EN ISO 12966-2:2011
(Main)Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters - Part 2: Preparation of methyl esters of fatty acids (ISO 12966-2:2011)
Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters - Part 2: Preparation of methyl esters of fatty acids (ISO 12966-2:2011)
ISO 12966-2:2011 specifies methods of preparing the methyl esters of fatty acids.
It includes methods for preparing fatty acid methyl esters from animal and vegetable fats and oils, fatty acids and soaps. To cover different requirements four methylation methods are specified, namely: a) a "rapid" transmethylation procedure under alkaline conditions; b) a "general" transmethylation/methylation procedure under sequential alkaline and acid conditions; c) a BF3 transmethylation procedure; d) an alternative procedure using acid-catalysed transmethylation of glycerides.
Methyl esters so produced are used in various analytical procedures requiring such derivatives, e.g. gas-liquid chromatography (GC), thin-layer chromatography (TLC), and infrared spectrometry (IR).
Tierische und pflanzliche Fette und Öle - Gaschromatographie von Fettsäuremethylestern - Teil 2: Herstellung von Fettsäuremethylestern (ISO 12966-2:2011)
Dieser Teil der ISO 12966 legt Verfahren zur Herstellung der Methylester von Fettsäuren fest.
Sie bezieht Verfahren für die Herstellung von Fettsäuremethylestern aus tierischen und pflanzlichen Fetten und Ölen, Fettsäuren und Seifen mit ein. Um den verschiedenen Anforderungen gerecht zu werden, sind vier Methylierungsverfahren festgelegt, insbesondere:
a) ein "Schnellverfahren" der Transmethylierung unter alkalischen Bedingungen;
b) ein "allgemeines" Transmethylierungs-/Methylierungsverfahren unter aufeinanderfolgenden alkalischen und sauren Bedingungen;
c) ein BF3-Transmethylierungsverfahren;
d) ein alternatives Verfahren mit säurekatalysierter Transmethylierung der Glyceride.
Die so gewonnenen Methylester werden für verschiedene analytische Verfahren verwendet, bei denen diese Derivate erforderlich sind, z. B. Gas flüssig Chromatographie (GC), Dünnschichtchromatographie (DC) und Infrarotspektrometrie (IR).
Corps gras d'origines animale et végétale - Chromatographie en phase gazeuse des esters méthyliques d'acides gras - Partie 2: Préparation des esters méthyliques d'acides gras (ISO 12966-2:2011)
L'ISO 12966-2:2011 spécifie des méthodes de préparation des esters méthyliques d'acides gras.
Elle comprend des méthodes de préparation des esters méthyliques d'acides gras à partir de corps gras d'origines animale et végétale, d'acides gras et de savons. Afin de répondre à différentes exigences, quatre méthodes de méthylation sont spécifiées, à savoir: a) une méthode de transméthylation «rapide» en conditions alcalines; b) une méthode de transméthylation/méthylation «générale» en conditions alcalines et acides séquentielles; c) une méthode de transméthylation au BF3; d) une méthode alternative reposant sur la transméthylation par catalyse acide de glycérides.
Les esters méthyliques ainsi obtenus sont utilisés dans diverses méthodes d'analyse exigeant des produits dérivés de ce type, par exemple la chromatographie en phase gazeuse, la chromatographie sur couche mince et la spectrométrie infrarouge.
Rastlinske in živalske maščobe in olja - Plinska kromatografija metilnih estrov maščobnih kislin - 2. del: Priprava metilnih estrov maščobnih kislin (ISO 12966-2:2011)
Ta del ISO 12966 določa metode priprave metilnih estrov maščobnih kislin. Zajema metode za pripravo metilnih estrov maščobnih kislin iz živalskih in rastlinskih maščob in olj, maščobnih kislin in mil. Da bi zajeli različne zahteve, so določene štiri metode metilacije, in sicer:
a) »hiter« postopek transmetilacije v alkalnih okoliščinah;
a) »splošni« postopek transmetilacije/metilacije v zaporednih alkalnih in kislih okoliščinah;
c) postopek transmetilacije BF3;
d) alternativni postopek z uporabo transmetilacije gliceridov, kataliziranih s kislino.
Metlini estri, proizvedeni tako, se uporabljajo v različnih analitičnih postopkih, kjer so potrebni taki derivati, npr. plinska-tekočinska kromatografija (GLC), tankoplastna kromatografija (TLC) in infrardeča spektrometrija (IR).
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-julij-2011
1DGRPHãþD
SIST EN ISO 5509:2001
5DVWOLQVNHLQåLYDOVNHPDãþREHLQROMD3OLQVNDNURPDWRJUDILMDPHWLOQLKHVWURY
PDãþREQLKNLVOLQGHO 3ULSUDYDPHWLOQLKHVWURYPDãþREQLKNLVOLQ ,62
Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters -
Part 2: Preparation of methyl esters of fatty acids (ISO 12966-2:2011)
Tierische und pflanzliche Fette und Öle - Gaschromatographie von
Fettsäuremethylestern - Teil 2: Herstellung von Fettsäuremethylestern (ISO 12966-
2:2011)
Corps gras d'origines animale et végétale - Chromatographie en phase gazeuse des
esters méthyliques d'acides gras - Partie 2: Préparation des esters méthyliques d'acides
gras (ISO 12966-2:2011)
Ta slovenski standard je istoveten z: EN ISO 12966-2:2011
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EUROPEAN STANDARD
EN ISO 12966-2
NORME EUROPÉENNE
EUROPÄISCHE NORM
February 2011
ICS 67.200.10 Supersedes EN ISO 5509:2000
English Version
Animal and vegetable fats and oils - Gas chromatography of
fatty acid methyl esters - Part 2: Preparation of methyl esters of
fatty acids (ISO 12966-2:2011)
Corps gras d'origines animale et végétale - Tierische und pflanzliche Fette und Öle -
Chromatographie en phase gazeuse des esters Gaschromatographie von Fettsäuremethylestern - Teil 2:
méthyliques d'acides gras - Partie 2: Préparation des Herstellung von Fettsäuremethylestern (ISO 12966-2:2011)
esters méthyliques d'acides gras (ISO 12966-2:2011)
This European Standard was approved by CEN on 14 February 2011.
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, 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 STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2011 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 12966-2:2011: E
worldwide for CEN national Members.
Contents Page
Foreword .3
Foreword
This document (EN ISO 12966-2:2011) 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 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 2011, and conflicting national standards shall be withdrawn at
the latest by August 2011.
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 ISO 5509:2000.
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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia,
Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain,
Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 12966-2:2011 has been approved by CEN as a EN ISO 12966-2:2011 without any
modification.
INTERNATIONAL ISO
STANDARD 12966-2
First edition
2011-02-15
Animal and vegetable fats and oils — Gas
chromatography of fatty acid methyl
esters —
Part 2:
Preparation of methyl esters of fatty acids
Corps gras d'origines animale et végétale — Chromatographie en
phase gazeuse des esters méthyliques d'acides gras —
Partie 2: Préparation des esters méthyliques d'acides gras
Reference number
ISO 12966-2:2011(E)
©
ISO 2011
ISO 12966-2:2011(E)
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ii © ISO 2011 – All rights reserved
ISO 12966-2:2011(E)
Contents Page
Foreword .iv
Introduction.v
1 Scope.1
2 Normative references.1
3 Reactions .1
4 Methodology .2
4.1 Preparation of test sample .2
4.2 Rapid method.2
4.3 General method .4
4.4 Transmethylation using boron trifluoride (BF ) catalyst .6
4.5 Acid-catalysed transmethylation of glycerides.9
Annex A (informative) Thin-layer chromatography method for testing the completeness of
derivatization .11
Bibliography.15
ISO 12966-2:2011(E)
Foreword
ISO (the International 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 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 standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
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.
ISO 12966-2 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
This first edition of ISO 12966-2 cancels and replaces ISO 5509:2000, of which it constitutes a technical
revision.
ISO 12966 consists of the following parts, under the general title Animal and vegetable fats and oils — Gas
chromatography of fatty acid methyl esters:
⎯ Part 2: Preparation of methyl esters of fatty acids
⎯ Part 3: Preparation of methyl esters using trimethylsulfonium hydroxide (TMSH)
The following part is under preparation:
⎯ Part 4: Determination of cis-, trans-, saturated, mono- and polyunsaturated fatty acids in vegetable or
non-ruminant oils and fats
The following part is planned:
⎯ Part 1: Guidelines on gas chromatography
iv © ISO 2011 – All rights reserved
ISO 12966-2:2011(E)
Introduction
General
Oils and fats (i.e. liquid and 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 aliphatic alcohols.
Due to the high molecular mass of the TAGs and their consequent low volatility, they are difficult to analyse
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, 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 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 required to maintain an equilibrium position which favours formation of the
FAMEs. Water can prevent the reaction going to completion, and its presence should therefore be minimized.
Alkali-catalysed procedures do not produce FAMEs from free fatty acids, due to the formation of soaps.
Esterification is an acid-catalysed mechanism which can be employed to form FAMEs from fatty acids. It is
possible that the fatty acids are naturally present in the sample of fat under examination. Formation of FAMEs
by this mechanism is commonly termed methylation. Again, an excess of methanol and the absence of water
are preconditions for the quantitative formation of FAMEs.
This part of ISO 12966 provides guidelines for the preparation of fatty acid methyl esters. In support of these
guidelines, various procedures to prepare fatty acid methyl esters are specified. These include:
a) “rapid” transmethylation under alkaline conditions;
b) “general” transmethylation/methylation under sequential alkaline and acid conditions;
c) boron trifluoride (BF ) transmethylation/methylation.
“Rapid” transmethylation method under alkali-catalysed conditions
This method is applicable to the routine analysis of edible fats and oils containing fatty acids down to butanoic
acid (C4:0) and/or for the determination of butanoic acid or hexanoic acid (C6:0) by GC using an internal
standard.
Alkaline catalysts transesterify neutral lipids in the presence of anhydrous methanol (trans
...
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