EN ISO 21150-2015 en Cosmetics - Microbiology - Detection of Escherichia coli《化妆品 微生物学 大肠杆菌的检测(ISO 21150 2015)》.pdf

1、BSI Standards PublicationBS EN ISO 21150:2015Cosmetics Microbiology Detection of Escherichia coliBS EN ISO 21150:2015 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO21150:2015. It supersedes BS EN ISO 21150:2009 which is withdrawn.The UK participation in its

2、 preparation was entrusted to TechnicalCommittee CW/217, Cosmetics.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication

3、. The British Standards Institution 2015. Published by BSI StandardsLimited 2015ISBN 978 0 580 89370 4ICS 07.100.99; 71.100.70Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Co

4、mmittee on 31 December 2015.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 21150 December 2015 ICS 07.100.99; 71.100.70 Supersedes EN ISO 21150:2009English Version Cosmetics - Microbiology - Detection of Escherichia coli (ISO 21150:2015)

5、 Cosmtiques - Microbiologie - Dtection dEscherichia coli (ISO 21150:2015) Kosmetische Mittel - Mikrobiologie - Nachweis von Escherichia coli (ISO 21150:2015) This European Standard was approved by CEN on 26 September 2015. CEN members are bound to comply with the CEN/CENELEC Internal Regulations whi

6、ch stipulate 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 Eu

7、ropean 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

8、are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,

9、Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey andUnited Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2015 CEN All rights of exploitation in any form and

10、 by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 21150:2015 EBS EN ISO 21150:2015EN ISO 21150:2015 (E) 3 European foreword This document (EN ISO 21150:2015) has been prepared by Technical Committee ISO/TC 217 “Cosmetics“ in collaboration with Technical Committee CEN/TC 392

11、“Cosmetics” the secretariat of which is held by AFNOR. 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 June 2016, and conflicting national standards shall be withdrawn at the latest by June 2016.

12、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. This document supersedes EN ISO 21150:2009. This document has been prepared under a ma

13、ndate given to CEN by the European Commission and the European Free Trade Association. 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

14、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 and the United Kingdom.

15、Endorsement notice The text of ISO 21150:2015 has been approved by CEN as EN ISO 21150:2015 without any modification. BS EN ISO 21150:2015ISO 21150:2015(E)Foreword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 25 Diluents and culture media . 25.1 General .

16、 25.2 Diluent for the bacterial suspension (tryptone sodium chloride solution) . 35.2.1 General 35.2.2 Composition . 35.2.3 Preparation . 35.3 Culture media . 35.3.1 General 35.3.2 Agar medium for the suitability test (see Clause 11) soybean-casein digest agar medium (SCDA) or tryptic soy agar (TSA)

17、 . 35.3.3 Enrichment broth 45.3.4 Selective agar medium for isolation of Escherichia coli . 45.3.5 Selective agar medium for confirmation of Escherichia coli 56 Apparatus and glassware 67 Strains of microorganisms . 68 Handling of cosmetic products and laboratory samples 69 Procedure. 69.1 General r

18、ecommendation 69.2 Preparation of the initial suspension in the enrichment broth 69.2.1 General 69.2.2 Water-miscible products. 79.2.3 Water-immiscible products 79.2.4 Filterable products . 79.3 Incubation of the inoculated enrichment broth . 79.4 Detection and identification of Escherichia coli . 7

19、9.4.1 Isolation . 79.4.2 Identification of Escherichia coli 710 Expression of the results (detection of Escherichia coli) 811 Neutralization of the antimicrobial properties of the product 811.1 General . 811.2 Preparation of inoculum . 811.3 Suitability of the detection method . 811.3.1 Procedure .

20、811.3.2 Interpretation of suitability test results 912 Test report . 9Annex A (informative) Other enrichment broths .10Annex B (informative) Neutralizers of antimicrobial activity of preservatives and rinsing liquids 13Bibliography .14 ISO 2015 All rights reserved iiiContents PageBS EN ISO 21150:201

21、5ISO 21150:2015(E)ForewordISO (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 subj

22、ect 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 take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IE

23、C) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be n

24、oted. 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 possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifyin

25、g 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 of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience o

26、f 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 information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supple

27、mentary informationThe committee responsible for this document is ISO/TC 217, Cosmetics.This second edition cancels and replaces the first edition (ISO 21150:2006), which has been technically revised.iv ISO 2015 All rights reservedBS EN ISO 21150:2015ISO 21150:2015(E)IntroductionMicrobiological exam

28、inations of cosmetic products are 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 the following: potential alteration of cosmetic products; pathogenicit

29、y of microorganisms; site of application of the cosmetic product (hair, skin, eyes, mucous membranes); type of users (adults, children under 3 years).For cosmetics and other topical products, the detection of skin pathogens such as Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans m

30、ay be relevant because they can cause skin or eye infections. The detection of other kinds of microorganism might be of interest since these microorganisms (including indicators of faecal contamination e.g. Escherichia coli) suggest hygienic failure during the manufacturing process. ISO 2015 All rig

31、hts reserved vBS EN ISO 21150:2015BS EN ISO 21150:2015Cosmetics Microbiology Detection of Escherichia coli1 ScopeThis International Standard gives general guidelines for the detection and identification of the specified microorganism Escherichia coli in cosmetic products. Microorganisms considered a

32、s 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 microbiological risk analysis, so as to determine the types of cosmeti

33、c products to which this International Standard is applicable. Products considered to present a low microbiological (see ISO 29621) risk include those with low water activity, hydro-alcoholic products, extreme pH values, etc.The method described in this International Standard is based on the detecti

34、on 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, subcultures can be performed on non-selective cultu

35、re 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-immiscible products). Other International Stand

36、ards (ISO 18415) 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 shown to be suitable.2 Normative referencesThe following documents, in whole or in part, are norma

37、tively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 21148:2005, Cosmetics Microbiology General instructions for

38、 microbiological examinationEN 12353, Chemical disinfectants and antiseptics Preservation of test organisms used for the determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal (including bacteriophages) activity3 Terms and definitionsFor the purp

39、oses of this document, the following terms and definitions apply.3.1productportion of an identified cosmetic product received in the laboratory for testing3.2sampleportion of the product (at least 1 g or 1 ml) that is used in the test to prepare the initial suspension3.3initial suspensionsuspension

40、(or solution) of the sample in a defined volume of an appropriate enrichment brothINTERNATIONAL STANDARD ISO 21150:2015(E) ISO 2015 All rights reserved 1BS EN ISO 21150:2015ISO 21150:2015(E)3.4sample dilutiondilution of the initial suspension3.5specified microorganismaerobic mesophilic bacterium or

41、yeast that is undesirable in a cosmetic product and is recognized as a skin pathogen species that may be harmful for human health or as an indication of hygienic failure in the manufacturing process3.6Escherichia coligram-negative rod, motile, smooth coloniesNote 1 to entry: The main characteristics

42、 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 to entry: Escherichia coli can be isolated from the moist environmental sources (air, water, soil) and is a

43、 faecal contamination indicator.3.7enrichment brothnon-selective liquid medium containing suitable neutralizers and/or dispersing agents and demonstrated to be suitable for the product under test4 PrincipleThe first step of the procedure is to perform an enrichment by using a non-selective broth med

44、ium 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 of the test (isolation) of the test is performed on a selective medium followed by identification tests.The possible inhibi

45、tion of microbial growth by the sample shall be neutralized to allow the detection of viable microorganisms.1In all cases and whatever the methodology, the neutralization of the antimicrobial properties of the product shall be checked and demonstrated (see Clause 11).5 Diluents and culture media5.1

46、GeneralGeneral instructions are given in ISO 21148. When water is mentioned in this International Standard, 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 population. It may contain neutralizer

47、s 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 enrichment broth (5.3.3.1) or any of the ones listed in Annex A is suitable for checking the pr

48、esence of Escherichia coli in accordance with this International Standard provided that they have been demonstrated to be suitable in accordance with Clause 11.Other diluents and culture media may be used if it has been demonstrated that they are suitable for use.2 ISO 2015 All rights reservedBS EN

49、ISO 21150:2015ISO 21150:2015(E)5.2 Diluent for the bacterial suspension (tryptone sodium chloride solution)5.2.1 GeneralThe diluent is used for the preparation of bacterial suspension used for the suitability test procedure (see Clause 11).5.2.2 Composition Tryptone, pancreatic digest of casein 1,0 g Sodium chloride 8,5 g Water 1 000 ml5.2.3 PreparationDissolve 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 sh