EN ISO 18363-4:2021

Animal and vegetable fats and oils - Determination of fatty-acid-bound chloropropanediols (MCPDs) and glycidol by GC/MS - Part 4: Method using fast alkaline transesterification and measurement for 2-MCPD, 3-MCPD and glycidol by GC-MS/MS (ISO 18363-4:2021)

This document specifies a rapid procedure for the simultaneous determination of 2-MCPD esters (bound 2-MCPD), 3‐MCPD esters (bound 3‐MCPD) and glycidyl esters (bound glycidol) in a single assay, based on alkaline catalysed ester cleavage and derivatization of cleaved (free) analytes with phenylboronic acid (PBA) prior to GC-MS/MS analysis. Glycidyl ester overestimation is corrected by addition of an isotopic labelled ester bound 3-MCPD which allows the quantification of 3-MCPD induced glycidol during the procedure.
This method is applicable to solid and liquid fats and oils. This document also applies to animal fats and used frying oils and fats, but these matrices were not included in the validation. For all three analytes the limit of quantification (LOQ) is 0,1 mg/kg and the limit of detection (LOD) is 0,03 mg/kg.
Milk and milk products (or fat coming from milk and milk products), infant formulas, emulsifiers, free fatty acids and other fats- and oils-derived matrices are excluded from the scope of this document.

Tierische und pflanzliche Fette und Öle - Bestimmung von fettsäuregebundenem Chlorpropandiol (MCPD) und Glycidol mittels GC/MS - Teil 4: Verfahren mittels schneller alkalischer Umesterung und Messung für 2-MCPD, 3-MCPD und Glycidol (ISO 18363-4:2021)

Dieses Dokument legt ein schnelles Verfahren für die gleichzeitige Bestimmung von 2 MCPD-Estern (gebundenes 2 MCPD), 3 MCPD-Estern (gebundenes 3 MCPD) und Glycidylestern (gebundenes Glycidol) in einer einzigen Prüfung auf der Grundlage einer alkalisch katalysierten Esterspaltung und Derivatisierung der abgespaltenen (freien) Analyten vor der GC MS/MS-Analyse mit Phenylboronsäure (PBA) fest. Die Überschätzung des Glycidylesters wird durch Zugabe eines isotopenmarkierten, estergebundenen 3-MCPD korrigiert, was die quantitative Bestimmung des durch 3-MCPD induzierten Glycidols während des Verfahrens ermöglicht.
Dieses Verfahren ist auf feste und flüssige Fette und Öle anwendbar. Dieses Dokument gilt auch für tierische Fette und gebrauchte Frittieröle und  fette, jedoch wurden diese Matrices nicht in die Validierungsstudie einbezogen. Für alle drei Analyten beträgt die Bestimmungsgrenze (LOQ, en: limit of quantification) 0,1 mg/kg und die Nachweisgrenze (LOD, en: limit of detection) 0,03 mg/kg.
Milch- und Milcherzeugnisse (oder Fett aus Milch und Milcherzeugnissen), Säuglings(anfangs)nahrung, Emulgatoren, freie Fettsäuren sowie andere aus Fetten und Ölen gewonnene Matrices sind vom Anwendungsbereich dieses Dokuments ausgeschlossen.

Corps gras d’origines animale et végétale - Détermination des esters de chloropropanediols (MCPD) et d’acides gras et des esters de glycidol et d’acides gras par CPG/SM - Partie 4: Méthode par transestérification alcaline rapide et mesure pour le 2-MCPD, le 3-MCPD et le glycidol par CPG-SM/SM (ISO 18363-4:2021)

Le présent document spécifie un mode opératoire rapide de détermination simultanée des esters de 2-MCPD (2-MCPD lié), des esters de 3-MCPD (3-MCPD lié) et des esters de glycidol (glycidol lié) en une seule analyse, reposant sur un clivage des esters en présence d’un catalyseur alcalin et une dérivation des analytes clivés (libres) avec de l’acide phénylboronique (PBA) avant l’analyse par CPG-SM/SM. La surestimation de la teneur en esters de glycidol est corrigée par l’ajout d’un ester de 3-MCPD marqué aux isotopes, ce qui permet la quantification du glycidol issu du 3-MCPD pendant le mode opératoire.
Cette méthode est applicable aux corps gras solides et liquides. Le présent document s’applique également aux graisses animales et aux corps gras de friture usagés, mais ces matrices n’étaient pas incluses dans la validation. Pour ces trois analytes, la limite de quantification (LQ) est de 0,1 mg/kg et la limite de détection (LOD) est de 0,03 mg/kg.
Le lait et les produits laitiers (ou les corps gras issus du lait et des produits laitiers), les formules infantiles, les émulsifiants, les acides gras libres et autres matrices dérivées de corps gras sont exclus du domaine d’application du présent document.

Živalske in rastlinske maščobe ter olja - Določevanje maščobno kislinsko vezanih kloropropandiolov (MCPDs) in glicidola z GC/MS - 4. del: Metoda z uporabo hitre alkalne transesterifikacije in meritev 2- MCPD, 3-MCPD in glicidolov z GC-MS/MS (ISO 18363-4:2021)

General Information

• Status: Published
• Publication Date: 17-Aug-2021
• Withdrawal Date: 27-Feb-2022
• ICS: 67.200.10 - Animal and vegetable fats and oils
• Technical Committee: CEN/TC 307 - Oilseeds, vegetables and animal fats and oils and their by-products - Methods of sampling and analys
• Drafting Committee: CEN/TC 307 - Oilseeds, vegetables and animal fats and oils and their by-products - Methods of sampling and analys
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 18-Aug-2021
• Completion Date: 18-Aug-2021
• Ref Project: SIST EN ISO 18363-4:2021 - Animal and vegetable fats and oils - Determination of fatty-acid-bound chloropropanediols (MCPDs) and glycidol by GC/MS - Part 4: Method using fast alkaline transesterification and measurement for 2-MCPD, 3-MCPD and glycidol by GC-MS/MS (ISO 18363-4:2021)

Standards Content (Sample)

SLOVENSKI STANDARD
01-november-2021
Živalske in rastlinske maščobe ter olja - Določevanje maščobno kislinsko vezanih
kloropropandiolov (MCPDs) in glicidola z GC/MS - 4. del: Metoda z uporabo hitre
alkalne transesterifikacije in meritev 2- MCPD, 3-MCPD in glicidolov z GC-MS/MS
(ISO 18363-4:2021)
Animal and vegetable fats and oils - Determination of fatty-acid-bound
chloropropanediols (MCPDs) and glycidol by GC/MS - Part 4: Method using fast alkaline
transesterification and measurement for 2-MCPD, 3-MCPD and glycidol by GC-MS/MS
(ISO 18363-4:2021)
Tierische und pflanzliche Fette und Öle - Bestimmung von fettsäuregebundenem
Chlorpropandiol (MCPD) und Glycidol mittels GC/MS - Teil 4: Verfahren mittels schneller
alkalischer Umesterung und Messung für 2-MCPD, 3-MCPD und Glycidol (ISO 18363-
4:2021)
Corps gras d’origines animale et végétale - Détermination des esters de
chloropropanediols (MCPD) et d’acides gras et des esters de glycidol et d’acides gras
par CPG/SM - Partie 4: Méthode par transestérification alcaline rapide et mesure pour le
2-MCPD, le 3-MCPD et le glycidol par CPG-SM/SM (ISO 18363-4:2021)
Ta slovenski standard je istoveten z: EN ISO 18363-4:2021
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.

EN ISO 18363-4
EUROPEAN STANDARD
NORME EUROPÉENNE
August 2021
EUROPÄISCHE NORM
ICS 67.200.10
English Version
Animal and vegetable fats and oils - Determination of fatty-
acid-bound chloropropanediols (MCPDs) and glycidol by
GC/MS - Part 4: Method using fast alkaline
transesterification and measurement for 2-MCPD, 3-MCPD
and glycidol by GC-MS/MS (ISO 18363-4:2021)
Corps gras d'origines animale et végétale - Tierische und pflanzliche Fette und Öle - Bestimmung
Détermination des esters de chloropropanediols von fettsäuregebundenem Chlorpropandiol (MCPD)
(MCPD) et d'acides gras et des esters de glycidol et und Glycidol mittels GC/MS - Teil 4: Verfahren mittels
d'acides gras par CPG/SM - Partie 4: Méthode par schneller alkalischer Umesterung und Messung für 2-
transestérification alcaline rapide et mesure pour le 2- MCPD, 3-MCPD und Glycidol (ISO 18363-4:2021)
MCPD, le 3-MCPD et le glycidol par CPG-SM/SM (ISO
18363-4:2021)
This European Standard was approved by CEN on 26 July 2021.

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 CEN-CENELEC 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 CEN-CENELEC Management
Centre has the same status as the official versions.

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. EN ISO 18363-4:2021 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 18363-4:2021) 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.
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 February 2022, and conflicting national standards
shall be withdrawn at the latest by February 2022.
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 implement this European Standard: 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 18363-4:2021 has been approved by CEN as EN ISO 18363-4:2021 without any
modification.
INTERNATIONAL ISO
STANDARD 18363-4
First edition
2021-08
Animal and vegetable fats and oils —
Determination of fatty-acid-bound
chloropropanediols (MCPDs) and
glycidol by GC/MS —
Part 4:
Method using fast alkaline
transesterification and measurement
for 2-MCPD, 3-MCPD and glycidol by
GC-MS/MS
Corps gras d’origines animale et végétale — Détermination des
esters de chloropropanediols (MCPD) et d’acides gras et des esters de
glycidol et d’acides gras par CPG/SM —
Partie 4: Méthode par transestérification alcaline rapide et mesure
pour le 2-MCPD, le 3-MCPD et le glycidol par CPG-SM/SM
Reference number
ISO 18363-4:2021(E)
©
ISO 2021
ISO 18363-4:2021(E)
© ISO 2021
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
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ISO 18363-4:2021(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Reagents . 2
5.1 General . 2
5.2 Standard and reference compounds . 2
5.3 Standard solutions . 3
5.3.1 General. 3
5.3.2 Stock solutions . 3
5.3.3 Working solutions . 3
5.4 Other reagents . 4
5.5 Reagent solutions . 4
6 Apparatus . 5
7 Sample and storage . 5
7.1 Sampling . 5
7.2 Preparation of the test sample . 5
7.3 Storage conditions . 6
8 Procedure. 6
8.1 Test sample preparation . 6
8.2 Preparation of the calibration curve . 7
8.3 Gas chromatography and mass spectrometry settings . 7
9 Expression of results . 8
9.1 General . 8
9.2 Quantification of 2-MCPD- and 3-MCPD esters . 8
9.3 Quantification of glycidyl esters .10
9.4 Quality control .12
10 Notes .13
11 Precision .14
11.1 General .14
11.2 Repeatability .14
11.3 Between-day reproducibility .14
Annex A (normative) Supporting tables .15
Annex B (i
...