1、Animal and vegetable fats and oils Gas chromatography of fatty acid methyl estersPart 2: Preparation of methyl esters of fatty acidsBS EN ISO 12966-2:2017BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 12966-2
2、April 2017 ICS 67.200.10 Supersedes EN ISO 12966-2:2011English Version Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters - Part 2: Preparation of methyl esters of fatty acids (ISO 12966-2:2017) Corps gras dorigines animale et vgtale - Chromatographie en phase gazeus
3、e des esters mthyliques dacides gras - Partie 2: Prparation des esters mthyliques dacides gras (ISO 12966-2:2017)Tierische und pflanzliche Fette und le - Gaschromatographie von Fettsuremethylestern - Teil 2: Herstellung von Fettsuremethylestern (ISO 12966-2:2017) This European Standard was approved
4、by CEN on 3 April 2017. 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 standa
5、rds 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 not
6、ified 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, I
7、celand, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CE
8、NELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2017 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 12966-2:2017 ENational forewordThis British Standard is the UK implementation of EN ISO 12966-2:2017. It supersedes
9、 BS EN ISO 12966-2:2011 which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee AW/307, Oilseeds, animal and vegetable fats and oils and their by-products.A list of organizations represented on this committee can be obtained on request to its secretary.This pu
10、blication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2017 Published by BSI Standards Limited 2017ISBN 978 0 580 95547 1ICS 67.200.10Compliance with a British Standard cannot confer immun
11、ity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 August 2017.Amendments/corrigenda issued since publicationDate Text affectedBRITISH STANDARDBS EN ISO 12966-2:2017EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM
12、EN ISO 12966-2 April 2017 ICS 67.200.10 Supersedes EN ISO 12966-2:2011English Version Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters - Part 2: Preparation of methyl esters of fatty acids (ISO 12966-2:2017) Corps gras dorigines animale et vgtale - Chromatographie
13、en phase gazeuse des esters mthyliques dacides gras - Partie 2: Prparation des esters mthyliques dacides gras (ISO 12966-2:2017)Tierische und pflanzliche Fette und le - Gaschromatographie von Fettsuremethylestern - Teil 2: Herstellung von Fettsuremethylestern (ISO 12966-2:2017) This European Standar
14、d was approved by CEN on 3 April 2017. 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
15、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 l
16、anguage 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, Gre
17、ece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR
18、 NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2017 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 12966-2:2017 EBS EN ISO 12966-2:2017EN ISO 12966-2:2017 (E) 2 European foreword This document (EN IS
19、O 12966-2:2017) has been prepared by Technical Committee ISO/TC 34 “Food products” in collaboration with Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products - Methods of sampling and analysis” the secretariat of which is held by AFNOR. This European Sta
20、ndard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by October 2017 and conflicting national standards shall be withdrawn at the latest by October 2017. Attention is drawn to the possibility that some of the elements of
21、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 ISO 12966-2:2011. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following co
22、untries 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,
23、Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12966-2:2017 has been approved by CEN as EN ISO 12966-2:2017 without any modification. ISO 12966-2:2017(E)Foreword ivIntroduction v1 Scope . 12 Normati
24、ve references 13 Terms and definitions . 14 Reactions 15 Methodology . 25.1 Preparation of test sample 25.2 Rapid method . 25.2.1 Applicability . 25.2.2 Principle 25.2.3 Reagents 35.2.4 Apparatus . 35.2.5 Procedure . 35.3 General method 45.3.1 Applicability . 45.3.2 Principle 45.3.3 Reagents 45.3.4
25、Apparatus and materials . 55.3.5 Procedure . 55.4 Transmethylation using boron trifluoride (BF3) catalyst 65.4.1 Principle 65.4.2 Applicability . 65.4.3 Reagents 65.4.4 Apparatus . 75.4.5 Procedure . 85.5 Acid-catalysed transmethylation of glycerides . 95.5.1 Principle 95.5.2 Reagents 95.5.3 Apparat
26、us 105.5.4 Preparation of test sample . 105.5.5 Procedure 10Annex A (informative) Thin-layer chromatography method for testing the completeness of derivatization .11Bibliography .15 ISO 2017 All rights reserved iiiContents PageBS EN ISO 12966-2:2017ISO 12966-2:2017(E)Foreword ivIntroduction v1 Scope
27、 . 12 Normative references 13 Terms and definitions . 14 Reactions 15 Methodology . 25.1 Preparation of test sample 25.2 Rapid method . 25.2.1 Applicability . 25.2.2 Principle 25.2.3 Reagents 35.2.4 Apparatus . 35.2.5 Procedure . 35.3 General method 45.3.1 Applicability . 45.3.2 Principle 45.3.3 Rea
28、gents 45.3.4 Apparatus and materials . 55.3.5 Procedure . 55.4 Transmethylation using boron trifluoride (BF3) catalyst 65.4.1 Principle 65.4.2 Applicability . 65.4.3 Reagents 65.4.4 Apparatus . 75.4.5 Procedure . 85.5 Acid-catalysed transmethylation of glycerides . 95.5.1 Principle 95.5.2 Reagents 9
29、5.5.3 Apparatus 105.5.4 Preparation of test sample . 105.5.5 Procedure 10Annex A (informative) Thin-layer chromatography method for testing the completeness of derivatization .11Bibliography .15 ISO 2017 All rights reserved iiiContents PageBS EN ISO 12966-2:2017ISO 12966-2:2017(E)ForewordISO (the In
30、ternational Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee h
31、as been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical
32、 standardization. The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in a
33、ccordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights.
34、Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www .iso .org/ patents).Any trade name used in this document is information given for the convenience of users and does not constitut
35、e an endorsement. For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the
36、 following URL: www .iso .org/ iso/ foreword .html.This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal and vegetable fats and oils.This second edition cancels and replaces the first edition (ISO 12966-2:2011), of which it constitutes a minor revisio
37、n.The changes compared to the previous edition are as follows: milk and milk fat products have been excluded from the scope.A list of all the parts in the ISO 12966 series can be found on the ISO website.iv ISO 2017 All rights reservedBS EN ISO 12966-2:2017ISO 12966-2:2017(E)ForewordISO (the Interna
38、tional Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has be
39、en established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical stan
40、dardization. The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accord
41、ance with the editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Detai
42、ls of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www .iso .org/ patents).Any trade name used in this document is information given for the convenience of users and does not constitute an
43、endorsement. For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the foll
44、owing URL: www .iso .org/ iso/ foreword .html.This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal and vegetable fats and oils.This second edition cancels and replaces the first edition (ISO 12966-2:2011), of which it constitutes a minor revision.The
45、 changes compared to the previous edition are as follows: milk and milk fat products have been excluded from the scope.A list of all the parts in the ISO 12966 series can be found on the ISO website.iv ISO 2017 All rights reserved ISO 12966-2:2017(E)IntroductionGeneralOils and fats (i.e. liquid and
46、solid lipids) are predominantly composed of fatty acid esters of glycerol (triacylglycerols, TAGs), with smaller amounts of fatty acid esters of sterols and long chain of aliphatic alcohols. Due to the high molecular mass of the TAGs and their consequent low volatility, they are difficult to analyse
47、 directly by gas chromatography (GC), especially if a detailed analysis of unsaturated fatty acids is required. Fatty acids themselves do not chromatograph well (except for short-chain-length fatty acids, e.g. butanoic and pentanoic acids). It is therefore better practice to form fatty acid esters,
48、usually the fatty acid methyl esters (FAMEs), prior to GC.The analysis of oils and fats has been extensively reviewed in Reference 9.The formation of FAMEs is a critical stage in the analysis of fatty acids. Non-quantitative conversion of fatty acids to FAMEs, modification of the structure of fatty
49、acids (e.g. changes in positional and geometric isomers present) and formation of non-FAME artefacts may all affect the quantitative determination of fatty acid composition.Transesterification is one mechanism which can be employed to form FAMEs from fatty acid esters in fats (i.e. triacylglycerol). Alkali- or acid-catalysed transesterification procedures can be used to form FAMEs in a methanolic medium; the procedure can be termed transmethylation. Transmethylation is a reversible process and a large excess of methanol is requir
