SIST EN ISO 20418-1:2018

Textiles - Qualitative and quantitative proteomic analysis of some animal hair fibres - Part 1: Peptide detection using LC-ESI-MS with protein reduction (ISO 20418-1:2018)

This document specifies a qualitative and quantitative testing method to determine the content of
wool, cashmere, yak fibres and their blends in textiles by microscope preliminary screening, protein
extraction, enzymatic digestion and specific peptides detection using a liquid chromatography-mass
spectrometer equipped with electrospray ionization source (LCI-ESI-MC).
This method can be applied to relevant textile products at each process stage (i.e. from raw material
to garment) with a homogeneous distribution of the components. It can be applied to different types of
textile materials (e.g. staples, tops, yarns and fabrics) that contain wool, cashmere or yak fibres and their
blends. The method is based on a preliminary identification of all fibres in the blend on the basis of their
morphology, by light microscopy. The proteins are then extracted by a thiourea/urea/dithiothreitol
(DTT) solution. An enzymatic digestion by trypsin of the protein extracted from the fibres is carried
out. Analysis of the specific markers is performed by LC-MS and the percent composition is calculated.
This method is applicable to samples containing other kinds of fibres than wool, cashmere and yak, by
combining its results with the results obtained using the ISO 1833 series and/or the ISO 17751 series.
This document does not apply if fibres of the same animal species are present (e.g. blends of cashmere
and mohair); in this case, the quantitative analysis can be performed using microscopic analysis (e.g.
ISO 17751 series).

Textilien - Qualitative und quantitative Proteomanalyse einiger Tierhaarfasern - Teil 1: Peptiddetektion mittels LC-ESI-MS mit Proteinreduktion (ISO 20418-1:2018)

Dieses Dokument legt ein qualitatives und quantitatives Prüfverfahren zur Bestimmung des Gehalts von Woll , Kaschmir  und Yakfasern sowie deren Mischungen in Textilien durch eine Vorabuntersuchung mittels Mikroskop, eine Proteinextraktion, einen enzymatischen Aufschluss sowie eine spezifische Peptiddetektion unter Anwendung eines mit einer Elektrosprayionisationsquelle ausgestatteten Flüssigkeitschromato¬graphie-Massenspektrometers (LC ESI MS) fest.
Dieses Verfahren kann bei entsprechenden textilen Erzeugnissen in jeder Prozessphase (d. h. vom Rohstoff bis zum Kleidungsstück) mit einer gleichmäßigen Verteilung der Bestandteile angewendet werden. Es kann bei verschiedenen Arten textiler Materialien angewendet werden (z. B. Stapelfasern, Kammzüge, Garne und Flächengebilde), die Woll , Kaschmir  oder Yakfasern und deren Mischungen enthalten. Das Verfahren beruht auf einer Vorabidentifizierung aller Fasern in der Mischung auf der Grundlage ihrer Morphologie, die mittels Lichtmikroskopie erfolgt. Die Proteine werden anschließend durch eine Thioharnstoff /Harn¬stoff /Dithiothreitol (DTT )Lösung extrahiert. Dann erfolgt der enzymatische Aufschluss des aus den Fasern extrahierten Proteins durch Trypsin. Danach wird mittels LC MS die Analyse der spezifischen Marker durchgeführt und die prozentuale Zusammensetzung berechnet.
Dieses Verfahren ist auf Proben anwendbar, die andere Arten von Fasern als Woll , Kaschmir  und Yakfasern enthalten, indem die Ergebnisse mit den Ergebnissen kombiniert werden, die unter Anwendung der Normenreihe ISO 1833 und/oder der Normenreihe ISO 17751 erhalten wurden.
Dieses Dokument gilt nicht, wenn Fasern von der gleichen Tierart vorliegen (z. B. Mischungen aus Kaschmir und Mohair); in diesem Fall kann die quantitative Analyse mittels mikroskopischer Analyse (z. B. Normenreihe ISO 17751) durchgeführt werden.

Textiles - Analyse protéomique qualitative et quantitative de certaines fibres animales - Partie 1: Détection des peptides par LC-ESI-MS avec réduction protéique (ISO 20418-1:2018)

Le présent document spécifie une méthode d'essai qualitative et quantitative pour la détermination de la composition des fibres de laine, de cachemire, de yack et leurs mélanges dans les textiles par contrôle préliminaire au microscope, extraction protéique, digestion enzymatique et détection de peptides spécifiques au moyen d'un système de chromatographie en phase liquide-spectrométrie de masse équipé d'une source d'ionisation par électropulvérisation (LC-ESI-MS).
Cette méthode peut être appliquée aux produits textiles pertinents à chaque étape de fabrication (c'est-à-dire, de la matière première au vêtement) avec une distribution homogène des composants. Elle peut être appliquée à différents types de matières textiles (par exemple, fibres à filer, rubans de peigné, fils et étoffes) qui contiennent des fibres de laine, de cachemire ou de yack et leurs mélanges. La méthode repose sur une identification préliminaire de toutes les fibres présentes dans le mélange sur la base de leur morphologie, par microscopie optique. Les protéines sont ensuite extraites par une solution de thiourée/urée/dithiothréitol (DTT). Une digestion enzymatique par la trypsine de la protéine extraite des fibres est réalisée. Une analyse des marqueurs spécifiques est réalisée par LC-MS et la composition en pourcentage est calculée.
La présente méthode est applicable aux échantillons contenant d'autres types de fibres que la laine, le cachemire et le yack, en combinant ses résultats avec les résultats obtenus en utilisant la série ISO 1833 et/ou la série ISO 17751.
Le présent document ne s'applique pas si des fibres de la même espèce animale sont présentes (par exemple, mélanges de cachemire et de mohair); dans ce cas, l'analyse quantitative peut être réalisée par microscopie (par exemple, série ISO 17751).

Tekstilije - Kvalitativna in kvantitativna proteomska analiza nekaterih vlaken dlak živali - 1. del: Peptidna detekcija z uporabo LC-ESI-MS z zmanjšanjem beljakovin (ISO 20418-1:2018)

Ta dokument določa kvalitativno in kvantitativno preskusno metodo za ugotavljanje vsebnosti volne, kašmira, jakovih vlaken in njihovih mešanic v tkaninah, pri kateri se uporabljajo predhodni pregled z mikroskopom, ekstrahiranje beljakovin, encimska razgradnja in detekcija specifičnih peptidov z masnim spektrometrom s tekočinsko kromatografijo in ionizacija z elektrorazprševanjem (LCI-ESI-MC).
To metodo je mogoče uporabiti za relevantne tekstilne izdelke v vseh stopnjah procesa (tj. od surovine do oblačila) s homogeno porazdelitvijo komponent. Uporabiti jo je mogoče za različne vrste tekstilnih materialov (npr. vlakna, sukano prejo in blago), ki vsebujejo volno, kašmir, jakova vlakna in njihove mešanice. Metoda temelji na predhodni identifikaciji vseh vlaken v mešanici na osnovi njihove morfologije s svetlobnim mikroskopom. Beljakovine so nato ekstrahirane z raztopino triosečnine/sečnine/ditiotreitola (DTT). Izvede se encimska razgradnja beljakovin, ekstrahiranih iz vlaken, s tripsinom. Opravi se analiza specifičnih označevalcev z LC-MS in izračuna se sestava v odstotkih.
Ta metoda se uporablja za vzorce, ki vsebujejo druge vrste vlaken razen volne, kašmira in jakove dlake, s kombiniranjem njenih rezultatov z rezultati, pridobljenimi v skladu s sklopom standardov ISO 1833 in/ali ISO 17751. Ta dokument se ne uporablja, če so prisotna vlakna iste živalske vrste (npr. mešanice kašmirske dlake in dlake moher); v tem primeru se lahko izvede kvantitativna analiza z mikroskopsko analizo (npr. sklop standardov ISO 17751).

General Information

• Status: Published
• Public Enquiry End Date: 30-Jul-2017
• Publication Date: 11-Jun-2018
• ICS: 59.060.01 - Textile fibres in general
• Technical Committee: ITEK - Textile and textile products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 04-Jun-2018
• Due Date: 09-Aug-2018
• Completion Date: 12-Jun-2018
• Ref Project: EN ISO 20418-1:2018 - Textiles - Qualitative and quantitative proteomic analysis of some animal hair fibres - Part 1: Peptide detection using LC-ESI-MS with protein reduction (ISO 20418-1:2018)

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 20418-1:2018
01-julij-2018
Tekstilije - Kvalitativna in kvantitativna proteomska analiza nekaterih vlaken dlak
živali - 1. del: Peptidna detekcija z uporabo LC-ESI-MS z zmanjšanjem beljakovin
(ISO 20418-1:2018)
Textiles - Qualitative and quantitative proteomic analysis of some animal hair fibres - Part
1: Peptide detection using LC-ESI-MS with protein reduction (ISO 20418-1:2018)
Textilien - Qualitative und quantitative Proteomanalyse einiger Tierhaarfasern - Teil 1:
Peptiddetektion mittels LC-ESI-MS mit Proteinreduktion (ISO 20418-1:2018)
Textiles - Analyse protéomique qualitative et quantitative de certaines fibres animales -
Partie 1: Détection des peptides par LC-ESI-MS avec réduction protéique (ISO 20418-
1:2018)
Ta slovenski standard je istoveten z: EN ISO 20418-1:2018
ICS:
59.060.01 Tekstilna vlakna na splošno Textile fibres in general
SIST EN ISO 20418-1:2018 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------

SIST EN ISO 20418-1:2018

---------------------- Page: 2 ----------------------

SIST EN ISO 20418-1:2018


EN ISO 20418-1
EUROPEAN STANDARD

NORME EUROPÉENNE

May 2018
EUROPÄISCHE NORM
ICS 59.060.01
English Version

Textiles - Qualitative and quantitative proteomic analysis
of some animal hair fibres - Part 1: Peptide detection using
LC-ESI-MS with protein reduction (ISO 20418-1:2018)
Textiles - Analyse protéomique qualitative et Textilien - Qualitative und quantitative
quantitative de certaines fibres animales - Partie 1: Proteomanalyse einiger Tierhaarfasern - Teil 1:
Détection des peptides par LC-ESI-MS avec réduction Peptiddetektion mittels LC-ESI-MS mit
protéique (ISO 20418-1:2018) Proteinreduktion (ISO 20418-1:2018)
This European Standard was approved by CEN on 16 March 2018.

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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, 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
© 2018 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 20418-1:2018 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------

SIST EN ISO 20418-1:2018
EN ISO 20418-1:2018 (E)
Contents Page
European foreword . 3
2

---------------------- Page: 4 ----------------------

SIST EN ISO 20418-1:2018
EN ISO 20418-1:2018 (E)
European foreword
This document (EN ISO 20418-1:2018) has been prepared by Technical Committee ISO/TC 38
“Textiles” in collaboration with Technical Committee CEN/TC 248 “Textiles and textile products” the
secretariat of which is held by BSI.
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 2018, and conflicting national standards
shall be withdrawn at the latest by November 2018.
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.
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, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
Endorsement notice
The text of ISO 20418-1:2018 has been approved by CEN as EN ISO 20418-1:2018 without any
modification.

3

---------------------- Page: 5 ----------------------

SIST EN ISO 20418-1:2018

---------------------- Page: 6 ----------------------

SIST EN ISO 20418-1:2018
INTERNATIONAL ISO
STANDARD 20418-1
First edition
2018-03
Textiles — Qualitative and
quantitative proteomic analysis of
some animal hair fibres —
Part 1:
Peptide detection using LC-ESI-MS
with protein reduction
Textiles — Analyse protéomique qualitative et quantitative de
certaines fibres animales —
Partie 1: Détection des peptides par LC-ESI-MS avec réduction
protéique
Reference number
ISO 20418-1:2018(E)
©
ISO 2018

---------------------- Page: 7 ----------------------

SIST EN ISO 20418-1:2018
ISO 20418-1:2018(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2018
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]
Fax: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii © ISO 2018 – All rights reserved

---------------------- Page: 8 ----------------------

SIST EN ISO 20418-1:2018
ISO 20418-1:2018(E)

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Abbreviated terms . 2
5 Reagents . 2
6 Apparatus . 3
7 Test method . 3
7.1 Sampling . 3
7.2 Preliminary identification . 3
7.3 Sample preparation . 3
7.4 Protein extraction . 4
7.5 Trypsin digestion of the extracted proteins . 4
7.6 Tryptic peptides analysis by LC-ESI-MS . 4
7.7 Calculation and expression of results. 4
7.8 Test report . 5
Annex A (informative) Parameters . 6
Annex B (informative) Markers . 8
Annex C (informative) Examples of interlaboratory results . 9
Bibliography .11
© ISO 2018 – All rights reserved iii

---------------------- Page: 9 ----------------------

SIST EN ISO 20418-1:2018
ISO 20418-1:2018(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 on 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 the following
URL: www .iso .org/ iso/ foreword .html.
This document was prepared by Technical Committee ISO/TC 38, Textiles.
A list of all parts in the ISO 20418 series can be found on the ISO website.
iv © ISO 2018 – All rights reserved

---------------------- Page: 10 ----------------------

SIST EN ISO 20418-1:2018
ISO 20418-1:2018(E)

Introduction
International producers of textiles in cashmere and other speciality fibres have to be aware of and be
able to guarantee the fibre content in order to protect consumers and defend themselves from common
frauds. Therefore, it is important to have a harmonized method of analysis at an international level to
avoid different interpretations of the results and related conflicts between stakeholders.
The innovations in the method described in this document are
— objective qualitative determination of the presence of fibres derived from animal species and
— quantitative assessment of the relative percentages present in blends.
© ISO 2018 – All rights reserved v

---------------------- Page: 11 ----------------------

SIST EN ISO 20418-1:2018

---------------------- Page: 12 ----------------------

SIST EN ISO 20418-1:2018
INTERNATIONAL STANDARD ISO 20418-1:2018(E)
Textiles — Qualitative and quantitative proteomic analysis
of some animal hair fibres —
Part 1:
Peptide detection using LC-ESI-MS with protein reduction
1 Scope
This document specifies a qualitative and quantitative testing method to determine the content of
wool, cashmere, yak fibres and their blends in textiles by microscope preliminary screening, protein
extraction, enzymatic digestion and specific peptides detection using a liquid chromatography-mass
spectrometer equipped with electrospray ionization source (LCI-ESI-MC).
This method can be applied to relevant textile products at each process stage (i.e. from raw material
to garment) with a homogeneous distribution of the components. It can be applied to different types of
textile materials (e.g. staples, tops, yarns and fabrics) that contain wool, cashmere or yak fibres and their
blends. The method is based on a preliminary identification of all fibres in the blend on the basis of their
morphology, by light microscopy. The proteins are then extracted by a thiourea/urea/dithiothreitol
(DTT) solution. An enzymatic digestion by trypsin of the protein extracted from the fibres is carried
out. Analysis of the specific markers is performed by LC-MS and the percent composition is calculated.
This method is applicable to samples containing other kinds of fibres than wool, cashmere and yak, by
combining its results with the results obtained using the ISO 1833 series and/or the ISO 17751 series.
This document does not apply if fibres of the same animal species are present (e.g. blends of cashmere
and mohair); in this case, the quantitative analysis can be performed using microscopic analysis (e.g.
ISO 17751 series).
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 refer
...