1、September 2017 English price group 13No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 07.100.40!%mPs“2744580www.din.
2、deDIN EN ISO 16212Cosmetics Microbiology Enumeration of yeast and mould (ISO 16212:2017);English version EN ISO 16212:2017,English translation of DIN EN ISO 16212:2017-09Kosmetische Mittel Mikrobiologie Zhlung von Hefen und Schimmelpilzen (ISO 16212:2017);Englische Fassung EN ISO 16212:2017,Englisch
3、e bersetzung von DIN EN ISO 16212:2017-09Cosmtiques Microbiologie Dnombrement des levures et des moisissures (ISO 16212:2017);Version anglaise EN ISO 16212:2017,Traduction anglaise de DIN EN ISO 16212:2017-09SupersedesDIN EN ISO 16212:201108www.beuth.deDocument comprises 26 pagesDTranslation by DIN-
4、Sprachendienst.In case of doubt, the German-language original shall be considered authoritative.08.17 DIN EN ISO 16212:2017-09 2 A comma is used as the decimal marker. National foreword This document (EN ISO 16212:2017) has been prepared by Technical Committee ISO/TC 217 “Cosmetics” (Secretariat: IS
5、IRI, Iran) in collaboration with Technical Committee CEN/TC 392 “Cosmetics” (Secretariat: AFNOR, France). The responsible German body involved in its preparation was DIN-Normenausschuss Lebensmittel und landwirtschaftliche Produkte (DIN Standards Committee Food and Agricultural Products), Working Co
6、mmittee NA 057-07-01 AA “Cosmetics”. The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 18415 DIN EN ISO 18415 ISO 18416 DIN EN ISO 18416 ISO 21148 DIN EN ISO 21148 ISO 29621 DIN EN ISO 29621 Amendments This standard differs from DIN EN IS
7、O 16212:2011-08 as follows: a) normative references have been updated; b) the term “validation” (in the German version Validierung) has been replaced throughout the text by “suitability” (in the German version Eignung); c) the standard has been editorially revised. Previous editions DIN EN ISO 16212
8、: 2011-08 National Annex NA (informative) Bibliography DIN EN ISO 18415, Cosmetics Microbiology Detection of specified and non-specified microorganisms DIN EN ISO 18416, Cosmetics Microbiology Detection of Candida albicans DIN EN ISO 21148, Cosmetics Microbiology General instructions for microbiolog
9、ical examination DIN EN ISO 29621, Cosmetics Microbiology Guidelines for the risk assessment and identification of microbiologically low-risk products EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 16212 July 2017 ICS 07.100.40 Supersedes EN ISO 16212:2011English Version Cosmetics - Microb
10、iology - Enumeration of yeast and mould (ISO 16212:2017) Cosmtiques - Microbiologie - Dnombrement des levures et des moisissures (ISO 16212:2017) Kosmetische Mittel - Mikrobiologie - Zhlung von Hefen und Schimmelpilzen (ISO 16212:2017) This European Standard was approved by CEN on 26 April 2017. CEN
11、 members are bound to comply with the CEN/CENELEC Internal Regulations 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 appl
12、ication to the CEN-CENELEC 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 translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Ma
13、nagement Centre has the same status as the official versions. CEN 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, Lat
14、via, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Av
15、enue Marnix 17, B-1000 Brussels 2017 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 16212:2017 EEuropean foreword .31 Scope . 62 Normative references 63 Terms and definitions . 64 Principles . 74.1 General . 74.2 Plate count 7
16、4.3 Membrane filtration . 75 Diluents, neutralizers and culture media . 85.1 General . 85.2 Neutralizing diluents and diluents 85.3 Diluent for yeast suspension (tryptone sodium chloride solution) 95.4 Culture media . 96 Apparatus and glassware . 107 Strain of microorganisms . 108 Handling of cosmet
17、ic products and laboratory samples . 119 Procedure 119.1 General recommendation . 119.2 Preparation of the initial suspension . 119.2.1 General. 119.2.2 Water-miscible products 119.2.3 Water-immiscible products . 119.3 Counting methods . 119.3.1 Dilutions for counting methods . 119.3.2 Plate-count m
18、ethods . 1210 Counting of colonies (plate counts and membrane filtration methods) . 1211 Expression of results . 1311.1 Method of calculation for plate count 1311.2 Interpretation . 1412 Neutralization of the antifungicidal properties of the product 1512.1 General 1512.2 Preparation of inoculum 1612
19、.3 Suitability of counting methods 1612.3.1 Principle . 1612.3.2 Suitability test of the pour-plate method 1612.3.3 Suitability of the surface spread method .1612.3.4 Suitability of the membrane filtration method .1613 Test report 17Annex A (informative) Other neutralizing diluents .18Annex B (infor
20、mative) Other diluents 20Annex C (informative) Other culture media .21Annex D (informative) Neutralizers of antifungicidal activity of preservatives and rinsing liquids 23Bibliography .24Contents PageDIN EN ISO 16212:2017-09 EN ISO 16212:2017 (E) 2Foreword .4European foreword This document (EN ISO 1
21、6212:2017) has been prepared by Technical Committee ISO/TC 217 “Cosmetics“ in collaboration with Technical Committee CEN/TC 392 “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 o
22、r by endorsement, at the latest by January 2018 and conflicting national standards shall be withdrawn at the latest by January 2018. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN shall not be held responsible for identifying
23、 any or all such patent rights. This document supersedes EN ISO 16212:2011. 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, D
24、enmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. End
25、orsement notice The text of ISO 16212:2017 has been approved by CEN as EN ISO 16212:2017 without any modification. DIN EN ISO 16212:2017-09 EN ISO 16212:2017 (E) 3 ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodi
26、es). 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 that committee. International organizations, governmental and
27、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 document and those intended for its further maintenance are des
28、cribed 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 Directives, Part 2 (see www .iso .org/ directives).Attention is dra
29、wn 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 development of the document will be in the Introduction and/or on
30、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 voluntary nature of standards, the meaning of ISO specific terms and expr
31、essions related to conformity assessment, as well as information about ISOs adherence to the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following URL: www .iso .org/ iso/ foreword .html.This document was prepared by Technical Committee ISO/TC 217, Cosm
32、etics.This second edition cancels and replaces the first edition (ISO 16212:2008), of which it constitutes a minor revision. The changes compared to the previous edition are as follows: in the Scope, “see ISO 29621” has been added and the reference has been added to the Bibliography; in the Scope, “
33、used” has been changed to “substituted” and “validated” has been changed to “shown to be suitable”; in 4.1, “validated” has been changed to “demonstrated”; in 4.3, “by a valid method” has been changed to “as described in Clause 12” and “validated procedure” has been replaced by “described procedure”
34、; in 5.1, “specifications” has been changed to “instructions”; in 5.2.3.1.2, “peptone” has been changed to “peptic digest of animal tissue”; in Clause 7, “validation” has been changed to “suitability”; in 9.3.2.1, “validated” has been changed to “demonstrated to be suitable”; in 9.3.2.3, “prepared a
35、s validated” has been changed to “demonstrated to be suitable”; in 11.2.1, “validated according to” has been changed to “demonstrated to be suitable for”; in 12.3, “validation” has been changed to “suitability”; in 12.3.2, instances of “validation” have been changed to “suitability test” and “valida
36、ted” has been changed to “satisfactory”; in 12.3.3, the first instance of “validation” has been changed to “suitability” and the second instance has been changed to “suitability test”; “validated” has been changed to “satisfactory”;DIN EN ISO 16212:2017-09 EN ISO 16212:2017 (E) 4 in 12.3.4, the firs
37、t instance of “validation” has been changed to “suitability” and the second instance has been changed to “suitability test”; “validated” has been changed to “satisfactory”; in Clause 13 f), “validation” has been changed to “suitability”; in A.1, B.1 and C.1, “validated” has been changed to “demonstr
38、ated to be suitable”.DIN EN ISO 16212:2017-09 EN ISO 16212:2017 (E) 5 1 ScopeThis document 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 con
39、sumers, it is advisable to perform an appropriate microbiological risk analysis to determine the types of cosmetic products to which this document is applicable. Products considered to present a low microbiological risk (see ISO 29621) include those with low water activity or extreme pH values, hydr
40、o-alcoholic products, etc.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 substituted for the tests presented here prov
41、ided that their equivalence has been demonstrated or the method has been otherwise shown to be suitable.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 approp
42、riate methods, if necessary.2 Normative referencesThe following documents are referred to in the text in such a way that some or all of their content constitutes requirements of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the ref
43、erenced document (including any amendments) applies.ISO 21148, Cosmetics Microbiology General instructions for microbiological examinationEN 12353, Chemical disinfectants and antiseptics Preservation of test organisms used for the determination of bactericidal (including Legionella), mycobactericida
44、l, sporicidal, fungicidal and virucidal (including bacteriophages) activity3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.ISO and IEC maintain terminological databases for use in standardization at the following addresses: ISO Online browsing plat
45、form: available at h t t p :/ www .iso .org/ obp IEC Electropedia: available at h t t p :/ www .electropedia .org/ 3.1yeastsingle-cell fungus, which multiplies mainly vegetatively by budding, able to grow under the test conditions specified in this document3.2mouldmycelium forming microfungus, inclu
46、ding spores and conidia, able to grow under the test conditions specified in this documentDIN EN ISO 16212:2017-09 EN ISO 16212:2017 (E) 6 3.3productportion of an identified cosmetic product received in the laboratory for testing3.4sampleportion of the product (3.3) (at least 1 g or 1 ml) that is us
47、ed in the test to prepare the initial suspension3.5initial suspensionsuspension (or solution) of the sample (3.4) in a defined volume of an appropriate enrichment broth3.6sample dilutiondilution of the initial suspension (3.5)4 Principles4.1 GeneralThis method involves enumeration of colonies on a s
48、elective agar medium. The possible inhibition of fungal 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 antifungicidal properties of the product shall be checked and demonstrated689.4.2 Plate countPlate count consists of the following steps. Preparation of poured plates, or spread plates, using a specified cultur
