FprEN ISO 25353

• 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
• Publication Date: 24-Jan-2027
• ICS: 59.060.01 - Textile fibres in general
• Technical Committee: CEN/TC 248 - Textiles and textile products
• Current Stage: 5020 - Submission to Vote - Formal Approval
• Start Date: 27-Aug-2026
• Completion Date: 27-Aug-2026
• Ref Project: kSIST FprEN ISO 25353:2026 - 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)

• Consolidated By: ISO/FDIS 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)
• Effective Date: 12-Feb-2026

Overview

FprEN ISO 25353 provides an internationally recognized, standardized procedure for the qualitative proteomic analysis of protein fibres from leather. This standard establishes a protocol for identifying the animal species present in leather protein fibres using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Developed by CEN and aligned with ISO practices, the method supports traceability and quality control in the textile and leather industries, particularly where animal-derived materials are present in products.

By applying advanced mass spectrometry techniques, FprEN ISO 25353 enables reliable, species-specific identification of protein fibres that are otherwise complex to determine via microscopy due to the processed nature of leather fibres. This standard integrates with existing fibre analysis methods, such as those documented in the ISO 1833 series, providing a comprehensive approach for composite textile products.

Key Topics

• Proteomic Analysis of Leather Fibres: The standard specifies the steps for extracting, digesting, and analyzing protein fibres using trypsin digestion and the MALDI-TOF MS technique. This approach identifies peptide markers that are unique to specific animal species.

• Species Identification: By detecting species-specific peptide markers in the mass spectra within the 1000-3000 m/z range, it is possible to determine whether fibres originate from animals such as cow, goat, sheep, pig, horse, or even exotic species like crocodile.

• Sample Preparation: The method describes accurate sample preparation, including drying, cutting, weighing, and chemical procedures per referenced standards (e.g., ISO 4044).

• Microscopy Integration: Prior to mass spectrometric analysis, preliminary identification via light microscopy is recommended, following ISO/TR 11827, to assess the morphological characteristics of all fibres in blended samples.

• Reporting and Documentation: The standard outlines requirements for comprehensive test reports detailing methodologies, results (including mass spectra), fibre origins, sample descriptions, and any procedural deviations.

Applications

• Raw Material Verification in Leather and Textile Manufacturing: Ensures the accurate declaration of animal-based fibre content in leather goods, fabrics, and composite yarns, supporting both regulatory compliance and consumer confidence.

• Quality Assurance and Traceability: Facilitates quality control measures by confirming the animal species origin of protein fibres, which can be critical for product labeling, authenticity verification, and supply chain transparency.

• Mixed Fibre Product Analysis: Used in conjunction with the ISO 1833 series, FprEN ISO 25353 enables comprehensive fibre composition analysis for blends containing leather protein and synthetic fibres such as polyester, as exemplified in Annex C.

• Research and Development: Supports innovation in applied textile science by providing a robust method for species differentiation in experimental or recycled materials.

Related Standards

• ISO 4044 – Leather - Chemical tests - Preparation of chemical test samples
• ISO/TR 11827 – Textiles - Composition testing - Identification of fibres using microscopy
• ISO 1833 Series – Textiles - Quantitative chemical analysis (especially Part 24 for blends including polyester)
• ISO 20418-2:2018 – Textiles - Qualitative and quantitative proteomic analysis of some animal hair fibres - Peptide detection using MALDI-TOF MS
• ISO 17131:2020 – Leather - Identification of leather with microscopy

By adopting FprEN ISO 25353, organizations can enhance authenticity testing, ensure compliance with international best practices for textile and leather analysis, and contribute to increased trust and reliability in the global textile market. The application of MALDI-TOF MS for protein fibre analysis as set by this standard marks a significant advancement in the identification of animal species in processed leather textiles.

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,
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
STANDARDS MAY ON OCCASION HAVE TO
ISO/CEN PARALLEL PROCESSING
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POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
NATIONAL REGULATIONS.
RECIPIENTS OF THIS DRAFT ARE INVITED
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NOTIFICATION OF ANY RELEVANT PATENT
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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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