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    SANS 21149-2008 Cosmetics - Microbiology - Enumeration and detection of aerobic mesophilic bacteria《化妆品 微生物学 好氧嗜温菌的计数和检测》.pdf

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    SANS 21149-2008 Cosmetics - Microbiology - Enumeration and detection of aerobic mesophilic bacteria《化妆品 微生物学 好氧嗜温菌的计数和检测》.pdf

    1、 Collection of SANS standards in electronic format (PDF) 1. Copyright This standard is available to staff members of companies that have subscribed to the complete collection of SANS standards in accordance with a formal copyright agreement. This document may reside on a CENTRAL FILE SERVER or INTRA

    2、NET SYSTEM only. Unless specific permission has been granted, this document MAY NOT be sent or given to staff members from other companies or organizations. Doing so would constitute a VIOLATION of SABS copyright rules. 2. Indemnity The South African Bureau of Standards accepts no liability for any

    3、damage whatsoever than may result from the use of this material or the information contain therein, irrespective of the cause and quantum thereof. ISBN 978-0-626-22065-5 SANS 21149:2008Edition 1ISO 21149:2006Edition 1SOUTH AFRICAN NATIONAL STANDARD Cosmetics Microbiology Enumeration and detection of

    4、 aerobic mesophilic bacteria This national standard is the identical implementation ISO 21149:2006 and is adopted with the permission of the International Organization for Standardization. Published by SABS Standards Division 1 Dr Lategan Road Groenkloof Private Bag X191 Pretoria 0001Tel: +27 12 428

    5、 7911 Fax: +27 12 344 1568 www.sabs.co.za SABS SANS 21149:2008 Edition 1 ISO 21149:2006 Edition 1 Table of changes Change No. Date Scope This South African standard was approved by National Committee SABS TC 217, Cosmetics, in accordance with procedures of the SABS Standards Division, in compliance

    6、with annex 3 of the WTO/TBT agreement. This SANS document was published in December 2008. Reference numberISO 21149:2006(E)ISO 2006INTERNATIONAL STANDARD ISO21149First edition2006-03-01Cosmetics Microbiology Enumeration and detection of aerobic mesophilic bacteria Cosmtiques Microbiologie Dnombremen

    7、t et dtection des bactries arobies msophiles SANS 21149:2008This s tandard may only be used and printed by approved subscription and freemailing clients of the SABS .ISO 21149:2006(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobes licensing policy, this file m

    8、ay be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In downloading this file, parties accept therein the responsibility of not infringing Adobes licensing policy. The ISO Central Secretariat acc

    9、epts no liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation parameters were optimized for printing. Every care has been taken to ensure that t

    10、he file is suitable for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below. ISO 2006 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any

    11、 form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISOs member body in the country of the requester. ISO copyright office Case postale 56 CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749

    12、 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2006 All rights reservedSANS 21149:2008This s tandard may only be used and printed by approved subscription and freemailing clients of the SABS .ISO 21149:2006(E) ISO 2006 All rights reserved iiiContents Page Foreword iv

    13、1 Scope . 1 2 Normative references . 1 3 Terms and definitions. 1 4 Principle. 2 4.1 General. 2 4.2 Plate count. 2 4.3 Membrane filtration. 2 4.4 Detection of bacteria by enrichment. 3 5 Diluents, neutralizers and culture media 3 5.1 General. 3 5.2 Neutralizing diluents and diluents 3 5.3 Diluent fo

    14、r the bacterial suspension (tryptone sodium chloride solution). 4 5.4 Culture media 4 6 Apparatus and glassware 6 7 Strains of microorganisms 7 8 Handling of cosmetic products and laboratory samples . 7 9 Procedure 7 9.1 General recommendation 7 9.2 Preparation of the initial suspension 7 9.3 Counti

    15、ng methods 8 9.4 Enrichment 9 10 Counting of colonies (plate counts and membrane filtration methods). 9 11 Detection of growth (enrichment method) . 9 12 Expression of results . 10 12.1 Method of calculation for plate count. 10 12.2 Interpretation. 10 12.3 Examples . 11 12.4 Detection after enrichme

    16、nt 13 13 Neutralization of the antimicrobial properties of the product 13 13.1 General. 13 13.2 Preparation of inoculum 14 13.3 Validation of counting methods 14 13.4 Validation of the detection method by enrichment . 15 13.5 Interpretation of validation results 15 14 Test report . 16 Annex A (infor

    17、mative) Other neutralizing diluents . 17 Annex B (informative) Other diluents. 19 Annex C (informative) Other culture media . 20 Annex D (informative) Neutralizers of antimicrobial activity of preservatives and rinsing liquids . 23 Bibliography . 24 SANS 21149:2008This s tandard may only be used and

    18、 printed by approved subscription and freemailing clients of the SABS .ISO 21149:2006(E) iv ISO 2006 All rights reservedForeword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International St

    19、andards 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 non-governmental, in liaison with ISO, also

    20、 take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committee

    21、s is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the 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 possib

    22、ility 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. ISO 21149 was prepared by Technical Committee ISO/TC 217, Cosmetics. SANS 21149:2008This s tandard may only be used and printed by ap

    23、proved subscription and freemailing clients of the SABS .INTERNATIONAL STANDARD ISO 21149:2006(E) ISO 2006 All rights reserved 1Cosmetics Microbiology Enumeration and detection of aerobic mesophilic bacteria 1 Scope This International Standard gives general guidelines for enumeration and detection o

    24、f mesophilic aerobic bacteria present in cosmetics, by counting the colonies on agar medium after aerobic incubation, or by checking the absence of bacterial growth after enrichment. Because of the large variety of cosmetic products within this field of application, this method may not be appropriat

    25、e for some products in every detail (e.g. certain water immiscible products). 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 validated. If needed, microorganisms enumerated or detec

    26、ted may be identified using suitable identification tests described in the standards given in the Bibliography. 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 cosmetic products to w

    27、hich 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. 2 Normative references The following referenced documents are indispensable for the application of this d

    28、ocument. 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 microbiological examination 3 Terms and definitions For the purpose

    29、s of this document, the following terms and definitions apply. 3.1 aerobic mesophilic bacteria mesophilic bacteria growing aerobically under the conditions specified in this International Standard NOTE In the described conditions, other types of microorganisms (e.g. yeast, mould) can be detected. 3.

    30、2 product portion of an identified cosmetic product received in the laboratory for testing SANS 21149:2008This s tandard may only be used and printed by approved subscription and freemailing clients of the SABS .ISO 21149:2006(E) 2 ISO 2006 All rights reserved3.3 sample portion of the product (at le

    31、ast 1 g or 1 ml) which is used in the test to prepare the initial suspension 3.4 initial suspension suspension (or solution) of a sample in a defined volume of an appropriate liquid (diluent, neutralizer, broth or combination of them) 3.5 sample dilution dilution of the initial suspension 4 Principl

    32、e 4.1 General This method involves enumeration of colonies on a non-selective agar medium or by the presence or absence of bacterial growth after enrichment. The possible inhibition of microbial growth by the sample shall be neutralized to allow the detection of viable microorganism 1. In all cases

    33、and whatever the methodology, the neutralization of the antimicrobial properties of the product shall be checked and validated 2 3 4. 4.2 Plate count Plate count consists of the following steps. Preparation of poured plates or spread plates, using a specified culture medium, and inoculation of the p

    34、lates using a defined quantity of the initial suspension or dilution of the product. Aerobic incubation of the plates at 32,5 C 2,5 C for 72 h 6 h. Counting the number of colony forming units (CFU) and calculation of the number of aerobic mesophilic bacteria per millilitre or per gram of product. 4.

    35、3 Membrane filtration Membrane filtration consists of the following steps. Transfer a suitable amount of the sample prepared as validated in the filtration apparatus wetted with a small volume of an appropriate sterile diluent, filter immediately and wash according to the validated procedure (see 13

    36、.3.4). Transfer the membrane filter onto the surface of the specified agar medium as specified in ISO 21148. Aerobic incubation of the membranes at 32,5 C 2,5 C for 72 h 6 h. Counting the number of colony forming units (CFU) and calculation of the number of aerobic mesophilic bacteria per millilitre

    37、 or per gram of product. SANS 21149:2008This s tandard may only be used and printed by approved subscription and freemailing clients of the SABS .ISO 21149:2006(E) ISO 2006 All rights reserved 34.4 Detection of bacteria by enrichment Detection of bacteria by enrichment consists of the following step

    38、s: Incubation at 32,5 C 2,5 C for at least 20 h of a defined quantity of the initial suspension in a non-selective liquid medium containing suitable neutralizers and/or dispersing agents. Transfer of a defined quantity of the previous suspension on non-selective solid agar medium. Aerobic incubation

    39、 at 32,5 C 2,5 C for 48 h to 72 h. Detection of growth and expression of results as “presence/absence” of aerobic mesophilic bacteria per sample S of product. 5 Diluents, neutralizers and culture media 5.1 General General specifications are given in ISO 21148. When water is used in a formula, use di

    40、stilled water or purified water as specified in ISO 21148. The following diluents, neutralizers and culture media are suitable for enumeration and detection of aerobic mesophilic bacteria. Other diluents, neutralizers and culture media may be used if they have been demonstrated to be suitable for us

    41、e. 5.2 Neutralizing diluents and diluents 5.2.1 General The diluent is used to disperse the sample. It may contain neutralizers if the specimen to be tested has antimicrobial properties. The efficacy of the neutralization shall be demonstrated before the determination of the count (see Clause 13). I

    42、nformation relative to suitable neutralizers is given in Annex D. 5.2.2 Neutralizing diluents 5.2.2.1 Fluid casein digest soy lecithin polysorbate 20 medium (SCDLP 20 broth) 5.2.2.1.1 Composition Pancreatic digest of casein 20,0 g Soy lecithin 5,0 g Polysorbate 20 40,0 ml Water 960,0 ml5.2.2.1.2 Pre

    43、paration Dissolve the polysorbate 20 in 960 ml of water by mixing while heating in a water bath at 49 C 2 C. Add pancreatic digest of casein and soy lecithin. Heat for about 30 min to obtain solution. Mix and dispense the medium into suitable containers. Sterilize in the autoclave at 121 C for 15 mi

    44、n. After sterilization, the pH shall be equivalent to 7,3 0,2 when measured at room temperature. SANS 21149:2008This s tandard may only be used and printed by approved subscription and freemailing clients of the SABS .ISO 21149:2006(E) 4 ISO 2006 All rights reserved5.2.2.2 Other neutralizing diluent

    45、s Other neutralizing diluents may be used as appropriate (see Annex A and Annex D). 5.2.3 Diluent 5.2.3.1 Fluid A 5.2.3.1.1 Composition Peptic digest of animal tissue 1,0 g Water 1 000 ml 5.2.3.1.2 Preparation Dissolve 1 g of peptone in water to make 1 l. Heat with frequent agitation. Dispense into

    46、suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilization, the pH shall be equivalent to 7,1 0,2 when measured at room temperature. 5.2.3.2 Other diluents Other diluents may be used as appropriate (see Annex B). 5.3 Diluent for the bacterial suspension (tryptone sodium

    47、 chloride solution) 5.3.1 Composition Tryptone, pancreatic digest of casein 1,0 g Sodium chloride 8,5 g Water 1 000 ml 5.3.2 Preparation Dissolve the components in the water by mixing while heating. Dispense into suitable containers. Sterilize in the autoclave at 121 C for 15 min. After sterilizatio

    48、n, the pH shall be equivalent to 7,0 0,2 when measured at room temperature. 5.4 Culture media 5.4.1 General Culture media may be prepared as follows, or from dehydrated culture media according to the instructions of the manufacturer. Ready-to-use media may be used when their composition and/or growt

    49、h yields are comparable to those of the formulas given herein. 5.4.2 Culture media for counting 5.4.2.1 Soybeancasein digest agar medium (SCDA) or tryptic soy agar (TSA) 5.4.2.1.1 Composition Pancreatic digest of casein 15,0 g Papaic digest of soybean meal 5,0 g SANS 21149:2008This s tandard may only be used and printed by approved subscription and freemailing clients of the SABS .ISO 21149:2006(E) ISO 2006 All rights reserved 5Sodium chloride 5,0 g Agar 1


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