1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS ISO 22196:2011Measurement of antibacterialactivity on plastics and othernon-porous surfacesBS ISO 22196:2011 BRITISH STANDARDNational forewordThis British Standard is the UK i
2、mplementation of ISO 22196:2011. Itsupersedes BS ISO 22196:2007 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee PRI/21, Testing of plastics.A list of organizations represented on this committee can beobtained on request to its secretary.This publication
3、 does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. BSI 2011ISBN 978 0 580 69420 2ICS 83.080.01Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority
4、 of theStandards Policy and Strategy Committee on 31 August 2011.Amendments issued since publicationDate Text affectedBS ISO 22196:2011Reference numberISO 22196:2011(E)ISO 2011INTERNATIONAL STANDARD ISO22196Second edition2011-08-01Measurement of antibacterial activity on plastics and other non-porou
5、s surfaces Mesurage de laction antibactrienne sur les surfaces en plastique et autres surfaces non poreuses BS ISO 22196:2011ISO 22196:2011(E) COPYRIGHT PROTECTED DOCUMENT ISO 2011 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form
6、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 09 47
7、 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2011 All rights reservedBS ISO 22196:2011ISO 22196:2011(E) ISO 2011 All rights reserved iiiContents Page Foreword iv Introduction.v 1 Scope1 2 Normative references1 3 Terms and definitions .2 4 Materials .2 4.1 Bacteria to be u
8、sed for the tests2 4.2 Reagents, culture media and solutions 3 5 Apparatus.4 6 Sterilization of apparatus and storage of stock cultures 5 6.1 Dry-heat sterilization.5 6.2 High-pressure steam sterilization5 6.3 Preparation of glassware5 6.4 Maintenance of stock cultures.5 7 Procedure.6 7.1 Pre-cultur
9、e of bacteria 6 7.2 Preparation of test specimens .6 7.3 Preparation of test inoculum6 7.4 Inoculation of test specimens6 7.5 Incubation of the inoculated test specimens .8 7.6 Recovery of bacteria from test specimens.8 7.7 Determining the viable bacteria count by the pour plate culture method.8 8 E
10、xpression of results9 8.1 Determination of the number of viable bacteria.9 8.2 Conditions for a valid test 9 8.3 Calculation of the antibacterial activity.9 8.4 Effectiveness of the antibacterial agent10 9 Repeatability and reproducibility.10 10 Test report10 Annex A (normative) Quality of biologica
11、l materials.11 A.1 General .11 A.2 Chemical composition of 1/500 nutrient broth (1/500 NB) 11 Annex B (informative) Repeatability and reproducibility12 B.1 Background12 B.2 Summary 12 B.3 Experiment .12 B.4 Results and discussion 13 Bibliography15 BS ISO 22196:2011ISO 22196:2011(E) iv ISO 2011 All r
12、ights reservedForeword ISO (the International Organization for Standardization) is a worldwide 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
13、 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 take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC)
14、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 committees is to prepare International Standards. Draft International Standards adopted by the technical committees
15、 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 possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be hel
16、d responsible for identifying any or all such patent rights. ISO 22196 was prepared by Technical Committee ISO/TC 61, Plastics, Subcommittee SC 6, Ageing, chemical and environmental resistance. This second edition cancels and replaces the first edition (ISO 22196:2007). The main change is the extens
17、ion of the scope of the standard to include non-porous surfaces other than plastics (for details, see the Introduction). BS ISO 22196:2011ISO 22196:2011(E) ISO 2011 All rights reserved vIntroduction Antibacterial materials and products have been widely and rapidly accepted by general consumers as fu
18、lfilling a relatively new function, which is distinguishable from the more traditional function of material protection. Antibacterial products created by incorporating an antibacterial agent (biocide) can suppress the growth of bacteria on the surfaces of products when conditions exist where growth
19、can occur. They can keep surfaces clean and sanitary and can also have an advantage in minimizing the impact on the environment by minimizing diffusion of the agent. This technology is significant for the quality of life, not only in developed countries but also in developing countries. Antibacteria
20、l products have been widely used in plastics, coating materials, ceramics, natural and artificial leather, stainless steel, rubber, etc. The products involved cover a variety of categories, such as electrical appliances, personal items, household goods, nursing-care articles, pet accessories and air
21、craft-interior fittings. The scope of the first edition of ISO 22196 was limited to plastics surfaces. In this second edition, the scope has been extended to include surfaces made of other non-porous materials, thus making the second edition applicable to products of the kinds listed above. The test
22、 method, which is based on JIS Z 280111, has remained unchanged. BS ISO 22196:2011BS ISO 22196:2011INTERNATIONAL STANDARD ISO 22196:2011(E) ISO 2011 All rights reserved 1Measurement of antibacterial activity on plastics and other non-porous surfaces 1 Scope WARNING Handling and manipulation of micro
23、organisms which are potentially hazardous requires a high degree of technical competence and may be subject to current national legislation and regulations. Only personnel trained in microbiological techniques should carry out such tests. Appropriate practices for disinfection, sterilization and per
24、sonal hygiene must be strictly observed. This International Standard specifies a method of evaluating the antibacterial activity of antibacterial-treated plastics, and other non-porous, surfaces of products (including intermediate products). It is not intended to be used to evaluate the effects and
25、propagation of bacteria on non-porous surfaces without antibacterial treatments. ISO 8461describes tests to evaluate the effects and propagation of bacteria on non-porous surfaces, which are different from those covered by this International Standard (see e.g. ISO 846:1997, method C). Secondary effe
26、cts of antibacterial treatments, such as the prevention of biodeterioration and odour, are not covered by this International Standard, which is not intended to be used or referenced as a method to document or claim biodegradability of, for instance, plastics materials. In the case of plastics, biode
27、gradation is covered in ISO 148512, ISO 148523and ISO 148554and related standards. Building materials are excluded, except where they are used in the same manner as treated articles. Antibacterial-treated textile products are excluded, even if the surfaces are coated or laminated (such products are
28、covered by ISO 207435). Photocatalytic materials and products are excluded (such materials and products are covered by ISO 274476). The results obtained should include a reference to this International Standard and the conditions used. Results obtained with this International Standard indicate antib
29、acterial activity under the specified experimental conditions used, and do not reflect activity under other circumstances where a variety of factors, such as temperature, humidity, different bacterial species, nutrient conditions, etc., have to be considered. A minimum diffusion of the antibacterial
30、 agents/chemicals into the test inoculum is necessary with this procedure. It is recommended that workers consult ISO 7218. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For unda
31、ted references, the latest edition of the referenced document (including any amendments) applies. ISO 7218, Microbiology of food and animal feeding stuffs General requirements and guidance for microbiological examinations BS ISO 22196:2011ISO 22196:2011(E) 2 ISO 2011 All rights reserved3 Terms and d
32、efinitions For the purposes of this document, the following terms and definitions apply. 3.1 antibacterial term describing a state where growth of bacteria on the surfaces of products is suppressed or describing the effect of an agent which suppresses the growth of bacteria on the surfaces of produc
33、ts 3.2 antibacterial agent agent that inhibits the growth of bacteria on the surfaces of products, used either as a surface treatment or as a compounded ingredient 3.3 antibacterial activity difference in the logarithm of the viable cell counts found on an antibacterial-treated product and an untrea
34、ted product after inoculation with and incubation of bacteria 3.4 antibacterial effectiveness ability of an antibacterial agent to inhibit the growth of bacteria on the surface of materials treated with an antibacterial agent, as determined by the value of the antibacterial activity 4 Materials 4.1
35、Bacteria to be used for the tests Both of the following species of bacteria shall be used: a) Staphylococcus aureus; b) Escherichia coli. The bacterial strains to be used are shown in Table 1. If bacterial strains obtained from culture collections other than those shown in Table 1 are used, they sha
36、ll be obtained from a member agency of the World Federation for Culture Collections (WFCC) or of the Japan Society for Culture Collections (JSCC) and shall be the same strains as those shown in Table 1. Prepare stock cultures of these species in accordance with the suppliers directions. Table 1 Bact
37、erial strains to be used Name Strain Staphylococcus aureus ATCC 6538P CIP 53.156 DSM 346 NBRC 12732 NCIB 8625 Escherichia coli ATCC 8739 CIP 53.126 DSM 1576 NBRC 3972 NCIB 8545 BS ISO 22196:2011ISO 22196:2011(E) ISO 2011 All rights reserved 3If required, other species can also be used, in which case
38、 the species and the reason for their use shall be included in the test report. 4.2 Reagents, culture media and solutions Water shall be distilled or deionized and have a conductivity of 1 S/cm. All reagents shall be of analytical grade and/or of a grade appropriate for microbiological purposes. 4.2
39、.1 Nonionic surfactant Polyoxyethylene sorbitan monooleate shall be used. 4.2.2 Biological materials The following biological materials are required: lecithin; D-glucose; yeast extract; meat extract (see Annex A); peptone (see Annex A); casein peptone; soybean peptone; tryptone. 4.2.3 Culture medium
40、 4.2.3.1 General The culture medium specified below shall be used. The medium may be obtained from commercial suppliers, in which case it shall be prepared for use in accordance with the manufacturers instructions. The quantity of the culture medium can be changed provided the same composition is re
41、tained. 4.2.3.2 Suspension medium 1/500 nutrient broth (1/500 NB) Prepare nutrient broth by dissolving 3,0 g of meat extract, 10,0 g of peptone and 5,0 g of sodium chloride in 1 000 ml of distilled or deionized water. Dilute the nutrient broth with distilled or deionized water to a 500-fold volume a
42、nd adjust the pH to a value between 6,8 and 7,2 with sodium hydroxide or hydrochloric acid. Sterilize by autoclaving (see 6.2). If it is not used immediately after preparation, store it at 5 C to 10 C. A 1/500 NB that has been kept for one week or longer after preparation shall not be used. 4.2.3.3
43、Nutrient agar Prepare nutrient agar by dissolving 5,0 g of meat extract, 10,0 g of peptone, 5,0 g of sodium chloride and 15,0 g of agar powder in 1 000 ml of distilled or deionized water. Heat, with stirring, on a hotplate or in a boiling-water bath until the agar has dissolved. Adjust the pH to a v
44、alue between 7,0 and 7,2 (at 25 C) with sodium hydroxide or hydrochloric acid. Sterilize by autoclaving (see 6.2). If it is not used immediately after preparation, store it at 5 C to 10 C. Nutrient agar that has been kept for one month or longer after preparation shall not be used. BS ISO 22196:2011
45、ISO 22196:2011(E) 4 ISO 2011 All rights reserved4.2.3.4 Plate count agar Prepare plate count agar by dissolving 2,5 g of yeast extract, 5,0 g of tryptone, 1,0 g of glucose and 15,0 g of agar powder in 1 000 ml of distilled or deionized water. Heat, with stirring, on a hotplate or in a boiling-water
46、bath until the agar has dissolved. Adjust the pH to a value between 7,0 and 7,2 (at 25 C) with sodium hydroxide or hydrochloric acid. Sterilize by autoclaving (see 6.2). If it is not used immediately after preparation, store it at 5 C to 10 C. Plate count agar that has been kept for one month or lon
47、ger after preparation shall not be used. 4.2.3.5 Slant culture medium Warm 6 ml to 10 ml of nutrient agar and pour into a screw-capped test tube. Sterilize by autoclaving (see 6.2). After sterilization, place the test tube at an angle of about 15 to the horizontal and allow the contents to solidify.
48、 If it is not used immediately after preparation, store it at 5 C to 10 C. Slant culture medium kept for one month or longer after preparation shall not be used. 4.2.3.6 Soybean casein digest broth with lecithin and polyoxyethylene sorbitan monooleate (SCDLP broth) Prepare SCDLP broth by dissolving
49、17,0 g of casein peptone, 3,0 g of soybean peptone, 5,0 g of sodium chloride, 2,5 g of disodium hydrogen phosphate, 2,5 g of glucose and 1,0 g of lecithin in 1 000 ml of distilled or deionized water. Mix thoroughly and add 7,0 g of nonionic surfactant. Adjust the pH to a value between 6,8 and 7,2 (at 25 C) with sodium hydroxide or hydrochloric acid. Sterilize by autoclaving (see 6.2). If it is not used immediately after preparation, store it at 5 C to 10 C. SCDLP broth kept for one month or longer
