EN 17644:2022
(Main)Foodstuffs - Detection of food allergens by liquid chromatography - mass spectrometry (LC-MS) methods - General considerations
Foodstuffs - Detection of food allergens by liquid chromatography - mass spectrometry (LC-MS) methods - General considerations
This document establishes an overall framework covering qualitative and quantitative methods for the determination of food allergens and allergenic ingredients using mass spectrometry-based methods for the determination of specific peptides/proteins. This document provides general guidelines and performance criteria applicable to this methodology. Guidelines, minimum requirements and performance criteria laid down in this document are intended to ensure that comparable and reproducible results are obtained by different analysts, instrumentation and laboratories.
Lebensmittel - Nachweis von Lebensmittelallergenen mit flüssigkeitschromatographisch-massenspektrometrischen (LC-MS) Verfahren - Allgemeine Betrachtungen
Dieses Dokument stellt einen Gesamtrahmen für die qualitativen und quantitativen Verfahren zur Bestimmung von Lebensmittelallergenen und allergenen Inhaltsstoffen unter Anwendung von auf der Massenspektrometrie basierenden Verfahren für die Bestimmung spezifischer Peptide/Proteine auf. Dieses Dokument bietet allgemeine Leitlinien und Leistungskriterien, die auf diese Verfahren anwendbar sind. Die in diesem Dokument angeführten Leitlinien, Mindestanforderungen und Leistungskriterien sind dafür vorgesehen sicherzustellen, dass von verschiedenen Analytikern, Geräten und Laboren vergleichbare und reproduzierbare Ergebnisse erhalten werden.
Produits alimentaires - Détection des allergènes alimentaires par des méthodes de chromatographie en phase liquide couplée à la spectrométrie de masse (CL-SM) - Considérations générales
Le présent document établit un cadre général couvrant les méthodes qualitatives et quantitatives pour la détermination des allergènes alimentaires et ingrédients allergisants par des méthodes de spectrométrie de masse pour la détermination de peptides/protéines spécifiques. Le présent document fournit les lignes directrices générales et les critères de performance applicables à cette méthode. Les lignes directrices, les exigences minimales et les critères de performance exposés dans le présent document ont pour but d’assurer l'obtention de résultats comparables et reproductibles, par différents analystes, instruments et laboratoires.
Živila - Odkrivanje prisotnosti alergenov v živilih s tekočinsko kromatografijo masno spektrometrijo (LC-MS) - Splošne ugotovitve
Ta dokument podaja splošen okvir kvalitativnih in kvantitativnih metod za določanje alergenov in alergenih sestavin z uporabo metod na osnovi masne spektrometrije za določitev specifičnih peptidov/proteinov. Ta dokument podaja splošne smernice in merila učinkovitosti za to metodologijo. Smernice, minimalne zahteve in merila učinkovitosti iz tega dokumenta so namenjeni zagotavljanju, da se pridobijo primerljivi ter ponovljivi rezultati s strani različnih analitikov, z različnimi instrumenti in v različnih laboratorijih.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-september-2022
Živila - Odkrivanje prisotnosti alergenov v živilih s tekočinsko kromatografijo
masno spektrometrijo (LC-MS) - Splošne ugotovitve
Foodstuffs - Detection of food allergens by liquid chromatography - mass spectrometry
(LC-MS) methods - General considerations
Lebensmittel - Nachweis von Lebensmittelallergenen mit flüssigkeitschromatographisch-
massenspektrometrischen (LC-MS) Verfahren - Allgemeine Betrachtungen
Produits alimentaires - Détection des allergènes alimentaires par des méthodes de
chromatographie en phase liquide couplée à la spectrométrie de masse (CL-SM) -
Considérations générales
Ta slovenski standard je istoveten z: EN 17644:2022
ICS:
67.050 Splošne preskusne in General methods of tests and
analizne metode za živilske analysis for food products
proizvode
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 17644
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2022
EUROPÄISCHE NORM
ICS 67.050
English Version
Foodstuffs - Detection of food allergens by liquid
chromatography - mass spectrometry (LC-MS) methods -
General considerations
Produits alimentaires - Détection des allergènes Lebensmittel - Nachweis von Lebensmittelallergenen
alimentaires par des méthodes de chromatographie en mit flüssigkeitschromatographisch-
phase liquide couplée à la spectrométrie de masse (CL- massenspektrometrischen (LC-MS) Verfahren -
SM) - Considérations générales Allgemeine Betrachtungen
This European Standard was approved by CEN on 22 May 2022.
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
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17644:2022 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 General laboratory requirements . 8
4.1 Principle . 8
4.2 Apparatus and equipment . 8
4.3 Material and reagents . 9
5 Method development . 9
5.1 General . 9
5.2 Sample preparation . 10
5.2.1 Grinding . 10
5.2.2 Extraction/purification . 10
5.2.3 Enzymatic digestion . 10
5.3 Detection . 11
5.3.1 General . 11
5.3.2 Selection of the target proteins/peptides . 11
5.3.3 Selection of measured MRM transitions . 12
5.3.4 Internal standard (IS) . 13
6 Method validation procedure . 13
6.1 General . 13
6.2 Method validation parameters . 14
6.2.1 Measurand . 14
6.2.2 Limit of detection and limit of quantification . 14
6.3 Selectivity . 15
6.4 Calibration curves . 15
6.5 Trueness . 15
6.6 Precision . 15
6.7 Measurement uncertainty . 16
6.8 Robustness. 16
6.9 Fit-for-purpose/applicability . 16
7 Routine analysis of allergenic food ingredients. 16
7.1 General . 16
7.2 Validation of an analytical run . 17
7.3 Test report . 18
Bibliography . 19
European foreword
This document (EN 17644:2022) has been prepared by Technical Committee CEN/TC 275 “Food
analysis - Horizontal methods”, the secretariat of which is held by DIN.
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 January 2023, and conflicting national standards shall
be withdrawn at the latest by January 2023.
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.
A complete listing of these bodies can be found on the CEN website.
According to the CEN-CENELEC Internal Regulations, the national standards organisations 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.
Introduction
Specific peptides or groups of peptides deriving from specific proteins can serve as markers for the
presence of food or food ingredients provoking allergic reactions. This document describes the procedure
for the development of qualitative and/or quantitative mass spectrometry-based methods for the
determination of protein-derived peptides as markers for potentially allergenic food ingredients or
constituents by analysing the protein/s extracted from a sample. Appropriate procedures for the
extraction of the targeted proteins are an essential part of each method. This document describes general
considerations for the application of liquid chromatography mass spectrometry-based methods in
qualitative or quantitative targeted analysis of specific peptides (derived from specific proteins) that are
representative for a food allergen. The document includes recommendations for method validation and
for the conversion of the analytical results to units of mg protein/kg food.
1 Scope
This document establishes an overall framework covering qualitative and quantitative methods for the
determination of food allergens and allergenic ingredients using mass spectrometry-based methods for
the determination of specific peptides/proteins. This document provides general guidelines and
performance criteria applicable to this methodology. Guidelines, minimum requirements and
performance criteria laid down in this document are intended to ensure that comparable and
reproducible results are obtained by different analysts, instrumentation and laboratories.
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.
EN 15842, Foodstuffs - Detection of food allergens - General considerations and validation of methods
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN 15842 and the following apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— IEC Electropedia: available at https://www.electropedia.org/
— ISO Online browsing platform: available at https://www.iso.org/obp
3.1
high-performance liquid chromatography–mass spectrometry
HPLC-MS
analytical chemistry technique that combines the separation capabilities of high-performance liquid
chromatography with the detection capabilities of mass spectrometry (MS)
Note 1 to entry: The abbreviation LC-MS is also used.
3.2
tandem mass spectrometry
MS/MS
MS
sequential combination of two mass analyses
Note 1 to entry: Different mass spectrometer instrument types exist, combining different principles of mass
detection, e.g. quadrupole, time-of-flight, ion trap, Fourier-Transform mass spectrometer.
3.3
multi-stage mass spectrometry
n
MS
sequential combination of more than two mass analyses
3.4
targeted mass spectrometry
mass spectrometry application analysing only specific ions (m/z) at specific retention times (RT)
Note 1 to entry: The targets are specified in an inclusion list.
Note 2 to entry: The opposite is untargeted MS measuring any ion present.
Note 3 to entry: In general, targeted MS increases method sensitivity.
3.5
peptide
amide that consists of two or more amino acids
Note 1 to entry: Peptides are formed by amide bonding of the amino group of one amino acid (AA) with the
carboxyl group of another AA.
Note 2 to entry: Peptides are usually obtained by enzymatic hydrolysis of proteins during sample preparation for
mass spectrometry-based methods.
3.6
marker peptide
peptide that is specific/unique for a definite protein and used as analyte in mass spectrometry-based
methods
Note 1 to entry: Portion of a pro
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