EN ISO 9936:2006
(Main)Animal and vegetable fats and oils - Determination of tocopherol and tocotrienol contents by high-performance liquid chromatography (ISO 9936:2006)
Animal and vegetable fats and oils - Determination of tocopherol and tocotrienol contents by high-performance liquid chromatography (ISO 9936:2006)
ISO 9936:2006 specifies a method for the determination of the contents of free alpha-, beta-, gamma- and delta-tocopherols and tocotrienols in animal and vegetable fats and oils by high-performance liquid chromatography.
For products containing tocopherol or tocotrienol esters, it is necessary to carry out a preliminary saponification.
Tierische und pflanzliche Fette und Öle - Bestimmung des Tocopherol- und Tocotrienol-Gehaltes mit Hochleistungsflüssigchromatographie (ISO 9936:2006)
Diese Internationale Norm legt ein Verfahren zur Bestimmung des Gehalts an freien a-, b-, g- und d Tocopherolen und Tocotrienolen (nachfolgend insgesamt als Tocole bezeichnet) in tierischen und pflanzlichen Fetten und Ölen (nachfolgend als Fette bezeichnet) durch Hochleistungsflüssigchromatographie (HPLC) fest.
Bei Produkten, die Ester von Tocopherolen oder Tocotrienolen enthalten, ist es erforderlich, eine vorausgehende Verseifung durchzuführen.
ANMERKUNG Ein geeignetes Verfahren mit einer Kaltverseifung wird in Anhang B nur zur Information beschrieben.
Corps gras d'origines animale et végétale - Détermination des teneurs en tocophérols et en tocotriénols par chromatographie en phase liquide à haute performance (ISO 9936:2006)
L'ISO 9936:2006 spécifie une méthode pour la détermination des teneurs en alpha-, bêta-, gamma- et delta-tocophérols et tocotriénols libres des graisses et des huiles d'origines animale et végétale, par chromatographie en phase liquide haute performance (CLHP).
Pour les produits contenant des esters de tocophérols ou de tocotriénols, il est nécessaire qu'ils subissent une saponification préalable.
Rastlinske in živalske maščobe in olja – Določevanje tokoferola in tokotrienola s tekočinsko kromatografijo visoke ločljivosti (ISO 9936:2006)
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-september-2006
5DVWOLQVNHLQåLYDOVNHPDãþREHLQROMD±'RORþHYDQMHWRNRIHURODLQWRNRWULHQRODV
WHNRþLQVNRNURPDWRJUDILMRYLVRNHORþOMLYRVWL ,62
Animal and vegetable fats and oils - Determination of tocopherol and tocotrienol contents
by high-performance liquid chromatography (ISO 9936:2006)
Tierische und pflanzliche Fette und Öle - Bestimmung des Tocopherol- und Tocotrienol-
Gehaltes mit Hochleistungsflüssigchromatographie (ISO 9936:2006)
Corps gras d'origines animale et végétale - Détermination des teneurs en tocophérols et
en tocotriénols par chromatographie en phase liquide a haute performance (ISO
9936:2006)
Ta slovenski standard je istoveten z: EN ISO 9936:2006
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EUROPEAN STANDARD
EN ISO 9936
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2006
ICS 67.200.10
English Version
Animal and vegetable fats and oils - Determination of tocopherol
and tocotrienol contents by high-performance liquid
chromatography (ISO 9936:2006)
Corps gras d'origines animale et végétale - Détermination Tierische und pflanzliche Fette und Öle - Bestimmung des
des teneurs en tocophérols et en tocotriénols par Tocopherol- und Tocotrienol-Gehaltes mit
chromatographie en phase liquide à haute performance Hochleistungsflüssigchromatographie (ISO 9936:2006)
(ISO 9936:2006)
This European Standard was approved by CEN on 13 April 2006.
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
versions.
CEN members are the national standards bodies of Austria, Belgium, 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: rue de Stassart, 36 B-1050 Brussels
© 2006 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 9936:2006: E
worldwide for CEN national Members.
Foreword
This document (EN ISO 9936:2006) 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 October 2006, and conflicting national
standards shall be withdrawn at the latest by October 2006.
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, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
Endorsement notice
The text of ISO 9936:2006 has been approved by CEN as EN ISO 9936:2006 without any
modifications.
INTERNATIONAL ISO
STANDARD 9936
Second edition
2006-04-15
Animal and vegetable fats and oils —
Determination of tocopherol and
tocotrienol contents by high-performance
liquid chromatography
Corps gras d'origines animale et végétale — Détermination des teneurs
en tocophérols et en tocotriénols par chromatographie en phase liquide
à haute performance
Reference number
ISO 9936:2006(E)
©
ISO 2006
ISO 9936:2006(E)
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ii © ISO 2006 – All rights reserved
ISO 9936:2006(E)
Contents Page
Foreword. iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions. 1
4 Principle. 1
5 Reagents. 2
6 Apparatus . 2
7 Sampling. 3
8 Preparation of test sample. 3
9 Procedure . 4
9.1 Preparation of calibration solutions . 4
9.2 Optimization of working parameters . 5
9.3 Preparation of test solution . 5
9.4 Determination. 5
10 Expression of results . 6
11 Precision. 7
11.1 Interlaboratory test . 7
11.2 Repeatability. 7
11.3 Reproducibility. 7
12 Test report . 7
Annex A (informative) Examples of chromatograms. 8
Annex B (informative) Saponification . 10
Annex C (informative) Results of interlaboratory tests. 12
Bibliography . 17
ISO 9936:2006(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 9936 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 9936:1997), which has been technically revised.
iv © ISO 2006 – All rights reserved
INTERNATIONAL STANDARD ISO 9936:2006(E)
Animal and vegetable fats and oils — Determination of
tocopherol and tocotrienol contents by high-performance liquid
chromatography
1 Scope
This International Standard specifies a method for the determination of the contents of free α-, β-, γ-, and
δ-tocopherols and tocotrienols (referred to jointly as tocols) in animal and vegetable fats and oils (referred to
hereinafter as fats) by high-performance liquid chromatography (HPLC).
For products containing tocopherol or tocotrienol esters, it is necessary to carry out a preliminary
saponification.
NOTE A suitable method involving a cold saponification procedure is described in Annex B for information only.
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 661, Animal and vegetable fats and oils — Preparation of test sample
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
tocol content
mass fraction of the individual tocols, determined using the method specified in this International Standard
NOTE The content is expressed in milligrams per kilogram as a whole number.
4 Principle
A test portion is dissolved in n-heptane and the individual tocols are separated by high-performance liquid
chromatography. The content of each tocol is calculated using calibration factors determined from calibration
solutions.
ISO 9936:2006(E)
5 Reagents
Use only reagents of HPLC grade or equivalent.
5.1 α-, β-, γ- and δ-tocopherol and tocotrienol standards.
If tocopherol standards are not available, a blend of wheat germ and soya bean oil may be used to identify
α-, β-, γ- and δ-tocopherols.
If tocotrienol standards are not available, palm oil may be used to identify α- and γ-tocotrienols. The
chromatograms obtained can be used to assist peak identification in test sample chromatograms, in which
case the calibration factors for the corresponding tocopherols should be used.
1)
NOTE α-, β-, γ- and δ-tocopherol and tocotrienol standards can be obtained from Merck ; α-tocopherol can be
obtained from various suppliers. It has been reported that the purity of some commercially available tocopherol standards
may vary between 85 % and 100 %. Thus, it is important to determine the concentration of prepared calibration solutions
by UV spectrometry (see 9.1.1).
5.2 Tetrahydrofuran, filtered through an HPLC nylon filter (0,45 µm).
5.3 n-Heptane, filtered through an HPLC nylon filter (0,45 µm).
5.4 HPLC mobile phase: any suitable mixture of solvents that has been proved to reach a
chromatographic resolution of peaks as good as the one presented in Table 2 (relative retention time of
tocopherols and tocotrienols) and in Annex A (chromatograms of a mixture of vegetable oils), should be used
(see Table C.3).
The preparation of a suitable mobile phase, 3,85 % (volume fraction) tetrahydrofuran solution in n-heptane, is
as follows. Using a 1 00
...
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