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1、BRITISH STANDARD BS EN ISO 13366-1:2008 Milk Enumeration of somatic cells Part 1: Microscopic method (Reference method) ICS 67.100.10 ? Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40
2、 MDTNo reproduction or networking permitted without license from IHS -,-,- BS EN ISO 13366-1:2008 This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 March 2008 BSI 2008 ISBN 978 0 580 57104 6 National foreword This British Standard is the UK
3、implementation of EN ISO 13366-1:2008. It supersedes BS EN ISO 13366-1:1997 which is withdrawn. The UK participation in its preparation was entrusted to Technical Committee AW/5, Chemical analysis of milk and milk products. A list of organizations represented on this committee can be obtained on req
4、uest to its secretary. This publication does not purport 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. Amendments/corrigenda issued since publication DateComm
5、ents Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS -,-,- EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO
6、 13366-1 February 2008 ICS 67.100.10Supersedes EN ISO 13366-1:1997 English Version Milk - Enumeration of somatic cells - Part 1: Microscopic method (Reference method) (ISO 13366-1:2008) Lait - Dnombrement des cellules somatiques - Partie 1: Mthode au microscope (Mthode de rfrence) (ISO 13366-1:2008)
7、 Milch - Zhlung somatischer Zellen - Teil 1: Mikroskopisches Verfahren (Referenzverfahren) (ISO 13366-1:2008) This European Standard was approved by CEN on 3 February 2008. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this Europe
8、an 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 Management Centre or to any CEN member. This European Standard exists in three official versions (English
9、, 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 Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, C
10、yprus, 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 United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION
11、 COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: rue de Stassart, 36 B-1050 Brussels 2008 CENAll rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 13366-1:2008: E Copyright British Standards Institution P
12、rovided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS -,-,- Foreword This document (EN ISO 13366-1:2008) has been prepared by Technical Committee ISO/TC 34 “Agric
13、ultural food products“ 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
14、endorsement, at the latest by August 2008, and conflicting national standards shall be withdrawn at the latest by August 2008. 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 id
15、entifying any or all such patent rights. This document supersedes EN ISO 13366-1:1997. 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, Cyprus, Czech Republic,
16、 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 ISO 13366-1:2008 has been
17、 approved by CEN as a EN ISO 13366-1:2008 without any modification. BS EN ISO 13366-1:2008 Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted w
18、ithout license from IHS -,-,- Reference numbers ISO 13366-1:2008(E) IDF 148-1:2008(E) INTERNATIONAL STANDARD ISO 13366-1 IDF 148-1 Second edition 2008-02-15 Milk Enumeration of somatic cells Part 1: Microscopic method (Reference method) Lait Dnombrement des cellules somatiques Partie 1: Mthode au mi
19、croscope (Mthode de rfrence) BS EN ISO 13366-1:2008 IDF 148-1:2008 Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS
20、-,-,- ii Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS -,-,- iii Contents Page Foreword iv Foreword. v 1 Scope 1
21、2 Terms and definitions .1 3 Principle1 4 Reagents.1 4.1 Dye solutions .1 4.2 Phosphate Buffer Solution (PBS).3 5 Apparatus .4 6 Sampling.4 7 Preparation of test sample4 7.1 Storage4 7.2 Preparation.5 8 Procedure .5 8.1 Preparation of the smear and staining5 8.2 Determination.6 9 Calculation and exp
22、ression of results.10 9.1 Rectangular shape counting in successive fields .10 9.2 Rectangular shape counting in bands 11 9.3 Circular shape counting in successive fields.11 9.4 Expression of results 12 10 Precision.12 10.1 Repeatability.12 10.2 Reproducibility.12 11 Test report13 Annex A (informativ
23、e) Collaborative trial14 Annex B (informative) Staining for goats milk 15 Annex C (informative) Poisson distribution .16 Bibliography17 BS EN ISO 13366-1:2008 IDF 148-1:2008 Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/59107700
24、01 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS -,-,- iv Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standa
25、rds 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-governmental, in liaison with ISO, also tak
26、e part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is
27、 to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibilit
28、y 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. ISO 13366-1|IDF 148-1 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products, and the In
29、ternational Dairy Federation (IDF). It is being published jointly by ISO and IDF. This second edition of ISO 13366-1|IDF 148-1 cancels and replaces the first edition (ISO 13366-1:1997), of which it constitutes a technical revision. ISO 13366 consists of the following parts, under the general title M
30、ilk Enumeration of somatic cells: Part 1: Microscopic method (Reference method) Part 2: Guidance on the operation of fluoro-opto-electronic counters BS EN ISO 13366-1:2008 IDF 148-1:2008 Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boein
31、g Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS -,-,- v Foreword IDF (the International Dairy Federation) is a non-profit organization representing the dairy sector worldwide. IDF membership comprises National Committees in ever
32、y member country as well as regional dairy associations having signed a formal agreement on cooperation with IDF. All members of IDF have the right to be represented at the IDF Standing Committees carrying out the technical work. IDF collaborates with ISO in the development of standard methods of an
33、alysis and sampling for milk and milk products. Draft International Standards adopted by the Action Teams and Standing Committees are circulated to the National Committees for voting. Publication as an International Standard requires approval by at least 50 % of the IDF National Committees casting a
34、 vote. 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. ISO 13366-1|IDF 148-1 was prepared by the International Dairy Federation (IDF) and Technical C
35、ommittee ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products. It is being published jointly by IDF and ISO. All work was carried out by the Joint ISO-IDF Action Team Automated methods of the Standing Committee on Quality assurance, statistics of analytical data and sampling under the
36、 aegis of its project leaders, Mrs. S. Orlandini (IT) and Mr. H.J.C.M. van den Bijgaart (NL). This edition of ISO 13366-1|IDF 148-1 cancels and replaces IDF 148A:1995. ISO 13366 consists of the following parts, under the general title Milk Enumeration of somatic cells: Part 1: Microscopic method (Re
37、ference method) Part 2: Guidance on the operation of fluoro-opto-electronic counters BS EN ISO 13366-1:2008 IDF 148-1:2008 Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reprod
38、uction or networking permitted without license from IHS -,-,- blank Copyright British Standards Institution Provided by IHS under license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS
39、 -,-,- 1 Milk Enumeration of somatic cells Part 1: Mthode au microscope (Mthode de rfrence) 1 Scope This part of ISO 13366|IDF 148 specifies a microscopic method (reference method) for the counting of somatic cells in both raw and chemically preserved milk. This part of ISO 13366|IDF 148 is applicab
40、le for the counting of somatic cells in cows milk, provided that the eventually mentioned prerequisites are met. This method is suitable for preparing standard test samples and determining reference method values that are required for calibrating mechanized and automated cell-counting methods. WARNI
41、NG The use of this standard may involve hazardous materials, operations and equipment. This standard does not purport to address all of the safety problems associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determin
42、e the applicability of regulatory limitations prior to use. 2 Terms and definitions For the purposes of this document, the following terms and definitions apply. 2.1 somatic cells those cells with nuclei, that is all leucocytes and epithelial cells, determined according to the procedure described in
43、 this part of ISO 13366|IDF 148 3 Principle A test portion of milk to be examined is spread over a slide to form a smear. The smear is dried. During this process, the cells are stained. Subsequently, the stained cells are counted using a microscope. The number of cells counted in a defined area are
44、multiplied by a working factor, to give the number of cells per millilitre. 4 Reagents Use only reagents of recognized analytical grade, unless otherwise specified, and distilled or deionized water or water of equivalent purity. 4.1 Dye solutions WARNING Tetrachloroethane is poisonous. Ethidium brom
45、ide is mutagenic. Proper actions for deactivation should be taken in case of spilling. Preparation and application of the dye solution shall be carried out in a fume cupboard, using protective equipment. BS EN ISO 13366-1:2008 IDF 148-1:2008 Copyright British Standards Institution Provided by IHS un
46、der license with BSI - Uncontrolled Copy Licensee=Boeing Co/5910770001 Not for Resale, 08/14/2008 21:46:40 MDTNo reproduction or networking permitted without license from IHS -,-,- 2 4.1.1 Modified Newman-Lampert stain solution (Levowitz-Weber modification) 4.1.1.1 Components Ethanol, 95 % (volume f
47、raction) 54,0 ml Tetrachloroethane a 40,0 ml Methylene blue 0,6 g Acetic acid, glacial 6,0 ml a Xylene can be used as an alternative in the same volume amount as mentioned for tetrachloroethane. 4.1.1.2 Preparation Mix the ethanol and the tetrachloroethane and stopper the bottle. Heat the mixture in a water bath (5.1) set at 65 C. Add the methylene blue under a fume cupboard and carefully mix. Cool the mixture in a refrigerator to 4 C. Then add the glacial acetic acid and carefully mix again. Pass the obtained solution through an appropriate filter (5.2) into an