SIST EN ISO 12966-1:2015
(Main)Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters - Part 1: Guidelines on modern gas chromatography of fatty acid methyl esters (ISO 12966-1:2014)
Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters - Part 1: Guidelines on modern gas chromatography of fatty acid methyl esters (ISO 12966-1:2014)
This part of ISO 12966 gives an overview of the gas chromatographic determination of fatty acids, free
and bound, in animal and vegetable fats and oils following their conversion to fatty acid methyl esters
(FAMEs).
The qualitative and quantitative determination of the composition of fatty acids by gas liquid
chromatography (GLC) is a widely used application in lipid analysis. It is used for the characterization of
fats and oils, or fatty foodstuffs after the extraction of the oil from the matrix. The bound fatty acids of
the triacylglycerols (TAGs) and, depending on the esterification method, the free fatty acids (FFA) and
other lipids, are converted to fatty acid methyl esters (FAMEs), which are determined by capillary gas
chromatography. Depending on the number of different fatty acids (theoretically more than 50 different
fatty acids can be present) capillary columns with a length of 10 m to 100 m are used for a separation.
The GLC of FAMEs is applicable to all natural and synthetic mixtures of tri-, di- and monoacylglycerols,
to fatty acid esters, free fatty acids, soaps and other fatty compounds. With this suite of standards,
FAMEs from C4 to C26 can be determined, including saturated fatty acid methyl esters, cis- and transmonounsaturated
fatty acid methyl esters, and cis- and trans-polyunsaturated fatty acid methyl esters.
For the determination of short chain fatty acids, isopropyl and butyl esters are often used so as to avoid
interferences with the solvent peak and in order to reduce differences in detector responses.
Tierische und pflanzliche Fette und Öle - Gaschromatographie von Fettsäuremethylestern - Teil 1: Leitfaden für die moderne Gaschromatographie von Fettsäuremethylestern (ISO 12966-1:2014)
Diese Internationale Norm gibt einen Überblick über die gaschromatographische Bestimmung von freien und gebundenen Fettsäuren in tierischen und pflanzlichen Fetten und Ölen, die deren Umwandlung zu Fettsäure-methylestern (FAME) folgt.
Die qualitative und quantitative Bestimmung der Zusammensetzung von Fettsäuren mit der Gas flüssig Chromatographie (GLC) ist ein häufig angewendetes Verfahren bei der Lipidanalyse. Es wird zur Charakterisierung von Fetten und Ölen oder fetthaltigen Lebensmitteln nach der Extraktion des Öls aus der Matrix angewendet. Die gebundenen Fettsäuren der Triacylglycerine (TAG) und, in Abhängigkeit vom Ver-fahren der Veresterung, die freien Fettsäuren (FFA) und andere Lipide werden zu Fettsäuremethylestern (FAME) umgewandelt, die mit der Kapillar-Gaschromatographie bestimmt werden. Abhängig von der Anzahl der unterschiedlichen Fettsäuren — theoretisch können mehr als 50 verschiedene Fettsäuren vorliegen — werden für eine Trennung Kapillarsäulen mit einer Länge von 10 m bis 100 m verwendet.
Die GLC von FAME ist für natürliche und synthetische Gemische von Tri-, Di- und Monoacylglycerinen, für Fettsäureester, freie Fettsäuren, Seifen und weitere Fettverbindungen geeignet. Mit dieser Normenreihe können FAME von C4 bis C26 einschließlich gesättigter Fettsäuremethylester, einfach ungesättigter cis- und trans-Fettsäuremethylester und mehrfach ungesättigter cis- und trans-Fettsäuremethylester bestimmt werden.
Für die Bestimmung kurzkettiger Fettsäuren werden häufig Isopropyl- und Buthylester verwendet, um damit Interferenzen mit dem Lösemittelpeak zu vermeiden und um die Differenzen in den Detektorresponsen zu reduzieren.
Corps gras d'origines animale et végétale - Chromatographie en phase gazeuse des esters méthyliques d'acides gras - Partie 1: Directive de chromatographie en phase gazeuse moderne des esters méthyliques d'acides gras (ISO 12966-1:2014)
L'ISO 12966-1:2014 donne un aperçu de la détermination par chromatographie en phase gazeuse des acides gras, libres et liés, dans les corps gras d'origines animale et végétale après leur conversion en esters méthyliques d'acides gras (EMAG).
Živalske in rastlinske maščobe in olja - Plinska kromatografija metilnih estrov maščobnih kislin - 1. del: Smernice za sodobno plinsko kromatografijo metilnih estrov maščobnih kislin (ISO 12966-1:2014)
Ta del standarda ISO 12966 podaja pregled določanja maščobnih kislin, prostih in vezanih, s plinsko kromatografijo v živalskih ter rastlinskih maščobah in oljih po njihovi pretvorbi v metilne estre maščobnih kislin (FAMEs).
Kvalitativno in kvantitativno določanje sestave maščobnih kislin s plinsko-tekočinsko kromatografijo (GLC) se pogosto uporablja pri analizi maščobnih kislin. Uporablja se za karakterizacijo maščob in olj ali maščobnih živil po ekstrakciji olja iz matrice. Vezane maščobne kisline triacilglicerolov (TAGs) in, odvisno od metode zaestrenja, proste maščobne kisline (FFA) ter druge maščobe so pretvorjene v metilne estre maščobnih kislin (FAMEs), ki se določijo s plinsko kromatografijo s kapilarnimi kolonami. Glede na število različnih maščobnih kislin (teoretično je lahko prisotnih več kot 50 različnih maščobnih kislin) se za separacijo uporabljajo kapilarne kolone z dolžino od 10 m do 100 m. Plinsko-tekočinska kromatografija metilnih estrov maščobnih kislin se uporablja pri vseh naravnih in sintetičnih mešanicah tri-, di- in monoacilglicerolov z estri maščobnih kislin, prostimi maščobnimi kislinami, mili in drugimi maščobnimi sestavinami. S temi standardi je mogoče določiti metilne estre maščobnih kislin od C4 do C26, vključno z nasičenimi metilnimi estri maščobnih kislin, mononenasičenimi metilnimi estri maščobnih kislin v cis in trans obliki ter večkrat nenasičenimi metilnimi estri maščobnih kislin v cis in trans obliki. Za določanje kratkoverižnih maščobnih kislin se pogosto uporabljajo izopropilni in butilni estri, da se prepreči vrhe topila in zmanjša razlike pri odzivih detektorja.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN ISO 12966-1:2015
01-marec-2015
1DGRPHãþD
SIST EN ISO 15304:2002
SIST EN ISO 15304:2002/AC:2005
SIST EN ISO 5508:1996
äLYDOVNHLQUDVWOLQVNHPDãþREHLQROMD3OLQVNDNURPDWRJUDILMDPHWLOQLKHVWURY
PDãþREQLKNLVOLQGHO 6PHUQLFH]DVRGREQRSOLQVNRNURPDWRJUDILMRPHWLOQLK
HVWURYPDãþREQLKNLVOLQ ,62
Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters -
Part 1: Guidelines on modern gas chromatography of fatty acid methyl esters (ISO 12966
-1:2014)
Tierische und pflanzliche Fette und Öle - Gaschromatographie von
Fettsäuremethylestern - Teil 1: Leitfaden für die moderne Gaschromatographie von
Fettsäuremethylestern (ISO 12966-1:2014)
Corps gras d'origines animale et végétale - Chromatographie en phase gazeuse des
esters méthyliques d'acides gras - Partie 1: Directive de chromatographie en phase
gazeuse moderne des esters méthyliques d'acides gras (ISO 12966-1:2014)
Ta slovenski standard je istoveten z: EN ISO 12966-1:2014
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
SIST EN ISO 12966-1:2015 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN ISO 12966-1:2015
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SIST EN ISO 12966-1:2015
EUROPEAN STANDARD
EN ISO 12966-1
NORME EUROPÉENNE
EUROPÄISCHE NORM
December 2014
ICS 67.200.10 Supersedes EN ISO 5508:1995
English Version
Animal and vegetable fats and oils - Gas chromatography of fatty
acid methyl esters - Part 1: Guidelines on modern gas
chromatography of fatty acid methyl esters (ISO 12966-1:2014)
Corps gras d'origines animale et végétale - Tierische und pflanzliche Fette und Öle -
Chromatographie en phase gazeuse des esters Gaschromatographie von Fettsäuremethylestern - Teil 1:
méthyliques d'acides gras - Partie 1: Lignes directrices Leitfaden für die moderne Gaschromatographie von
relatives à la chromatographie en phase gazeuse moderne Fettsäuremethylestern (ISO 12966-1:2014)
des esters méthyliques d'acides gras (ISO 12966-1:2014)
This European Standard was approved by CEN on 20 October 2014.
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
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 12966-1:2014 E
worldwide for CEN national Members.
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SIST EN ISO 12966-1:2015
EN ISO 12966-1:2014 (E)
Contents Page
Foreword .3
2
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SIST EN ISO 12966-1:2015
EN ISO 12966-1:2014 (E)
Foreword
This document (EN ISO 12966-1:2014) has been prepared by Technical Committee ISO/TC 34 "Food
products" in collaboration 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 June 2015, and conflicting national standards shall be withdrawn at
the latest by June 2015.
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 5508:1995.
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.
Endorsement notice
The text of ISO 12966-1:2014 has been approved by CEN as EN ISO 12966-1:2014 without any modification.
3
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SIST EN ISO 12966-1:2015
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SIST EN ISO 12966-1:2015
INTERNATIONAL ISO
STANDARD 12966-1
First edition
2014-12-01
Animal and vegetable fats and oils —
Gas chromatography of fatty acid
methyl esters —
Part 1:
Guidelines on modern gas
chromatography of fatty acid methyl
esters
Corps gras d’origines animale et végétale — Chromatographie en
phase gazeuse des esters méthyliques d’acides gras —
Partie 1: Lignes directrices relatives à la chromatographie en phase
gazeuse moderne des esters méthyliques d’acides gras
Reference number
ISO 12966-1:2014(E)
©
ISO 2014
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SIST EN ISO 12966-1:2015
ISO 12966-1:2014(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2014
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of
the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2014 – All rights reserved
---------------------- Page: 8 ----------------------
SIST EN ISO 12966-1:2015
ISO 12966-1:2014(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Principle . 1
4 Preparation of FAME . 2
5 Columns . 3
6 GLC of FAMEs . 4
7 Evaluation of the chromatograms . 4
7.1 Peak area and area per cent . 4
7.2 Evaluation by means of an internal standard or correction factors . 4
8 Test report . 5
Bibliography . 6
© ISO 2014 – All rights reserved iii
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SIST EN ISO 12966-1:2015
ISO 12966-1:2014(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.
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 accordance 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. 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
constitute an endorsement.
For an explanation on the meaning of ISO specific terms and expressions related to conformity
assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers
to Trade (TBT) see the following URL: Foreword - Supplementary information
The committee responsible for this document is ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
This first edition of ISO 12966-1, together with ISO 12966-4, cancels and replaces ISO 5508:1990 and
ISO 15304:2002 which have been technically revised.
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 1: Guidelines on modern 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)
— Part 4: Capillary gas chromatographic method
iv © ISO 2014 – All rights reserved
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SIST EN ISO 12966-1:2015
ISO 12966-1:2014(E)
Introduction
This part of ISO 12966 is one of a suite of four International Standards for the preparation and
determination of fatty acid methyl esters by gas chromatography in animal and vegetable fats and oils.
ISO 12966 (all parts) is applicable to crude, refined, partially hydrogenated or fully hydrogenated fats,
oils and fatty acids derived from animal and vegetable sources.
ISO 12966 (all parts) is not suitable for the analysis of dairy, ruminant fats and oils (including milk and
milk products or fat coming from milk and milk products), or products supplemented with conjugated
linoleic acid (CLA). Furthermore it is not intended to be applied to polymerized and oxidized fats and
oils.
This part of ISO 12966 is a guideline to the modern gas chromatography of fatty acid methyl esters, while
ISO 12966-2 and ISO 12966-3 cover the preparation of fatty acid methyl esters by different methods. In
ISO 12966-4, the conditions for the analysis of fatty acid methyl esters by capillary gas chromatography
are given.
This suite of International Standards replaces the following International Standards:
— ISO 5508:1990 is replaced by ISO 12966-1 and ISO 12966-4
— ISO 15304:2002 is replaced by ISO 12966-1 and ISO 12966-4
— ISO 5509:2000 is replaced by ISO 12966-2 and ISO 12966-3
© ISO 2014 – All rights reserved v
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SIST EN ISO 12966-1:2015
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SIST EN ISO 12966-1:2015
INTERNATIONAL STANDARD ISO 12966-1:2014(E)
Animal and vegetable fats and oils — Gas chromatography
of fatty acid methyl esters —
Part 1:
Guidelines on modern gas chromatography of fatty acid
methyl esters
1 Scope
This part of ISO 12966 gives an overview of the gas chromatographic determination of fatty acids, free
and bound, in animal and vegetable fats and oils following their conversion to fatty acid methyl esters
(FAMEs).
The qualitative and quantitative determination of the composition of fatty acids by gas liquid
chromatography (GLC) is a widely used application in lipid analysis. It is used for the characterization of
fats and oils, or fatty foodstuffs after the extraction of the oil from the matrix. The bound fatty acids of
the triacylglycerols (TAGs) and, depending on the esterification method, the free fatty acids (FFA) and
other lipids, are converted to fatty acid methyl esters (FAMEs), which are determined by capillary gas
chromatography. Depending on the number of different fatty acids (theoretically more than 50 different
fatty acids can be present) capillary columns with a length of 10 m to 100 m are used for a separation.
The GLC of FAMEs is applicable to all natural and synthetic mixtures of tri-, di- and monoacylglycerols,
to fatty acid esters, free fatty acids, soaps and other fatty compounds. With this suite of standards,
FAMEs from C4 to C26 can be determined, including saturated fatty acid methyl esters, cis- and trans-
monounsaturated fatty acid methyl esters, and cis- and trans-polyunsaturated fatty acid methyl esters.
For the determination of short chain fatty acids, isopropyl and butyl esters are often used so as to avoid
interferences with the solvent peak and in order to reduce differences in detector responses.
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.
ISO 12966-2, Animal and vegetable fats and oils — Gas chromatography of fatty acid methyl esters — Part 2:
Preparation of methyl esters of fatty acids
ISO 12966-3, Animal and vegetable fats and oils — Gas chromatography of fatty acid methyl esters —
Part 3: Preparation of methyl esters using trimethylsulfonium hydroxide (TMSH)
1)
ISO 12966-4:— , Animal and vegetable fats and oils — Determination of methyl esters of fatty acids —
Part 4: Capillary gas chromatographic method
3 Principle
Gas liquid chromatography (GLC) is used for the qualitative and quantitative analysis of FAMEs. The
FAMEs are prepared in accordance with ISO 12966-2 or ISO 12966-3 and the dissolved FAMEs are
then injected into and vaporized within the injector. The separation of FAMEs is achieved on analytical
1) Under preparation. To be publishe
...
SLOVENSKI STANDARD
oSIST prEN ISO 12966-1:2013
01-december-2013
äLYDOVNHLQUDVWOLQVNHPDãþREHLQROMD3OLQVNDNURPDWRJUDILMDPHWLOHVWURY
PDãþREQLKNLVOLQGHO 6PHUQLFH]DVRGREQRSOLQVNRNURPDWRJUDILMRPHWLO
HVWURYPDãþREQLKNLVOLQ ,62',6
Animal and vegetable fats and oils - Gas chromatography of fatty acid methyl esters -
Part 1: Guidelines on modern gas chromatography of fatty acid methy esters (ISO/DIS
12966-1:2013)
Tierische und pflanzliche Fette und Öle - Gaschromatographie von
Fettsäuremethylestern - Teil 1: Leitfaden für die moderne Gaschromatographie von
Fettsäuremethylestern (ISO/DIS 12966-1:2013)
Corps gras d'origines animale et végétale - Chromatographie en phase gazeuse des
esters méthyliques d'acides gras - Partie 1: Directive de chromatographie en phase
gazeuse moderne des esters méthyliques d'acides gras (ISO/DIS 12966-1:2013)
Ta slovenski standard je istoveten z: prEN ISO 12966-1
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
oSIST prEN ISO 12966-1:2013 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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oSIST prEN ISO 12966-1:2013
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oSIST prEN ISO 12966-1:2013
DRAFT INTERNATIONAL STANDARD
ISO/DIS 12966-1
ISO/TC 34/SC 11 Secretariat: BSI
Voting begins on: Voting terminates on:
2013-09-26 2014-02-16
Animal and vegetable fats and oils — Gas chromatography
of fatty acid methyl esters —
Part 1:
Guidelines on modern gas chromatography of fatty acid
methy esters
Corps gras d’origines animale et végétale — Chromatographie en phase gazeuse des esters méthyliques
d’acides gras —
Partie 1: Directive de chromatographie en phase gazeuse moderne des esters méthyliques d’acides gras
[Revision of second edition (ISO 5508:1990) and first edition ISO 15304:2002]
ICS: 67.200.10
ISO/CEN PARALLEL PROCESSING
This draft has been developed within the International Organization for
Standardization (ISO), and processed under the ISO lead mode of collaboration
as defined in the Vienna Agreement.
This draft is hereby submitted to the ISO member bodies and to the CEN member
bodies for a parallel five month enquiry.
Should this draft be accepted, a final draft, established on the basis of comments
received, will be submitted to a parallel two-month approval vote in ISO and
THIS DOCUMENT IS A DRAFT CIRCULATED
formal vote in CEN.
FOR COMMENT AND APPROVAL. IT IS
THEREFORE SUBJECT TO CHANGE AND MAY
NOT BE REFERRED TO AS AN INTERNATIONAL
STANDARD UNTIL PUBLISHED AS SUCH.
To expedite distribution, this document is circulated as received from the
IN ADDITION TO THEIR EVALUATION AS
committee secretariat. ISO Central Secretariat work of editing and text
BEING ACCEPTABLE FOR INDUSTRIAL,
composition will be undertaken at publication stage.
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
STANDARDS MAY ON OCCASION HAVE TO
BE CONSIDERED IN THE LIGHT OF THEIR
POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
Reference number
NATIONAL REGULATIONS.
ISO/DIS 12966-1:2013(E)
RECIPIENTS OF THIS DRAFT ARE INVITED
TO SUBMIT, WITH THEIR COMMENTS,
NOTIFICATION OF ANY RELEVANT PATENT
RIGHTS OF WHICH THEY ARE AWARE AND TO
©
PROVIDE SUPPORTING DOCUMENTATION. ISO 2013
---------------------- Page: 3 ----------------------
oSIST prEN ISO 12966-1:2013
ISO/DIS 12966-1:2013(E)
Copyright notice
This ISO document is a Draft International Standard and is copyright-protected by ISO. Except as
permitted under the applicable laws of the user’s country, neither this ISO draft nor any extract
from it may be reproduced, stored in a retrieval system or transmitted in any form or by any means,
electronic, photocopying, recording or otherwise, without prior written permission being secured.
Requests for permission to reproduce should be addressed to either ISO at the address below or ISO’s
member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Reproduction may be subject to royalty payments or a licensing agreement.
Violators may be prosecuted.
ii © ISO 2013 – All rights reserved
---------------------- Page: 4 ----------------------
oSIST prEN ISO 12966-1:2013
ISO/DIS 12966-1
Contents Page
Foreword . iv
Introduction . vii
1 Scope . 1
2 Normative references . 1
3 Principle . 1
4 Preparation of FAME . 2
5 Columns . 3
6 GLC of FAMEs . 4
7 Evaluation of the chromatograms . 4
7.1 Peak area and area percent . 4
7.2 Evaluation by means of an internal standard or correction factors . 4
8 Test report . 4
Bibliography . 6
© ISO 2013 – All rights reserved iii
---------------------- Page: 5 ----------------------
oSIST prEN ISO 12966-1:2013
ISO/DIS 12966-1
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 12966-1 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
This second/third/. edition cancels and replaces the first/second/. edition (), [clause(s) / subclause(s) /
table(s) / figure(s) / annex(es)] of which [has / have] been technically revised.
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 1: Guidelines on modern gas chromatography of fatty acid methy esters
iv © ISO 2013 – All rights reserved
---------------------- Page: 6 ----------------------
oSIST prEN ISO 12966-1:2013
ISO/DIS 12966-1
Introduction
This International Standard is part 1 of a suite of four standards for the preparation and determination of fatty
acid methyl esters by gas chromatography in animal and vegetable fats and oils, fat spreads and in fats and
oils isolated from foodstuffs. The standards are not applicable to polymerized and oxidized fats or fatty acids.
Part 1 is a guideline to the modern gas chromatography of fatty acid methyl esters, while parts 2 and 3 cover
the preparation of fatty methyl esters by different methods. In part 4 of this International Standard, the
conditions for the analysis of fatty acid methyl esters by GLC are given.
This suite of standards replaces the following standards:
ISO 5508:1990 is replaced by ISO 12996-1 and ISO 12996-4
ISO 15304:2002 is replaced by ISO 12996-4
ISO 5509:2000 is replaced by ISO 12996-2 and ISO 12996-3
© ISO 2013 – All rights reserved v
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oSIST prEN ISO 12966-1:2013
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oSIST prEN ISO 12966-1:2013
DRAFT INTERNATIONAL STANDARD ISO/DIS 12966-1
Animal and vegetable fats and oils — Gas chromatography of
fatty acid methyl esters — Part 1: Guidelines on modern gas
chromatography of fatty acid methy esters
1 Scope
This International Standard gives an overview of the gas chromatographic determination of fatty acids, free
and bound, in animal and vegetable fats and oils following their conversion to fatty acid methyl esters
(FAMEs).
The qualitative and quantitative determination of the composition of fatty acids by gas liquid chromatography
(GLC) is a widely used application in lipid analysis. It is used for the characterisation of fats and oils, or fatty
foodstuffs after the extraction of the oil from the matrix. The bound fatty acids of the triacylglycerols (TAGs)
and, depending on the esterification method, the free fatty acids (FFA) and other lipids, are converted to fatty
acid methyl esters (FAMEs), which are determined by capillary gas chromatography. Depending on the
number of different fatty acids - theoretically more than 50 different fatty acids can be present - capillary
columns with a length of 10 meters to 100 meters are used for a separation.
The GLC of FAMEs is applicable to all natural and synthetic mixtures of tri-, di- and monoacylglycerols, to fatty
acid esters, free fatty acids, soaps and other fatty compounds. With this suite of standards, FAMEs from C4 to
C26 can be determined, including saturated fatty acid methyl esters, cis- and trans-monounsaturated fatty
acid methyl esters, and cis- and trans-polyunsaturated fatty acid methyl esters.
For the determination of short chain fatty acids, isopropyl and butyl esters are often used so as to avoid
interferences with the solvent peak and in order to reduce differences in detector responses.
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.
ISO 12966-2, Animal and vegetable fats and oils — Gas chromatography of fatty acid methyl esters — Part 2:
Preparation of methyl ester of fatty acids
ISO 12966-3, Animal and vegetable fats and oils — Gas chromatography of fatty acid methyl esters — Part 3:
Preparation of methyl esters using trimethylsulfonium hydroxide (TMSH)
ISO 12966-4, Animal and vegetable fats and oils — Gas chromatography of fatty acid methyl esters — Part 4:
Determination of cis-, trans-, saturated, mono- and polyunsaturated fatty acids in vegetable or non-ruminant
oils and fats
3 Principle
Gas liquid chromatography (GLC) is used for the qualitative and quantitative analysis of FAMEs. The FAMEs
are prepared according to ISO 12996 Part 2 or 3 and the dissolved FAMEs are then injected into and
vaporized within the injector. The separation of FAMEs is achieved on analytical columns of different polarity
and lengths. For the detection of the FAMEs, a Flame Ionisation Detector (FID) is used.
In the gas chromatography of FAMEs with FID hydrogen shall be used as the carrier gas (mobile phase), for
MSD applications helium shall be used. The separation can be done in a shorter time with sharper peaks by
© ISO 2013 – All rights reserved 1
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oSIST prEN ISO 12966-1:2013
ISO/DIS 12966-1
using hydrogen. The stationary phase is a microscopic layer of a thin liquid film on an inert solid surface,
made of steel, glass or fused silica.
The volatilised compounds being analysed interact, during their passage through the capillary tubing, with the
stationary phase coating the inner surface of the column. Due to this different interaction of different
compounds, they exit the column, or elute, at a different time, which is called the retention time of the
compound at a given set of analysis parameters. The comparison of retention times is used for the
identification of the different compounds.
NOTE When a mass selective detector is used, special care must be taken in order to achieve quantitative results
due to differences in fragmentation patterns of individual FAMEs. Also, for mass spectrometry, derivatives other than the
FAMEs discussed here are used, as for example, picolinyl esters or dimethyl oxazolidines and others.
4 Preparation of FAME
The preparation of the fatty acid methyl esters shall be carried out according to part 2 and 3 of ISO 12966. An
overview is giv
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