《BS-ISO-21150-2006.pdf》由会员分享，可在线阅读，更多相关《BS-ISO-21150-2006.pdf（24页珍藏版）》请在三一文库上搜索。
1、BRITISH STANDARD BS ISO 21150:2006 Cosmetics Microbiology Detection of Escherichia coli ICS 07.100.99; 71.100.70 ? Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS ISO 21150:2006 This British Standard was published under the authority of t
2、he Standards Policy and Strategy Committee on 31 May 2006 BSI 2006 ISBN 0 580 48338 X National foreword This British Standard reproduces verbatim ISO 21150:2006 and implements it as the UK national standard. The UK participation in its preparation was entrusted to Technical Committee CW/217, Cosmeti
3、cs, which has the responsibility to: aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep UK interests informed; monitor related international and European developments and promulgate
4、them in the UK. A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international publications referred to in this document may be found in the BSI Catalogue under the section entitled “Internationa
5、l Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online. 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 S
6、tandard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, the ISO title page, pages ii to v, a blank page, pages 1 to 15 and a back cover. The BSI copyright notice displayed in this document indicates when the do
7、cument was last issued. Amendments issued since publication Amd. No. DateComments Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI Reference number ISO 21150:2006(E) INTERNATIONAL STANDARD ISO 21150 First edition 2006-04-15 Cosmetics Microbio
8、logy Detection of Escherichia coli Cosmtiques Microbiologie Dtection dEscherichia coli BS ISO 21150:2006 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ii Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncon
9、trolled Copy, (c) BSI BS ISO 21150:2006 iii Contents Page Foreword iv Introduction v 1 Scope . 1 2 Normative references . 1 3 Terms and definitions. 1 4 Principle . 2 5 Diluents and culture media 2 5.1 General. 2 5.2 Diluent for the bacterial suspension (tryptone sodium chloride solution). 3 5.3 Cul
10、ture media 3 6 Apparatus and glassware 6 7 Strains of microorganisms 6 8 Handling of cosmetic products and laboratory samples . 6 9 Procedure 6 9.1 General recommendations 6 9.2 Preparation of the initial suspension in the enrichment broth 6 9.3 Incubation of the inoculated enrichment broth . 7 9.4
11、Detection and identification of Escherichia coli 7 10 Expression of the results (detection of Escherichia coli) 8 11 Neutralization of the antimicrobial properties of the product 8 11.1 General. 8 11.2 Preparation of inoculum. 9 11.3 Validation of the detection method. 9 12 Test report . 10 Annex A
12、(informative) Other enrichment broths. 11 Annex B (informative) Neutralizers of antimicrobial activity of preservatives and rinsing liquids . 14 Bibliography . 15 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS ISO 21150:2006 iv Foreword
13、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
14、committee has been 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 elect
15、rotechnical 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 to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the
16、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 possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for ident
17、ifying any or all such patent rights. ISO 21150 was prepared by Technical Committee ISO/TC 217, Cosmetics. Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI v Introduction Microbiological examinations of cosmetic products are to be carried out
18、 according to an appropriate microbiological risk analysis in order to ensure their quality and safety for consumers. Microbiological risk analysis depends on several parameters such as: potential alteration of cosmetic products; pathogenicity of microorganisms; site of application of the cosmetic p
19、roduct (hair, skin, eyes, mucous membranes, etc.); type of users (adults, children under 3 years, etc.). For cosmetics and other topical products, the detection of skin pathogens such as Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans may be relevant. The detection of other kinds
20、of microorganisms might be of interest since these microorganisms (including indicators of faecal contamination, e.g. Escherichia coli) suggest hygienic failure during manufacturing process. BS ISO 21150:2006 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolle
21、d Copy, (c) BSI blank Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 1 Cosmetics Microbiology Detection of Escherichia coli 1 Scope This International Standard gives general guidelines for the detection and identification of the specified m
22、icroorganism Escherichia coli in cosmetic products. Microorganisms considered as specified in this International Standard might differ from country to country according to national practices or regulations. In order to ensure product quality and safety for consumers, it is advisable to perform an ap
23、propriate 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, extreme pH values, etc. This Inter
24、national Standard specifies a method that is based on the detection of Escherichia coli in a non-selective liquid medium (enrichment broth), followed by isolation on a selective agar medium. Other methods may be appropriate depending on the level of detection required. NOTE For the detection of Esch
25、erichia coli, subcultures can be performed on non-selective culture media followed by suitable identification steps (e.g. using identification kits). 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 (
26、e.g. certain water-immiscible products). Other International Standards may be appropriate. Other methods (e.g. automated) can be substituted for the test presented here provided that their equivalence has been demonstrated or the method has been otherwise validated. 2 Normative references The follow
27、ing 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 21148:1), Cosmetics Microbiology General instructions f
28、or microbiological examination 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 product portion of an identified cosmetic product received in the laboratory for testing 3.2 sample portion of the product (at least 1 g or 1 ml) which is used in
29、the test to prepare the initial suspension 1) To be published. BS ISO 21150:2006 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 2 3.3 initial suspension suspension (or solution) of the sample in a defined volume of an appropriate enrichment
30、 broth 3.4 sample dilution dilution of the initial suspension 3.5 specified microorganism aerobic mesophilic bacteria or yeast which is undesirable in a cosmetic product because it can cause skin or eye infection, or it can be recognized as an indicator of hygienic failure in the manufacturing proce
31、ss 3.6 Escherichia coli Gram-negative rod, motile, smooth colonies NOTE 1 The main characteristics for identification are catalase positive, oxidase negative, fermentation of lactose, production of indole, growth on selective medium containing bile salts with characteristic colonies. NOTE 2 Escheric
32、hia coli can be isolated from the moist environmental sources (air, water, soil) and is a faecal contamination indicator. 3.7 enrichment broth non-selective liquid medium containing suitable neutralizers and/or dispersing agents and validated for the product under test 4 Principle The first step of
33、the procedure is to perform an enrichment by using a non-selective broth medium to increase the number of microorganisms without the risk of inhibition by the selective ingredients that are present in selective/differential growth media. The second step (isolation) of the test is performed on a sele
34、ctive medium followed by identification tests. The possible inhibition of microbial growth by the sample shall be neutralized to allow the detection of viable microorganisms 5. In all cases and whatever the methodology, the neutralization of the antimicrobial properties of the product shall be check
35、ed and validated 6 7 8. 5 Diluents and culture media 5.1 General Use the general instructions given in ISO 21148. When water is mentioned in this document, use distilled water or purified water as specified in ISO 21148. The enrichment broth is used to disperse the sample and to increase the initial
36、 microbial population. It may contain neutralizers if the specimen to be tested has antimicrobial properties. The efficacy of the neutralization shall be demonstrated (see Clause 11). Information relative to suitable neutralizers is given in Annex B. The following enrichment broth is suitable for ch
37、ecking the presence of Escherichia coli according to this International Standard provided that it is validated according to Clause 11. Other diluents and culture media may be used if they have been demonstrated to be suitable for use. BS ISO 21150:2006 Licensed Copy: sheffieldun sheffieldun, na, Sun
38、 Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 3 5.2 Diluent for the bacterial suspension (tryptone sodium chloride solution) The diluent is used for the preparation of bacterial suspension used for the validation procedure (see Clause 11). 5.2.1 Composition Tryptone, pancreatic digest
39、of casein 1,0 g Sodium chloride 8,5 g Water 1 000 ml 5.2.2 Preparation Dissolve the components in water by mixing while heating. Dispense into suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilization and cooling down, the pH shall be equivalent to 7,0 0,2 when measure
40、d at room temperature. 5.3 Culture media 5.3.1 General Culture media may be prepared using the descriptions provided below or from dehydrated culture media, according to the instructions from the manufacturer. The instructions provided by the supplier of the media should be followed. NOTE Ready-to-u
41、se media may be used when their composition and/or growth yields are comparable to those of the formulas given herein. 5.3.2 Agar medium for validation soybean-casein digest agar medium (SCDA) or tryptic soy agar (TSA) 5.3.2.1 Composition Pancreatic digest of casein 15,0 g Papaic digest of soybean m
42、eal 5,0 g Sodium chloride 5,0 g Agar 15,0 g Water 1 000 ml 5.3.2.2 Preparation Dissolve the components or the dehydrated complete medium in the water by mixing while heating. Dispense the medium into suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilization and cooling
43、 down, the pH shall be equivalent to 7,3 0,2 when measured at room temperature. BS ISO 21150:2006 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2006, Uncontrolled Copy, (c) BSI 4 5.3.3 Enrichment broth 5.3.3.1 Eugon LT 100 broth 5.3.3.1.1 General This medium contains ingr
44、edients which neutralize inhibitory substances present in the sample: lecithin and polysorbate 80, dispersing agent: octoxynol 9. 5.3.3.1.2 Composition Pancreatic digest of casein 15,0 g Papaic digest of soybean meal 5,0 g L-cystine 0,7 g Sodium chloride 4,0 g Sodium sulfite 0,2 g Glucose 5,5 g Egg
45、lecithin 1,0 g Polysorbate 80 5,0 g Octoxynol 9 1,0 g Water 1 000 ml 5.3.3.1.3 Preparation Dissolve the components, polysorbate 80, octoxynol 9 and egg lecithin, successively into boiling water until their complete dissolution. Dissolve the other components by mixing while heating. Dispense the medi
46、um into suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilization and cooling down, the pH shall be equivalent to 7,0 0,2 when measured at room temperature. 5.3.3.2 Other enrichment broths Other enrichment broths may be used as appropriate (see Annex A). 5.3.4 Selectiv
47、e agar medium for isolation of Escherichia coli 5.3.4.1 MacConkey agar medium 5.3.4.1.1 Composition Pancreatic digest of gelatin 17,0 g Pancreatic digest of casein 1,5 g Peptic digest of animal tissue 1,5 g BS ISO 21150:2006 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 26 02:56:59 GMT+00:00 2
48、006, Uncontrolled Copy, (c) BSI 5 Lactose 10,0 g Bile salts mixture 1,5 g Sodium chloride 5,0 g Agar 13,5 g Neutral red 30,0 mg Crystal violet 1,0 mg Water 1 000 ml 5.3.4.1.2 Preparation Dissolve all solid components in the water and boil for 1 min to effect solution. Dispense in suitable containers
49、 and sterilize at 121 C for 15 min. The pH, after sterilization and cooling down, shall be equivalent to 7,1 0,2 when measured at room temperature. 5.3.5 Selective agar medium for confirmation of Escherichia coli 5.3.5.1 Levine eosin-methylene blue agar medium 5.3.5.1.1 Composition Pancreatic digest of gelatin 10,0 g Potassium dihydrogen phosphate (KH2PO4) 2,0 g Agar 15,0 g Lactose 10,0 g Eosine Y 400 mg Methylene blue 65 mg Water 1 000 ml 5.3.5.1.2 Preparatio