EN ISO 16140-2:2016

Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method (ISO 16140-2:2016)

ISO 16140-2:2016 specifies the general principle and the technical protocol for the validation of alternative, mostly proprietary, methods for microbiology in the food chain. Validation studies according to ISO 16140-2:2016 are intended to be performed by organizations involved in method validation.
It is applicable to the validation of methods for the analysis (detection or quantification) of microorganisms in
-      products intended for human consumption,
-      products intended for animal feeding,
-      environmental samples in the area of food and feed production, handling, and
-      samples from the primary production stage.
It is in particular applicable to bacteria and fungi. Some clauses of ISO 16140-2:2016 could be applicable to other (micro) organisms or their metabolites on a case-by-case-basis. In the future, guidance for other organisms (e.g. viruses and parasites) will be included in ISO 16140:2016 (all parts).

Mikrobiologie der Lebensmittelkette - Verfahrensvalidierung - Teil 2: Arbeitsvorschrift für die Validierung von alternativen (urheberrechtlich geschützten) Verfahren anhand eines Referenzverfahrens (ISO 16140-2:2016)

Dieser Teil von ISO 16140 legt die allgemeinen Benennungen und Definitionen in Bezug auf die Verfahrensvalidierung auf dem Gebiet der Mikrobiologie der Lebensmittelkette fest.
Der vorliegende Teil von ISO 16140 ist anwendbar auf die Validierung von Verfahren zur Untersuchung (Nachweis oder quantitative Bestimmung) von Mikroorganismen in:
   Erzeugnissen, die für den menschlichen Verzehr und als Futtermittel vorgesehen sind;
   Umgebungsproben im Bereich der Herstellung und Handhabung von Lebensmitteln;
   Proben aus dem Bereich der Primärproduktion.

Microbiologie de la chaîne alimentaire - Validation des méthodes - Partie 2: Protocole pour la validation de méthodes alternatives (commerciales) par rapport à une méthode de référence (ISO 16140-2:2016)

L'ISO 16140-2 :2016 établit le principe général ainsi que le protocole technique de validation des méthodes alternatives, qui sont pour la plupart commerciales, dans le domaine de la microbiologie de la chaîne alimentaire. Les études de validation conformément à l'ISO 16140-2 :2016 sont destinées à être effectuées par des organismes impliqués dans la validation de méthode.
Elle est applicable à la validation de méthodes pour l'analyse (détection ou quantification) de micro-organismes présents dans:
-      les produits destinés à la consommation humaine,
-      les produits destinés à l'alimentation animale,
-      les échantillons environnementaux dans le domaine de la production et de la manutention de produits alimentaires, et
-      les échantillons au stade de la production primaire.
L'ISO 16140-2 :2016 est notamment applicable aux bactéries et aux champignons. Certains paragraphes de l'ISO 16140-2 :2016 peuvent être applicables à d'autres micro-organismes ou à leurs métabolites au cas par cas. À l'avenir, des préconisations concernant d'autres organismes (par exemple, virus et parasites) seront incluses dans l'ISO 16140 (toutes les parties).

Mikrobiologija v prehranski verigi - Validacija metode - 2. del: Protokol za validacijo alternativnih (lastniških) metod glede na referenčno metodo (ISO 16140-2:2016)

Ta del standarda ISO 16140 določa splošno načelo in tehnični protokol za validacijo
alternativnih, večinoma lastniških metod za mikrobiologijo v prehranski verigi. Študije validacije
v skladu s tem delom standarda ISO 16140 so namenjene organizacijam, ki se ukvarjajo z validacijo metod.
Ta del standarda ISO 16140 se uporablja za validacijo metod za analizo (odkrivanje ali kvantifikacijo) mikroorganizmov v:
– proizvodih, namenjenih za prehrano ljudi;
– proizvodih, namenjenih za živalsko krmo;
– okoljskih vzorcih na področju proizvodnje hrane in krme; ter
– vzorcih iz faze primarne proizvodnje.
Ta del standarda ISO 16140 se uporablja zlasti za bakterije in glive. Nekatere točke tega dela
standarda ISO 16140 se lahko uporabljajo za druge (mikro-) organizme ali njihove metabolite za vsak posamezen primer.
V prihodnosti bodo v ta del ali ločen del standarda ISO 16140 vključeni napotki za druge organizme (npr. viruse in parazite).

General Information

• Status: Published
• Publication Date: 19-Jul-2016
• ICS: 07.100.30 - Food microbiology
• Technical Committee: CEN/TC 463 - Microbiology of the food chain
• Drafting Committee: CEN/TC 463 - Microbiology of the food chain
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 20-Jul-2016
• Completion Date: 20-Jul-2016
• Ref Project: SIST EN ISO 16140-2:2016 - Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method (ISO 16140-2:2016)

Relations

• Replaces: EN ISO 16140:2003 - Microbiology of food and animal feeding stuffs - Protocol for the validation of alternative methods (ISO 16140:2003)
• Effective Date: 08-Jun-2022
• Replaces: EN ISO 16140:2003/A1:2011 - Microbiology of food and animal feeding stuffs - Protocol for the validation of alternative methods - Amendment 1 (ISO 16140:2003/AMD 1:2011)
• Effective Date: 27-Jul-2016
• Amended By: EN ISO 16140-2:2016/A1:2024 - Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method - Amendment 1:
• Effective Date: 23-Jun-2021

Standards Content (Sample)

SLOVENSKI STANDARD
01-oktober-2016
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SIST EN ISO 16140:2003
SIST EN ISO 16140:2003/A1:2012
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Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of
alternative (proprietary) methods against a reference method (ISO 16140-2:2016)
Mikrobiologie der Lebensmittelkette - Verfahrensvalidierung - Teil 2: Arbeitsvorschrift für
die Validierung von alternativen (urheberrechtlich geschützten) Verfahren anhand eines
Referenzverfahrens (ISO 16140-2:2016)
Microbiologie de la chaîne alimentaire - Validation des méthodes - Partie 2: Protocole
pour la validation de méthodes alternatives (commerciales) par rapport à une méthode
de référence (ISO 16140-2:2016)
Ta slovenski standard je istoveten z: EN ISO 16140-2:2016
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 16140-2
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2016
EUROPÄISCHE NORM
ICS 07.100.30 Supersedes EN ISO 16140:2003
English Version
Microbiology of the food chain - Method validation - Part 2:
Protocol for the validation of alternative (proprietary)
methods against a reference method (ISO 16140-2:2016)
Microbiologie de la chaîne alimentaire - Validation des Mikrobiologie der Lebensmittelkette -
méthodes - Partie 2: Protocole pour la validation de Verfahrensvalidierung - Teil 2: Arbeitsvorschrift für die
méthodes alternatives (commerciales) par rapport à Validierung von alternativen (urheberrechtlich
une méthode de référence (ISO 16140-2:2016) geschützten) Verfahren anhand eines
Referenzverfahrens (ISO 16140-2:2016)
This European Standard was approved by CEN on 12 May 2016.

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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, 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: Avenue Marnix 17, B-1000 Brussels
© 2016 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16140-2:2016 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
European foreword
This document (EN ISO 16140-2:2016) has been prepared by Technical Committee ISO/TC 34 "Food
products" in collaboration with 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 January 2017, and conflicting national standards shall
be withdrawn at the latest by January 2017.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent
rights.
This document supersedes EN ISO 16140:2003.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
Endorsement notice
The text of ISO 16140-2:2016 has been approved by CEN as EN ISO 16140-2:2016 without any
modification.
INTERNATIONAL ISO
STANDARD 16140-2
First edition
2016-06-15
Microbiology of the food chain —
Method validation —
Part 2:
Protocol for the validation of
alternative (proprietary) methods
against a reference method
Microbiologie de la chaîne alimentaire — Validation des méthodes —
Partie 2: Protocole pour la validation de méthodes alternatives
(commerciales) par rapport à une méthode de référence
Reference number
ISO 16140-2:2016(E)
©
ISO 2016
ISO 16140-2:2016(E)
© ISO 2016, Published in Switzerland
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of
the requester.
ISO copyright office
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ii © ISO 2016 – All rights reserved

ISO 16140-2:2016(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 General principles for the validation of alternative methods . 1
5 Qualitative methods — Technical protocol for validation . 2
5.1 Method comparison study . 2
5.1.1 General considerations . 2
5.1.2 Paired or unpaired study . 2
5.1.3 Sensitivity study . 2
5.1.4 Relative level of detection study . 7
5.1.5 Inclusivity and exclusivity study . 9
5.2 Interlaboratory study .10
5.2.1 General considerations .10
5.2.2 Measurement protocol .10
5.2.3 Calculations and summary of data .12
5.2.4 Interpretation of data .15
6 Quantitative methods — Technical protocol for validation .16
6.1 Method comparison study .16
6.1.1 General considerations .16
6.1.2 Relative trueness study .16
6.1.3 Accuracy profile study.20
6.1.4 Limit of quantification study .24
6.1.5 Inclusivity and exclusivity study .24
6.2 Interlaboratory study .26
6.2.1 General considerations .26
6.2.2 Measurement protocol .26
6.2.3 Calculations, summary, and interpretation of data .27
Annex A (informative) Classification of sample types and suggested target combinations for
validation studies .30
Annex B (normative) Order of preference for use of naturally and artificially contaminated
samples in validation studies .46
Annex C (informative) General protocols for contamination by mixture and artificial
contamination of foods .47
Annex D (informative) Models for RLOD calculations using data from the method
comparison study .50
Annex E (normative) Points to be considered when selecting strains for testing inclusivity
and exclusivity .52
Annex F (informative) Considerations for calculations of the relative level of detection
(RLOD) between laboratories as obtained in an interlaboratory study .54
Annex G (informative) Principle of the accuracy profile for validation of quantitative models .57
Annex H (informative) Application of the accuracy profile in the method comparison study .59
Annex I (informative) Example of the application of the accuracy profile for an
interlaboratory study .62
Bibliography .66
ISO 16140-2:2016(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out
through ISO technical committees. Each member body interested in a subject for which a technical
committee has been established has the right to be represented on t
...