SIST EN 12393-1:2014
(Main)Foods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 1: General considerations
Foods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 1: General considerations
This European Standard gives general considerations for the determination of pesticide residues in foods of plant origin. Each method specified in this European Standard is suitable for identifying and quantifying a definite range of those organohalogen, and/or organophosphorus and/or organonitrogen pesticides which occur as residues in foodstuffs of plant origin. This European Standard contains the following 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 with gel permeation chromatography [6]. The applicability of the three methods M, N and P for residue analysis of organohalogen, organophosphorus and organonitrogen pesticides, respectively, is given for each method.
Pflanzliche Lebensmittel - Multiverfahren zur Bestimmung von Pestizidrückständen mit GC oder LC-MS/MS - Teil 1: Allgemeines
Diese Europäische Norm gibt allgemeine Hinweise zur Bestimmung von Pestizidrückständen in pflanzlichen Lebensmitteln.
Jedes in dieser Europäischen Norm festgelegte Verfahren ist zur Identifizierung und Quantifizierung einer jeweils definierten Anzahl an Organohalogen- und/oder Organophosphor- und/oder Organostickstoff-Pesti-ziden geeignet, die als Rückstände in pflanzlichen Lebensmitteln vorkommen.
Diese Europäische Norm enthält die folgenden Verfahren, 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 mit Florisil ) [1], [2], [3];
- Verfahren N: Extraktion mit Aceton, Flüssig/flüssig-Verteilung mit Dichlormethan oder Cyclo-hexan/Ethylacetat und Reinigung durch Gelpermeations- und Kieselgelchromatographie [4], [5];
- Verfahren P: Extraktion mit Ethylacetat und, falls erforderlich, Reinigung durch Gelpermeations-Chroma-tographie [6].
Die Anwendbarkeit der drei Verfahren M bis P zur Rückstandsanalyse von Organohalogen-, Organophosphor- und Organostickstoff-Pestiziden ist für jedes Verfahren angegeben.
Aliments d'origine végétale - Méthodes multirésidus de détermination de résidus de pesticides par CPG ou CL-SM/SM - Partie 1: Généralités
La présente Norme européenne donne des considérations générales concernant la détermination des résidus de pesticides dans les aliments d’origine végétale.
Chaque méthode spécifiée dans la présente Norme européenne est adaptée à l’identification et à la quantification d’une gamme bien définie de pesticides organohalogénés et/ou organophosphorés et/ou organoazotés qui sont présents à l’état de résidus dans les produits alimentaires d’origine végétale.
La présente Norme européenne contient les méthodes suivantes qui ont été soumises à des études interlaboratoires et/ou qui sont adoptées dans toute l’Europe :
- méthode M : extraction par acétone, partage liquide-liquide par dichlorométhane/éther de pétrole et purification sur colonne de Florisil® 1) si nécessaire [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 acétate d’éthyle et, si nécessaire, purification par chromatographie par perméation de gel [6].
L’applicabilité des trois méthodes M à P pour l’analyse des résidus de pesticides organohalogénés, organophosphorés et organoazotés, respectivement, est précisée pour chaque méthode.
Živila rastlinskega izvora - Multirezidualne metode za določevanje ostankov pesticidov s plinsko kromatografijo (GC) ali tekočinsko kromatografijo (LC) z masno selektivno detekcijo (MS/MS) - 1. del: Splošno
Ta evropski standard določa splošne vidike določevanja ostankov pesticidov v živilih rastlinskega izvora. Vsaka metoda iz tega evropskega standarda je primerna za določitev in količinsko opredelitev določenega razpona organohalogenskih in/ali organofosfornih in/ali organodušikovih pesticidov, ki se pojavljajo kot ostanki v živilih rastlinskega izvora. Ta evropski standard zajema naslednje metode, 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]. Uporabnost vseh treh metod M, N in P za analizo ostankov organohalogenskih, organofosfornih in organodušikovih pesticidov je navedena za vsako metodo.
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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 1: AllgemeinesAliments d'origine végétale - Méthodes multirésidus de détermination de résidus de pesticides par CPG ou CL-SM/SM - Partie 1: GénéralitésFoods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 1: General considerations67.080.01Sadje, 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 12393-1:2013SIST EN 12393-1:2014en,fr,de01-januar-2014SIST EN 12393-1:2014SLOVENSKI
STANDARDSIST EN 12393-1:20091DGRPHãþD
SIST EN 12393-1:2014
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 12393-1
November 2013 ICS 67.050 Supersedes EN 12393-1:2008English Version
Foods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS - Part 1: General considerations
Aliments d'origine végétale - Méthodes multirésidus de détermination de résidus de pesticides par CPG ou CL-SM/SM - Partie 1: Généralités
Pflanzliche Lebensmittel - Multiverfahren zur Bestimmung von Pestizidrückständen mit GC oder LC-MS/MS - Teil 1: Allgemeines 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 12393-1:2013: ESIST EN 12393-1:2014
EN 12393-1:2013 (E) 2 Contents Page Foreword .3 Introduction .4 1 Scope .5 2 Normative references .5 3 Principle .5 4 Reagents .6 5 Apparatus .9 6 Procedure . 11 7 Determination . 13 8 Confirmatory tests . 14 9 Evaluation of results . 14 10 Test report . 15 Annex A (informative)
Purification of solvents and reagents . 16 Bibliography . 17
SIST EN 12393-1:2014
EN 12393-1:2013 (E) 3 Foreword This document (EN 12393-1:2013) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”, the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by May 2014, and conflicting national standards shall be withdrawn at the latest by May 2014. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights. This document supersedes EN 12393-1:2008. The following significant technical changes have been made: a) separation of analytes by liquid chromatography with MS/MS-detection in methods N and P; b) incorporation of information on GC-MS/MS detection; c) deletion of method L as no longer in use; d) editorial updating of the document according to references, etc. EN 12393, Foods of plant origin — Multiresidue methods for the determination of pesticide residues by GC or LC-MS/MS
is divided into three parts: Part 1 "General considerations" provides general considerations with regard to reagents, apparatus, gas chromatography, etc., applying to each of the selected analytical methods; Part 2 "Methods for extraction and clean-up" presents methods M, N and P for the extraction and clean-up using techniques such as liquid-liquid partition, adsorption column chromatography or gel permeation column chromatography, etc.; Part 3 "Determination and confirmatory tests" gives some recommended techniques for the qualitative and the quantitative measurements of residues and the confirmation of the results. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: 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 the United Kingdom. SIST EN 12393-1:2014
EN 12393-1:2013 (E) 4
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 3.1. SIST EN 12393-1:2014
EN 12393-1:2013 (E) 5 1 Scope This European Standard gives general considerations for the determination of pesticide residues in foods of plant origin. Each method specified in this European Standard is suitable for identifying and quantifying a definite range of those organohalogen, and/or organophosphorus and/or organonitrogen pesticides which occur as residues in foodstuffs of plant origin. This European Standard contains the following 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 with gel permeation chromatography [6]. The applicability of the three methods M, N and P for residue analysis of organohalogen, organophosphorus and organonitrogen pesticides, respectively, is given for each method. 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 12393-2:2013, 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 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 Principle 3.1 General 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 always be clearly documented and demonstrated to give valid results. The methods described in this European Standard are based on a four-stage process (in some cases two stages may be combined, in whole or in part), as given in 3.2 to 3.5.
1)
Florisil® is an example of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. SIST EN 12393-1:2014
EN 12393-1:2013 (E) 6 Quality control procedures for pesticide residue analysis, e.g. published by the European Commission [7], should be followed in its updated versions. The principles laid down in this guidance document such as initial method validation, on-going performance verification and calibration should be followed carefully in order to ensure a proper use of different combinations of separation and detection methods. Also, further information such as in [8] might be helpful. 3.2 Extraction Extraction of the residues from the sample matrix by the use of appropriate solvents, so as to obtain the maximum efficiency of extraction of the residues and minimum co-extraction of any substances which can give rise to interferences in the determination. 3.3 Clean-up Removal of interfering materials from the sample extract to obtain a solution of the extracted residue in a solvent which is suitable for determination by the selected method of determination. 3.4 Determination Gas chromatography (GC) with selective detectors may be used: electron-capture detection (ECD) for organohalogen, thermionic detector (NPD, P-mode or N/P mode) for organophosphorus and organonitrogen compounds and flame-photometric detector (FPD) for organophosphorus and organosulfurous pesticides. Hall detector (ECHD), atomic emission detector (AED) and mass spectrometry (MS) and tandem mass spectrometry may also be used for a large class of pesticides. As alternative, liquid chromatography (LC) with MS-MS-detection has been proven to be successful.
3.5 Confirmation Procedures to confirm the identity and quantity of observed residues should be used, particularly in those cases where it would appear that the maximum residue limit (MRL) has been exceeded.
3.6 Use of internal standards An internal standard may be added in a constant amount to samples, the blank and calibration standards. This substance can be used for checking critical points of methods (e.g. volatile or unstable compounds) and/or for quantification purposes, if appropriate. This is done to correct for the loss of analyte during sample preparation and sample clean-up or to check reproducibility of final determination steps by GC or LC. The internal standard should be preferably a compound that has very similar physico-chemical and chemical properties compared to the analyte.
4 Reagents 4.1 General Use reagents of purity suitable for pesticide residue analysis and check their purity (see 4.2). If required, purify water and solvents used, e.g. as described in Annex A, and check their purity (see 4.2). Purify and periodically activate adsorbents according to the requirements of the different analytical methods; check their purity (see 4.2). Take every precaution to avoid possible contamination of water, solvents, adsorbents, etc. from plastics and rubber materials. SIST EN 12393-1:2014
EN 12393-1:2013 (E) 7 4.2 Check for purity of reagents 4.2.1 Solvents Concentrate solvents by the factor involved in the respective method to be used. Test for purity under the same conditions as used in the method. The chromatogram should not show any interfering impurity.
4.2.2 Water Extract 10 parts by volume of water with one part by volume of n-hexane or light petroleum, dichloromethane or any other non-water miscible solvent used in the method. Separate the organic phase, concentrate by the factor involved in the respective method and test for purity under the same conditions as used in the method. The chromatogram should not show any interfering impurity. 4.2.3 Inorganic salts Extract inorganic salts, for example sodium chloride, after purification according to Annex A or the requirements of the different analytical methods. Extract the salts and any aqueous solution used, with n-hexane or light petroleum, dichloromethane or any other non-water miscible solvent used in the method. Concentrate the extract by the factor involved in the respective method and test the purity under the same conditions as used in the method. The chromatogram should not show any interfering impurity. 4.2.4 Adsorbents Elute an amount of adsorbent equal to that used in the analytical method with the corresponding type and volume of solvent or solvent mixture. Concentrate the eluate as indicated in the analytical method and test for purity. The chromatogram should not show any interfering impurity. Check the activity of adsorbents regularly as described in the methods M to P (see EN 12393-2). 4.2.5 Standard materials and solutions Use standard materials of at least 95 % purity and traceable quality as standards for residue analysis. Ensure dilute solutions are prepared and checked frequently, and that standard solutions are stored in glass bottles in a refrigerator and every precaution is taken to avoid possible contamination from plastics or rubber materials. Ensure that the standard solutions are not directly exposed to sunlight or ultraviolet light for prolonged periods of time. Examine analytical standards for impurities. When stored at -20 °C, standard materials are generally stable for at least a year. To allow equilibration, it is recommended to allow the standards to come up to room temperature before the containers are opened. Stock solutions of concentration 1 mg/ml, if kept in a freezer (at about -20 °C), are usually stable for 6 months, but the influence of repeated uses has to be checked. Changes in volume due to solvent evaporation, for example through the space between a glass stopper and the neck of a flask, can be a source of error. Therefore, the use of polytetrafluoroethylene (PTFE) screw-cap flasks is recommended for the storage of stock and standard solutions. Experience has shown that errors introduced in the preparation, handling and storage of standards and standard solutions are major sources of inaccuracies. Experiences obtained by other national, European and international bodies should be observed [7], [8]. 4.3 Safety aspects associated with reagents 4.3.1 General The analysis of pesticide residues in a food matrix includes the use of several hazardous chemicals. Safety precautions as given in 4.3.2 and 4.3.3 shall be observed at all times. SIST EN 12393-1:2014
EN 12393-1:2013 (E) 8 4.3.2 Pesticides Many pesticides are extremely toxic by various routes of exposure, especially in their concentrated forms. As an example, the family of organophosphorus pesticides is consistently highly toxic, not only by oral ingestion, but dermally and by inhalation as well. When working with standard materials, standard solutions, etc., observe the following minimal precautions at all times (consult safety data sheets or labels for additional information): a) Perform all laboratory sampling, mixing, weighing, etc., under an effective fume removal device in an area having a good forced ventilation of non-recirculated air; or wear a gas mask of the proper type. If the mask is used, replace cartridges as recommended, since using a contaminated mask could be worse than wearing no mask at all. b) Keep pesticides off the skin. Wear clean protective clothing and non-permeable gloves (such as polyethylene gloves) as necessary. Wash hands thoroughly with soap and water to avoid contaminating food. c) Label clearly all containers with the name and concentration of the appropriate pesticide. d) Study and have readily available information on symptoms of poisoning and first aid treatment for each type of pesticide being handled. e) Consult a physician about preventive measures and antidotes for use in emergencies when pesticide poisoning is suspected. f) Follow your organisation's procedures when disposing of waste pesticides. The manufacturer can be contacted for advice on disposal problems. g) Do not enter laboratories working with pesticide residues or other laboratories after handling pesticide formulations until protective clothing and gloves have been removed and hands thoroughly washed with soap and water. 4.3.3 Hazardous reagents Do not let vapours concentrate to a flammable level in the work area, since it is impossible to eliminate all chance of sparks from static electricity even though electrical equipment is earthed (e.g. use of spark-proof refrigerators or freezers). When working with flammable solvents, use an effective fume removal device to remove these vapours as they are released. Vapours from certain volatile solvents are highly toxic. Several of these solvents can easily be absorbed through the skin. Use an effective fume removal device to remove vapours of these solvents as they are released. A list of some hazardous reagents is given in Table 1. The use of hazardous solvents mentioned in the Montreal Protocol [9] (such as chlorinated solvents) should be minimized as far as possible. SIST EN 12393-1:2014
EN 12393-1:2013 (E) 9 Table 1 — Hazardous reagents, their effects and ways of containment Name of reagent Problem Comment Solution Acetone
Highly flammable Forms explosive peroxides with oxidising agents. Use an effective fume removal device. Acetonitrile Toxic Avoid contact with skin and eyes. Use an effective fume removal device. Cyclohexane Highly flammable
Use an effective fume removal device. Dichloromethane Toxic Avoid contact with eyes and avoid breathing vapours. Use an effective fume removal device. Diethyl ether Extremely flammable Unstable peroxides can form upon long standing or exposure to sunlight in bottles. Store protected from light. Use an effective fume removal device. See also the warning on peroxides. Ethanol
Flammable
Use an effective fume removal device when heating or evaporating. Ethyl acetate Flammable, especially when being evaporated Irritating to eyes and respiratory tract. Use an effective fume removal device. n-Hexane Highly flammable Toxic
Use an effective fume removal device. Iso-octane Highly flammable
Use an effective fume removal device. Light petroleum Extremely flammable
Use an effective fume removal device. Methanol Flammable
Toxic Avoid contact with eyes. Avoid breathing the vapours. Use an effective fume removal device.
SAFETY MEASURES: Peroxides form in diethyl ether, dioxane, and other ethers during storage. They are explosive and have to be destroyed before distillation or evaporation. Exposure to light increases peroxide formation in ethers. Filtration through activated aluminium oxide is reported to be effective in removing peroxides. 5 Apparatus 5.1 Glassware: General Thoroughly clean glassware shall be used for residue analysis. There is a risk of carry-over of pesticides through use of glassware and other laboratory equipment. This should be taken into account, particularly when employing laboratory washing machines. Hot detergent solution (with no interfering compounds) may be used for cleaning, but afterwards the glassware shall be well rinsed with distilled water and acetone before drying. Before being washed in a washing machine, the glassware shall be rinsed with acetone, then with water. Wash it in the machine with non-chlorinated detergent, rinse with water and dry. In both cases, verify that the detergent does not leave any interfering impurity. It is also advisable to rinse glassware again with the solvent to be used immediately before use. Common laboratory glassware or equipment such as beakers, round-bottomed flasks, watch glasses, pipettes, filter papers, glass wool, glass rods and glass beads, etc. are not listed in the apparatus clause of each method in detail. SIST EN 12393-1:2014
EN 12393-1:2013 (E) 10 5.2 Speci
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