SIST EN ISO 15304:2002
(Main)Animal and vegetable fats and oils - Determination of the content of trans fatty acid isomers of vegetable fats and oils - Gas chromatographic method (ISO 15304:2002)
Animal and vegetable fats and oils - Determination of the content of trans fatty acid isomers of vegetable fats and oils - Gas chromatographic method (ISO 15304:2002)
This International Standard specifies a gas chromatographic method using capillary columns for the determination of the content of trans fatty acid isomers of vegetable oils and fats.
The method is specially designed to evaluate, by a single capillary gas chromatographic (GC) procedure, the level of trans isomers as formed during (high temperature) refining, or during hydrogenation of vegetable oils or fats.
The method may also be used to report all other fatty acids (e.g. to obtain the full fatty acid composition and total amounts of saturated fatty acids, mono-unsaturated fatty acids and poly-unsaturated fatty acids) from the same sample and same analysis.
Tierische und pflanzliche Fette und Öle - Bestimmung des Gehaltes an trans-Fettsäure-Isomeren in pflanzlichen Fetten und Ölen - Gaschromatographisches Verfahren (ISO 15304:2002)
Diese Internationale Norm legt ein kapillargaschromatographisches Verfahren zur Bestimmung des Gehaltes an trans-Fettsäure-Isomeren in pflanzlichen Ölen und Fetten fest.
Das Verfahren ist speziell dazu bestimmt, die Menge der bei (Hochtemperatur-)Raffination oder bei der Hydrierung pflanzlicher Öle und Fette gebildeten trans-Isomere mit Kapillar-Gaschromatographie (Kapillar-GC) in einem Schritt zu bestimmen.
Das Verfahren ist ebenfalls zur Analyse aller anderen Fettsäuren anwendbar (z. B. um die Gesamt-Fettsäurezusammensetzung und die Gesamtmenge gesättigter Fettsäuren, einfach-ungesättigter Fettsäuren und mehrfach-ungesättigter Fettsäuren) in derselben Probe in einem Analysengang zu bestimmen.
ANMERKUNG 1 Der mit diesem Verfahren erhaltene Gehalt an trans-Isomeren muss nicht mit dem Gehalt an trans-Isomeren übereinstimmen, der mit anderen Verfahren ermittelt wurde.
ANMERKUNG 2 Während der (Hochtemperatur-)Raffination (Entsäuerung und Desodorierung) werden nur geometrische Isomere der einfach- und mehrfach-ungesättigten Fettsäuren gebildet. Die Doppelbindungen verbleiben in der ursprünglichen Position. Während der Hydrierung werden beide Formen, die Stellungs- und die geometrischen Isomere, gebildet.
ANMERKUNG 3 Einige spezielle cis- und trans-Isomere, die während der Hydrierung gebildet werden, können gleichzeitig eluieren. Dies kann die Genauigkeit des Ergebnisses beeinflussen. Der Anteil dieser Isomere ist üblicherweise in partiell hydrierten Ölen und Fetten unbedeutend.
Corps gras d'origines animale et végétale - Détermination de la teneur en isomeres trans d'acides gras de corps gras d'origine végétale - Méthode par chromatographie en phase gazeuse (ISO 15304:2002)
La présente Norme internationale spécifie une méthode par chromatographie en phase gazeuse utilisant des colonnes capillaires pour la détermination de la teneur en isomères trans d'acides gras de corps gras d'origine végétale.
Cette méthode est tout particulièrement conçue pour évaluer la teneur en isomères trans obtenus lors du raffinage (à haute température) ou de l'hydrogénation des corps gras d'origine végétale et ce, selon la méthode de chromatographie en phase gazeuse avec une seule colonne capillaire.
La méthode peut également être utilisée pour réaliser la détermination de tous les autres acides gras, par exemple pour obtenir la composition complète des acides gras et les quantités totales d'acides gras saturés, monoinsaturés et polyinsaturés d'un même échantillon à partir d'une même analyse.
Živalske in rastlinske maščobe in olja - Določevanje trans izomer maščobnih kislin rastlinskih maščob in olj - Plinska kromatografska metoda (ISO 15304:2002)
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN ISO 15304:2002
01-september-2002
äLYDOVNHLQUDVWOLQVNHPDãþREHLQROMD'RORþHYDQMHWUDQVL]RPHUPDãþREQLKNLVOLQ
UDVWOLQVNLKPDãþRELQROM3OLQVNDNURPDWRJUDIVNDPHWRGD ,62
Animal and vegetable fats and oils - Determination of the content of trans fatty acid
isomers of vegetable fats and oils - Gas chromatographic method (ISO 15304:2002)
Tierische und pflanzliche Fette und Öle - Bestimmung des Gehaltes an trans-Fettsäure-
Isomeren in pflanzlichen Fetten und Ölen - Gaschromatographisches Verfahren (ISO
15304:2002)
Corps gras d'origines animale et végétale - Détermination de la teneur en isomeres trans
d'acides gras de corps gras d'origine végétale - Méthode par chromatographie en phase
gazeuse (ISO 15304:2002)
Ta slovenski standard je istoveten z: EN ISO 15304:2002
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
SIST EN ISO 15304:2002 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN ISO 15304:2002
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SIST EN ISO 15304:2002
EUROPEAN STANDARD
EN ISO 15304
NORME EUROPÉENNE
EUROPÄISCHE NORM
March 2002
ICS 67.200.10
English version
Animal and vegetable fats and oils - Determination of the
content of trans fatty acid isomers of vegetable fats and oils -
Gas chromatographic method (ISO 15304:2002)
Corps gras d'origines animale et végétale - Détermination Tierische und pflanzliche Fette und Öle - Bestimmung des
de la teneur en isomères trans d'acides gras de corps gras Gehaltes an trans-Fettsäure-Isomeren in pflanzlichen
d'origine végétale - Méthode par chromatographie en phase Fetten und Ölen - Gaschromatographisches Verfahren
gazeuse (ISO 15304:2002) (ISO 15304:2002)
This European Standard was approved by CEN on 8 March 2002.
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 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 Management Centre has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,
Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2002 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 15304:2002 E
worldwide for CEN national Members.
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SIST EN ISO 15304:2002
EN ISO 15304:2002 (E)
CORRECTED 2002-04-17
Foreword
This document (ISO 15304:2002) has been prepared by Technical Committee ISO/TC 34
"Agricultural 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 September 2002, and conflicting
national standards shall be withdrawn at the latest by September 2002.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of
the following countries are bound to implement this European Standard: Austria, Belgium,
Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy,
Luxembourg, Malta, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the
United Kingdom.
Endorsement notice
The text of the International Standard ISO 15304:2002 has been approved by CEN as a
European Standard without any modifications.
NOTE Normative references to International Standards are listed in annex ZA (normative).
2
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SIST EN ISO 15304:2002
EN ISO 15304:2002 (E)
Annex ZA
(normative)
Normative references to international publications
with their relevant European publications
This European Standard incorporates by dated or undated reference, provisions from other
publications. These normative references are cited at the appropriate places in the text and the
publications are listed hereafter. For dated references, subsequent amendments to or revisions
of any of these publications apply to this European Standard only when incorporated in it by
amendment or revision. For undated references the latest edition of the publication referred to
applies (including amendments).
NOTE Where an International Publication has been modified by common modifications,
indicated by (mod.), the relevant EN/HD applies.
Publication Year Title EN Year
ISO 661 1989 Animal and vegetable fats and EN ISO 661 1995
oils — Preparation of test sample
ISO 5509 2000 Animal and vegetable fats and EN ISO 5509 2000
oils — Preparation of methyl
esters of fatty acids.
3
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SIST EN ISO 15304:2002
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SIST EN ISO 15304:2002
INTERNATIONAL ISO
STANDARD 15304
First edition
2002-03-15
Corrected version
2003-05-15
Animal and vegetable fats and oils —
Determination of the content of trans fatty
acid isomers of vegetable fats and oils —
Gas chromatographic method
Corps gras d'origines animale et végétale — Détermination de la teneur en
isomères trans d'acides gras de corps gras d'origine végétale — Méthode
par chromatographie en phase gazeuse
Reference number
ISO 15304:2002(E)
©
ISO 2002
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SIST EN ISO 15304:2002
ISO 15304:2002(E)
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ii © ISO 2002 – All rights reserved
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SIST EN ISO 15304:2002
ISO 15304:2002(E)
Contents
Foreword . iv
1 Scope. 1
2 Normative references. 1
3 Terms and definitions. 1
4 Principle . 2
5 Reagents and materials. 2
6 Apparatus. 2
7 Sampling . 3
8 Preparation of test sample . 3
9 Preparation of methyl esters. 3
10 Procedure. 3
11 Calculations . 5
12 Precision . 7
13 Test report. 7
Annex A (informative) Optimum conditions. 8
Annex B (informative) Examples of typical chromatograms obtained under the recommended
conditions . 11
Annex C (informative) Equivalent chain length (ECL) values . 16
Annex D (informative) FID response factor and FID correction factor. 17
Annex E (informative) Results of interlaboratory test . 18
Bibliography. 20
© ISO 2002 – All rights reserved iii
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SIST EN ISO 15304:2002
ISO 15304:2002(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 3.
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 International Standard may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 15304 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal and
vegetable fats and oils.
Annexes A to E of this International Standard are for information only.
In this corrected version, the identification of the main C18:2 cis isomer peak in Figure B.2 (the central peak in the
figure) has been corrected from
C18:1 12cis
to
C18:2 9cis, 12cis
iv © ISO 2002 – All rights reserved
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SIST EN ISO 15304:2002
INTERNATIONAL STANDARD ISO 15304:2002(E)
Animal and vegetable fats and oils — Determination of the content
of trans fatty acid isomers of vegetable fats and oils — Gas
chromatographic method
1 Scope
This International Standard specifies a gas chromatographic method using capillary columns for the determination
of the content of trans fatty acid isomers of vegetable oils and fats.
The method is specially designed to evaluate, by a single capillary gas chromatographic (GC) procedure, the level
of trans isomers as formed during (high temperature) refining, or during hydrogenation of vegetable oils or fats.
The method may also be used to report all other fatty acids (e.g. to obtain the full fatty acid composition and total
amounts of saturated fatty acids, mono-unsaturated fatty acids and poly-unsaturated fatty acids) from the same
sample and same analysis.
NOTE 1 The trans-isomer content as obtained with this method may not agree with the trans-isomer content as obtained
using other methods.
NOTE 2 During (high temperature) refining (deacidification and deodorization), only geometrical isomers are formed of the
mono- and poly-unsaturated fatty acids; i.e. the double bond(s) remain(s) at the same natural position. During hydrogenation,
both positional and geometrical isomers are formed.
NOTE 3 For some specific cis- and trans-isomers formed during hydrogenation, co-elution is possible. This could influence
the accuracy of the result. The level of these isomers is usually negligible in normal partially hydrogenated oils and fats.
2 Normative references
The following normative documents contain provisions which, through reference in this text, constitute provisions of
this International Standard. For dated references, subsequent amendments to, or revisions of, any of these
publications do not apply. However, parties to agreements based on this International Standard are encouraged to
investigate the possibility of applying the most recent editions of the normative documents indicated below. For
undated references, the latest edition of the normative document referred to applies. Members of ISO and IEC
maintain registers of currently valid International Standards.
ISO 661, Animal and vegetable fats and oils — Preparation of test sample
ISO 5509, Animal and vegetable fats and oils — Preparation of methyl esters of fatty acids
3 Terms and definitions
For the purposes of this International Standard, the following terms and definitions apply.
3.1
content of trans fatty acid isomers of (high temperature) refined oils and fats
sum of the C18:1 trans, C18:2 trans and C18:3 trans fatty acid methyl esters, expressed as a mass fraction of all
fatty acid methyl esters
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SIST EN ISO 15304:2002
ISO 15304:2002(E)
3.2
content of trans fatty acid isomers of partially hydrogenated oils and fats
sum of all trans double-bond-containing fatty acid methyl esters, expressed as a mass fraction of all fatty acid
methyl esters
NOTE The content of trans fatty acid isomers is expressed in percent.
4 Principle
The methylated fatty acids of the sample are separated on a capillary gas chromatography column with a high polar
stationary phase, with respect to their chain length, degree of (un)saturation and geometry and position of the
double bonds.
5 Reagents and materials
Use only reagents of recognized analytical grade, unless otherwise specified.
5.1 Carrier gas, preferentially helium or hydrogen, or otherwise nitrogen, of gas chromatographic quality, dried
and with oxygen removed by suitable filters.
WARNING — Hydrogen, which is used only with capillary columns, can double the speed of the analysis
(in comparison with helium) but is hazardous. Safety devices are available and it is essential that a suitable
device be incorporated into the apparatus.
5.2 Certified Reference Material (CRM), BCR 162 (soya/maize blend), European Commission, Community
1)
Bureau of Reference.
NOTE In addition to the use of CRM from the EC, the use of other calibration standards from reputable suppliers such as
Supelco, Larodan, Nuchek and Sigma may be accepted. Perhaps different standards will be necessary for different
hydrogenated oils (e.g. lauric, non-lauric oils and palm oil).
6 Apparatus
Usual laboratory equipment and, in particular, the following.
6.1 Gas chromatograph, equipped with a capillary injection system (preferred split mode, operated at a split
ratio of approximately 1:100) and flame ionization detector (FID).
TM
2) 3) 4)
6.2 Capillary column, with a high polar stationary phase (e.g. CP -Sil 88 , SP-2340 , BPX-70 or similar
highly polar cyanopropyl phases such as SP-2380 and SP-2560 which can give similar resolution of the various
geometrical isomers).
TM
NOTE For improved separations, a 100 m SP-2560 or CP -Sil 88 column and hydrogen as carrier gas are
recommended.
1) European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Geel,
Belgium.
2) Available from Chrompack, Middelburg, The Netherlands.
3) Available from Supelco, Bellafonte, PA, USA.
4) Available from SGE Inc., Austin, Texas, USA.
These types of columns are examples of suitable products which are available commercially. This information is given for the
convenience of users of this International Standard and does not constitute an endorsement by ISO of these products.
2 © ISO 2002 – All rights reserved
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SIST EN ISO 15304:2002
ISO 15304:2002(E)
Examples of dimensions:
TM
CP -Sil 88, (50 to 100) m × 0,25 mm i.d., 0,20 µm film;
SP-2360, (50 to 100) m × 0,25 mm i.d., 0,20 µm film;
SP-2340, 60 m × 0,25 mm i.d., 0,20 µm film;
BPX-70, 50 m × 0,22 mm i.d., 0,25 µm film.
Optimum conditions shall be defined by the user following the instructions in 10.3. (See also annex A.)
7 Sampling
Sampling is not part of the method specified in this International Standard. A recommended sampling method is
given in ISO 5555 [1].
It is important that the laboratory receive a sample which is truly representative and has not been damaged or
changed during transport or storage.
8 Preparation of test sample
Prepare the test sample in accordance with ISO 661.
Before taking the test portion from the sample, mix the sample thoroughly. Melt solid samples completely to ensure
proper mixing.
9 Preparation of methyl esters
Prepare the methyl esters from the triglycerides of the prepared test sample in accordance with ISO 5509.
Methods specified in AOCS Official Method Ce 2-66 [2] or IUPAC 2.301 [3] may also be used.
For trans-isomer determination in, for example, virgin olive oils, the trans-esterification routine as specified in
ISO 5509 is recommended to avoid any heating of the samples.
10 Procedure
10.1 General
In conjunction with the analysis of the test sample (or a series of test samples), analyse a blank sample (n-heptane)
and a reference sample of CRM (5.2).
10.2 GC conditions
10.2.1 Set up the gas chromatograph with one of the recommended combinations of temperature and column as
described in Table 1.
Measure the average carrier gas linear velocity as indicated in Table 1, with a split ratio of approximately 1:100.
See annex B for typical chromatograms obtained with the conditions given in Table 1.
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SIST EN ISO 15304:2002
ISO 15304:2002(E)
Table 1 — Recommended GC conditions for identification and quantification of trans isomers in refined
and hydrogenated vegetable oil samples
Parameter Proposed optimum conditions
TM
Stationary phase SP-2340 CP -Sil 88 BPX-70
Temperature conditions, °C isotherm 192 isotherm 175 isotherm 198
Column head pressure, kPa 125 130 155
15 19
...
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