EN 16274:2021
(Main)Method for analysis of allergens - Quantification of an extended list of 57 suspected allergens in ready to inject fragrance materials by gas chromatography mass spectrometry
Method for analysis of allergens - Quantification of an extended list of 57 suspected allergens in ready to inject fragrance materials by gas chromatography mass spectrometry
The present method permits the identification and quantification of the volatile compounds suspected as allergens, which are present in the fragrance compounds and fragrance raw materials used in cosmetic products. The analysis is performed by gas chromatography and mass spectrometry (GC-MS) on matrix samples which are "ready to be injected" and which are compatible with gas chromatography.
The analytes covered by this procedure are based on the contents of Tables 13.1 and 13.2 in the SCCS 1459/11 opinion document (1) and as listed in the legislation proposed by the European Commission. The rationale behind the final choice of procedure analytes is given in the table found in Annex J.
The method was validated at IFRA and CEN level.
Analyseverfahren für Allergene - Quantifizierung einer erweiterten Liste von 57 zu vermutenden Allergenen in einspritzfertigen Duftstoffen mittels Gaschromatographie/Massenspektrometrie
Das vorliegende Verfahren ermöglicht die Identifizierung und Quantifizierung der als Allergene vermuteten flüchtigen Verbindungen, die in den in kosmetischen Mitteln verwendeten Verbindungen von Duftstoffen und Rohstoffen von Duftstoffen enthalten sind. Die Analyse wird mithilfe der Gaschromatographie und Massenspektrometrie (GC MS) an Matrixproben durchgeführt, die „einspritzfertig“ und mit der Gaschromatographie kompatibel sind.
Die von diesem Verfahren erfassten Analyten basieren auf den Inhalten der in der Stellungnahme des SCCS 1459/11 (1) enthaltenen Tabelle 13.1 und Tabelle 13.2 und den von der Europäischen Kommission vorgeschlagenen aufgeführten Rechtsvorschriften. Die Begründung für die endgültige Auswahl der während des Verfahrens angewendeten Analyten ist in der in Anhang J enthaltenen Tabelle aufgeführt.
Das Verfahren wurde auf IFRA und CEN Ebene validiert.
Méthode d’analyse des allergènes - Quantification de la liste étendue des 57 allergènes suspectés dans les matières premières de parfumerie et les compositions parfumantes prêtes à être injectées, par chromatographie en phase gazeuse/spectrométrie de masse
La présente méthode permet l’identification et la quantification des composés volatils suspectés allergènes, qui sont présents dans les matières premières de parfumerie et les compositions parfumantes utilisées dans les produits cosmétiques. L’analyse est réalisée par chromatographie en phase gazeuse couplée à la spectrométrie de masse (CG-SM) sur des échantillons de matrice qui sont « prêts à être injectés » et qui sont compatibles avec la chromatographie en phase gazeuse.
Les analytes couverts par ce mode opératoire sont basés sur le contenu des Tableaux 13.1 et 13.2 du document d’opinion CSSC 1459/11 (1) et sont mentionnés dans la législation proposée par la Commission européenne. Les raisons qui justifient le choix final des analytes pour ce mode opératoire sont données dans le tableau figurant à l’Annexe J.
La méthode a été validée au niveau de l’IFRA et du CEN.
Analizne metode za alergene - Kvantitativno določevanje 57 domnevnih alergenov z razširjenega seznama v dišavnih izdelkih s plinsko kromatografsko analizo vzorcev, ki so pripravljeni za injeciranje, in masno spektrometrijo
Namen predlaganega standarda je opisati metodo za analizo 57 kemično določenih domnevnih alergenov (nekateri od njih obstajajo v več izomernih oblikah ali kot mešanice) v vzorcih dišav in surovin, ki so pripravljeni za injiciranje v skladu z mnenjem SCCS (SCCS/1459/11).
Ta nova analitična metoda uporablja plinsko kromatografijo in masno spektrometrijo (GC-MS) za odkrivanje in kvantitativno določevanje 57 dišavnih snovi in njihovih pomembnih izomerov s koncentracijo, ki presega 0,0002 % (2 mg/kg), v vzorcih dišav in surovin, ki so pripravljeni za injiciranje. Z zagotavljanjem te metode bo omogočeno presejanje (kompleksnih) vzorcev dišav in surovin, ki so pripravljeni za injiciranje, s čimer se ugotavlja prisotnost katerega koli od teh kemično določenih domnevnih alergenov. To bo torej osnova za izračun ustreznih informacij, na podlagi katerih bo lahko kozmetična industrija zagotovila ustrezno obveščanje potrošnikov. Ta analitična metoda uporablja plinsko kromatografijo in masno spektrometrijo s kombinacijo dveh kolon GC različne polarnosti s posebno metodologijo za kvantitativno določevanje.
General Information
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Standards Content (Sample)
SLOVENSKI STANDARD
01-januar-2022
Nadomešča:
SIST EN 16274:2012
Analizne metode za alergene - Kvantitativno določevanje 57 domnevnih alergenov
z razširjenega seznama v dišavnih izdelkih s plinsko kromatografsko analizo
vzorcev, ki so pripravljeni za injeciranje, in masno spektrometrijo
Method for analysis of allergens - Quantification of an extended list of 57 suspected
allergens in ready to inject fragrance materials by gas chromatography mass
spectrometry
Analyseverfahren für Allergene - Quantifizierung einer erweiterten Liste von 57 zu
vermutenden Allergenen in einspritzfertigen Duftstoffen mittels
Gaschromatographie/Massenspektrometrie
Méthode d’analyse des allergènes - Quantification de la liste étendue des 57 allergènes
suspectés dans les matières premières de parfumerie et les compositions parfumantes
prêtes à être injectées, par chromatographie en phase gazeuse/spectrométrie de masse
Ta slovenski standard je istoveten z: EN 16274:2021
ICS:
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
71.100.70 Kozmetika. Toaletni Cosmetics. Toiletries
pripomočki
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 16274
EUROPEAN STANDARD
NORME EUROPÉENNE
May 2021
EUROPÄISCHE NORM
ICS 71.100.60 Supersedes EN 16274:2012
English Version
Method for analysis of allergens - Quantification of an
extended list of 57 suspected allergens in ready to inject
fragrance materials by gas chromatography mass
spectrometry
Méthode d'analyse des allergènes - Quantification de la Analyseverfahren für Allergene - Quantifizierung einer
liste étendue des 57 allergènes suspectés dans les erweiterten Liste von 57 zu vermutenden Allergenen
matières premières de parfumerie et les compositions in einspritzfertigen Duftstoffen mittels
parfumantes prêtes à être injectées, par Gaschromatographie/Massenspektrometrie
chromatographie en phase gazeuse/spectrométrie de
masse
This European Standard was approved by CEN on 8 February 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 16274:2021 E
worldwide for CEN national Members.
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 6
5 Reagents . 7
5.1 Solvents . 7
5.1.1 Methyl pivalate . 7
5.1.2 Tert-Butyl Methyl Ether (MTBE) . 7
5.2 Reference samples (suspected allergens) . 7
5.2.1 General. 7
6 Apparatus . 11
6.1 Gas chromatograph equipped with flame ionization detector (GC-FID) . 11
6.2 Gas chromatograph coupled to a mass spectrometer (GC-MS) . 11
6.2.1 General. 11
6.2.2 GC-MS system. 11
6.3 Capillary columns for GC . 11
6.4 Analytical balance . 12
7 Stock and sample solutions - preparation and storage . 12
7.1 General information . 12
7.1.1 Choice of the solvent . 12
7.1.2 Miscellaneous . 13
7.2 Preparation of stock solutions from reference samples . 13
7.3 Preparation of internal standard solutions and calibration solutions . 13
7.3.1 General. 13
7.3.2 Stock Solutions of Internal Standards (1g/kg equivalent) . 13
7.3.3 Calibration Solutions Concentrations . 13
7.3.4 Preparation of Calibration solutions . 14
8 Mass spectrometer acquisition conditions . 16
8.1 Establishment of retention times and SIM chromatograph windows . 16
8.2 SIM Window – Set up and criteria . 17
8.3 GC-MS scan mode verification . 17
9 Sample analysis . 17
9.1 General. 17
9.2 Sample preparation for analysis . 17
9.3 Sequence . 18
9.3.1 Blank . 18
9.3.2 Calibration solutions . 18
9.3.3 Check solution . 18
9.3.4 Samples . 19
10 Data validation and treatment . 19
10.1 General. 19
10.2 Examination of Q values . 19
10.3 Relative intensity of characteristic ions in SIM . 20
10.4 Relative ion intensity using scan data . 20
10.5 Data verification scheme and reporting of final concentration . 21
10.6 GC-MS scan mode verification . 21
10.7 Mass spectrum evaluation in SCAN mode . 21
11 Test report . 21
Annex A (informative) Determination of calibrant and reference sample purity . 22
Annex B (informative) GC capillary column parameters . 29
Annex C (informative) SIM Ions for SIM or SIM-SCAN Use. 30
Annex D (informative) Example of SIM windows . 37
Annex E (informative) Example of chromatograms . 46
Annex F (normative) Decisional tree for quantification of suspected allergens . 48
Annex G (informative) Preparation of stock solutions from reference samples . 49
Annex H (informative) Calibration methods and approach . 54
Annex I (informative) Quantification of allergens . 56
Annex J (informative) Suspected allergens selected as analytes targets – Rationale . 59
Annex K (informative) Other general information. 69
Bibliography . 70
European foreword
This document (EN 16274:2021) has been prepared by Technical Committee CEN/TC 347 “Methods for
analysis of allergens”, the secretariat of which is held by SNV.
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 November 2021, and conflicting national standards shall
be withdrawn at the latest by November 2021.
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.
This document supersedes EN 16274:2012.
The most significant technical changes made in comparison to the previous version are as follows:
— Number of allergens has been extended from 26 to 57 chemically defined molecules.
— Solvents: moves from methyl pivalate and ortho fluorotoluene and acetone to methyl pivalate and
MTBE and other solvents provided they are tested prior being used.
— Sample preparation: allows to use weight/volume approach rather than only weight/weight.
— Data processing: provides explanations for a calibration curve using a forced through zero approach.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association, and supports essential requirements of EU Directive(s).
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
Directive 2003/15/EC amending Council Directive 76/768/EEC, relating to cosmetic produ
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