1、December 2016 English price group 13No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 67.100.30!%b7“2596320www.din.de
2、DIN EN ISO 11816-2Milk and milk products Determination of alkaline phosphatase activity Part 2: Fluorimetric method for cheese (ISO 118162:2016);English version EN ISO 118162:2016,English translation of DIN EN ISO 11816-2:2016-12Milch und Milchprodukte Bestimmung der Aktivitt der alkalischen Phospha
3、tase Teil 2: Fluorimetrisches Verfahren fr Kse (ISO 118162:2016);Englische Fassung EN ISO 118162:2016,Englische bersetzung von DIN EN ISO 11816-2:2016-12Lait et produits laitiers Dtermination de lactivit de la phosphatase alcaline Partie 2: Mthode fluorimtrique pour le fromage (ISO 118162:2016);Vers
4、ion anglaise EN ISO 118162:2016,Traduction anglaise de DIN EN ISO 11816-2:2016-12SupersedesDIN EN ISO 118162:200306www.beuth.deDocument comprises 22 pagesDTranslation by DIN-Sprachendienst.In case of doubt, the German-language original shall be considered authoritative.12.16 DIN EN ISO 11816-2:2016-
5、12 2 A comma is used as the decimal marker. National foreword This document (EN ISO 11816-2:2016) has been prepared by Technical Committee ISO/TC 34 “Food products” in collaboration with Technical Committee CEN/TC 302 “Milk and milk products Methods of sampling and analysis” (Secretariat: NEN, Nethe
6、rlands). The responsible German body involved in its preparation was DIN-Normenausschuss Lebensmittel und landwirtschaftliche Produkte (DIN Standards Committeee Food and Agricultural Products), Working Committee NA 057-05-13 AA Milch und Milcherzeugnisse Probenahme- und Untersuchungsverfahren. The D
7、IN Standards corresponding to the International Standards referred to in this document are as follows: ISO 707 DIN EN ISO 707 ISO 5725-1 DIN ISO 5725-1 ISO 5725-2 DIN ISO 5725-2 ISO 11816-1 DIN EN ISO 11816-1 Amendments This standard differs from DIN EN ISO 11816-2:2003-06 as follows: a) the standar
8、d has been technically revised and brought in line with the current rules of presentation. Previous editions DIN EN ISO 11816-2: 2003-06 National Annex NA (informative) Bibliography DIN EN ISO 707, Milk and milk products Guidance on sampling DIN EN ISO 11816-1, Milk and milk products Determination o
9、f alkaline phosphatase activity Part 1: Fluorimetric method for milk and milk-based drinks DIN ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions DIN ISO 5725-2, Accuracy (trueness and precision) of measurement methods and resu
10、lts Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 11816-2 August 2016 ICS 67.100.30 Supersedes EN ISO 11816-2:2003English Version Milk and milk products - Determination of alka
11、line phosphatase activity - Part 2: Fluorimetric method for cheese (ISO 11816-2:2016) Lait et produits laitiers - Dtermination de lactivit de la phosphatase alcaline - Partie 2: Mthode fluorimtrique pour le fromage (ISO 11816-2:2016) Milch und Milchprodukte - Bestimmung der Aktivitt der alkalischen
12、Phosphatase - Teil 2: Fluorimetrisches Verfahren fr Kse (ISO 11816-2:2016) This European Standard was approved by CEN on 29 July 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 s
13、tandard 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 i
14、n 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
15、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 andUnited Kingdom. EUROP
16、EAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR 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 worldwide for CEN national Members. Ref. No. EN ISO 11816-2:2016
17、EEuropean foreword .31 Scope . 62 Normative reference . 63 Terms and definitions . 64 Principle 65 Reagents 76 Apparatus . 87 Sampling 98 Preparation of test sample . 99 Procedure. 99.1 Verification of instrument performance .99.1.1 General99.1.2 Daily instrument tests 109.1.3 Controls . 109.2 Reage
18、nt controls to test the suitability of ready to use working substrate (5.3) . 109.3 Calibration . 119.4 Determination 119.5 Test sample related controls 129.5.1 Recommended negative and positive control tests 129.5.2 Interfering substance test . 139.5.3 Heat-stable microbial alkaline phosphatase con
19、trol test 1310 Calculation and expression of results 1310.1 Calibration ratio 1310.2 Calculation . 1410.2.1 Supernatant . 1410.2.2 Cheese 1410.3 Expression of results . 1511 Precision 1511.1 Interlaboratory test 1511.2 Repeatability 1511.3 Reproducibility 1512 Test report 15Annex A (informative) Int
20、erlaboratory trial .16Annex B (informative) Examples of preparation of a test sample 18Bibliography .20Contents PageDIN EN ISO 11816-2:2016-12 EN ISO 11816-2:2016 (E)2Forewords .4European foreword This document (EN ISO 11816-2:2016) has been prepared by Technical Committee ISO/TC 34 “Food products”
21、in collaboration with Technical Committee CEN/TC 302 “Milk and milk products - Methods of sampling and analysis” the secretariat of which is held by NEN. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the late
22、st by February 2017, and conflicting national standards shall be withdrawn at the latest by February 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
23、 such patent rights. This document supersedes EN ISO 11816-2:2003. 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, E
24、stonia, 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 T
25、he text of ISO 11816-2:2016 has been approved by CEN as EN ISO 11816-2:2016 without any modification. DIN EN ISO 11816-2:2016-12 EN ISO 11816-2:2016 (E)3 ForewordsISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The
26、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 that committee. International organizations, governmental and non-gover
27、nmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for its further maintenance are described in
28、the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).Attention is drawn to the pos
29、sibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list
30、of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment, as well as
31、 information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for this document is ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products and the International Dairy F
32、ederation (IDF). It is being published jointly by ISO and IDF.This second edition of ISO 11816-2|IDF 155-2 cancels and replaces the first edition (ISO 11816-2|IDF 155-2:2003), which has been technically revised.ISO 11816|IDF 155 consists of the following parts, under the general title Milk and milk
33、products Determination of alkaline phosphatase activity: Part 1: Fluorimetric method for milk and milk-based drinks Part 2: Fluorimetric method for cheeseDIN EN ISO 11816-2:2016-12 EN ISO 11816-2:2016 (E)4 IDF (the International Dairy Federation) is a non-profit private sector organization represent
34、ing the interests of various stakeholders in dairying at the global level. IDF members are organized in National Committees, which are national associations composed of representatives of dairy-related national interest groups including dairy farmers, dairy processing industry, dairy suppliers, acad
35、emics and governments/food control authorities. ISO and IDF collaborate closely on all matters of standardization relating to methods of analysis and sampling for milk and milk products. Since 2001, ISO and IDF jointly publish their International Standards using the logos and reference numbers of bo
36、th organizations.Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. IDF shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will
37、be in the Introduction and/or on the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute and endorsementISO 11816|IDF 155 was prepared by the IDF Standing Committee on Analyt
38、ical Methods for Processing Aids and Indicators and ISO Technical Committee ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products.The work was carried out by the IDF/ISO Project Group on Alkaline phosphatase activity in cheese (P06), of the Standing Committee on Analytical Methods for
39、Processing Aids and Indicators, under the aegis of its project leader Mrs. M. Nicolas (FR).This ISO/IDF International Standard cancels and replaces ISO 11816-2|IDF 155-2:2003, which has been technically revised.ISO 11816|IDF 155 consists of the following parts, under the general title Milk and milk
40、products Determination of alkaline phosphatase activity: Part 1: Fluorimetric method for milk and milk-based drinks Part 2: Fluorimetric method for cheeseDIN EN ISO 11816-2:2016-12 EN ISO 11816-2:2016 (E)5 1 ScopeThis part of ISO 11816|IDF 155 specifies a fluorimetric method for the determination of
41、 alkaline phosphatase (ALP, EC 3.1.3.1) activity in cheese.This method is applicable to soft cheeses, semi-hard and hard cheeses provided that the mould is only on the surface of the cheese and not also in the inner part (e.g. blue veined cheeses). For large hard cheeses, specific conditions of samp
42、ling apply (see Clause 7).The instrument can read activities in the supernatant up to 7 000 milliunits per litre (mU/l).2 Normative referenceThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only
43、 the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitionsISO 5725-2, Accuracy (trueness and precision
44、) of measurement methods and results Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1alkaline phosphatase activityALP activityactivit
45、y of the alkaline phosphatase present in the product, determined by the specified procedure Note 1 to entry: The alkaline phosphatase activity is expressed as milliunits of enzyme activity per gram of sample (mU/g).3.2unit of alkaline phosphatase activityamount of alkaline phosphatase enzyme that ca
46、talyses the transformation of 1 mol of substrate per minute4 PrincipleThe alkaline phosphatase activity of the sample is measured by a continuous fluorimetric direct kinetic assay. A non-fluorescent aromatic monophosphoric ester substrate, 2-2-benzothiazolyl-6-hydroxybenzothiazole phosphate, in the
47、presence of any alkaline phosphatase derived from the sample, undergoes hydrolysis of its phosphate radical, producing a highly fluorescent product. Fluorimetric DIN EN ISO 11816-2:2016-12 EN ISO 11816-2:2016 (E)6 measurement of alkaline phosphatase (ALP) activity is measured at 38 C over a 3 min pe
48、riod when using the Fluorophos1). This includes pre-incubation of substrate and sample, followed by multiple kinetic readings of the reaction rate.NOTE Although this is a 3 min test, the first minute is an equilibration period to ensure that the sample is at 38 C. Measurements of activity are actually made from the beginning of the second minute to the end of the third minute (i.e. over a 2 min period).5 ReagentsUse o
