1、June 2009DEUTSCHE NORM English price group 13No part of this standard 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).ICS 65.120!$X5|“1531889www.di
2、n.deDDIN EN 15741Animal feeding stuffs Determination of OC-pesticides and PCBs by GC/MSEnglish version of DIN EN 15741:2009-06Futtermittel Bestimmung der OC-Pestizide und PCBs mittels GC/MS-VerfahrenEnglische Fassung DIN EN 15741:2009-06www.beuth.deDocument comprises 27 pagesDIN EN 15741:2009-06 2 N
3、ational foreword This standard has been prepared by Technical Committee CEN/TC 327 “Animal feeding stuffs Methods of sampling and analysis” (Secretariat: NEN, Netherlands). The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (
4、Food and Agricultural Products Standards Committee), Technical Committee NA 057-03-03 AA Futtermittel. The DIN Standard corresponding to the International Standard referred to in this document is as follows: ISO 6497 DIN EN ISO 6497 National Annex NA (informative) Bibliography DIN EN ISO 6497, Anima
5、l feeding stuffs Sampling EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN 15741February 2009ICS 65.120English VersionAnimal feeding stuffs - Determination of OC-pesticides andPCBs by GC/MSAliments des animaux - Dtermination des pesticidesorganochlors (OC) et des polychlorobiphnyles (PCB) parGC/MSF
6、uttermittel - Bestimmung der OC-Pestizide und PCBsmittels GC/MS-VerfahrenThis European Standard was approved by CEN on 24 January 2009.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national st
7、andard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the CEN Management Centre or to any CEN member.This European Standard exists in three official versions (English, French, German). A version in any other
8、 language made by translationunder the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Fin
9、land,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMIT
10、EE FR NORMUNGManagement Centre: Avenue Marnix 17, B-1000 Brussels 2009 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 15741:2009: EEN 15741:2009 (E) 2 Contents Page Foreword . 3 1 Scope 4 2 Normative references 4 3 Terms and definit
11、ions . 5 4 Principle 5 5 Reagents and materials 5 6 Apparatus . 10 7 Sampling . 13 8 Preparation of test sample . 13 9 Procedure . 13 10 Calculation and expression of results. 15 11 Precision . 18 12 Test report 20 13 Important considerations . 21 Annex A (informative) Results of interlaboratory tes
12、ts . 22 Annex B Description of PTV injection system (in case this is available). 23 Bibliography 25 DIN EN 15741:2009-06 EN 15741:2009 (E) 3 Foreword This document (EN 15741:2009) has been prepared by Technical Committee CEN/TC 327 “Animal feeding stuffs”, the secretariat of which is held by NEN. Th
13、is 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 August 2009, and conflicting national standards shall be withdrawn at the latest by August 2009. Attention is drawn to the possibility that some of th
14、e 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 has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association, and suppor
15、ts essential requirements of EC Directive(s). 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, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germa
16、ny, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. DIN EN 15741:2009-06 EN 15741:2009 (E) 4 1 Scope This European Standard specifies a gas chromatog
17、raphic/mass spectrometric method for the determination of organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs) in animal feeding stuffs and oil. The method is applicable to animal feeding stuffs with a water content up to about 20 wt% and oil/fatty samples containing residues of one
18、or more of the following OCs and PCBs and some of their isomers and degradation products: Aldrin; Dieldrin; Chlordane (= sum of Chlordane isomers and Oxychlordane); DDT (= sum of isomers op-DDT, pp-DDT, pp-TDE (pp-DDD), and pp-DDE); Endosulfan (sum of -/-isomers and Endosulfan-sulphate); Endrin; Hep
19、tachlor (= sum of Heptachlor and -Heptachlorepoxide); Hexachlorobenzene (HCB); Hexachlorocyclohexane isomers -HCH (-BHC), -HCH (-BHC), -HCH (-BHC or lindane); PCB 28, 52, 101, 138, 153 and 180 (“Indicator PCBs”) and PCB 198, 209. The method is not yet applicable to Chlorocamphene (Toxaphene), a comp
20、lex mixture of polychlorinated camphenes. Chlorocamphene has a very distinctive chromatographic profile and is easily recognisable by GC/ECD. Positive identification of the toxaphene isomers can be performed by negative chemical ionisation mass spectrometry (NCI-MS), electron impact tandem mass spec
21、trometry (EI-MSxMS) or electron impact high resolution mass spectrometry (EI-HRMS) 1, which is not within the scope of this method. The limit of quantification for the mentioned organochlorine pesticides and PCBs is 5 ng/g in general. However, 10 ng/g applies for Heptachlor Aldrin, Endrin, Dieldrin,
22、 and Endosulfan (-, - and sulphate). Individual laboratories are responsible to ensure that the equipment they used will achieve these limits of quantifications. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, on
23、ly the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 6498, Animal feeding stuffs Preparation of test samples DIN EN 15741:2009-06 EN 15741:2009 (E) 5 3 Terms and definitions For the purpose of this document, the f
24、ollowing terms and definitions apply. 3.1 limit of detection smallest measured content, from which it is possible to deduce the presence of the analyte with reasonable statistical certainty NOTE The limit of detection is numerically equal to three times the standard deviation of the mean of blank de
25、terminations (n10). 3.2 limit of quantification lowest content of the analyte which can be measured with reasonable statistical certainty NOTE If both accuracy and precision are constant over a concentration range around the limit of detection, then the limit of quantification is numerically equal t
26、o 6 times the standard deviation of the mean of blank determinations (n10). 3.3 feed Additives substances are feed additives when they comply with the definition of feed additives given in the Regulation 1831/2003 4 Principle A test portion of animal feeding stuff is fortified with internal standard
27、 (PCB 198), and is extracted with ethylacetate. The extract is concentrated and subsequently purified by: Gel permeation chromatography (GPC), with cyclohexane/ethylacetate as eluting solvent chromatography on partially deactivated silica gel. The collected fraction containing the compounds of inter
28、est is concentrated and re-dissolved in a solution containing another internal standard (PCB 209) as a reference standard. After concentration an aliquot of the extract is injected into a GC-MS, using a splitless injector (an alternative here is PTV injection, see Note below). NOTE In case more sens
29、itivity is necessary or less volume reduction is wanted, injection of a larger volume by PTV is possible (an example is described in Annex B). 5 Reagents and materials 5.1 General Use only reagents of recognized analytical grade and with a purity suitable for OC and PCB residue analysis. Check the p
30、urity of the reagents by performing a blank test under the same conditions as used in the method. The chromatogram should not show any interfering impurity at the retention time of compounds of interest. WARNING The use of this European Standard can involve hazardous materials, operations and equipm
31、ent. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this European Standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. DIN EN 15741:200
32、9-06 EN 15741:2009 (E) 6 5.2 Ethylacetate 5.3 Cyclohexane 5.4 Ethylacetate/Cyclohexane = 1+1 parts by volume Mix 500 ml of ethylacetate (5.2) with 500 ml of cyclohexane (5.3) and mix thoroughly. Store at room temperature in a tightly closed glass bottle. 5.5 Hexane 5.6 Decane 5.7 Hexane/Decane = 95+
33、5 part by volume Mix 950 ml of hexane (5.5) with 50 ml of decane (5.6) and mix thoroughly. Store at room temperature in a tightly closed glass bottle. 5.8 Iso-octane 5.9 Toluene 5.10 Silica gel, deactivated with 3,5% water Heat silica gel 60 (63m to 200m = 70 mesh to 230 mesh), at 130C for at least
34、5 h, allow to cool in a desiccator, and store in a tightly stopped container in the desiccator. To 96,5 g dried silica gel in a 300 ml Erlenmeyer flask with a ground joint, add 3,5 ml water dropwise from a burette, with continuous swirling. Immediately stopper the flask with a ground stopper and sha
35、ke vigorously for 5 min until all lumps have disappeared. Next shake for 2 h on a mechanical shaker, and then store in a tightly stoppered container. Deactivated silica gel is tenable during approximately 2 weeks if carefully stored. 5.11 Hexane/toluene = 3+7 parts by volume Mix 30 ml of n-hexane (5
36、.5) with 70 ml of toluene (5.9) and mix thoroughly. Store at room temperature in a tightly closed glass bottle. 5.12 Internal standard (PCB 209) 5.12.1 PCB 209 Stock solution 1, 100 g/ml Weigh 5-10 mg ( 0,01 mg) of PCB 209 (5.12) in a brown medicine glass bottle of 100 ml and add iso-octane (5.8) to
37、 achieve a concentration of 100 g/ml. Store the solution in a refrigerator at 4C ( 3C). The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. Or use a commercially available standard solution of 100 g/ml. 5.12.2 PCB 209 Stock solution 2, 10,0 g/
38、ml Dilute 10,0 ml of PCB 209 Stock solution 1 (5.12.1) to 100,0 ml with hexane (5.5). Store the solution in a refrigerator at 4C ( 3C). The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. 5.12.3 PCB 209 Work solution, concentration 1 000 ng/ml
39、 Dilute 10 ml of PCB 209 Stock solution 2 (5.12.2) to 100,0 ml with hexane (5.5). Store the solution in a refrigerator at 4C ( 3C). The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. DIN EN 15741:2009-06 EN 15741:2009 (E) 7 5.13 Internal stan
40、dard (PCB 198) 5.13.1 PCB 198 Stock solution 1, 100 g/ml Weigh 5-10 mg ( 0,01 mg) of PCB 198 (5.13) in a brown medicine glass bottle of 100 ml and add iso-octane (5.8) to achieve a concentration of 100 g/ml. Store the solutions in a refrigerator at 4C ( 3C). The solutions are tenable under these con
41、ditions during at least 5 years if the weight is carefully controled. Or use a commercially available standard solution of 100 g/ml. 5.13.2 PCB 198 Stock solution 2, 5,0 g/ml Pipet 5,0 ml ofPCB 198 Stock solution 1 (5.13.1) to graduated flask of 100,0 ml with hexane (5.5). Store the solution in a re
42、frigerator at 4C ( 3C). The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. 5.13.3 PCB 198 Work solution, 1 000 ng/ml Pipet 2,0 ml of the PCB 198 Stock solution 2 (5.13.2) into a 10,0 ml graduated flask and dilute with hexane (5.5) to 10,0 ml.
43、 Store the solution in a refrigerator at 4C ( 3C). The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. 5.14 PCB congeners Stock standard solution, 10 g/ml PCB 28 (2,4,4 trichlorobiphenyl); CAS Number: 7012-37-5 PCB 52 (2,2,5,5 tetrachlorobiphe
44、nyl); CAS Number: 35693-99-3 PCB 101 (2,2,4,5,5 pentachlorobiphenyl); CAS Number: 37680-73-2 PCB 138 (2,2,3,4,4,5 hexachlorobiphenyl); CAS Number: 35065-28-2 PCB 153 (2,2,4,4,5,5 hexachlorobiphenyl); CAS Number: 35065-27-1 PCB 180 (2,2,3,4,4,5,5 heptachlorobiphenyl); CAS Number: 35065-29-3 Or a Cert
45、ified Mixture at a concentration of 10 g/ml. 5.15 PCB congeners Work standard solution, 2,0 g/ml Dilute 2,0 ml of PCB congeners Stock standard solution (5.14) to 10,0 ml with hexane (5.5). Store the solution in a refrigerator at 4C ( 3C). The solution is tenable under these conditions during at leas
46、t 5 years if the weight is carefully controled. 5.16 OC-pesticide reference standards, as follows: Each with a purity of not less than 99%. Aldrin (1R,4S,4aS,5S,8R,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5,8-imethanonaphthalene); CAS Number: 309-00-2 Dieldrin (1R,4S,4aS,5R,6R,7S,8S
47、,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,6,7,8,8a-octahydro-6,7epoxy-1,4:5,8-dimethanonaphthalene) CAS Number: 60-57-1 DIN EN 15741:2009-06 EN 15741:2009 (E) 8 Chlordane (1,2,4,5,6,7,8,8-octachloro-2,3,3a,4,7,7a-hexahydro-4,7-ethano-1H-indene); and isomer; CAS Numbers: 5103-71-9 and 5103-74-2 Oxychlo
48、rdane (4,7-Methanoindan, 1,2,4,5,6,7,8,8-octachloro-2,3-epoxy-3a,4,7,7a-tetrahydro-, exo,endo-); CAS Number: 27304-13-8 op-DDT o,p-(1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane) CAS Number: 789-02-6 pp-DDT p,p-(1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane) CAS Number: 50-29-3 pp-TDE (pp-DDD) p,p
49、-1,1-dichloro-2,2-bis(4-chlorophenyl) ethane CAS Number: 72-54-8 pp-DDE p,p-(1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene) CAS Number: 72-55-9 Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin 3-oxide); two stereoisomers, , (I), CAS Number: 959-98-8 and , (II) CAS Number: 33213-65-9. Endosulfan-sulphate; CAS Number: 1031-07-8 Endrin (1R,4S,4aS,5S,6S,7R,8R,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,6,7
