EN 12393-2:2013
(Main)Foods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 2: Methods for extraction and clean-up
Foods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 2: Methods for extraction and clean-up
This European Standard specifies methods for the extraction and clean-up of food samples of plant origin for quantitative determination of pesticide residues.
Different solvents can be used for this purpose. These pesticide residues are generally associated with other co-extracted compounds which would interfere in the analysis. To purify the crude extracts to be analysed, several techniques can be used.
This European Standard contains the following extraction and clean-up methods that have been subjected to interlaboratory studies and/or are adopted throughout Europe:
- method M: Extraction with acetone and liquid-liquid partition with dichloromethane/light petroleum, if necessary clean-up on Florisil® ) [1], [2], [3];
- method N: Extraction with acetone, liquid-liquid partition with dichloromethane or cyclohexane/ethyl acetate and clean-up with gel permeation and silica gel chromatography [4], [5];
- method P: Extraction with ethyl acetate, and if necessary, clean-up by gel permeation chromatography [6].
This European Standard specifies the details of methods M, N and P for the extraction and the clean-up of food samples of plant origin. Several solvents at different volumes are used for extraction. Techniques of clean-up are listed such as liquid-liquid partition, liquid chromatography on various adsorbents and gel permeation chromatography.
A table providing the couples (matrix/pesticide) which have been submitted to collaborative studies and a list of indicative applicability of the method to different pesticides are given for each method, wherever possible.
Pflanzliche Lebensmittel - Multiverfahren zur Bestimmung von Pestizidrückständen mit GC oder LC-MS/MS - Teil 2: Verfahren zur Extraktion und Reinigung
Diese Europäische Norm legt Verfahren für die Extraktion und Reinigung pflanzlicher Lebensmittel zur Bestimmung von Pestizidrückständen fest.
Zu diesem Zweck können verschiedene Lösemittel verwendet werden. Die Pestizidrückstände liegen im Allgemeinen gemeinsam mit mitextrahierten Begleitstoffen vor, die bei der Analyse stören können. Zur Reinigung dieser rohen Extrakte können unterschiedliche Verfahren angewendet werden.
Diese Europäische Norm enthält die folgenden Extraktions und Reinigungsverfahren, die in Ringversuchen erfolgreich geprüft worden und/oder in ganz Europa anerkannt sind:
- Verfahren M: Extraktion mit Aceton und Flüssig/Flüssig Verteilung mit Dichlormethan/Petroleumbenzin, falls erforderlich, Reinigung an Florisil® ) [1], [2], [3];
- Verfahren N: Extraktion mit Aceton, Flüssig/Flüssig Verteilung mit Dichlormethan oder Cyclohexan/Ehtyl-acetat und Reinigung durch Gelpermeations und Kieselgel Chromatographie [4], [5];
- Verfahren P: Extraktion mit Ethylacetat und, falls erforderlich, Reinigung durch Gelpermeations-Chromatographie [6].
Diese Europäische Norm legt Einzelheiten der Verfahren M, N und P zur Extraktion und Reinigung von Proben pflanzlicher Lebensmittel fest. Zur Extraktion werden mehrere Lösemittel in unterschiedlichen Mengen verwendet. Als Reinigungsverfahren werden z. B. die Flüssig/Flüssig Verteilung, die Flüssigkeitschromatogra-phie an verschiedenen Adsorptionsmitteln und die Gelpermeations Chromatographie aufgeführt.
Soweit möglich, ist in jedem Verfahren der jeweilige Anwendungsbereich beschrieben, und die Pestizid/ Matrix-Kombinationen, die in Ringversuchen getestet wurden, sind tabellarisch aufgelistet, siehe Tabelle 3.
Aliments d'origine végétale - Méthodes multirésidus de détermination de résidus de pesticides par CPG ou CL-SM/SM - Partie 2: Méthodes d'extraction et de purification
La présente Norme européenne spécifie des méthodes pour l'extraction et la purification d'échantillons d'aliments d'origine végétale en vue du dosage des résidus de pesticides.
Différents solvants peuvent ainsi être utilisés. Ces résidus de pesticides sont généralement associés à d'autres composés co-extraits pouvant interférer avec l'analyse. Plusieurs méthodes peuvent être utilisées pour purifier les extraits bruts devant être analysés.
La présente Norme européenne contient les méthodes d'extraction et de purification suivantes qui ont été soumises à des études interlaboratoires et/ou qui sont adoptées dans toute l'Europe :
- méthode M : extraction à l'acétone, partage liquide-liquide au dichlorométhane/éther de pétrole et purification au Florisil®, le cas échéant ) [1], [2], [3] ;
- méthode N : extraction par acétone, partage liquide-liquide par dichlorométhane ou cyclohexane/acétate d'éthyle et purification par chromatographie par perméation de gel et de gel de silice [4], [5] ;
- méthode P : extraction avec l'acétate d'éthyle et, si nécessaire, purification par chromatographie par perméation de gel [6].
La présente Norme européenne détaille les méthodes M, N et P pour l'extraction et la purification d'échantillons d'aliments d'origine végétale. L'extraction fait appel à plusieurs solvants, à différents volumes. Les techniques de purification telles que le partage liquide-liquide, la chromatographie liquide sur divers adsorbants et la chromatographie par perméation de gel sont mentionnées.
Un tableau contenant les couples (matrice/pesticide) qui ont été soumis à des études collaboratives, ainsi qu'une liste indicative d'applicabilité de la méthode aux différents pesticides, sont donnés pour chaque méthode, lorsque c'est possible.
Živila rastlinskega izvora - Multirezidualne metode za določevanje ostankov pesticidov s plinsko kromatografijo ali tekočinsko kromatografijo z masno selektivno detekcijo (LC-MS/MS) - 2. del: Metode za ekstrakcijo in čiščenje
Ta evropski standard določa metode za ekstrakcijo in čiščenje vzorcev živil rastlinskega izvora za kvantitativno določevanje ostankov pesticidov. V ta namen se lahko uporabljajo različna topila. Ti ostanki pesticidov so običajno povezani z drugimi sočasno ekstrahiranimi spojinami, ki bi motile analizo. Za purifikacijo surovih ekstraktov za analizo se lahko uporabi več tehnik. Ta evropski standard zajema naslednje metode za ekstrakcijo in čiščenje, ki so bile predmet medlaboratorijskih študij in/ali so sprejete po vsej Evropi: – metoda M: ekstrakcija z acetonom in ločitev tekoče-tekoče z diklorometanom/lahkim bencinom, po potrebi čiščenje s Florisil® 1) [1], [2], [3]; – metoda N: ekstrakcija z acetonom, ločitev tekoče-tekoče z diklorometanom ali cikloheksanom/etil acetatom ter čiščenje z gelsko permeacijsko in silikagelsko kromatografijo [4], [5]; – metoda P: ekstrakcija z etil acetatom in po potrebi čiščenje z gelsko permeacijsko kromatografijo [6]. Ta evropski standard določa podrobnosti metod M, N in P za ekstrakcijo ter čiščenje vzorcev živil rastlinskega izvora. Za ekstrakcijo se uporablja več topil v različnih količinah. Navedene so tehnike čiščenja, kot so ločitev tekoče tekoče, tekočinska kromatografija za različne adsorbente in gelska permeacijska kromatografija. Preglednica s pari (matrika/pesticid), ki so bili predloženi za medlaboratorijske študije, in seznam indikativne uporabnosti metode za različne pesticide sta navedena za vsako metodo, kadar je to mogoče.
General Information
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Standards Content (Sample)
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Pflanzliche Lebensmittel - Multiverfahren zur Bestimmung von Pestizidrückständen mit GC oder LC-MS/MS - Teil 2: Verfahren zur Extraktion und ReinigungAliments d'origine végétale - Méthodes multirésidus de détermination de résidus de pesticides par CPG ou CL-SM/SM - Partie 2: Méthodes d'extraction et de purificationFoods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 2: Methods for extraction and clean-up67.080.01Sadje, zelenjava in njuni proizvodi na splošnoFruits, vegetables and derived products in general67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 12393-2:2013SIST EN 12393-2:2014en,fr,de01-januar-2014SIST EN 12393-2:2014SLOVENSKI
STANDARDSIST EN 12393-2:20091DGRPHãþD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 12393-2
November 2013 ICS 67.050 Supersedes EN 12393-2:2008English Version
Foods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 2: Methods for extraction and clean-up
Aliments d'origine végétale - Méthodes multirésidus de détermination de résidus de pesticides par CPG ou CL-SM/SM - Partie 2: Méthodes d'extraction et de purification Pflanzliche Lebensmittel - Multiverfahren zur Bestimmung von Pestizidrückständen mit GC oder LC-MS/MS - Teil 2: Verfahren zur Extraktion und Reinigung This European Standard was approved by CEN on 21 September 2013.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre:
Avenue Marnix 17,
B-1000 Brussels © 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 12393-2:2013: ESIST EN 12393-2:2014
Average water content of crops and foods . 41 Bibliography . 42
Introduction This European Standard comprises a range of multi-residue methods of equal status: no single method can be identified as the prime method because, in this field, methods are continuously developing. The selected methods included in this European Standard have been validated and/or are widely used throughout Europe. Because these methods can be applied to the very wide range of food commodities/pesticide combinations, using different systems for determination, there are occasions when variations in equipment used, extraction, clean-up and chromatographic conditions are appropriate to improve method performance, see Clause 3. SIST EN 12393-2:2014
Different solvents can be used for this purpose. These pesticide residues are generally associated with other co-extracted compounds which would interfere in the analysis. To purify the crude extracts to be analysed, several techniques can be used. This European Standard contains the following extraction and clean-up methods that have been subjected to interlaboratory studies and/or are adopted throughout Europe: method M: Extraction with acetone and liquid-liquid partition with dichloromethane/light petroleum, if necessary clean-up on Florisil® 1) [1], [2], [3]; method N: Extraction with acetone, liquid-liquid partition with dichloromethane or cyclohexane/ethyl acetate and clean-up with gel permeation and silica gel chromatography [4], [5]; method P: Extraction with ethyl acetate, and if necessary, clean-up by gel permeation chromatography [6]. This European Standard specifies the details of methods M, N and P for the extraction and the clean-up of food samples of plant origin. Several solvents at different volumes are used for extraction. Techniques of clean-up are listed such as liquid-liquid partition, liquid chromatography on various adsorbents and gel permeation chromatography. A table providing the couples (matrix/pesticide) which have been submitted to collaborative studies and a list of indicative applicability of the method to different pesticides are given for each method, wherever possible. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. EN 12393-1:2013, Foods of plant origin — Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS — Part 1: General considerations EN 12393-3:2013, Foods of plant origin — Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS — Part 3: Determination and confirmatory tests 3 Principles As already described in the introduction, in certain occasions it is possible to improve the method performance by variations in equipment used, extraction, clean-up and chromatographic conditions. Such variations shall be always clearly documented and demonstrated to give valid results. The pesticide residues are extracted from the sample by the use of appropriate solvents, so as to obtain the maximum efficiency of extraction of the pesticide residues and minimum co-extracted substances which can give rise to interferences in the determination. Any interfering materials are removed from the sample extract to obtain a solution of the extracted pesticide residues in a solvent which is suitable for quantitative examination by the selected method of determination.
1) Florisil®,is an example of a suitable product available commercially from U.S. Silica company. This information is given for convenience of users of this standard and does not constitute an endorsement by CEN of this product. SIST EN 12393-2:2014
g ml g/ml M 100 Acetone: 200 ½
N
100 a Acetone: 200 ½
P 10 Ethyl acetate: 20 ½
a Only relevant if the water content of the matrix is greater than 70 %. 4.2 Clean-up 4.2.1 Liquid-liquid partition The liquid-liquid partition procedures are summarized in Table 2. Table 2 — Liquid-liquid partition Method Aliquot portion of extract Volume of added water Volume of solvent Ratio
(AE) ml (VW) ml (VS) ml AE / VW M 80 0 200 - a N 200 x a 100 - a a Depends on the water content of the matrix.
Two techniques of liquid-liquid partition are proposed: with added water (method N); no added water (method M). 4.2.2 Adsorption column chromatography Methods: M, N with different adsorbents: silica gel, charcoal, Florisil®, used pure or in mixture. 4.2.3 Gel permeation chromatography with Bio-Beads® S-X32) Method N, and, if needed, method P.
2) BioBeads® S-X3 is an example of a suitable product available commercially. This information is given for convenience of users of this standard and does not constitute an endorsement by CEN of this product. SIST EN 12393-2:2014
Heat at 500 °C for at least 4 h, allow to cool, and store in a stoppered bottle. 5.2.5 Dichloromethane 5.2.6 Acetonitrile 5.2.7 Sodium sulfate
Heat at 500 °C for at least 4 h, allow to cool, and store in a stoppered bottle. 5.2.8 Florisil® (or equivalent), 150 µm to 250 µm (60 mesh to 100 mesh) Activate by heating at 130 °C to 135 °C for at least 5 h, allow to cool in a desiccator and transfer to an airtight stoppered jar. The adsorbent thus treated keeps its activity only for four days. It can subsequently be reactivated by the same treatment. The activity of the adsorbent should be checked from time to time by eluting pesticide standard materials as described in the method. 5.2.9 Diethyl ether, peroxide-free, containing 2 % (V+V) ethanol 5.2.10 Eluting solvent A: diethyl ether/light petroleum 6+94 (V/V) 5.2.11 Eluting solvent B: diethyl ether/light petroleum 15+85 (V/V) 5.2.12 Eluting solvent C: diethyl ether/light petroleum 50+50 (V/V) 5.2.13 Eluting solvent D: dichloromethane/light petroleum 20+80 (V/V) 5.2.14 Eluting solvent E: dichloromethane/light petroleum/acetonitrile 50+49,65+0,35 (V/V/V) 5.2.15 Eluting solvent F: dichloromethane/light petroleum/acetonitrile 50+48,5+1,5 (V/V/V) SIST EN
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