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 978-0-626-22301-4 SANS 5261:2009Edition 3.1Any reference to SABS SM 261 is deemedto be a reference to this standard(Government Notice No. 1373 of 8
4、November 2002SOUTH AFRICAN NATIONAL STANDARD Bactericidal efficacy of anti-bacterial liquid toilet soap 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 5261:2009 Edition 3.1 Table of ch
5、anges Change No. Date Scope Amdt 1 2009 Amended to change the designation of SABS standards to SANSstandards, and to update reference numbers for test organisms (3.5). Foreword This South African standard was approved by National Committee SABS TC 1022, Antiseptics, disinfectants and detergent-disin
6、fectants, in accordance with procedures of the SABS Standards Division, in compliance with annex 3 of the WTO/TBT agreement. This document was published in February 2009. This document supersedes SABS SM 261:2001 (edition 3). A vertical line in the margin shows where the text has been technically mo
7、dified by amendment No. 1. SANS 5261:2009 Edition 3.1 1Bactericidal efficacy of anti-bacterial liquid toilet soap 1 Scope This standard specifies a method to determine the bactericidal efficacy of anti-bacterial liquid toilet soap. 2 Normative references The following referenced documents are indisp
8、ensable 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 S
9、ABS Standards Division. SANS 7218/ISO 7218, Microbiology of food and animal feeding stuffs General requirements and guidance for microbiological examinations. 3 Methods of microbiological testing 3.1 General The tests shall be undertaken by persons experienced in microbiological techniques, using as
10、eptic techniques. Media, reagents and equipment shall comply with the requirements and guidelines given in SANS 7218. NOTE 1 In order to ensure accuracy of these tests, it is recommended that the tests be repeated. NOTE 2 For the purpose of checking the resistance of the test organisms and other tes
11、t conditions, it is advisable to include a reference standard. It is essential that the reference standard is a liquid toilet soap, but, because a universal standard is difficult to select, each laboratory should make its own choice of material. 3.2 Accuracy Except where otherwise specified, allow t
12、he following tolerances: a) on temperatures 2 C b) on masses. 1,0 % c) on volumes 1,0 % d) on pH value. 0,1 pH unit 3.3 Laboratory ware 3.3.1 Glassware Ensure that all glassware is resistant to repeated heat sterilization and the glass is free from inhibitory substances such as heavy metals and free
13、 alkali. Borosilicate glass with an expansion coefficient of less than 6 10-6K-1is recommended. SANS 5261:2009 Edition 3.1 2 3.3.2 Universal container culture bottles Bottles made of glass, fitted with standard screwed metal caps with rubber liners and that have nominal capacities of a) 30 mL, and b
14、) 100 mL. Do not use plastics containers or glass containers fitted with plastics tops. 3.3.3 Culture tubes Rimless cylindrical tubes with hemispherical ends and a nominal wall thickness of 1,5 mm, of internal diameter 16 mm and of length 160 mm. 3.3.4 Graduated pipettes Total delivery pipettes for
15、bacteriological purposes only, that have an outflow opening of diameter 2 mm to 3 mm, are graduated in units of 0,1 mL and are of sizes to deliver 1,0 mL, 5,0 mL and 10,0 mL. 3.3.5 Measuring cylinders Graduated measuring cylinders with or without stoppers and of capacities 5 mL, 10 mL, 100 mL, 500 m
16、L and 1 000 mL. 3.3.6 Erlenmeyer flasks Erlenmeyer flasks of capacities 250 mL, 2 L and 3 L. 3.3.7 Petri dishes Petri dishes of diameter 90 mm and of height 20 mm and made of glass or of wettable polystyrene. 3.3.8 Reagent bottles Bottles of capacities 50 mL and 100 mL and that have polypropylene or
17、 other plastics stoppers of such design that they can be used to deliver drops of the reagent. 3.4 Test media 3.4.1 Reconstitution of media Many of the media required are obtainable commercially in dehydrated form and, for uniformity of results, the use of such media is recommended. If these are use
18、d, follow the manufacturers instructions strictly regarding the reconstitution and sterilization. The water used for reconstitution of media shall be distilled sterile water. 3.4.2 Nutrient agar 3.4.2.1 Ingredients Agar . 15,0 g Peptone . 5,0 g Sodium chloride. 5,0 g SANS 5261:2009 Edition 3.1 3Yeas
19、t extract 2,0 g Beef extract . 1,0 g Water . 1 000 mL 3.4.2.2 Preparation Dissolve the ingredients in the water and adjust the pH value to 7,1. Dispense in 10 mL and 15 mL volumes into bottles (see 3.3.2) and sterilize by autoclaving at (121 ) C for 15 min. Allow only the 10 mL volumes to solidify i
20、n a sloped position. 30+3.4.3 Nutrient broth 3.4.3.1 Ingredients Peptone . 5,0 g Sodium chloride. 5,0 g Yeast extract 2,0 g Beef extract . 1,0 g Water . 1 000 mL 3.4.3.2 Preparation Dissolve the ingredients in the water and adjust the pH value to 7,1. Dispense in 10 mL and 100 mL volumes into contai
21、ners (see 3.3.2) and sterilize by autoclaving at (121 ) C for 15 min. 30+3.5 Test organisms1) 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. Amdt 1 NOTE 1 Additional organisms may b
22、e used if so desired. NOTE 2 The extreme importance of using the standard strain is emphasized. 3.6 Maintenance of test organisms 3.6.1 From a newly opened freeze-dried culture or recently received agar culture, subculture the test organisms into bottles that contain nutrient broth (see 3.4.3). 3.6.
23、2 Incubate the bottles at 37 C for 24 h. Prepare subcultures from the cultures in the bottles onto slopes of nutrient agar (see 3.4.2). Incubate the slopes at 37 C for 24 h. 3.6.3 From each of these slope cultures, prepare four subcultures (stock cultures) of each test organism on nutrient agar (see
24、 3.4.2). Incubate the stock cultures at 37 C for 24 h and then store them in a refrigerator maintained at 4 C except for Pseudomonas aeruginosa, which is stored at room temperature. 1) Test organisms are obtainable from the South African Bureau of Standards, Private Bag X191, Pretoria, 0001. SANS 52
25、61:2009 Edition 3.1 4 NOTE Not more than six serial subcultures shall be taken from each stock culture before resorting to a new freeze-dried culture. 3.7 Preparation of cultures for test suspensions 3.7.1 For each of the test organisms inoculate a nutrient agar slope (see 3.4.2) from a stock cultur
26、e (see 3.6.3) and incubate at 37 C for 24 h. 3.7.2 For the test use a 24 h culture that has been subcultured for two successive days. After six subcultures, restart the process using a fresh stock culture (see 3.6.3). NOTE The physiological condition of the test organisms is important and might infl
27、uence inter-laboratory and intra-laboratory variations in test results. 3.7.3 Using 10 mL of sterile water, wash the bacterial growth resulting from 24 h incubation from the slope (see 3.7.1), scraping the agar surface if necessary. Carefully decant the suspended growth into an Erlenmeyer flask and
28、shake the flask vigorously to suspend all growth in the water. So standardize the suspension, by using a spectrophotometer in conjunction with a standard curve, a haemocytometer, a Petroff-Husser counting chamber or any other suitable means, that it contains 0,9 107organisms per millilitre to 2,0 10
29、7organisms per millilitre. Use the suspension within 3 h of preparation. 3.8 Procedure NOTE Use each test organism to test the sample and carry out each set of tests in triplicate. 3.8.1 Measure 4,8 mL of the test sample and 4,8 mL of distilled water for the control, into suitable containers and pla
30、ce them in a water bath maintained at 22 C 2 C. When temperature equilibrium has been reached (after approximately 30 min), add to the test sample 0,2 mL of the Staphylococcus aureus organism suspension (see 3.7.3), and mix well. 3.8.2 Sixty seconds after adding the test organism suspension, withdra
31、w 0,1 mL of the mixture and transfer it to 100 mL of the nutrient broth (see 3.4.3) in another container. Place the second container with the inoculated nutrient broth in an incubator maintained at 37 C. After at least 48 h, examine the broth for the presence of bacterial growth. 3.8.3 If no growth
32、is visible, further inoculate the broth (see 3.8.2) with one drop only of the first (freshly prepared) test organism suspension (see 3.7.3) and replace the container with the inoculated broth in the incubator. After at least 24 h, examine the broth for the presence of bacterial growth. 3.8.4 If bact
33、erial growth is clearly visible, deem the test sample to have killed the bacteria in the original test organism suspension (see 3.8.1). 3.8.5 If bacterial growth is not present, eliminate the possibility of bacteriostases having occurred in step 3.8.2 by determining what increased volume of nutrient
34、 broth shall be used or suitable inactivator added in step 3.8.2 to give bacterial growth in step 3.8.3. Discard the results of the first test, and repeat the test but use the increased volume of nutrient broth or add a suitable inactivator. Assess the result in terms of 3.8.4. 3.8.6 Repeat the test
35、 procedure in 3.8.1 to 3.8.5 (inclusive) using, successively, the Escherichia coli and Pseudomonas aeruginosa test suspensions. SABS SABS Standards Division The objective of the SABS Standards Division is to develop, promote and maintain South African National Standards. This objective is incorporat
36、ed in the Standards Act, 2008 (Act No. 8 of 2008). Amendments and Revisions South African National Standards are updated by amendment or revision. Users of South African National Standards should ensure that they possess the latest amendments or editions. The SABS continuously strives to improve the
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