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 978-0-626-22939-9 SANS 6321:2009Edition 1 SOUTH AFRICAN NATIONAL STANDARD Determination of the microbial inhibition of cosmetic soap bars and liquid
4、 hand and body washes Published by SABS Standards Division 1 Dr Lategan Road Groenkloof Private Bag X191 Pretoria 0001Tel: +27 12 428 7911 Fax: +27 12 344 1568 www.sabs.co.za SABS SANS 6321:2009 Edition 1 Table of changes Change No. Date Scope Foreword This South African standard was approved by Nat
5、ional Committee SABS TC 217, Cosmetics, in accordance with procedures of the SABS Standards Division, in compliance with annex 3 of the WTO/TBT agreement. This document was published in September 2009. SANS 6321:2009 Edition 1 1 Contents Page Foreword 1 Scope . 3 2 Normative references . 3 3 Princip
6、le 3 4 Test method . 3 4.1 General. 3 4.2 Accuracy. 4 4.3 Culture medium, reagents, reference cultures and controls 4 4.4 Procedure. 5 5 Interpretation of results 6 Bibliography 6 SANS 6321:2009 Edition 1 2 This page is intentionally left blank SANS 6321:2009 Edition 1 3 Determination of the microbi
7、al inhibition of cosmetic soap bars and liquid hand and body washes 1 Scope This standard describes a method for testing and comparing the microbial inhibition properties of cosmetic soap bars and liquid hand and body washes. 2 Normative references The following referenced documents are indispensabl
8、e 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. Information on currently valid national and international standards can be obtained from the SABS St
9、andards Division. SANS 636, Disinfectants based on quaternary ammonium compounds. SANS 7218/ISO 7218, Microbiology of food and animal feeding stuffs General requirements and guidance for microbiological examinations. 3 Principle When soap bars and liquid hand and body washes that have antimicrobial
10、properties are inoculated into medium No. 1 (see 4.3.1.1), specific active ingredients diffuse into the surrounding agar thereby creating a zone of inhibition around the product, which is measured as an indication of the microbial inhibition properties of the product. NOTE The organisms referenced i
11、n this standard are the indicator organisms used when testing for hygiene purposes. 4 Test method 4.1 General Sampling and testing shall be carried out by personnel familiar with microbiological procedures. Only media, reagents, equipment and test organisms that comply with the requirements and guid
12、elines given in SANS 636 and SANS 7218 shall be used. SANS 6321:2009 Edition 1 4 4.2 Accuracy Except where otherwise specified, allow the following tolerances: a) on temperatures 2 C; b) on masses. 1,0 %; c) on volumes 1,0 %; and d) on pH values . 0,1 pH unit. 4.3 Culture medium, reagents, reference
13、 cultures and controls 4.3.1 Culture medium NOTE 1 The culture medium listed in 4.3.1.1 is commercially available in dehydrated form and is made up in accordance with the manufacturers instructions. NOTE 2 Glass distilled water or deionised water should be used. 4.3.1.1 Medium No. 1 Ingredients Agar
14、 15,0 g Beef extract 15,0 g Glucose 1,0 g Pancreatic digest of casein 4,0 g Peptone 6,0 g Yeast extract. 3,0 g Water 1 000 mL 4.3.1.2 Preparation 4.3.1.2.1 Dissolve the ingredients in approximately 900 mL of water and mix. 4.3.1.2.2 Make up to 1 L. 4.3.1.2.3 Adjust the pH so that after sterilization
15、 the pH value is 6,6 0,05. 4.3.1.2.4 Dispense 20 mL 0,2 mL volumes into suitable containers and sterilize in an autoclave for 15 min 0,5 min at 121 C. 4.3.2 Horse serum Sterile inactivated horse serum that is free from preservatives. SANS 6321:2009 Edition 1 5 4.3.3 Reference cultures 4.3.3.1 Test o
16、rganisms Use the following test organisms: a) Staphylococcus aureus Sta 10 ATCC 6538; b) Escherichia coli Esc 20 ATCC 8739; and c) Pseudomonas aeruginosa . Pse 16 ATCC 15442. NOTE Other reference organisms may also be used in addition to the ones specified in 4.3.3.1. 4.3.3.2 Preparation of test org
17、anism suspensions Maintain the test organisms and prepare the cultures as described in SANS 636. Using sterile water as the diluent, prepare suspensions of the test organisms as described in SANS 636, but containing 105cfu/mL 104cfu/mL. 4.3.4 Controls 4.3.4.1 Positive control Natural honey with no a
18、dditives, e.g. colourants. 4.3.4.2 Negative control Sterile deinonised water. 4.4 Procedure 4.4.1 Melt the contents of a bottle of the medium No. 1 (see 4.3.1.1) and cool to 45 C 2 C. Add 1 mL of the sterile inactivated horse serum (see 4.3.2) and 1 mL of S. aureus test organism suspension prepared
19、in accordance with 4.3.3.2. Mix well and avoid the formation of air bubbles, then pour the mixture into a Petri dish of 90 mm diameter and allow to solidify. 4.4.2 Using a sterile cutter that produces cylindrical wells (holes) of 8 mm diameter, make five evenly spaced straight cylindrical wells in t
20、he solidified medium. Remove and discard the plugs of medium. Seal the bottom of each well using one or two drops of the molten medium in 4.3.1.1 and allow to solidify. 4.4.3 Introduce the test sample into the wells in such a way that each well is completely filled with the sample, taking care not t
21、o form bubbles. When the test sample is a soap bar, grind a sufficient amount to a powder, using a sterile pestle and mortar, and introduce the powder into the wells. Ensure that the surface of the agar remains free from the sample. 4.4.4 Incubate the Petri dish at 37 C 1 C for 18 h to 24 h. 4.4.5 A
22、t the end of this period (see 4.4.4), remove the Petri dish from the incubator. Measure the diameter of the zone of inhibition diagonally across the well to the nearest millimetre. 4.4.6 Take the average of the five diameters and record this to the nearest millimetre. 4.4.7 Repeat the procedure desc
23、ribed in 4.4.1 to 4.4.6 (inclusive) using the E. coli and P. aeruginosa test suspensions successively. SANS 6321:2009 Edition 1 6 4.4.8 Repeat the procedure described in 4.4.1 to 4.4.6 (inclusive) for the positive control and the negative control. 5 Interpretation of results 5.1 For the product to p
24、ass the test, the average diameter of the zone of inhibition for each of the test organisms shall be at least 10 mm. 5.2 If the zones are not clearly defined for one of the reference organism (e.g. hazy, incomplete zone, or distorted shape), the test shall be repeated for the specified reference org
25、anism. Bibliography Janssen A.M, Scheffer J.J.C, and Svendsen A.B. 1987. Antimicrobial Activity of Essential Oils: A 1976 1986 Literature Review. Aspect of the Test Methods. Planta medica, 53:395 398. Mangena T, and Muyima N.Y.O. 1999. Comparative Evaluation of the Antimicrobial Activities of Essent
26、ail Oils of Artemisia afra, Pteronia incana and Rosmarinus officinalis on selected bacteria and yeast strains. Letters in Applied Microbiology, 28:291 296. Prabuseenivasan S, Jayakumar, M and Ignacimuthu S. 2006. In vitro antibacterial activity of some plant essential oils. BMC Complementary and Alt
27、ernative Medicine, 6:33. SABS SABS Standards Division The objective of the SABS Standards Division is to develop, promote and maintain South African National Standards. This objective is incorporated in the Standards Act, 2008 (Act No. 8 of 2008). Amendments and Revisions South African National Stan
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