EN ISO 21150-2009 0000 Cosmetics - Microbiology - Detection of Escherichia coli《化妆品 微生物学 大肠杆菌的测定》.pdf

1、BRITISH STANDARDBS EN ISO 21150:2009Cosmetics Microbiology Detection of Escherichia coliICS 07.100.99; 71.100.70g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g5

2、5g3g47g36g58Provided by IHSNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-National forewordThis British Standard is the UK implementation of EN ISO 21150:2009. It is identical to ISO 21150:2006. It supersedes BS ISO 21150:2006 which is withdrawn.The UK participati

3、on in its preparation was entrusted to Technical Committee CW/217, Cosmetics.A list of organizations 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 corre

4、ct application.Compliance with a British Standard cannot confer immunity from legal obligations.BS EN ISO 21150:2009This British Standard waspublished under the authorityof the Standards Policy andStrategy Committeeon 31 May 2006 BSI 2010Amendments/corrigenda issued since publicationDate Comments 31

5、 August 2010 This corrigendum renumbers BS ISO 21150:2006 as BS EN ISO 21150:2009ISBN 978 0 580 66943 9Provided by IHSNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 21150June 2009ICS 07.100.99; 71.100.70English

6、 VersionCosmetics - Microbiology - Detection of Escherichia coli (ISO21150:2006)Cosmtiques - Microbiologie - Dtection dEscherichia coli(ISO 21150:2006)Kosmetik - Mikrobiologie - Nachweis von Escherichia coli(ISO 21150:2006)This European Standard was approved by CEN on 30 May 2009.CEN members are bou

7、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 CEN

8、 Management 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 CEN Management Centre has the same status

9、as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slova

10、kia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: Avenue Marnix 17, B-1000 Brussels 2009 CEN All rights of exploitation in any form and by any means reservedworldwide for C

11、EN national Members.Ref. No. EN ISO 21150:2009: EProvided by IHSNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-Foreword The text of ISO 21150:2006 has been prepared by Technical Committee ISO/TC 217 “Cosmetics” of the International Organization for Standardization

12、 (ISO) and has been taken over as EN ISO 21150:2009. 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 2009, and conflicting national standards shall be withdrawn at the latest by December

13、2009. 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 the CEN/CENELEC Internal Regulations, the national standards organi

14、zations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Rom

15、ania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 21150:2006 has been approved by CEN as a EN ISO 21150:2009 without any modification. BS EN ISO 21150:2009EN ISO 21150:2009 (E)ii Provided by IHSNot for ResaleNo reproduction or networking

16、permitted without license from IHS-,-,-iiiContents Page Introduction . iv 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 Culture media

17、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 Detection a

18、nd 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 (informativ

19、e) Other enrichment broths. 11 Annex B (informative) Neutralizers of antimicrobial activity of preservatives and rinsing liquids . 14 Bibliography . 15 BS EN ISO 21150:2009EN ISO 21150:2009 (E)Provided by IHSNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-ivIntrodu

20、ction Microbiological examinations of cosmetic products are to be carried out 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 pro

21、ducts; pathogenicity of microorganisms; site of application of the cosmetic product (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 a

22、eruginosa and Candida albicans may be relevant. The detection of other kinds 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 EN ISO 21150:2009EN ISO 21150

23、:2009 (E)Provided by IHSNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-1Cosmetics Microbiology Detection of Escherichia coli 1 Scope This International Standard gives general guidelines for the detection and identification of the specified microorganism Escherichi

24、a 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 appropriate microbiologic

25、al 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 International Standard speci

26、fies 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 Escherichia coli, subcultur

27、es 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 (e.g. certain water-immi

28、scible 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 following referenced document

29、s 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 for microbiological exam

30、ination 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 the test to prepare the

31、 initial suspension 1)To be published. BS EN ISO 21150:2009EN ISO 21150:2009 (E)Provided by IHSNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-2 3.3 initial suspension suspension (or solution) of the sample in a defined volume of an appropriate enrichment broth 3.4

32、 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 process 3.6 Esc

33、herichia 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 Escherichia coli c

34、an 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 the proced

35、ure 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 selective medi

36、um followed by identification tests. The possible inhibition of microbial growth by the sample shall be neutralized to allow the detection of viable microorganisms5. In all cases and whatever the methodology, the neutralization of the antimicrobial properties of the product shall be checked and vali

37、dated6 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 microbial p

38、opulation. 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 checking the p

39、resence 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 EN ISO 21150:2009EN ISO 21150:2009 (E)Provided by IHSNot for ResaleNo repr

40、oduction or networking permitted without license from IHS-,-,-35.2 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

41、 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 measur

42、ed 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-

43、use 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

44、meal 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 coolin

45、g down, the pH shall be equivalent to 7,3 0,2 when measured at room temperature. BS EN ISO 21150:2009EN ISO 21150:2009 (E)Provided by IHSNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-4 5.3.3 Enrichment broth 5.3.3.1 Eugon LT 100 broth 5.3.3.1.1 General This mediu

46、m contains ingredients 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 Glu

47、cose 5,5 g Egg 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. Di

48、spense the medium 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 Selective 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 EN ISO 21150:2009EN ISO 21150:2009 (E)Provided by IHSNo