kSIST FprEN ISO 25353:2026

• Abstract:

  This document specifies a qualitative procedure to qualifying leather animal species of (fibrous) protein from leather by MALDI-TOF mass spectrometer (MS).; The composition of other fibres can be measured by methods described in the ISO 1833 series. Both results are then combined to determine the whole composition of fibres (see Annex C as an example of mixtures of (fibrous) protein from leather and polyester).; The method is based on a preliminary identification, by light microscopy, of all fibres in a blend on the basis of their morphology, according to ISO/TR 11827.

• Status: Not Published
• Public Enquiry End Date: 09-Dec-2025
• ICS: 59.060.01 - Textile fibres in general
• Technical Committee: ITEK - Textile and textile products
• Current Stage: 5020 - Formal vote (FV) (Adopted Project)
• Start Date: 04-Sep-2026
• Due Date: 23-Oct-2026
• Completion Date: 09-Sep-2026
• Ref Project: FprEN ISO 25353 - Textiles - Qualitative proteomic analysis of protein fibres from leather - Peptide detection using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (ISO/FDIS 25353:2026)

Overview

kSIST FprEN ISO 25353:2026 specifies the procedure for the qualitative proteomic analysis of protein fibres obtained from leather, focusing on the identification of animal species. This is achieved through peptide detection using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The standard is relevant for the textile and leather industries, providing reliable identification when traditional microscopic methods fall short, especially when leather fibres are pulverized or blended.

This standard is published by SIST and aligns with international best practices for textile fibre analysis. It forms part of a suite of standards enabling comprehensive fibre composition analysis when combined with ISO 1833 series methods for other fibres.

Key Topics

• Qualitative Proteomic Analysis: Establishes a systematic approach for identifying the species of origin of protein fibres in leather, leveraging advanced mass spectrometry technology.
• Peptide Detection: Utilizes enzymatic digestion of protein fibres, followed by MALDI-TOF MS to detect animal species-specific peptide markers.
• Sample Preparation and Testing: Outlines sample preparatory steps, including initial light microscopy as per ISO/TR 11827, drying, cutting, enzymatic digestion, and spectrometric analysis.
• Animal Species Identification: Provides procedures for confirming the presence of specific animal-derived fibres, such as cow, goat, sheep, pig, horse, and crocodile, based on unique peptide mass markers.
• Reporting Requirements: Specifies the minimum details to include in test reports, ensuring traceability and reproducibility.

Applications

The use of MALDI-TOF MS for leather protein fibre analysis offers several practical benefits across the textile, leather, and quality assurance sectors:

• Leather Supply Chain Integrity: Ensures accurate identification of animal species, supporting legal compliance and building consumer confidence in product labeling.
• Blend Analysis: When used in conjunction with ISO 1833 series, laboratories can determine the full fibre composition of blends (such as leather and polyester mixtures), aiding manufacturers in process control and product specification.
• Counterfeit and Authenticity Testing: Supports forensic and quality assurance activities to detect mislabeled or counterfeit leather products.
• Research & Development: Provides textile and material science researchers with robust tools for species differentiation, which can impact product innovation and sustainability assessments.

Related Standards

The scope and reliability of fibre identification in complex textile blends are greatly enhanced when kSIST FprEN ISO 25353:2026 is applied alongside other international standards:

• ISO 1833 Series: Methods for quantitative chemical analysis of textile fibre mixtures; particularly relevant for determining proportions of non-protein fibres.
• ISO/TR 11827: Offers a method for identifying all fibres within a blend by light microscopy before proceeding to proteomic analysis.
• ISO 17131:2020: Describes microscopy techniques for leather fibre identification, helpful as a preliminary step.
• ISO 20418-2:2018: Provides protocols for qualitative and quantitative proteomic analysis of animal hair fibres using MALDI-TOF MS.

Summary of Benefits

Implementing kSIST FprEN ISO 25353:2026 enables:

• Accurate, reproducible animal species identification in protein fibres from leather
• Greater confidence in product claims and improved traceability in the textile industry
• Effective resolution of complex fibre blends, supporting quality control and research efforts

By adopting this standard, organizations can ensure best practice in the detection and analysis of protein-based textile fibres, reinforcing both product quality and regulatory compliance.

SLOVENSKI STANDARD
oSIST prEN ISO 25353:2025
01-december-2025
Tekstilije - Kvalitativna proteomska analiza beljakovinskih vlaken iz usnja -
Odkrivanje peptidov z uporabo MALDI-TOF MS (ISO/DIS 25353:2025)
Textiles - Qualitative analysis proteomic analysis of protein fibres from leather - Peptide
detection using MALDI-TOF MS (ISO/DIS 25353:2025)
Textilien - Qualitative Analyse‑Proteomanalyse von Proteinfasern aus Leder -
Peptidnachweis mittels MALDI-TOF-MS (ISO/DIS 25353:2025)
Textiles - Analyse protéomique qualitative de fibres protéiques issues du cuir - Détection
des peptides par MALDI-TOF MS (ISO/DIS 25353:2025)
Ta slovenski standard je istoveten z: prEN ISO 25353
ICS:
59.060.01 Tekstilna vlakna na splošno Textile fibres in general
oSIST prEN ISO 25353:2025 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

oSIST prEN ISO 25353:2025
oSIST prEN ISO 25353:2025
DRAFT
International
Standard
ISO/DIS 25353
ISO/TC 38
Textiles — Qualitative analysis
Secretariat: JISC
proteomic analysis of protein fibres
Voting begins on:
from leather — Peptide detection
2025-09-25
using MALDI-TOF MS
Voting terminates on:
2025-12-18
Textiles — Analyse protéomique qualitative de fibres de protéines
issues de cuirs — Détection des peptides par MALDI-TOF MS
ICS: 59.060.01
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document has not been edited by the ISO Central Secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
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USER PURPOSES, DRAFT INTERNATIONAL
STANDARDS MAY ON OCCASION HAVE TO
ISO/CEN PARALLEL PROCESSING
BE CONSIDERED IN THE LIGHT OF THEIR
POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
NATIONAL REGULATIONS.
RECIPIENTS OF THIS DRAFT ARE INVITED
TO SUBMIT, WITH THEIR COMMENTS,
NOTIFICATION OF ANY RELEVANT PATENT
RIGHTS OF WHICH THEY ARE AWARE AND TO
PROVIDE SUPPORTING DOCUMENTATION.
Reference number
ISO/DIS 25353:2025(en)
oSIST prEN ISO 25353:2025
DRAFT
ISO/DIS 25353:2025(en)
International
Standard
ISO/DIS 25353
ISO/TC 38
Textiles — Qualitative analysis
Secretariat: JISC
proteomic analysis of protein fibres
Voting begins on:
from leather — Peptide detection
using MALDI-TOF MS
Voting terminates on:
Textiles — Analyse protéomique qualitative de fibres de protéines
issues de cuirs — Détection des peptides par MALDI-TOF MS
ICS: 59.060.01
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document has not been edited by the ISO Central Secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
© ISO 2025
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
STANDARDS MAY ON OCCASION HAVE TO
ISO/CEN PARALLEL PROCESSING
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
BE CONSIDERED IN THE LIGHT OF THEIR
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
or ISO’s member body in the country of the requester.
NATIONAL REGULATIONS.
ISO copyright office
RECIPIENTS OF THIS DRAFT ARE INVITED
CP 401 • Ch. de Blandonnet 8
TO SUBMIT, WITH THEIR COMMENTS,
CH-1214 Vernier, Geneva
NOTIFICATION OF ANY RELEVANT PATENT
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RIGHTS OF WHICH THEY ARE AWARE AND TO
PROVIDE SUPPORTING DOCUMENTATION.
Email: [e-mail]
Website: www.iso.org
Published in Switzerland Reference number
ISO/DIS 25353:2025(en)
ii
oSIST prEN ISO 25353:2025
ISO/DIS 25353:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
3.1 protein .1
3.2 peptide .1
3.3 marker .1
4 Abbreviated terms . 2
5 Principle . 2
6 Reagents . 2
6.1 Water, Grade 3 according to ISO 3696. .2
6.2 Ammonium bicarbonate, NH HCO , 96 % or higher in purity. .2
4 3
6.3 Trypsin, digestive enzymes break down peptide bonds following alginine and lysine. .2
6.4 2-mercapto ethanol .2
6.5 Ammonium bicarbonate solution (100 mmol/l). .2
6.6 Buffer solution (5 % of 2-mercapto ethanol, 25 mmol ammonium bicarbonate solution). .2
6.7 2,5mg/ml Trypsin, 25 mmol Ammonium bicarbonate solution. .2
6.8 Acetonitrile, 99,8 % or higher in purity. .3
6.9 Trifluoroacetic acid (TFA), 99,8 % or higher in purity. .3
6.10 α-cyano-4-hyroxycinnamic acid (CHCA), 99,5 % or higher in purity, used as a matrix
for peptides in MALDI MS analyses. .3
6.11 Matrix Solution.3
7 Apparatus . 3
7.1 Conical flask, of minimum capacity 250 ml, glass stoppered. .3
7.2 Heating apparatus, suitable for maintaining the temperature of the conical flask at (50
± 2) °C .3
7.3 MALDI-TOF mass spectrometer, for measuring molecular weight of peptides. .3
7.4 Aluminium block bath, to keep the reaction temperature constant, 30 °C to 100 °C. .3
7.5 Pipettes, capable of measuring and taking (0 to 20) μl (±0,20 μl), (20 to 200) μl (±1,60
μl), (200 to 1 000) μl (±8 μl). .3
7.6 Thermo mixer, to mix while keeping the reaction temperature constant, 35 °C to 100 °C .3
7.7 Analytical balance, with a resolution of 1 mg or better.3
8 Preparation of test specimens . 3
9 Procedure . 3
9.1 Protein extraction .3
9.2 Animal species identification of protein fibres from leather .4
10 Test report . 4
Annex A (normative) Animal species-specific marker in MALDI-TOF MS analysis . 5
Annex B (informative) Animal species-specific peaks in MALDI-TOF MS analysis . 6
Annex C (informative) Quantitative analysis of mixtures of protein fibres from leather and
polyester .10
Bibliography .11

iii
oSIST prEN ISO 25353:2025
ISO/DIS 25353:2025(en)
Foreword
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