BS EN ISO 16212-2011 Cosmetics Microbiology Enumeration of yeast and mould《化妆品 微生物学 酵母与霉菌的计数》.pdf

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1、BS EN ISO16212:2011ICS 07.100.99; 71.100.70NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBRITISH STANDARDCosmetics Microbiology Enumeration of yeastand mouldIncorporating corrigendumJune 2011Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled

2、 Copy, (c) BSINational forewordThis British Standard is the UK implementation of EN ISO 16212:2011. It is identical with ISO 16212:2008. It supersedes BS ISO 16212:2008, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee CW/217, Cosmetics.A list of organi

3、zations represented on this committee can be obtained on request 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 obligation

4、s.BS EN ISO 16212:2011This British Standard was published under the authority of the StandardsPolicy and Strategy Committee on 31 October 2008 BSI 2011Amendments/corrigenda issued since publicationDate Comments 31 July 2011 This corrigendum renumbers BS ISO 16212:2008 as BS EN ISO 16212:2011ISBN 978

5、 0 580 72865 5Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSIEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 16212 June 2011 ICS 71.100.70 English Version Cosmetics - Microbiology - Enumeration of yeast and mould (ISO 16212:2008) Cosmtique

6、s - Microbiologie - Dnombrement des levures et des moisissures (ISO 16212:2008) Kosmetik - Mikrobiologie - Zhlung von Hefen und Schimmelpilzen (ISO 16212:2008) This European Standard was approved by CEN on 12 May 2011. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which s

7、tipulate 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 Europe

8、an 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

9、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, Romania, Slovakia, Slovenia, Spain, Sweden,

10、 Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2011 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref

11、. No. EN ISO 16212:2011: ELicensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSI3 Foreword The text of ISO 16212:2008 has been prepared by Technical Committee ISO/TC 217 “Cosmetics” of the International Organization for Standardization (ISO) and has been

12、taken over as EN ISO 16212:2011 by Technical Committee CEN/TC 392 “Cosmetics” 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 latest by December 2011, and conflictin

13、g national standards shall be withdrawn at the latest by December 2011. 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. According to t

14、he 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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvi

15、a, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 16212:2008 has been approved by CEN as a EN ISO 16212:2011 without any modification. BS EN ISO16212:2011EN ISO162

16、12:2011 (E)Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSI ISO 201 iiiContents Page Foreword iv 1 Scope . 1 2 Normative references . 1 3 Terms and definitions. 1 4 Principles. 2 4.1 General. 2 4.2 Plate count. 2 4.3 Membrane filtration. 2 5 Diluent

17、s, neutralizers and culture media 3 5.1 General. 3 5.2 Neutralizing diluents and diluents 3 5.3 Diluent for yeast suspension (tryptone sodium chloride solution). 4 5.4 Culture media 4 6 Apparatus and glassware 5 7 Strain of microorganisms 5 8 Handling of cosmetic products and laboratory samples . 6

18、9 Procedure 6 9.1 General recommendation 6 9.2 Preparation of the initial suspension 6 9.3 Counting methods 6 10 Counting of colonies (plate counts and membrane filtration methods). 7 11 Expression of results . 8 11.1 Method of calculation for plate count. 8 11.2 Interpretation. 8 12 Neutralization

19、of the antifungicidal properties of the product. 10 12.1 General. 10 12.2 Preparation of inoculum 10 12.3 Validation of counting methods 11 13 Test report . 12 Annex A (informative) Other neutralizing diluents . 13 Annex B (informative) Other diluents. 15 Annex C (informative) Other culture media .

20、16 Annex D (informative) Neutralizers of antifungicidal activity of preservatives and rinsing liquids 18 Bibliography . 19 1BS EN ISO16212:2011EN ISO16212:2011 (E)Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSIiv ISO 2011Foreword ISO (the Internati

21、onal 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

22、 established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standa

23、rdization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for

24、voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility 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

25、such patent rights. ISO 16212 was prepared by Technical Committee ISO/TC 217, Comestics. BS EN ISO16212:2011EN ISO16212:2011 (E)Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSIINTERNATIONAL STANDARD ISO 201 1Cosmetics Microbiology Enumeration of yea

26、st and mould 1 Scope This International Standard gives general guidelines for enumeration of yeast and mould present in cosmetics by counting the colonies on selective agar medium after aerobic incubation. In order to ensure product quality and safety for consumers, it is advisable to perform an app

27、ropriate microbiological risk analysis so as to determine the types of cosmetic products to which this International Standard is applicable. Products considered to present a low microbiological risk include those with low water activity, hydro-alcoholic products, products with extreme pH values, etc

28、. Because of the large variety of cosmetic products within this field of application, this method might not be suited to some products in every detail (e.g. certain water-immiscible products). Other methods (e.g. automated) can be used for the test presented here provided that their equivalence has

29、been demonstrated or the method has been otherwise validated. Yeast enumerated can be identified using suitable identification tests, for example tests described in the standards listed in the Bibliography. Mould enumerated can be identified by other appropriate methods, if necessary. 2 Normative re

30、ferences The following referenced documents are indispensable for the application 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. ISO 21148, Cosmetics Microbiology Genera

31、l instructions for microbiological examination EN 12353, Chemical disinfectants and antiseptics Preservation of test organisms used for the determination of bactericidal, mycobactericidal, sporicidal and fungicidal activity 3 Terms and definitions For the purposes of this document, the following ter

32、ms and definitions apply. 3.1 yeast single-cell fungus, which multiplies mainly vegetatively by budding, able to grow under the test conditions specified in this International Standard 3.2 mould mycelium forming microfungus, including spores and conidia, able to grow under the test conditions specif

33、ied in this International Standard 1EN ISO16212:2011 (E)BS EN ISO16212:2011Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSI2 ISO 20113.3 product portion of an identified cosmetic product received in the laboratory for testing 3.4 sample portion of t

34、he product (at least 1 g or 1 ml) which is used in the test to prepare the initial suspension 3.5 initial suspension suspension (or solution) of a sample in a defined volume of an appropriate liquid (diluent, neutralizer, broth or combination thereof) 3.6 sample dilution(s) dilution(s) of the initia

35、l suspension 4 Principles 4.1 General This method involves enumeration of colonies on a selective agar medium. The possible inhibition of fungal growth by the sample shall be neutralized to allow the detection of viable microorganism (see Reference 2). In all cases and whatever the methodology, the

36、neutralization of the antifungicidal properties of the product shall be checked and validated (see References 3, 5, 6). 4.2 Plate count Plate count consists of the following steps. a) Preparation of poured plates or spread plates, using a specified culture medium, and inoculation of the plates using

37、 a defined quantity of the initial suspension or dilution of the product. b) Aerobic incubation of the plates at 25 C 2,5 C for 3 d to 5 d. c) Counting of the number of colony-forming units (CFU) and calculation of the amount of yeast and mould per millilitre or per gram of product. NOTE An alternat

38、ive condition for incubation is 22,5 C 2,5 C for 5 d to 7 d using the culture medium without antibiotic. 4.3 Membrane filtration Membrane filtration consists of the following steps. a) Transfer of a suitable amount of the sample, prepared by a valid method, in the filtration apparatus, wetted with a

39、 small volume of an appropriate sterile diluent. Immediate filtration and washing according to the validated procedure. Transfer of the membrane filter on to the surface of the specified agar medium as specified in ISO 21148. b) Aerobic incubation of the membranes at 25 C 2,5 C for 3 d to 5 d. c) Co

40、unting of the number of colony-forming units (CFU) and calculation of the amount of yeast and mould per millilitre or per gram of product. NOTE An alternative condition for incubation is 22,5 C 2,5 C for 5 d to 7 d using the culture medium without antibiotic. BS EN ISO16212:2011EN ISO16212:2011 (E)L

41、icensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSI ISO 201 35 Diluents, neutralizers and culture media 5.1 General General specifications are given in ISO 21148. When water is mentioned in a formula, use distilled water or purified water as specified i

42、n ISO 21148. The following diluents, neutralizers and culture media are suitable for enumeration of yeasts and moulds. Other diluents, neutralizers and culture media may be used if they have been demonstrated to be suitable for use. 5.2 Neutralizing diluents and diluents 5.2.1 General The diluent is

43、 used to disperse the sample. It may contain neutralizers if the sample to be tested has antifungicidal properties. The efficacy of the neutralization shall be demonstrated before the determination of the count (see Clause 12). Information relative to suitable neutralizers is given in Annex D. 5.2.2

44、 Neutralizing diluent 5.2.2.1 Fluid casein digest-soy lecithin-polysorbate 20 medium (SCDLP 20 broth) 5.2.2.1.1 Composition pancreatic digest of casein 20,0 g soy lecithin 5,0 g polysorbate 20 40 ml water 960 ml 5.2.2.1.2 Preparation Dissolve the polysorbate 20 in 960 ml of water by mixing while hea

45、ting in a water bath at 49 C 2 C. Add pancreatic digest of casein and soy lecithin. Heat for about 30 min to effect solution. Mix and dispense the medium into suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilization, the pH shall be equivalent to 7,3 0,2 when measured

46、 at room temperature. 5.2.2.2 Other neutralizing diluents Other neutralizing diluents may be used as appropriate (see Annex A and Annex D). 5.2.3 Diluent 5.2.3.1 Fluid A 5.2.3.1.1 Composition peptic digest of animal tissue 1,0 g water 1 000 ml 1BS EN ISO16212:2011EN ISO16212:2011 (E)Licensed Copy: W

47、ang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/11/2011 06:29, Uncontrolled Copy, (c) BSI4 ISO 20115.2.3.1.2 Preparation Dissolve 1 g of peptone in water to make 1 l. Heat with frequent agitation. Dispense into suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilization the pH

48、 shall be equivalent to 7,1 0,2 when measured at room temperature. 5.2.3.2 Other diluents Other diluents may be used as appropriate (see Annex B). 5.3 Diluent for yeast suspension (tryptone sodium chloride solution) 5.3.1 Composition tryptone, pancreatic digest of casein 1,00 g sodium chloride 8,50

49、g water 1 000 ml 5.3.2 Preparation Dissolve the components in the water by mixing while heating. Dispense into suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilization the pH shall be equivalent to 7,0 0,2 when measured at room temperature. 5.4 Culture media 5.4.1 General Culture media may be prepared as follows, or from dehydrated culture media according to the manufacturers instructions. Ready-to-use media may be used when their composition and/or growth yields are compa