EN ISO 15788-2:2004

SLOVENSKI STANDARD
01-junij-2005
Živalske in rastlinske maščobe in olja - Ugotavljanje stigmastadienov v rastlinskih
oljih - 2. del: Metoda s tekočinsko kromatografijo visoke ločljivosti (HPLC) (ISO
15788-2:2003)
Animal and vegetable fats and oils - Determination of stigmastadienes in vegetable oils -
Part 2: Method using high-performance liquid chromatography (HPLC) (ISO 15788-
2:2003)
Tierische und pflanzliche Fette und Öle - Bestimmung der Stigmastadiene in
Pflanzenölen - Teil 2: Verfahren mit Hochleistungsflüssigchromatographie (HPLC) (ISO
15788-2:2003)
Corps gras d'origines animale et végétale - Dosage des stigmastadienes dans les huiles
végétales - Partie 2: Méthode par chromatographie liquide a haute performance (CLHP)
(ISO 15788-2:2003)
Ta slovenski standard je istoveten z: EN ISO 15788-2:2004
ICS:
67.200.10 Rastlinske in živalske Animal and vegetable fats
maščobe in olja and oils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN ISO 15788-2
NORME EUROPÉENNE
EUROPÄISCHE NORM
November 2004
ICS 67.200.10
English version
Animal and vegetable fats and oils - Determination of
stigmastadienes in vegetable oils - Part 2: Method using high-
performance liquid chromatography (HPLC) (ISO 15788-2:2003)
Corps gras d'origines animale et végétale - Dosage des Tierische und pflanzliche Fette und Öle - Bestimmung der
stigmastadiènes dans les huiles végétales - Partie 2: Stigmastadiene in Pflanzenölen - Teil 2: Verfahren mit
Méthode par chromatographie liquide à haute performance Hochleistungsflüssigchromatographie (HPLC) (ISO 15788-
(CLHP) (ISO 15788-2:2003) 2:2003)
This European Standard was approved by CEN on 13 February 2003.
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 Central Secretariat 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 Central Secretariat has the same status as the official
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CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,
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Slovenia, Spain, Sweden, Switzerland and United Kingdom.
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© 2004 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 15788-2:2004: E
worldwide for CEN national Members.

Foreword
The text of ISO 15788-2:2003 has been prepared by Technical Committee ISO/TC 34
"Agricultural food products" of the International Organization for Standardization (ISO) and has
been taken over as EN ISO 15788-2:2004 by 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 May 2005, and conflicting national standards
shall be withdrawn at the latest by May 2005.

According to the CEN/CENELEC Internal Regulations, the national standards organizations of
the following countries are bound to implement this European Standard: Austria, Belgium,
Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary,
Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.

Endorsement notice
The text of ISO 15788-2:2003 has been approved by CEN as EN ISO 15788-2:2004 without any
modifications.
INTERNATIONAL ISO
STANDARD 15788-2
First edition
2003-02-15
Animal and vegetable fats and oils —
Determination of stigmastadienes
in vegetable oils —
Part 2:
Method using high-performance liquid
chromatography (HPLC)
Corps gras d'origines animale et végétale — Dosage des
stigmastadiènes dans les huiles végétales —
Partie 2: Méthode par chromatographie liquide à haute performance
(CLHP)
Reference number
ISO 15788-2:2003(E)
©
ISO 2003
ISO 15788-2:2003(E)
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ii © ISO 2003 — All rights reserved

ISO 15788-2:2003(E)
Contents Page
Foreword. iv
1 Scope. 1
2 Normative references. 1
3 Terms and definitions. 1
4 Principle. 2
5 Reagents. 2
6 Apparatus. 3
7 Sampling. 4
8 Preparation of test sample. 4
8.1 General. 4
8.2 External standard method. 4
8.3 Internal standard method. 4
9 Procedure. 5
9.1 High-pressure liquid chromatography (HPLC) .5
9.2 Identification of steradienes . 5
10 Expression of results. 6
10.1 External standard. 6
10.2 Internal standard. 6
11 Precision. 7
11.1 Interlaboratory test. 7
11.2 Repeatability. 7
11.3 Reproducibility. 7
12 Test report. 7
Annex A (informative) Results of interlaboratory test . 8
Annex B (informative) Examples of chromatograms. 9
Bibliography . 10

ISO 15788-2:2003(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 15788-2 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
ISO 15788 consists of the following parts, under the general title Animal and vegetable fats and oils —
Determination of stigmastadienes in vegetable oils:
 Part 1: Method using capillary-column gas chromatography (Reference method)
 Part 2: Method using high-performance liquid chromatography (HPLC)
iv © ISO 2003 — All rights reserved

INTERNATIONAL STANDARD ISO 15788-2:2003(E)

Animal and vegetable fats and oils — Determination
of stigmastadienes in vegetable oils —
Part 2:
Method using high-performance liquid chromatography (HPLC)
1 Scope
This part of ISO 15788 specifies a method for the determination of steradienes, especially stigmastadienes.
Steradienes are formed by dehydration of sterols during bleaching and also partially during steam washing
and deodorization. The method is also suitable as a screening method to detect the presence of refined
vegetable oils in virgin oils such as virgin olive oil.
NOTE ISO 15788-1 is the reference method for the determination of stigmastadienes in vegetable oils whilst this part
of ISO 15788 can be used as a rapid screening method. In view of the precision of this method (see Annex A), samples of
virgin olive oils close to the limit adopted by international regulations (IOOC, EC) can be verified by the GLC method given
in ISO 15788-1.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
ISO 3696:1987, Water for analytical use — Specification and test method
ISO 12228, Animal and vegetable fats and oils — Determination of individual and total sterols content — Gas
chromatographic method
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
stigmastadienes content
that part of the stigmastadienes separated by liquid chromatography under the conditions specified in this
International Standard
NOTE It is expressed in milligrams per kilogram.
3.2
steradienes content
that part of all the steradienes separated by liquid chromatography under the conditions specified in this
International Standard
NOTE It is expressed in milligrams per kilogram.
ISO 15788-2:2003(E)
4 Principle
Steradienes are separated from the major part of other lipids as non-polar fat compounds, using petroleum
ether on a silica gel column. The petroleum ether eluate is concentrated then analysed by RP-18-HPLC and
UV-detection at 235 nm. They are quantified, depending on the kind of sample, using an internal or external
standard.
5 Reagents
WARNING — Attention is drawn to the regulations which prescribe the handling of dangerous matter.
Technical, organizational and personal
...