kSIST FprEN ISO 25353:2026
(Main)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)
- 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
- 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
kSIST FprEN ISO 25353:2026 is a CEN standard for textiles that uses MALDI-TOF MS to identify the animal species of protein fibres from leather. It specifies a qualitative test procedure for laboratories that need to confirm which leather-derived fibres are present in a sample or blend.
The standard matters when leather fibres are mixed with other fibres and microscopy alone is not enough to identify the animal origin. It also connects the leather result with the separate fibre-analysis methods used for the other components.
What does kSIST FprEN ISO 25353:2026 specify?
kSIST FprEN ISO 25353:2026 specifies a qualitative procedure for identifying the animal species of protein fibres from leather by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). It is organized as a short laboratory standard: scope, references, terms, principle, reagents, apparatus, specimen preparation, procedure, test report, and three annexes.
The standard is designed to be used together with microscopy for the initial blend check, and with fibre-composition methods for the non-leather part of the sample.
| Annex | What it covers |
|---|---|
| Annex A | Normative animal species-specific markers for MALDI-TOF MS analysis |
| Annex B | Informative animal species-specific peaks shown as figures |
| Annex C | Informative example for quantitative analysis of mixtures of protein fibres from leather and polyester |
What are the key requirements of kSIST FprEN ISO 25353:2026?
kSIST FprEN ISO 25353:2026 starts from a blend that has already been examined by light microscopy, using fibre morphology as the first sorting step. That preliminary step matters because the MALDI-TOF MS method is then focused on the leather-derived protein fibres rather than on the whole blend.
Sample preparation and extraction
Clause 8 requires test specimens of protein fibres from leather to be prepared by cutting and drying before testing, in line with ISO 4044. In practice, this gives the lab a defined sample condition before chemical extraction starts.
Clause 9.1 sets out protein extraction and enzymatic digestion. The method uses 2-mercaptoethanol and trypsin before MALDI-TOF MS analysis, which means the laboratory is looking for peptide patterns rather than intact leather fibres.
Reagents and apparatus
Clause 6 lists the reagents needed for biochemical analysis, including water of the grade specified in ISO 3696, ammonium bicarbonate, trypsin, 2-mercaptoethanol, acetonitrile, trifluoroacetic acid (TFA), and α-cyano-4-hyroxycinnamic acid (CHCA) as the matrix. These materials matter because peptide extraction and ionization depend on reagent quality and matrix preparation.
Clause 7 lists the equipment needed: basic glassware, controlled heating and temperature equipment, pipettes, an analytical balance, and a MALDI-TOF mass spectrometer. In practice, the lab needs routine protein-analysis equipment plus access to MALDI-TOF MS.
Identification of animal species
Clause 9.2 says identification is made by checking for species-specific peaks and markers in the mass spectrum. Annex A provides the normative marker set for the listed animal species, while Annex B shows the corresponding peaks and peptide sequences as examples.
Reporting
Clause 10 requires the test report to identify the standard used, describe the specimen if possible, state the date, include the test results and mass spectra, name the animal species found, and record any deviations or unusual observations. That makes the result traceable for quality control, supplier checks, or product documentation.
What terms does kSIST FprEN ISO 25353:2026 define?
- Protein - A polymer of amino acids; in this standard it is the material class being analyzed in leather fibres.
- Peptide - A small protein fragment; the method detects species-linked peptides after enzymatic digestion.
- Marker - A monoisotopic m/z peak used to identify the animal species.
- MALDI - Matrix-assisted laser desorption/ionization, the ionization method used before mass analysis.
- TOF - Time of flight, the mass-analyzing principle used to separate ions by how long they take to reach the detector.
- m/z - The mass-to-charge-related quantity used to read peaks in the mass spectrum.
Who uses kSIST FprEN ISO 25353:2026?
kSIST FprEN ISO 25353:2026 is used by textile and leather testing laboratories, quality teams, and technical buyers who need to identify the animal species in protein fibres from leather. It is also relevant when a leather spun yarn or another blend needs composition checking, especially when the leather component cannot be identified confidently by microscopy alone.
Which standards are used with kSIST FprEN ISO 25353:2026?
- ISO 3696 - Provides the water specification used for laboratory reagents.
- ISO 4044 - Provides the preparation of chemical test samples for leather.
- ISO/TR 11827 - Provides the microscopy-based fibre identification step used before MALDI-TOF MS.
- ISO 1833 series - Provides methods for measuring the other fibres in a blend so the full fibre composition can be combined.
- ISO 1833-24 - Provides the polyester-removal method used in Annex C.
- ISO 20418-2:2018 - Supplies related proteomic terminology for protein, peptide, and marker.
What does the kSIST FprEN ISO 25353:2026 document contain?
The document contains a reagent list, apparatus list, specimen-preparation instructions, and a two-step analytical procedure for extraction and species identification. Annex A gives the species-specific marker table, and Annex B presents the marker peaks and amino acid sequences as figures for the listed leather species.
Annex C gives an example workflow for mixtures of protein fibres from leather and polyester, including dissolution of the polyester fraction and recovery of the leather fibres for weighing or follow-up analysis. This is the part readers use when they need to relate the MALDI-TOF MS result to overall fibre composition.
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Frequently Asked Questions
kSIST FprEN ISO 25353:2026 is a draft published by the Slovenian Institute for Standardization (SIST). Its full title is "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)". This standard covers: 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.
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.
kSIST FprEN ISO 25353:2026 is classified under the following ICS (International Classification for Standards) categories: 59.060.01 - Textile fibres in general. The ICS classification helps identify the subject area and facilitates finding related standards.
kSIST FprEN ISO 25353:2026 is available in PDF format for immediate download after purchase. The document can be added to your cart and obtained through the secure checkout process. Digital delivery ensures instant access to the complete standard document.
Standards Content (Sample)
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
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
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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.
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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
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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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