with Ultra-Turrax) Filtration Multiple Partitioning Steps Separation/Transfers of Entire Extract Use of a Lot of Glassware Evaporation/Reconstitution Classical SPE with Columns & Manifold Simplified Alternatives No Way Around this Shaking Centrifugation Single Partitioning ( On-line -Approach) Take Aliquots (Use ISTD) Extraction/Partitioning in Single Vessel Large Volume Injection; Sensitive Instr. Dispersive SPE The analytical procedure A schematic view of the analytical procedure is shown below. The entire method is published under [1]. Weigh 10 g of Sample (50 ml Teflon-Tube) 1 Add 10 ml Acetonitrile Add ISTD-Solution Shake Vigorously 1 min Add 4 g MgSO 4 and 1 g NaCl Shake Vigorously 1 min Shake 30 s and Centrifuge Take Aliquot and Add MgSO 4 (and Sorbent) Shake 30 s and Centrifuge (Add 0.1% HAc and Analyte Protectants ) MGPR, May 2003, GC-MSD Aix en Provence, and LC-MS France , 6, 7 8 9, Picture No.
3 Procedure
4 Procedure
5 VALIDATION STUDIES Recovery studies: Following the above procedure, recovery studies for more than 250 pesticides have been performed. The recoveries achieved from various matrices lay between 70 and 110 % (usually between 90 and 100 %). RSD s lay mostly between 2-7 %. Some examples: Matrix Cucumber, GC-MSD or LC-MS, n=5 Level 0.1 mg/kg Pesticide Group Rec. %RSD Pesticide Acephate 96.3 % 7.8 Imidacloprid Azoxystrobin Strobilurine 97.9 % 1.7 Lufenuron Carbendazim Benzimidazole 98.1 % 5.7 Methamidophos Cyfluthrin Pyrethroide 96.7 % 3.3 Mevinphos Cypermethrin Pyrethroide 96.7 % 2.7 Parathion Dichlorvos % 5.7 Propamocarb Diclobutrazole Azole % 3.1 Pyriproxyfen Dimethomorph Cinnamic acid der % 3.3 Tebufenozide Fenbutatin oxide Organotin 96.5 % 4.4 Tebufenpyrad Flufenoxuron Benzoyl urea 99.6 % 5.9 Thiabendazole Lindan Organochlorine 97.2 % 3.4 Triflumizole Imazalil Imidazole 91.2 % 4.6 Vinclozolin Group Rec. Neonicotionoide % Benzoyl urea 96.0 % 86.9 % 96.7 % % % % Diacylhydrazine 97.1 % Pyrazole 95.6 % Benzimidazole 96.3 % Imidazole 95.8 % Dicarboximide 97.5 % %RSD A great advantage of this method is the ability to recover basic and acidic pesticides even at ph values in which they are expected to exist predominantly in ionised form. This is probably related to the relative high water content of the organic phase after the partitioning step. Basic Pesticides: Matrix Apple, LC-MS, n=5 Level 0.1 mg/kg P ropam ocarb Im a z a lil Prochloraz T h ia b e n d a z o le C arbendazim Fenpropim orph S p iro x a m in e pka of corresp. acid 9.5 pka of corresp. acid 6.5 pka of corresp. acid 3.8 pka of corresp. acid 4.7 pka of corresp. acid 4.2 pka of corresp. acid 6.9 pka of corresp. acid Recovery % Note: The pka indicates the ph value above which the acids lay predominantly in ionised form.
6 Acidic Pesticides: Matrix Apple, LC-MS/MS, n=2 Level 0.1 mg/kg 120 Recovery % Im azapyr Clopyralid Picloram Benazolin 4-C PA Im azethapyr Dicamba MCPA 2,4-D Im aza quin Lower pka values N aphthylacetic acid Fluoxypyr 2,4,5-T Mecoprop Triclopyr 2,4,5-TP 2,4-DP General trend Propyzamid Io xy n il Bentazon Bromoxynil Fluazifop Bromacil 2,4-DB MCPB ph ph 6 ph ph 5 Higher pka values ph ph p H 3 SOLVENT CONSUMPTION The introduction of this method has led to a dramatic reduction of the total solvent consumption in pesticide residue analysis. This is demonstrated in the figure below. 655 more pesticides than Becker 535 including basic & acidic pesticides, but low recoveries for very polar ones more pesticides included sum other solv organochlorine Becker, up to 1990 Becker, Mini, Specht, Anastassiade s, MRM = multiresidue method QuEChERS; since 2002
7 CONCLUSIONS Rapid (8 samples in less than 30 min) Simple (no laborious steps, minimal sources of errors) Cheap (ca. 1 per sample for the sample preparation) Low Solvent Consumption (10 ml acetonitrile) Practically No Glassware Needed Wide Pesticide Range (polars, ph-dependent compounds) Extract in Acetonitrile (GC- and LC-amenable) Quick, Easy, Cheap, Effective, Rugged, Safe REFERENCE [1] Anastassiades, M., S. Lehotay, D. Stajnbaher and F. Schenck (2003). Fast and easy multiresidue method employing acetonitrile extraction/partitioning and "dispersive solid-phase extraction" for the determination of pesticide residues in produce. J AOAC Int 86(2): [2] Anastassiades, M., K. Mastovska and S. Lehotay (2003). Evaluation of analyte protectants to improve gas chromatographic analysis of pesticides. J. Chromgr. A, in press.
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