1、BRITISH STANDARDBS EN ISO 16140:2003Microbiology of food and animal feeding stuffs Protocol for the validation of alternative methodsICS 07.100.30g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3
2、g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58+A1:2011Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-National forewordThis British Standard is the UK implementation of EN ISO 16
3、140:2003+A1:2011. It is identical to ISO 16140:2003, incorporating amendment 1:2011. It supersedes BS EN ISO 16140:2003, which is withdrawn.The start and finish of text introduced or altered by amendment is indicated in the text by tags. Tags indicating changes to ISO text carry the number of the IS
4、O amendment. For example, text altered by ISO amendment 1 is indicated by !“.The UK participation in its preparation was entrusted to Technical Committee AW/9, Microbiology.A list of organizations represented on this committee can be obtained on request to its secretary. This publication does not pu
5、rport to include all the necessary provisions of a contract. Users are responsible for its correct application.Compliance with a British Standard cannot confer immunity from legal obligations.BS EN ISO 16140:2003+A1:2011This British Standard was published under the authority of the Standards Policy
6、and Strategy Committee on23 May2003 BSI 2011ISBN 978 0 580 67570 6Amendments/corrigenda issued since publicationDate Comments31 October 2011 Implementation of ISO amendment 1:2011 with CEN endorsement A1:2011Copyright European Committee for Standardization Provided by IHS under license with CENNot f
7、or ResaleNo reproduction or networking permitted without license from IHS-,-,-EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 16140:2003+A1August 2011ICS 07.100.30English versionMicrobiology of food and animal feeding stuffs - Protocol for thevalidation of alternative methods (ISO 16140:2003)M
8、icrobiologie des aliments - Protocole pour la validationdes mthodes alternatives (ISO 16140:2003)Mikrobiologie von Lebensmitteln und Futtermitteln -Arbeitsvorschrift fr die Validierung alternativer Verfahren(ISO 16140:2003)This European Standard was approved by CEN on 6 July 2002.CEN members are bou
9、nd to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the Man
10、agement 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 translationunder the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the o
11、fficialversions.CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,Hungary, Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Slovakia, Spain, Sweden, Switzerland and UnitedKingdom.EUROPEAN COMMITTEE F
12、OR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2003 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 16140:2003 ECopyright European Committee fo
13、r Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-ContentspageForeword. 3Introduction 41 Scope 52 Normative references . 53 Terms and definitions. 64 General principles for the validation and the certification o
14、f alternative methods 75 Qualitative methods - Technical protocol for their validation86 Quantitative methods -Technical protocol for their validation 18Annex A (normative) Specific rules for the acceptance of external results already obtained in a priorvalidation scheme . 34Annex B (informative) Cl
15、assification of sample types for validation studies . 36Annex C (normative) Use of naturally contaminated samples and preparation of artificiallycontaminated samples in validation studies 39Annex D (normative) Duplication of samples for the determination of relative accuracy and of relativedetection
16、 level for qualitative methods 41Annex E (normative) Calculation of the confidence intervals associated with the number of samplestested 43Annex F (normative) Test applied to the examination of discordant results 44Annex G (normative) Points to be considered when selecting strains for testing select
17、ivity 45Annex H (normative) Guidelines for the organisation and conducting collaborative studies 47Annex I (normative) Determination that negative controls are free of target analyte 50Annex J (normative) Replication of samples for interlaboratory studies of qualitative methods 51Annex K (normative)
18、 Consideration of data .53Annex L (informative) Interlaboratory study of qualitative methods: criteria of accordance,concordance and concordance odds ratio 54Annex M (normative) Replication of samples for the determination of relative accuracy of quantitativemethods . 59Annex N (normative) Examples
19、of acceptable and unacceptable situations and range ofmeasurements for the estimation of the regression line for quantitative methods. 61Annex O (normative) Assessment of the linearity of quantitative methods by graphical representation. 63Annex P (normative) Detection and quantification limits for
20、counts. 64Annex Q (normative) 66Annex R (normative) Calculations with the regression method. 68Annex S (normative) Examples of calculations for quantitative methods 73Annex T (normative) Collaborative study Ring test results with duplicates 78Annex U (informative) List of symbols and abbreviations 7
21、9Bibliography . 87BS EN ISO 16140:2003+A1:2011EN ISO 16140:2003+A1:2011 (E)2Calculation of Rousseeuws scale estimator QnIndicators for Mandels robust h- and k-statisticsAnnex V (informative) 80Annex W (informative) . 81Example: Determination of E. coli.Copyright European Committee for Standardizatio
22、n Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-ForewordThis document (EN ISO 16140:2003) has been prepared by Technical Committee CEN/TC 275 “Food analysis -Horizontal methods“, the secretariat of which is held by DIN, in co
23、llaboration with Technical Committee ISO/TC 34“Agricultural food products“.This European Standard shall be given the status of a national standard, either by publication of an identical text orby endorsement, at the latest by November 2003, and conflicting national standards shall be withdrawn at th
24、e latestby November 2003.The annexes A, C to K and M to T are normative. The annexes B, L and U are informative.This document contains also a Bibliography.According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following coun-tries are bound to implement this E
25、uropean Standard: Austria, Belgium, Czech Republic, Denmark, Finland,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal,Slovakia, Spain, Sweden, Switzerland and the United Kingdom.Foreword to amendment A1 This document (EN ISO 16140:2003/A1:20
26、11) 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 Amendment to the European Standard EN ISO 16140:2003 shall be given the status of a national
27、standard, either by publication of an identical text or by endorsement, at the latest by February 2012, and conflicting national standards shall be withdrawn at the latest by February 2012. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
28、rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. 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, Croati
29、a, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of
30、 ISO 16140:2003/AMD 1:2011 has been approved by CEN as EN ISO 16140:2003/A1:2011 withoutany modification. BS EN ISO 16140:2003+A1:2011EN ISO 16140:2003+A1:2011 (E)3Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permi
31、tted without license from IHS-,-,-IntroductionThe need for the food industry to rapidly assess the microbiological quality of raw materials and finished productsand the microbiological status of manufacturing procedures, has led to the development and refinement of alterna-tive microbiological metho
32、ds of analysis that are quicker and/or easier to perform than the corresponding referencemethod; some can also be automated.Among these alternative methods, some can yield results that are equivalent to those provided by the referencemethod, while others can lead to results that differ appreciably.T
33、he suppliers/producers of the alternative methods, the food and drink industry, the public health services andother authorities need a reliable common protocol for the validation of such alternative methods. The data gener-ated can also be the basis for the certification of a method by an independen
34、t organisation.Because of the extent of the methods comparative study described in this standard for use by the organising labo-ratory, the procedure is sometimes not appropriate for use as an “in house“ method for the validation of an alterna-tive method by an individual laboratory.BS EN ISO 16140:
35、2003+A1:2011EN ISO 16140:2003+A1:2011 (E)4Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-1 ScopeThis document establishes the general principle and the technical protocol for th
36、e validation of alter-native methods in the field of microbiological analysis of food, animal feeding stuff and environmentaland veterinary samples (see 5.1.1.2.1) for:the validation of alternative methods which can be used in particular in the framework of the offi-cial control;the international ac
37、ceptance of the results obtained by the alternative method.It also establishes the general principles of certification of these alternative methods, based on thevalidation protocol defined in this EN ISO 16140 (see 4.2).Where an alternative method is used on a routine basis for internal laboratory u
38、se without the re-quirement to meet (higher) external criteria of quality assurance, a less stringent comparative valida-tion of the alternative method than that set in this standard may be appropriate.2 Normative referencesThis European Standard incorporates by dated or undated reference, provision
39、s from other publica-tions. These normative references are cited at the appropriate places in the text, and the publicationsare listed hereafter. For dated references, subsequent amendments to or revisions of any of thesepublications apply to this European Standard only when incorporated in it by am
40、endment or revision.For undated references the latest edition of the publication referred to applies (including amend-ments).ISO 3534-1, Statistics Vocabulary and symbols Part 1: Probability and general statistical terms.ISO 5725, Accuracy (trueness and precision) of measurement methods and results.
41、BS EN ISO 16140:2003+A1:2011EN ISO 16140:2003+A1:2011 (E)5Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-3 Terms and definitionsFor the purposes of this European Standard, the f
42、ollowing terms and definitions apply:3.1alternative methodmethod of analysis that demonstrates or estimates, for a given category of products, the same analyte(3.4) as is measured using the corresponding reference method (3.2).NOTE 1 The method can be proprietary or non commercial, and does not need
43、 to cover an entire analysis pro-cedure, that is from the preparation of samples to the test report.NOTE 2 The alternative method exhibits attributes appropriate to the users needs, for example:speed of analysis and/or response;ease of execution and/or automation;analytical properties (precision, ac
44、curacy, limit of detection, etc.);miniaturisation;reduction of cost.NOTE 3 The term “alternative“ is used to refer to the entire “test procedure and reaction system“. This termincludes all ingredients whether material or otherwise, required for implementing the method.3.2reference methodinternationa
45、lly recognised method and widely accepted.NOTE For the purpose of this standard, these are International and European Standards and if not existing,certain national standards of equivalent standing.3.3validation of an alternative methoddemonstration that adequate confidence is provided that the resu
46、lts obtained by the alternativemethod are comparable to those obtained using the reference methodNOTE The word “comparable“ is defined in this EN ISO 16140 by a technical protocol adapted to each typeof method (see clauses 5 and 6).3.4analytecomponent measured by the method of analysis. It may be th
47、e microorganism3.5qualitative methodmethod of analysis whose response is either the presence or absence of the analyte (3.4) detectedeither directly or indirectly in a certain amount of sample3.6quantitative methodmethod of analysis whose response is the amount of the analyte (3.4) measured either d
48、irectly (enu-meration in a mass or a volume), or indirectly (colour absorbance, impedance, etc.) in a certainamount of sampleBS EN ISO 16140:2003+A1:2011EN ISO 16140:2003+A1:2011 (E)6Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction
49、or networking permitted without license from IHS-,-,-3.7methods comparison studystudy, performed by the organising laboratory of the alternative method against the reference method3.8inter-laboratory studystudy of the methods performance using common samples in several laboratories and under the con-trol of the organisin
