kSIST FprEN 18179:2026

SLOVENSKI STANDARD
oSIST prEN 18179:2025
01-junij-2025
Pristnost živil - Metoda za odkrivanje predhodno zamrznjenega perutninskega
mesa z določanjem aktivnosti HADH
Food authenticity - Method for detecting previously frozen poultry meat by determination
of HADH activity
Lebensmittelauthentizität - Verfahren zum Nachweis von zuvor gefrorenem
Geflügelfleisch durch Bestimmung der HADH-Aktivität
Authenticité des aliments - Méthode de détection de la viande de volaille préalablement
congelée par détermination de l'activité HADH
Ta slovenski standard je istoveten z: prEN 18179
ICS:
67.120.20 Perutnina in jajca Poultry and eggs
oSIST prEN 18179:2025 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

oSIST prEN 18179:2025
oSIST prEN 18179:2025
DRAFT
EUROPEAN STANDARD
prEN 18179
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2025
ICS 67.120.20
English Version
Food authenticity - Method for detecting previously frozen
poultry meat by determination of HADH activity
Authenticité des aliments - Méthode de détection de la Lebensmittelauthentizität - Verfahren zum Nachweis
viande de volaille préalablement congelée par von zuvor gefrorenem Geflügelfleisch durch
détermination de l'activité HADH Bestimmung der HADH-Aktivität
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 460.
If this draft becomes a European Standard, 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.

This draft European Standard was established by CEN 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, Türkiye and
United Kingdom.
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.

Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.

EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2025 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 18179:2025 E
worldwide for CEN national Members.

oSIST prEN 18179:2025
prEN 18179 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 Symbols and Abbreviations . 5
5 Principle . 6
6 Reagents and materials . 6
7 Apparatus . 7
8 Procedure . 7
8.1 General . 7
8.2 Sub-sampling . 7
8.3 Sample pressing . 8
8.4 Spectrophotometry . 9
9 Accept/Reject criteria calculations . 10
9.1 General . 10
9.2 Data analysis . 10
9.2.1 Calculate the rate of decrease in absorbance ΔE/min as: . 10
9.2.2 Calculate the HADH activity (U/mL) using the following formula . 10
9.2.3 Calculate the R value for each sample as follows: . 10
10 Test report Analytical quality assurance . 11
Annex A (informative) Validation status and performance criteria as determined through
the application of a collaborative trial . 12
A.1 General . 12
A.2 Repeatability . 12
Annex B (informative) Threshold values . 13
Annex C (informative) Specifications for “Juice” extraction Press . 14
Bibliography . 15
oSIST prEN 18179:2025
prEN 18179 (E)
European foreword
This document (prEN 18179:2025) has been prepared by Technical Committee CEN/TC 460 “Food
Authenticity”, the secretariat of which is held by DIN.
This document is currently submitted to the CEN Enquiry.
This document contains public sector information licensed under the UK Open Government Licence
v3.0 (https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/)
oSIST prEN 18179:2025
prEN 18179 (E)
Introduction
The EC Poultry meat Marketing Standards Regulations (Commission Regulation (EC) No 543/2008)
[1], which are now incorporated into Council Regulation (EU) No 1308/2013 [2] of the European
Parliament and of the Council , require that poultry meat (whole birds and portions) is labelled to reflect
the storage conditions that have been employed (fresh, frozen, or rapidly frozen), and defines the
temperature conditions employed for each category. However, refrigeration technologies are now
available which do not comply with the conditions specified in the regulations and can result in the
mislabelling of poultry meat. There is therefore a need for the implementation of a robust analytical
method that is capable of distinguishing between fresh and previously frozen poultry for the
enforcement of the relevant regulatory legislation.
oSIST prEN 18179:2025
prEN 18179 (E)
1 Scope
This document specifies a procedure that can be used to determine the HADH activity in poultry meat
samples by spectrophotometry. The results can provide an indication whether poultry breast meat has
been previously frozen based on the ratio of relative HADH activity, and can be used to verify the labelling
of poultry breast meat sold as chilled poultry. When meat is frozen and thawed, the muscle mitochondria
are damaged and the HADH enzyme is released into the intracellular fluid. The relative increase in a
result above a value of 0,5 for the amount ratio of HADH found in fluid pressed activity from a sample
before and after laboratory freezing can be used to indicate whether it freezing, indicates that the sample
has been previously frozen. The HADH activity is determined using a spectrophotometric procedure.
This protocol document is applicable specifically to chicken and turkey breast meat but can be used for
other cuts and/or species with appropriate limit values. Additional validation can be required.
The method document is not applicable to minced meat or to poultry preparations.
The compliance assessment process is not part of this document.
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 references, the latest edition of the referenced document (including any amendments)
applies.
EN ISO 20813:2022, Molecular biomarker analysis - Methods of analysis for the detection and
identification of animal species in foods and food products (nucleic acid-based methods) - General
requirements and definitions (ISO 20813:2019)
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https://www.iso.org/obp
— IEC Electropedia: available at https://www.electropedia.org/
3.1
HADH activity
activity expressed in international units per ml of meat press juice (U/ml), 1 U representing 1 micromole
of substrate converted per minute at pH 6,0 and 25 °C
Note 1 to entry: The ratio R is the ratio of the HADH activities obtained for samples analysed before (X ) and after
1 0
(X ) a laboratory freezing step. R = X /X .
1 1 0 1
4 Symbols and Abbreviations
Acetyl Co A Acetyl coenzyme A
EDTA Ethylenediamine tetra acetic acid
HADH Hydroxyacyl-Coenzyme A dehydrogena

oSIST prEN 18179:2025
prEN 18179 (E)
5 Principle
Two sub-samples are prepared for each poultry breast sample using the procedure described. The first
sub-sample is analysed directly and the second sub-sample is analysed after freezing for at least 2 days
at (-20 ± 2) °C. After thawing, where required, the meat juice is expressed from the sample and diluted
with a phosphate buffer. The HADH activity (3.1) is measured by the rate of decrease in the absorption
of a treated sample extract at 340 nm using a UV spectrophotometer. The ratio (R ) of the HADH activity
(3.1) for the original and laboratory-frozen sub-samples is then used to determine whether the meat
sample has previously been frozen by comparison against threshold limits. (Annex A)
6 Reagents and materials
6.1 During the analysis, unless otherwise stated, only reagents of recognised laboratory reagent
grade (GPR) or analytical reagent grade (AR) shall be used and distilled or demineralized water or water
of equivalent purity according to EN ISO 20813:2022. Regarding laboratory organization, see [3]. Water
should be de-ionised or of similar quality.
6.2 Potassium dihydrogen orthophosphate (KH PO ), 0,1 mol/L solution. CAS No 7778-77-0.
2 4
Weigh (13,6 ± 0,1) g into a volumetric flask and make to 1 L with water.
6.3 Dipotassium hydrogen orthophosphate trihydrate (K HPO *3H O), 0,1 mol/L solution. CAS
2 4 2
No 16788-57-1.
Weigh (22,8 ± 0,1) g into a volumetric flask and make to 1 L with water.
6.4 Potassium phosphate buffer solution, pH 6,0 ± 0,05, CAS No 7778-77-0.
To 1 L of potassium di-hydrogen phosphate solution, (6.2), slowly add the dipotassium hydrogen
phosphate solution (6.3) until a pH of 6,0 ± 0,05 is obtained using a pH meter. The solution can be stored
under refrigeration (approximately 4 °C) for at least one month. The pH should be checked before use.
6.5 Ethylenediamine tetraacetic acid, disodium salt (EDTA) solution (26,9 mmol/L) CAS No
6381-92-6.
Weigh out (0,500 ± 0,005) g EDTA, disodium salt into a 50 mL beaker, and add approximately 40 mL of
water. Warm on a hotplate, with stirring, until dissolved. Transfer quantitatively to a 50 mL volumetric
flask with water, make up to the mark and mix. This solution can be stored under refrigeration for at
least one month.
6.6 ß-nicotinamide adenine dinucleotide, reduced-form, disodium salt hydrate (NADH)
solution (7,05 mmol/L) CAS No 606-68-8.
Weigh (25 ± 2) mg of NADH into a 5 mL volumetric flask. Make up to the mark with water and mix. The
solution can be stored for up to a week a refrigerator.
The weight taken should be corrected for the % purity, and
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