1、BS EN ISO 18416:2015Cosmetics Microbiology Detection of Candida albicansIncorporating corrigendum February 2017(ISO 18416:2015)BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06BS EN ISO 18416:2015 BRITISH STANDARDNational forewordThis British Standard is the UK implem
2、entation of EN ISO 18416:2015. It supersedes BS EN ISO 18416:2009 which iswithdrawn.The UK participation 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 n
3、ot purport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2017.Published by BSI Standards Limited 2017ISBN 978 0 580 97526 4 ICS 07.100.40 Compliance with a British Standard cannot confer immunity from legal
4、 obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 December 2015.Amendments/corrigenda issued since publicationDate T e x t a f f e c t e dImplementation of ISO corrected text 15 December28 February 20172016: See ISO Foreword for
5、 detailsEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 18416 December 2015 ICS 07.100.40 Supersedes EN ISO 18416:2009English Version Cosmetics - Microbiology - Detection of Candida albicans (ISO 18416:2015, Corrected version 2016-12-15) Cosmtiques - Microbiologie - Dtection de Candida albi
6、cans (ISO 18416:2015, Version corrige 2016-12-15) Kosmetische Mittel - Mikrobiologie - Nachweis von Candida albicans (ISO 18416:2015, korrigierte Fassung 2016-12-15) This European Standard was approved by CEN on 26 September 2015. CEN members are bound to comply with the CEN/CENELEC Internal Regulat
7、ions which 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.
8、 This European 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
9、members 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, Po
10、rtugal, 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
11、form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 18416:2015 EEN ISO 18416:2015 (E) European foreword This document (EN ISO 18416:2015) has been prepared by Technical Committee ISO/TC 217 “Cosmetics“ in collaboration with Technical Committee CEN/TC 392 “Cosmetics” th
12、e 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 juin 2016, and conflicting national standards shall be withdrawn at the latest by juin 2016. Attention is d
13、rawn 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 18416:2009. This document has been prepared under a mandate given to
14、 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 Republic, Denm
15、ark, 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. Endorsement no
16、tice The text of ISO 18416:2015, Corrected version 2016-12-15 has been approved by CEN as EN ISO 18416:2015 without any modification. BS EN ISO 18416:2015iiISO 18416:2015(E)Foreword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 25 Diluents and culture medi
17、a . 25.1 General . 25.2 Diluent for the yeast 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 suitability test (see Clause 11) 35.3.3 Enrichment broth 45.3.4 Selective agar medium for i
18、solation of Candida albicans 55.3.5 Corn meal agar with 1 % polysorbate 80 . 56 Apparatus and glassware 57 Strains of microorganisms . 58 Handling of cosmetic products and laboratory samples 69 Procedure. 69.1 General recommendation 69.2 Preparation of the initial suspension in the enrichment broth
19、69.2.1 General 69.2.2 Water-miscible products. 69.2.3 Water-immiscible products 79.2.4 Filterable products . 79.3 Incubation of the inoculated enrichment broth . 79.4 Detection and identification of Candida albicans 79.4.1 Isolation . 79.4.2 Identification of Candida albicans . 710 Expression of the
20、 results (detection of Candida albicans) 811 Neutralization of the antimicrobial properties of the product 811.1 General . 811.2 Preparation of inoculum . 811.3 Suitability of the detection method . 911.3.1 Procedure . 911.3.2 Interpretation of suitability test results 912 Test report . 9Annex A (in
21、formative) Other media .11Annex B (informative) Neutralizers of antimicrobial activity of preservatives and rinsing liquids 16Bibliography .17 ISO 2015 All rights reserved iiiContents PageBS EN ISO 18416:2015ISO 18416:2015(E)ForewordISO (the International Organization for Standardization) is a world
22、wide 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 established has the right to be represented on t
23、hat 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 standardization.The procedures used to develop this doc
24、ument 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 noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directi
25、ves, 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 identifying any or all such patent rights. Details of any patent rights identified during the developme
26、nt 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 of users and does not constitute an endorsement.For an explanation on the meaning of ISO speci
27、fic 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 - Supplementary informationThe committee responsible for this document is ISO/TC 217, Cosmetics.This
28、second edition cancels and replaces the first edition (ISO 18416:2007), of which it constitutes a minor revision.This corrected version of ISO 18416:2015 incorporates the following correction: in the second paragraph of 11.2, “108CFU per ml” has been replaced by “106CFU per ml”. iv ISO 2015 All righ
29、ts reservedBS EN ISO 18416:2015ISO 18416:2015(E)IntroductionMicrobiological examinations 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 parameter
30、s such as the following: potential alteration of cosmetic products; pathogenicity 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 pathog
31、ens such as Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans may 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 c
32、oli) suggest hygienic failure during the manufacturing process. ISO 2015 All rights reserved vBS EN ISO 18416:2015Cosmetics Microbiology Detection of Candida albicans1 ScopeThis International Standard gives general guidelines for the detection and identification of the specified microorganism Candid
33、a albicans 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 microbiolo
34、gical risk analysis to determine the types of cosmetic product 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
35、 this International Standard is based on the detection of Candida albicans in a non-selective liquid medium (enrichment broth), followed by isolation on a selective agar medium. Other methods may be appropriate dependent on the level of detection required.NOTE For the detection of Candida albicans,
36、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 may not be appropriate in every detail for some products (e.g. certai
37、n water immiscible products). Other International Standards (ISO 18415) may be appropriate. Other methods (e.g. automated) may be substituted for the tests presented here provided that their equivalence has been demonstrated or the method has been otherwise shown to be suitable.2 Normative reference
38、sThe following documents, in whole or in part, are normatively 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 211
39、48:2005, Cosmetics Microbiology General instructions for 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 bac
40、teriophages) activity3 Terms and definitionsFor the purposes 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 prepar
41、e the initial suspension3.3initial suspensionsuspension (or solution) of the sample in a defined volume of an appropriate enrichment brothINTERNATIONAL STANDARD ISO 18416:2015(E) ISO 2015 All rights reserved 1BS EN ISO 18416:2015ISO 18416:2015(E)3.4sample dilutiondilution of the initial suspension3.
42、5specified microorganismaerobic mesophilic bacterium or 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 process3.6Candida albicansyeast that form white to be
43、ige, creamy and convex colonies on the surface of a selective mediumNote 1 to entry: The main characteristic for identification is the production of germ tube and/or pseudomycelium and chlamydospore when the test is performed following the method specified in this International Standard.3.7enrichmen
44、t 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 medium to increase the number of microorganisms
45、 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 inhibition of microbial growth by the sample shall
46、 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 GeneralGeneral instructions are given in ISO
47、 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 neutralizers if the specimen to be tested has antimicro
48、bial 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 presence of Candida albicans in accordance w
49、ith this International Standard provided that it has 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 ISO 18416:2015ISO 18416:2015(E)5.2 Diluent for the yeast suspension (tryptone sodium chloride solution)5.2.1 GeneralThe diluent is used for the preparation of yeast suspension used for the suitability test procedure (see Clause 11).5.2.2 Composition tryptone, pancreatic digest of casein 1,
