SIST EN 17254:2019

Foodstuffs - Minimum performance requirements for determination of gluten by ELISA

This document specifies minimum method performance requirements for enzyme-linked immunosorbent assays that quantify non-fragmented or fragmented gluten from wheat (e.g. Triticum aestivum), rye, and barley in raw and processed foodstuffs.
This document is intended to be used in addition to EN 15842.

Lebensmittel - Minimale Leistungsanforderungen für die Glutenbestimmung mit ELISA

In diesem Dokument werden Mindestanforderungen an die Verfahrensleistung für Enzym-gekoppelte Immunosorbent-Tests festgelegt, die nicht-fragmentiertes oder fragmentiertes Gluten aus Weizen (z. B. Triticum aestivum), Roggen und Gerste in rohen und verarbeiteten Lebensmitteln quantifizieren.
Dieses Dokument ist für die Verwendung in Ergänzung zu EN 15842 vorgesehen.

Produits alimentaires - Exigences de performances minimales pour la détermination du gluten par une méthode ELISA

Le présent document spécifie les exigences de performances minimales pour les méthodes ELISA permettant de quantifier le gluten de blé (par exemple, Triticum aestivum), de seigle et d’orge non fragmenté ou fragmenté, dans des produits alimentaires crus et transformés.
Le présent document est destiné à être utilisé conjointement avec l’EN 15842.

Živila - Minimalne zahteve za ugotavljanje glutena z metodo ELISA

V tem dokumentu so opredeljene minimalne zahteve glede učinkovitosti encimskih imunosorbentnih testov, ki količinsko opredeljujejo nefragmentiran ali fragmentiran gluten v pšenici (npr. Triticum aestivum), rži in ječmenu v surovih in predelanih živilih.

General Information

• Status: Published
• Public Enquiry End Date: 24-Jul-2018
• Publication Date: 06-Nov-2019
• ICS: 67.050 - General methods of tests and analysis for food products
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 16-Oct-2019
• Due Date: 21-Dec-2019
• Completion Date: 07-Nov-2019
• Directive: Not Harmonized882/2004 - Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 onofficial controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules
• Ref Project: EN 17254:2019 - Foodstuffs - Minimum performance requirements for determination of gluten by ELISA

Relations

• Refers: SIST EN 15842:2019 - Foodstuffs - Detection of food allergens - General considerations and validation of methods
• Effective Date: 28-Jan-2026
• Refers: SIST EN 15633-1:2019 - Foodstuffs - Detection of food allergens by immunological methods - Part 1: General considerations
• Effective Date: 28-Jan-2026

Overview

SIST EN 17254:2019 sets the minimum performance requirements for determining gluten in foodstuffs using enzyme-linked immunosorbent assays (ELISA). This European standard ensures that both non-fragmented and fragmented gluten from wheat, rye, and barley are reliably quantified in raw and processed foods. By specifying clear method performance parameters, it supports food safety for populations such as those with celiac disease and helps food producers comply with regulatory gluten limits. This document is applicable alongside EN 15842, providing a robust framework for gluten analysis in the food industry.

Key Topics

• Gluten Detection by ELISA: ELISA methods are the primary means of detecting and quantifying gluten in foods due to their sensitivity, specificity, and adaptability to different food matrices.
• Scope and Applicability:
  • Covers gluten from wheat (including common Triticum aestivum), rye, and barley.
  • Applies to both raw ingredients and processed food products.
• Minimum Performance Requirements:
  • Applicability Range: 10–40 mg/kg for both non-fragmented and fragmented gluten.
  • Limit of Detection (LOD): 5 mg/kg.
  • Limit of Quantification (LOQ): 10 mg/kg.
  • Precision and Recovery: Clearly defined criteria for repeatability, reproducibility, and recovery rates for reliable gluten analysis.
• Regulatory Aspects:
  • Supports compliance with the Commission Implementing Regulation (EU) No 828/2014.
  • Assists in labeling products as “gluten-free” (<20 mg/kg) or “very low gluten” (≤100 mg/kg).
• Cross-Reactivity and Specificity: Addresses testing against a broad range of possible interfering substances to ensure method specificity.
• Test Robustness: Includes testing the analytical process under varying conditions to confirm consistent results.

Applications

• Food Safety Assurance: Key for manufacturers, testing laboratories, and regulatory bodies to ensure food products meet gluten labeling requirements for sensitive populations, especially those with celiac disease.
• Product Labeling: Enables accurate gluten quantification for trustworthy “gluten-free” and “very low gluten” labeling in compliance with EU regulations.
• Quality Control: Provides a standardized approach for industry laboratories and quality assurance teams to validate their ELISA methodologies.
• Regulatory Compliance: Essential reference for food operators to avoid cross-contamination, maintain consumer trust, and meet international gluten safety standards.
• Research & Product Development: Supports R&D teams in developing and validating new food products designed for gluten-intolerant consumers.

Related Standards

Several complementary and referenced standards support SIST EN 17254:2019:

• EN 15842: General considerations and validation of methods for detecting food allergens.
• EN 15633-1: General considerations for immunological detection of food allergens.
• EN ISO/IEC 17025: General requirements for the competence of testing and calibration laboratories.
• CEN/TR 16338: Templates for information supply on immunological and molecular biological methods for food allergens.
• ISO 5725-1: Accuracy (trueness and precision) of measurement methods and results - General principles.
• AOAC International guidelines: Validation procedures for quantitative gluten ELISA methods and collaborative study procedures.

Practical Value

By implementing SIST EN 17254:2019, organizations in the food industry can:

• Achieve reliable and comparable gluten analysis results.
• Minimize the risk of accidental gluten exposure in gluten-sensitive individuals.
• Enhance international marketability of food products by aligning with EU and global standards.
• Build confidence in food safety management protocols for gluten detection.

Utilizing standardized gluten quantification methods benefits consumer safety, business compliance, and overall food quality in an increasingly allergen-aware market.

Standards Content (Sample)

SLOVENSKI STANDARD
01-december-2019
Živila - Minimalne zahteve za ugotavljanje glutena z metodo ELISA
Foodstuffs - Minimum performance requirements for determination of gluten by ELISA
Lebensmittel - Minimale Leistungsanforderungen für die Glutenbestimmung mit ELISA
Produits alimentaires - Exigences de performances minimales pour la détermination du
gluten par une méthode ELISA
Ta slovenski standard je istoveten z: EN 17254:2019
ICS:
67.050 Splošne preskusne in General methods of tests and
analizne metode za živilske analysis for food products
proizvode
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN 17254
EUROPEAN STANDARD
NORME EUROPÉENNE
October 2019
EUROPÄISCHE NORM
ICS 67.050
English Version
Foodstuffs - Minimum performance requirements for
determination of gluten by ELISA
Produits alimentaires - Exigences de performances Lebensmittel - Minimale Leistungsanforderungen für
minimales pour la détermination du gluten par une die Glutenbestimmung mit ELISA
méthode ELISA
This European Standard was approved by CEN on 12 August 2019.

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
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17254:2019 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
Introduction . 3
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 General information on the test system. 7
5 Performance requirements . 7
5.1 General . 7
5.2 Applicability range . 7
5.3 Limit of Detection (LOD) . 7
5.4 Limit of Quantification (LOQ) . 7
5.5 Recovery . 8
5.6 Precision . 8
5.7 Specificity . 8
5.8 Cross-Reactivity . 8
5.9 Robustness. 8
5.10 Overview on minimum performance requirements . 8
Bibliography . 10

European foreword
This document (EN 17254:2019) has been prepared by Technical Committee CEN/TC 275 “Food
analysis - horizontal methods”, the secretariat of which is held by DIN.
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 April 2020, and conflicting national standards shall be
withdrawn at the latest by April 2020.
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 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
0.1 Coeliac Disease
Coeliac disease (CD) is an autoimmune disease of the small intestine, primarily affecting genetically
susceptible individuals. Its prevalence has been estimated to be 1 % of the population worldwide. CD
becomes manifest in a chronic enteropathy, caused by an irreversible intolerance for gluten. Upon
ingestion of gluten, the incomplete gastrointestinal digestion of these proteins leads to the appearance
of gluten-derived peptides. It contains overlapping T-cell epitopes, and its deamidated form is a potent
T-cell stimulator. Toxic gluten peptides cause the stimulation of the innate and the adaptive immune
system. This leads to histological changes in the small intestine mucosa of coeliac patients, resulting in
severe symptoms including chronic diarrhoea, abdominal distension, and malabsorption of nutrients.
Coeliac disease, if untreated, is associated with increased morbidity and the only accepted treatment is
a strict and lifelong adherence to a gluten free diet, which interrupts the immune response triggered by
gluten [1].
0.2 Gluten
The definition of gluten slightly differs depending on the context in which it is used. In this document,
the gluten definition (see 3.1) according to the Osborne fractionation is used, similar to the definition
stated in the Codex Alimentarius.
Gluten in starch industry refers to the protein rich fraction that is separated from starch during the
starch production process. The term is therefore applied to different cereals, mainly wheat and corn,
which are predominantly used for starch production. In the baking industry, gluten refers to a protein
rich ingredient, which leads to an increase in volume and “fluffiness” of baking products such as bread.
In this context, only wheat contains gluten since only the protein rich fraction from this cereal leads to
an improvement of the physicochemical properties of baking products. The definition of gluten in the
context of coeliac disease refers to the protein fraction, which is toxic for CD patients. This definition is
based on the Osbourne fractionation of cereal proteins (e.g. from wheat and its crossbred varieties),
precisely the proteins insoluble in water and 0,5 mol/l sodium chloride solution.
Due to thermal processing and enzymatic hydrolysis during food production, gluten in food is often not
present in its native form. The above-mentioned processing can lead to the denaturation and/or to the
fragmentation of gluten proteins into peptides. Gluten peptides have partly different properties in
regards to solubility and detectability by immuno-analytical techniques, but are still able to trigger
immune reactions in coeliac disease patients [2].
Commission Implementing Regulation 828/2014 [3] states that oats contained in foodstuffs for people
intolerant to gluten shall have been specially produced, prepared and/or processed in a way to avoid
contamination by wheat, rye, barley, or their crossbred varieties and the gluten content of such oats
may not exceed 20 mg/kg.
0.3 Regulatory limits
Commission Implementing Regulation (EU) No 828/2014 of the European Union states that there are
two different threshold levels for gluten in foodstuffs.
The term “gluten-free” may be used for food products if the gluten content does not exceed
20 mg gluten/kg. On the other hand, if the gluten content does not exceed 100 mg gluten/kg products
may bear the term “very low gluten”.
Oats can be tolerated by most but not all people who are intolerant to gluten. Therefore, the allowance
of oats that are not contaminated with wheat, rye or barley in foods covered by this standard can be
determined at the national level [4], [5].
0.4 Measurement of the gluten content
The gluten concen
...