DIN EN 12393-3-2014 Foods of plant origin - Multiresidue methods for the determination of pesticide residues by GC or LC-MS MS - Part 3 Determination and confirmatory tests German .pdf

1、January 2014 Translation by DIN-Sprachendienst.English price group 12No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).IC

2、S 67.050!%+KE“2084034www.din.deDDIN EN 12393-3Foods of plant origin Multiresidue methods for the determination of pesticide residues by GCor LC-MS/MS Part 3: Determination and confirmatory tests;English version EN 12393-3:2013,English translation of DIN EN 12393-3:2014-01Pflanzliche Lebensmittel Mul

3、tiverfahren zur Bestimmung von Pestizidrckstnden mit GC oder LC-MS/MS Teil 3: Verfahren zur Bestimmung und Absicherung;Englische Fassung EN 12393-3:2013,Englische bersetzung von DIN EN 12393-3:2014-01Aliments dorigine vgtale Mthodes multirsidus de dtermination de rsidus de pesticides par CPG ouCL-SM

4、/SM Partie 3: Dtermination et essais de confirmation;Version anglaise EN 12393-3:2013,Traduction anglaise de DIN EN 12393-3:2014-01SupersedesDIN EN 12393-3:2009-02www.beuth.deDocument comprises 22 pagesIn case of doubt, the German-language original shall be considered authoritative.01.14 DIN EN 1239

5、3-3:2014-01 2 A comma is used as the decimal marker. National foreword This document (EN 12393-3:2013) has been prepared by Technical Committee CEN/TC 275 “Food analysis Horizontal methods” (Secretariat: DIN, Germany), Working Group WG 4 “Pesticides in food of plant origin” (Secretariat: DIN, German

6、y). The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-01-08 AA Pestizide. Amendments This standard differs from DIN EN 12393-3:2009-02 as follows

7、: a) method L: “Extraction with acetone, liquid-liquid partition with dichloromethane and clean-up on a silica gel/charcoal column” has been deleted; b) methods N and P now also allow the separation of analytes by liquid chromatography with MS/MS-detection; c) the scope of method N has been extended

8、 to include a definite range of pesticides and their validation data; d) the former Annex B “Analysis of pesticide residues using mass spectrometry” has been replaced by a new Annex B “Typical GC-MS/MS-operating conditions”; e) Annex C “Typical LC operating conditions” has been added; f) the standar

9、d has been editorially revised and references have been updated. Previous editions DIN EN 12393-3: 1998-12, 2009-02 EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 12393-3 November 2013 ICS 67.050 Supersedes EN 12393-3:2008English Version Foods of plant origin - Multiresidue methods for the det

10、ermination of pesticide residues by GC or LC-MS/MS -Part 3: Determination and confirmatory tests Aliments dorigine vgtale - Mthodes multirsidus de dtermination de rsidus de pesticides par CPG ou CL-SM/SM - Partie 3: Dtermination et essais de confirmationPflanzliche Lebensmittel - Multiverfahren zur

11、Bestimmung von Pestizidrckstnden mit GC oder LC-MS/MS - Teil 3: Verfahren zur Bestimmung und Absicherung 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

12、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 (En

13、glish, 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

14、, 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, Switzerla

15、nd, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR 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 Me

16、mbers. Ref. No. EN 12393-3:2013: EEN 12393-3:2013 (E) 2 Contents Page Foreword 3 Introduction .4 1 Scope 5 2 Normative references 5 3 General 5 4 Determination .5 5 Confirmatory tests .8 Annex A (informative) Typical GC operating conditions 10 Annex B (informative) Typical GC-MS/MS-operating conditi

17、ons 12 Annex C (informative) Typical LC operating conditions 16 Bibliography . 20 DIN EN 12393-3:2014-01 EN 12393-3:2013 (E) 3 Foreword This document (EN 12393-3:2013) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”, the secretariat of which is held by DIN. T

18、his 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 ele

19、ments 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 12393-3:2008. This document will supersede EN 12393-3:2008 with the following significant technical changes: a) intr

20、oduction of the LC-MS/MS as a recommended technique for the determination of pesticide residues; b) deletion of method L as no longer in use; c) deletion of old Annex B with considerations concerning MS confirmation; d) addition of a new Annex B with suitable GC-MS/MS operating conditions; e) additi

21、on of new Annex C with typical LC-MS/MS operating conditions. 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, appa

22、ratus, gas chromatography, etc., applying to each of the analytical selected 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 c

23、hromatography, 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 f

24、ollowing 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, Netherland

25、s, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. DIN EN 12393-3:2014-01 EN 12393-3:2013 (E) 4 Introduction This European Standard comprises a range of multi-residue methods of equal status: no single method can be identified as the

26、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, usin

27、g 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. DIN EN 12393-3:2014-01 EN 12393-3:2013 (E) 5 1 Scope This European Standard gives guidance on

28、 some recommended techniques for the determination of pesticide residues in foods of plant origin and on confirmatory tests. The identity of any observed pesticide residue is confirmed, particularly in those cases in which it would appear that the maximum residue limit has been exceeded. 2 Normative

29、 references Not applicable. 3 General The methods specified in this European Standard permit identification and quantification of pesticide residues by gas chromatography using selective detectors or liquid chromatography with tandem-mass spectrometric detector (LC-MS/MS). All relevant results requi

30、re confirmation of identity and quantity. The procedures listed for confirmation such as alternative GC columns, alternative GC detectors, high-performance liquid chromatography (HPLC), column fractionation, derivatisation, spectral measurements, etc. are all of value. Results obtained using mass sp

31、ectrometry (MS) present the most definitive evidence for confirma-tion/identification purpose. 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 va

32、riations shall be always clearly documented and demonstrated to give valid results. 4 Determination 4.1 General 4.1.1 Identification A number of parameters can be employed to determine the identity of an analyte present in the sample extract. This includes: a) retention time of the analyte in questi

33、on (RT) or, even better, the retention time ratio against the ISTD (Rt(A)/Rt(ISTD) obtained from the same run (the simultaneous use of columns of different polarity improves this type of identification); b) in case of MS or MS/MS detection, the relative abundance of simultaneously recorded signals (

34、in general 3 ions are required in MS applications and 2 SRM transitions in MS/MS); c) the application of high resolution mass spectrometry; d) in case of MS with electron impact ionisation the comparison of the full scan mass spectrum of a suspected peak (when indicated after subtraction of backgrou

35、nd) with spectral libraries; DIN EN 12393-3:2014-01 EN 12393-3:2013 (E) 6 e) the quantification of equivalent amounts with different specific GC detectors, such as electron capture (ECD), nitrogen-phosphorous (NPD) or flame photometric (FPD) detector. The parameters obtained for the analyte to be id

36、entified in the sample extract are compared with those obtained for the pesticides in the calibration solution(s). Should a higher degree of certainty be required for the confirmation of the analyte identity, additional measures may be necessary, such as the use of different chromatographic separati

37、on conditions or the evaluation of additional m/z or SRM-transitions. The occurrence of several stable isotopes of certain elements (e.g. Cl, Br, S) may be very helpful to identify substances by MS techniques. For more information about the required identification criteria, see 1. 4.1.2 Quantificati

38、on For quantification, a chromatographic system calibrated with an sufficient number of appropriately distributed calibration points has to be used. The precision of calibration has to fulfil minimum requirements. Make sure that all the measurements are performed within the calibrated range of the s

39、ystem. In exceptional cases only, single-level calibration may be used. It has to be checked that the response of analytes present in complex mixtures does not differ from the response of separate analytes. Mixtures of isomers, degradation products and derivatives of analytes may require special con

40、ditions during calibration. For calibration, either standards in solvents or standards prepared in blank matrix (matrix-matched standards) may be used. If matrix effects during GC injection or atmospheric pressure ionisation cannot be excluded, the use of matrix-matched standards or, even better, a

41、quantification by standard addition has to be preferred. To detect instable detector response or such errors, which influence the amount of the analyte in the final extract, one or more internal standards should be added either to extracts or before extraction. To consider specific losses of individ

42、ual analytes or their matrix effects, stable isotope labelled standards (if available) may be added to the sample before extraction. All signals automatically identified by software tools may be considered as potential pesticide residues. However, any final quantification of relevant pesticide resid

43、ues should be based on visual inspection of chromatograms. Before this European Standard can be used to quantify pesticides which are not tested before, a complete initial method validation is required. In all other cases, an on-going performance verification is sufficient to demonstrate the accurac

44、y of the analytical method in a given laboratory. For more information about the required quantification criteria, see 1 in its current version. 4.2 Gas chromatography (GC) 4.2.1 General The detectors (see EN 12393-1:2013, 3.4) should be properly adjusted, according to the manufacturers instructions

45、. Variations in detector sensitivity should be checked periodically by verifying the linearity of the calibration curves using standard solutions of pesticides. The measurement may be performed using various instruments, instrument parameters and columns. Some suitable instrument parameters and colu

46、mns are listed in Annexes A and B. For suitable experimental conditions of GC-MS measurement, see 2. For suitable experimental conditions of GC-MS/MS measurement, see 3. It has been found in practice that equivalent results can be achieved despite the adoption of different GC conditions, and differe

47、nt vendors of instruments. On the other hand, specifying standard GC parameters does not guarantee that the quality of the results generated will be identical. DIN EN 12393-3:2014-01 EN 12393-3:2013 (E) 7 4.2.2 GC columns Columns should be conditioned for at least 24 h at a temperature near the maxi

48、mum recommended operating temperature with the type of stationary phase used and should then be tested for their efficiency and selectivity at the required operating temperature using standard mixtures of pesticides. The end of the column should always be disconnected from the detector during conditioning. Pure (oxygen-free) and dry (water-free) nitrogen, hydrogen or helium should be used as carrier gas. The flow rate depends on the size and type of column used. Generally, ensure that gas flow rates are controlled a