SIST-TS CEN ISO/TS 22115:2021
(Main)Animal and vegetable fats and oils - Separation of lipid classes by capillary gas chromatography (fingerprint method) (ISO/TS 22115:2021)
Animal and vegetable fats and oils - Separation of lipid classes by capillary gas chromatography (fingerprint method) (ISO/TS 22115:2021)
This method is suitable for the semi-quantitative analysis of oils, fats and oil/fat-related samples
(deodistillates).
Screening of oils, fats and oil/fat-related samples to obtain main (e.g. TAGs) and minor component (e.g.
sterols, sterol esters, tocopherols, squalene, wax esters, fatty alcohols, and glycerol) information in one
single analysis. For a truly quantitative analysis of pre-identified compound classes specific methods
are more appropriate.
Beside the (semi-)quantitative determination of the oil/fat composition mentioned above, the method
can also be used as a useful qualitative screening tool for the relative comparison of sample
compositions.
Tierische und pflanzliche Fette und Öle - Relative Zusammensetzung von Ölen und Derivaten mittels Kapillargaschromatographie (Fingerprint-Verfahren) (ISO/TS 22115:2021)
Dieses Dokument legt ein Verfahren für die semiquantitative Analyse von Ölen, Fetten und öl-/fettbezogenen Proben (Deodestillaten) fest.
Es ist anwendbar für das Screening von Ölen, Fetten und öl-/fettbezogenen Proben, um Informationen über die Hauptbestandteile (z. B. Triglyceride) und Nebenbestandteile (z. B. Sterole, Sterolester, Tocopherole, Wachsester, Fettalkohole und Glycerol) in einer einzigen Analyse zu erhalten. Für eine wirklich quantitative Analyse von vorab identifizierten Verbindungsklassen sind spezifische Verfahren zweckmäßiger.
Das Verfahren kann auch als ein nützliches qualitatives Screening-Werkzeug für den relativen Vergleich von Probenzusammensetzungen angewendet werden.
Corps gras d'origines animale et végétale - Séparation des classes lipidiques par chromatographie en phase gazeuse sur colonne capillaire (méthode fingerprint) (ISO/TS 22115:2021)
Le présent document spécifie une méthode d’analyse semi-quantitative des corps gras et échantillons dérivés de corps gras (déodistillats).
Il est applicable à l’analyse des corps gras et échantillons dérivés de corps gras pour obtenir des informations sur les composés majoritaires (par exemple triglycérides) et minoritaires (par exemple stérols, esters de stérol, tocophérols, esters de cire, alcools gras, glycérol) au cours d'une seule analyse. Pour une analyse réellement quantitative des classes de composés préalablement identifiées, les méthodes spécifiques sont plus appropriées.
La méthode peut également être utilisée comme outil d’analyse qualitative en vue d'effectuer une comparaison relative des compositions d'échantillons.
Živalske in rastlinske maščobe in olja - Ločevanje lipidnih razredov s kapilarno plinsko kromatografijo (metoda fingerprint) (ISO/TS 22115:2021)
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST-TS CEN ISO/TS 22115:2021
01-september-2021
Živalske in rastlinske maščobe in olja - Ločevanje lipidnih razredov s kapilarno
plinsko kromatografijo (metoda fingerprint) (ISO/TS 22115:2021)
Animal and vegetable fats and oils - Separation of lipid classes by capillary gas
chromatography (fingerprint method) (ISO/TS 22115:2021)
Tierische und pflanzliche Fette und Öle - Relative Zusammensetzung von Ölen und
Derivaten mittels Kapillargaschromatographie (Fingerprint-Verfahren) (ISO/TS
22115:2021)
Corps gras d'origines animale et végétale - Séparation des classes lipidiques par
chromatographie en phase gazeuse sur colonne capillaire (méthode fingerprint) (ISO/TS
22115:2021)
Ta slovenski standard je istoveten z: CEN ISO/TS 22115:2021
ICS:
67.200.10 Rastlinske in živalske Animal and vegetable fats
maščobe in olja and oils
SIST-TS CEN ISO/TS 22115:2021 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST-TS CEN ISO/TS 22115:2021
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SIST-TS CEN ISO/TS 22115:2021
CEN ISO/TS 22115
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
July 2021
TECHNISCHE SPEZIFIKATION
ICS 67.200.10
English Version
Animal and vegetable fats and oils - Separation of lipid
classes by capillary gas chromatography (fingerprint
method) (ISO/TS 22115:2021)
Corps gras d'origines animale et végétale - Séparation Tierische und pflanzliche Fette und Öle - Relative
des classes lipidiques par chromatographie en phase Zusammensetzung von Ölen und Derivaten mittels
gazeuse sur colonne capillaire (méthode fingerprint) Kapillargaschromatographie (Fingerprint-Verfahren)
(ISO/TS 22115:2021) (ISO/TS 22115:2021)
This Technical Specification (CEN/TS) was approved by CEN on 29 January 2021 for provisional application.
The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.
CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.
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, Republic of North Macedonia, Romania, Serbia, 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: Rue de la Science 23, B-1040 Brussels
© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN ISO/TS 22115:2021 E
worldwide for CEN national Members.
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SIST-TS CEN ISO/TS 22115:2021
CEN ISO/TS 22115:2021 (E)
Contents Page
European foreword . 3
2
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SIST-TS CEN ISO/TS 22115:2021
CEN ISO/TS 22115:2021 (E)
European foreword
This document (CEN ISO/TS 22115:2021) 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.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN websites.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to announce this Technical Specification: 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, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the
United Kingdom.
Endorsement notice
The text of ISO/TS 22115:2021 has been approved by CEN as CEN ISO/TS 22115:2021 without any
modification.
3
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SIST-TS CEN ISO/TS 22115:2021
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SIST-TS CEN ISO/TS 22115:2021
TECHNICAL ISO/TS
SPECIFICATION 22115
First edition
2021-06
Animal and vegetable fats and oils —
Separation of lipid classes by capillary
gas chromatography (fingerprint
method)
Corps gras d'origines animale et végétale — Séparation des classes
lipidiques par chromatographie en phase gazeuse sur colonne
capillaire (méthode fingerprint)
Reference number
ISO/TS 22115:2021(E)
©
ISO 2021
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SIST-TS CEN ISO/TS 22115:2021
ISO/TS 22115:2021(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2021
All rights reserved. Unless otherwise specified, or required in the context of its implementation, 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
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2021 – All rights reserved
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SIST-TS CEN ISO/TS 22115:2021
ISO/TS 22115:2021(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 . 3
7 Sample . 3
7.1 Sampling . 3
7.2 Preparation of test sample . 3
8 Procedure. 3
8.1 Preparation of the internal standard, tridecanoylglycerol (5.2.5), c = 20 mg/ml . 3
8.2 Preparation of the individual standard solutions for determination of response factors . 3
8.3 Silylation of the standards . 4
8.4 Preparation of the sample . 4
8.5 Sample solution . 4
8.6 Silylation of the sample . 4
8.7 Gas chromatography . 4
8.8 Peak identification and integration . 5
9 Result of the determination . 6
9.1 Calculation of the response factor . 6
9.2 Quantitative determination . 6
10 Precision of the method . 7
10.1 Interlaboratory test. 7
10.2 Repeatability . 7
10.3 Reproducibility . 7
11 Test report . 7
Annex A (informative) Typical chromatograms . 8
Annex B (informative) Results of an interlaboratory test .18
Bibliography .22
© ISO 2021 – All rights reserved iii
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SIST-TS CEN ISO/TS 22115:2021
ISO/TS 22115:2021(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 of the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO’s adherence to the
World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www .iso .org/
iso/ foreword .html.
This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11,
Animal and vegetable fats and oils, in collaboration with the European Committee for Standardization
(CEN) Technical Committee CEN/TC 307, Oilseeds, vegetable and animal fats and oils and their by-
products — Methods of sampling and analysis, in accordance with the Agreement on technical cooperation
between ISO and CEN (Vienna Agreement).
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www .iso .org/ members .html.
iv © ISO 2021 – All rights reserved
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SIST-TS CEN ISO/TS 22115:2021
TECHNICAL SPECIFICATION ISO/TS 22115:2021(E)
Animal and vegetable fats and oils — Separation of lipid
classes by capillary gas chromatography (fingerprint
method)
1 Scope
This document specifies a method for the semi-quantitative analysis of oils, fats and oil/fat-related
samples (deodistillates).
It is applicable to the screening of oils, fats and oil/fat-related samples to obtain main (e.g. triglycerides)
and minor (e.g. sterols, sterol esters, tocopherols, wax esters, fatty alcohols, glycerol) component
information in one single analysis. For a truly quantitative analysis of pre-identified compound classes,
specific methods are more appropriate.
The method can also be used as a useful qualitative screening tool for the relative comparison of sample
compositions.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements 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
No terms and definitions are listed in this document.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
4 Principle
The hydroxylated compounds are transformed into silyl derivatives. This operation has no effect on the
apolar (non-hydroxylated) compounds also present in the sample. The sample prepared is analysed by
gas chromatography (GC) on a high-temperature capillary column with a low film thickness, with an
on-column injector and flame-ionization detector.
For quantitative purposes, the compounds are quantified in the presence of an internal standard
(1,2,3-tridecanoylglycerol) and the response factors are determined from a reference standard from
each class.
© ISO 2021 – All rights reserved 1
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SIST-TS CEN ISO/TS 22115:2021
ISO/TS 22115:2021(E)
5 Reagents
WARNING — Attention is drawn to the regulations which specify the handling of hazardous
substances. Technical, organizational and personal safety measures shall be followed.
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and
distilled or demineralized water or water of equivalent purity.
5.1 Silylation reagent mixture: Methylimidazole (CAS 616-47-7) and MSHFBA (CAS 53296-64-3).
Prepare a mixture of silylation reagents: 1 ml of MSHFBA + 50 µl of Methylimidazole.
NOTE This mixture cannot be stored more than one week due to moisture absorption. Indeed, silyl
derivatives are moisture sensitive.
1)
5.2 Reference substances .
5.2.1 Oleic acid (CAS 112-80-1).
5.2.2 1-Monoolein (CAS 111-03-5).
5.2.3 1,3-Diolein (CAS 2465-32-9).
5.2.4 Triolein (CAS 122-32-7).
5.2.5 Tridecanoylglycerol (CAS 621-71-6).
5.2.6 Eicosanol (CAS 629-96-9).
5.2.7 α-Tocopherol (CAS 10191-41-0), γ tocopherol (CAS 54-28-4), δ tocopherol (CAS 119-13-1).
5.2.8 Cholesterol (CAS 57-88-5).
5.2.9 Cholesterol palmitate (CAS 601-34-3).
5.2.10 Squalene (CAS 111-02-4).
2)
5.2.11 Plant sterol mix (CAS 474-67-9, 474-62-4, 83-48-7, 83-46-5).
5.3 Isooctane, trace organic analysis grade, purity 99 % min.
5.4 Chloroform, chromatographic quality.
1) Suitable suppliers are Sigma-Aldrich (www .sigmaaldrich .com) or Larodan (www .larodan .com). This
information is given for the convenience of users of this document and does not constitute an endorsement by ISO
of this supplier.
2) A suitable supplier is Larodan (www .larodan .com). This information is given for the convenience of users of
this document and does not constitute an endorsement by ISO of this supplier.
2 © ISO 2021 – All rights reserved
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SIST-TS CEN ISO/TS 22115:2021
ISO/TS 22115:2021(E)
6 Apparatus
Usual laboratory equipment and, in particular, the following.
6.1 Volumetric flask, 100 ml and 10 ml.
6.2 Oven, at 103 °C ± 1 °C
6.3 Conical glass sample vials, 10 ml capacity.
6.4 Gas chromatograph for capillary columns, equipped with an on-column injector or equivalent
device, a temperature-programmable oven and a flame ionization detector (FID).
6.5 Fused silica capillary column, capable of being programmed up to 400 °C (“high temperature”
type) for which the following characteristics are advised: 100 % dimethylpolysiloxane or 95 % dimethyl/
5 % diphenyl polysiloxane stationary phase, length 30 m, internal diameter 0,32 mm or 0,25 mm, film
thickness 0,1 µm. Other columns of similar polarity and selectivity may also be used.
6.6 Microsyringe, 5 µl to 10 µl capacity, suitable for on-column injection in gas chromatography.
6.7 Analytical balance, reading accuracy 0,001 g.
6.8 Ultrasonic bath.
6.9 Nitrogen evaporator.
7 Sample
7.1 Sampling
Sampling is not part of the method specified in this document. A recommended sampling method is
[1]
given in ISO 5555 .
It is important that the laboratory receive a sample which is truly representative and has not been
damaged or changed during transport or storage.
7.2 Preparation of test sample
Prepare the test sample in accordance with ISO 661.
8 Procedure
8.1 Preparation of the internal standard, tridecanoylglycerol (5.2.5), c = 20 mg/ml
Weigh 200 mg tridecanoylglycerol (5.2.5) into a 10 ml volumetric flask, fill it up with isooctane (5.3).
8.2 Preparation of the individual standard solutions for determination of response
factors
A 5 mg/ml standard solution in isooctane (5.3) is prepared for each compound (see Table 1) to determine
the response factor for each component group.
© ISO 2021 – All rights reserved 3
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SIST-TS CEN ISO/TS 22115:2021
ISO/TS 22115:2021(E)
Table 1 — List of the standards for the calculation of the response factors
Class determined Standard
Alcohols Eicosanol (5.2.6)
Free fatty acids Oleic acid (5.2.1)
Hydrocarbons Squalene (5.2.10)
Monoglycerides Monoolein (5.2.2)
Tocopherols α-Tocopherol (5.2.7)
Sterols Cholesterol (5.2.8)
Diglycerides Diolein (5.2.3)
Sterol esters Cholesterol palmitate (5.2.9)
Triglycerides Triolein (5.2.4)
8.3 Silylation of the standards
Introduce 100 µl of the internal standard solution, tridecanoylglycerol (5.2.5), c = 20 mg/ml, and
200 µl of the individual standard solution (each component separately) into a silylation tube (6.3) and
evaporate the solvent to dryness under nitroge
...
SLOVENSKI STANDARD
kSIST-TS FprCEN ISO/TS 22115:2020
01-december-2020
Živalske in rastlinske maščobe in olja - Določanje relativne sestave olj in derivatov
s kapilarno plinsko kromatografijo (metoda fingerprint)
Animal and vegetable fats and oils - Relative composition of oils and derivatives by
capillary gas chromatography (fingerprint method)
Tierische und pflanzliche Fette und Öle - Relative Zusammensetzung von Ölen und
Derivaten mittels Kapillargaschromatographie (Fingerprint-Verfahren)
Corps gras d'origines animale et végétale - Composition relative des corps gras et
produits dérivés par chromatographie gazeuse capillaire (méthode fingerprint)
Ta slovenski standard je istoveten z: FprCEN ISO/TS 22115
ICS:
67.200.10 Rastlinske in živalske Animal and vegetable fats
maščobe in olja and oils
kSIST-TS FprCEN ISO/TS 22115:2020 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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kSIST-TS FprCEN ISO/TS 22115:2020
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kSIST-TS FprCEN ISO/TS 22115:2020
TECHNICAL ISO/TS
SPECIFICATION 22115
First edition
Animal and vegetable fats and
oils — Relative composition of oils
and derivatives by capillary gas
chromatography (fingerprint method)
Corps gras d'origines animale et végétale — Composition relative
des corps gras et produits dérivés par chromatographie gazeuse
capillaire (méthode fingerprint)
PROOF/ÉPREUVE
Reference number
ISO/TS 22115:2020(E)
©
ISO 2020
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kSIST-TS FprCEN ISO/TS 22115:2020
ISO/TS 22115:2020(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2020
All rights reserved. Unless otherwise specified, or required in the context of its implementation, 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
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii PROOF/ÉPREUVE © ISO 2020 – All rights reserved
---------------------- Page: 4 ----------------------
kSIST-TS FprCEN ISO/TS 22115:2020
ISO/TS 22115:2020(E)
Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 1
5 Reagents . 1
6 Apparatus . 2
7 Sample . 3
7.1 Sampling . 3
7.2 Preparation of test sample . 3
8 Procedure. 3
8.1 Preparation of the internal standard, Tridecanoylglycerol (5.2.5), c = 20 mg/ml . 3
8.2 Preparation of the individual standard solutions for determination of response factors . 3
8.3 Silylation of the standards . 4
8.4 Preparation of the sample . 4
8.5 Sample solution . 4
8.6 Silylation of the sample . 4
8.7 Gas chromatography . 4
8.8 Peak identification and integration . 5
9 Result of the determination . 5
9.1 Calculation of the response factor . 5
9.2 Quantitative determination . 6
10 Precision of the method . 7
10.1 Interlaboratory test. 7
10.2 Repeatability . 7
10.3 Reproducibility . 7
11 Test report . 7
Annex A (informative) Typical chromatograms . 8
Annex B (informative) Results of an interlaboratory test .18
Bibliography .22
© ISO 2020 – All rights reserved PROOF/ÉPREUVE iii
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kSIST-TS FprCEN ISO/TS 22115:2020
ISO/TS 22115:2020(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 of the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO’s adherence to the
World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www .iso .org/
iso/ foreword .html.
The committee responsible for this document is ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils, in collaboration with the European Committee for Standardization (CEN)
Technical Committee CEN/TC 307, Oilseeds, vegetable and animal fats and oils and their by-products —
Methods of sampling and analysis, in accordance with the Agreement on technical cooperation between
ISO and CEN (Vienna Agreement).
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www .iso .org/ members .html.
iv PROOF/ÉPREUVE © ISO 2020 – All rights reserved
---------------------- Page: 6 ----------------------
kSIST-TS FprCEN ISO/TS 22115:2020
TECHNICAL SPECIFICATION ISO/TS 22115:2020(E)
Animal and vegetable fats and oils — Relative composition
of oils and derivatives by capillary gas chromatography
(fingerprint method)
1 Scope
This document specifies a method for the semi-quantitative analysis of oils, fats and oil/fat-related
samples (deodistillates).
It is applicable to the screening of oils, fats and oil/fat-related samples to obtain main (e.g. triglycerides)
and minor (e.g. sterols, sterol esters, tocopherols, wax esters, fatty alcohols, glycerol) component
information in one single analysis. For a truly quantitative analysis of pre-identified compound classes,
specific methods are more appropriate.
The method can also be used as a useful qualitative screening tool for the relative comparison of sample
compositions.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements 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
No terms and definitions are listed in this document.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
4 Principle
The hydroxylated compounds are transformed into silyl derivatives. This operation has no effect on the
apolar (non-hydroxylated) compounds also present in the sample. The sample prepared is analysed by
gas chromatography (GC) on a high-temperature capillary column with a low film thickness, with an
on-column injector and flame-ionization detector.
For quantitative purposes, the compounds are quantified in the presence of an internal standard
(1,2,3-Tridecanoylglycerol) and the response factors are determined from a reference standard from
each class.
5 Reagents
WARNING — Attention is drawn to the regulations which specify the handling of hazardous
substances. Technical, organizational and personal safety measures shall be followed.
© ISO 2020 – All rights reserved PROOF/ÉPREUVE 1
---------------------- Page: 7 ----------------------
kSIST-TS FprCEN ISO/TS 22115:2020
ISO/TS 22115:2020(E)
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and
distilled or demineralized water or water of equivalent purity.
5.1 Silylation reagent mixture: Methylimidazole (CAS 616-47-7) and MSHFBA (CAS 53296-64-3).
Prepare a mixture of silylation reagents: 1 ml of MSHFBA + 50 µl of Methylimidazole.
NOTE This mixture cannot be stored more than one week due to moisture absorption. Indeed, silyl
derivatives are moisture sensitive.
1)
5.2 Reference substances .
5.2.1 Oleic acid (CAS 112-80-1).
5.2.2 1-Monoolein (CAS 111‑03‑5).
5.2.3 1,3-Diolein (CAS 2465-32-9).
5.2.4 Triolein (CAS 122‑32‑7).
5.2.5 Tridecanoylglycerol (CAS 621-71-6).
5.2.6 Eicosanol (CAS 629-96-9).
5.2.7 α-Tocopherol (CAS 10191-41-0), γ tocopherol (CAS 54-28-4), δ tocopherol (CAS 119‑13‑1).
5.2.8 Cholesterol (CAS 57-88-5).
5.2.9 Cholesterol palmitate (CAS 601-34-3).
5.2.10 Squalene (CAS 111‑02‑4).
2)
5.2.11 Plant sterol mix (CAS 474-67-9, 474-62-4, 83-48-7, 83-46-5).
5.3 Isooctane, trace organic analysis grade, purity 99 % min.
5.4 Chloroform, chromatographic quality.
6 Apparatus
Usual laboratory equipment and, in particular, the following.
6.1 Volumetric flask, 100 ml and 10 ml.
6.2 Oven, at 103 °C ± 1 °C
6.3 Conical glass sample vials, 10 ml capacity.
1) Suitable suppliers are Sigma-Aldrich (www .sigmaaldrich .com) or Larodan (www .larodan .com). This
information is given for the convenience of users of this document and does not constitute an endorsement by ISO
of this supplier.
2) A suitable supplier is Larodan (www .larodan .com). This information is given for the convenience of users of
this document and does not constitute an endorsement by ISO of this supplier.
2 PROOF/ÉPREUVE © ISO 2020 – All rights reserved
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ISO/TS 22115:2020(E)
6.4 Gas chromatograph for capillary columns, equipped with an on-column injector or equivalent
device, a temperature-programmable oven and a flame ionization detector (FID).
6.5 Fused silica capillary column, capable of being programmed up to 400 °C (“high temperature”
type) for which the following characteristics are advised: 100 % dimethylpolysiloxane or 95 % dimethyl/
5 % diphenyl polysiloxane stationary phase, length 30 m, internal diameter 0,32 mm or 0,25 mm, film
thickness 0,1 µm. Other columns of similar polarity and selectivity may also be used.
6.6 Microsyringe, 5 µl to 10 µl capacity, suitable for on-column injection in gas chromatography.
6.7 Analytical balance, reading accuracy 0,001 g.
6.8 Ultrasonic bath.
6.9 Nitrogen evaporator.
7 Sample
7.1 Sampling
Sampling is not part of the method specified in this document. A recommended sampling method is
[1]
given in ISO 5555 .
It is important that the laboratory receive a sample which is truly representative and has not been
damaged or changed during transport or storage.
7.2 Preparation of test sample
Prepare the test sample in accordance with ISO 661.
8 Procedure
8.1 Preparation of the internal standard, Tridecanoylglycerol (5.2.5), c = 20 mg/ml
Weigh 200 mg Tridecanoylglycerol (5.2.5) into a 10 ml volumetric flask, fill it up with isooctane (5.3).
8.2 Preparation of the individual standard solutions for determination of response
factors
A 5 mg/ml standard solution in isooctane (5.3) is prepared for each compound (see Table 1) to determine
the response factor for each component group.
Table 1 — List of the standards for the calculation of the response factors
Class determined Standard
Alcohols Eicosanol (5.2.6)
Free fatty acids Oleic acid (5.2.1)
Hydrocarbons Squalene (5.2.10)
Monoglycerides Monoolein (5.2.2)
Tocopherols α-Tocopherol (5.2.7)
Sterols Cholesterol (5.2.8)
Diglycerides Diolein (5.2.3)
© ISO 2020 – All rights reserved PROOF/ÉPREUVE 3
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Table 1 (continued)
Class determined Standard
Sterol esters Cholesterol palmitate (5.2.9)
Triglycerides Triolein (5.2.4)
8.3 Silylation of the standards
Introduce 100 µl of the internal standard solution, Tridecanoylglycerol (5.2.5), c = 20 mg/ml, and
200 µl of the individual standard solution (each component separately) into a silylation tube (6.3) and
evaporate the solvent to dryness under nitrogen (6.9). Introduce 100 µl of the silylation mixture (5.1).
Keep at 103 °C for 15 min (6.2), then take up into 5 ml isooctane (5.3).
It is recommended when preparing the following solution to check the method precision (optional).
In a 100 ml volumetric flask (6.1), exactly weigh about 50 mg of each standard (5.2), including 50 mg of
the internal standard solution, Tridecanoylglycerol (5.2.5), and dilute to 100 ml with chloroform (5.4).
This solution can be stored at ambient temperature for six months.
To keep the solution homogenous over time, the solution has to be placed into an ultrasonic bath for
10 min before use.
Introduce 0,5 ml of the solution into a silylation tube (6.3) and dry under nitrogen (6.9). Introduce 100 µl
of the silylation mixture (5.1). Keep at 103 °C for 15 min (6.2), then take up into 10 ml isooctane (5.3).
8.4 Preparation of the sample
If the sample is solid at room temperature, completely melt the sample in a 103 °C oven (6.2) and
homogenize it.
8.5 Sample solution
In a 10 ml volumetric flask (6.1), exactly weigh about 100 mg of sample. Introduce exactly 1 ml of
internal standard solution (see 8.1) and dilute to 10 ml with isooctane (5.3).
If the sample is not totally soluble in isooctane, chloroform or toluene may be used instead of isooctane.
8.6 Silylation of the sample
Introduce 0,5 ml of the sample solution (see 8.5) into a silylation tube (6.3) and evaporate the solvent to
dryness under nitrogen (6.9.). Introduce 100 µl of the silylation mixture (5.1). Keep at 103 °C for 15 min
(6.2), then take up into 10 ml isooctane (5.3).
8.7 Gas chromatography
Install the column in the gas chromatograph and check the working conditions by injecting the solvent,
isooctane (5.3). The baseline should be straight with a small positive drift. If the drift is high, proceed
to condition the column. For a negative drift, check the connections of the column.
If the column is being used for the first time, it is necessary to condition the column by heating it in the
oven using a temperature gradient up to 370 °C (depending on the oven t
...
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