SIST-TS CEN 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 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.

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

• Status: Published
• Public Enquiry End Date: 14-Dec-2020
• Publication Date: 16-Aug-2021
• ICS: 67.200.10 - Animal and vegetable fats and oils
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 20-Jul-2021
• Due Date: 24-Sep-2021
• Completion Date: 17-Aug-2021
• Ref Project: CEN ISO/TS 22115:2021 - Animal and vegetable fats and oils - Separation of lipid classes by capillary gas chromatography (fingerprint method) (ISO/TS 22115:2021)

Overview

CEN ISO/TS 22115:2021 - "Animal and vegetable fats and oils - Separation of lipid classes by capillary gas chromatography (fingerprint method)" - defines a semi-quantitative capillary GC fingerprinting method for oils, fats and oil/fat-related samples (including deodistillates). The technique provides a single-analysis overview of main lipid classes (e.g. triglycerides) and minor components (e.g. sterols, sterol esters, tocopherols, wax esters, fatty alcohols, glycerol). It is intended for screening and relative comparison of sample compositions rather than fully quantitative determination of pre‑identified compound classes.

Key topics and technical requirements

• Analytical principle: Hydroxylated compounds are converted to silyl derivatives and analyzed by capillary gas chromatography (GC) on a high‑temperature column; non‑hydroxylated (apolar) compounds remain unaffected.
• Instrumentation: Requires a GC for capillary columns with an on‑column injector (or equivalent), temperature‑programmable oven and flame ionization detector (FID); a fused silica capillary column rated up to ~400 °C with low film thickness is recommended.
• Reagents and standards:
  • Silylation reagent (example mixture: MSHFBA + methylimidazole, 1 ml + 50 µL).
  • Internal standard: 1,2,3‑tridecanoylglycerol.
  • Reference compounds used for response factor determination include oleic acid, mono/di/triglycerides, tocopherols, cholesterol, cholesterol esters, squalene, plant sterol mix, etc.
  • Solvents: isooctane, chloroform (chromatographic grade).
• Sample preparation: Follow ISO 661 for test‑sample preparation; perform silylation of sample and standards prior to GC.
• Data treatment: Peak identification and integration, calculation of response factors from reference standards, and semi‑quantitative reporting. The method includes criteria for repeatability, reproducibility and interlaboratory precision (informative annexes with example chromatograms and interlab results).
• Limitations: Designed as a fingerprint/screening method-use dedicated, validated quantitative methods when absolute quantification of a given class is required.

Applications and users

• Food and oil testing laboratories performing quality control, compositional screening and batch comparison.
• Producers of edible oils and fats and processors of fat‑derived ingredients who need rapid profiling of major and minor lipid classes.
• Regulatory or research labs conducting authenticity screening, compositional surveys or comparative studies of deodistillates and refined streams.
• Useful for preliminary screening prior to targeted quantitative analyses.

Related standards

• ISO 661 - Animal and vegetable fats and oils - Preparation of test sample (normative reference cited in CEN ISO/TS 22115:2021).
• CEN/TC 307 and ISO/TC 34 collaboration context - this document is a CEN‑endorsed ISO Technical Specification, initially provisional for three years.

Keywords: CEN ISO/TS 22115:2021, capillary gas chromatography, lipid classes, fingerprint method, oils and fats screening, silylation, GC‑FID, semi‑quantitative analysis.

Standards Content (Sample)

SLOVENSKI STANDARD
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
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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.

Contents Page
European foreword . 3

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.
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
ISO/TS 22115:2021(E)
© 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: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii © ISO 2021 – All rights reserved

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/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

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/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

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/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
...