EN ISO 16187-2013 en Footwear and footwear components - Test method to assess antibacterial activity《鞋和鞋类组件 评估抗菌活性的试验方法》.pdf

1、BSI Standards PublicationBS EN ISO 16187:2013Footwear and footwearcomponents Test methodto assess antibacterial activity(ISO 16187:2013)BS EN ISO 16187:2013 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO16187:2013.The UK participation in its preparation was

2、 entrusted to TechnicalCommittee TCI/69, Footwear, leather and coated fabrics.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correct

3、application. The British Standards Institution 2013. Published by BSI StandardsLimited 2013ISBN 978 0 580 73488 5ICS 61.060Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Commi

4、ttee on 31 August 2013.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 16187 August 2013 ICS 61.060 English Version Footwear and footwear components - Test method to assess antibacterial activity (ISO 16187:2013) Chaussure et composants d

5、e chaussure - Mthode dessai pour valuer lactivit antibactrienne (ISO 16187:2013) Schuhe und Schuhbestandteile - Prfverfahren zur Bestimmung der antibakteriellen Wirkung (ISO 16187:2013) This European Standard was approved by CEN on 12 July 2013. CEN members are bound to comply with the CEN/CENELEC I

6、nternal 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 application to the CEN-CENELEC Management Centre or to

7、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 Management Centre has the same status as the official

8、 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, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Nor

9、way, Poland, Portugal, Romania, 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: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of expl

10、oitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 16187:2013: EBS EN ISO 16187:2013EN ISO 16187:2013 (E) 3 Foreword Le prsent document (EN ISO 16187:2013) a t labor par le Comit Technique ISO/TC 216 “Chaussure” en collaboration avec le Comit Technique

11、 CEN/TC 309 “Chaussure”, dont le secrtariat est tenu par AENOR. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by February 2014, and conflicting national standards shall be withdrawn at the latest b

12、y February 2014. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN-CENELEC Internal Regulations, the national stand

13、ards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Lu

14、xembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 16187:2013 has been approved by CEN as EN ISO 16187:2013 without any modification. BS EN ISO 16187:2013ISO 16187:2013(E)

15、ISO 2013 All rights reserved iiiContents PageForeword iv1 Scope . 12 Normative references 13 Terms and definitions . 14 Safety 15 Apparatus and materials 26 Reagents and culture medium . 27 Test microorganisms 47.1 Test strains . 47.2 Storage of strains 48 Preparation of test inoculums 49 Preparatio

16、n of test samples 49.1 General . 49.2 Test specimen . 59.3 Pre-treatment of the test specimen 510 Test procedure 511 Expression of results 512 Test report . 6Annex A (normative) Static challenge test . 7Annex B (normative) Film contact method 9Annex C (normative) Dynamic challenge test 12Annex D (in

17、formative) Summarized results of round robin tests .14Bibliography .16BS EN ISO 16187:2013ISO 16187:2013(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is nor

18、mally 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 take part in

19、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 described in the ISO/IEC Directives, Part 1. In particular th

20、e 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. www.iso.org/directivesAttention is drawn to the possibility that some of the elements of this document ma

21、y 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 the ISO list of patent declarations received. www.iso.org/patentsAn

22、y trade name used in this document is information given for the convenience of users and does not constitute an endorsement.The committee responsible for this document is ISO/TC 216, Footwear.iv ISO 2013 All rights reservedBS EN ISO 16187:2013INTERNATIONAL STANDARD ISO 16187:2013(E)Footwear and foot

23、wear components Test method to assess antibacterial activityCAUTION Test methods specified herein require the use of bacteria. These tests are only to be carried out in facilities with containment techniques for handling microorganisms and by persons with training and experience in the use of microb

24、iological techniques. Appropriate safety precautions are to be observed with due consideration given to country-specific regulations.1 ScopeThis International Standard specifies quantitative test methods to evaluate the antibacterial activity of footwear and components.This International Standard is

25、 applicable to all types of footwear and components employing non-diffusing antibacterial treatments.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited

26、 applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 3696, Water for analytical laboratory use Specification and test methodsISO 19952, Footwear Vocabulary3 Terms and definitionsFor the purposes of this document, the terms and definit

27、ions given in ISO 19952 and the following apply.3.1antibacterial activityefficacy of a material or finish used to prevent or mitigate the growth of bacteria, to reduce the number of bacteria or to kill bacteria3.2control samplematerial identical to the test material but without antibacterial treatme

28、nt4 SafetyHandling of microorganisms which are potentially hazardous requires a high degree of technical competence and can be subject to current national legislation and regulations. Only personnel trained in microbiological techniques should carry out such tests. Codes of practice for disinfection

29、, sterilization and personal hygiene shall be strictly observed.NOTE It is recommended that workers consult IEC 600682-10, appendix A “Danger to personnel”, and ISO 7218. ISO 2013 All rights reserved 1BS EN ISO 16187:2013ISO 16187:2013(E)5 Apparatus and materials5.1 GeneralStandard laboratory equipm

30、ent and the following.5.2 Biological safety cabinet.5.3 Incubator, capable of maintaining a temperature of (37 2) C.5.4 Autoclave.5.5 Humidity chamber, capable of maintaining a temperature of (37 2) C and a relative humidity not less than 90 %.5.6 Ultraviolet lamp.5.7 Wide mouth jars, with cap, 100

31、ml, capable of being used with an autoclave (5.4).5.8 Cover film that does not affect bacterial growth or absorb water, which can be made of either polyethylene, polypropylene or polyester poly (ethylene terephthalate). Film that is 0,05 mm to 0,10 mm thick is recommended. For example, disposal bag

32、suitable for use with an autoclave (5.4).5.9 Vortex mixer.5.10 Dimensional shaker, two dimensional or three dimensional, capable of adjusting to 50 rpm.5.11 Shaking incubator, capable of maintaining a temperature of (37 2) C and a rotational frequency of (120 10) rpm.6 Reagents and culture medium6.1

33、 PrincipleThe preparation and test shall be freshly prepared in order to ensure the culture quality.NOTE This can be done according to ISO/TS 11133-1, ISO/TS 11133-2, or according to national standards or regulations.Reagents used in tests shall be of analytical grade and/or suited for microbiologic

34、al purposes.Use only water Grade 3 according to ISO 3696.6.2 Nutrient broth (NB)6.2.1 CompositionBeef extract, 3,0 g.Peptone, 5,0 g.Sodium chloride (NaCl), 5,0 g.Water, 1 000 ml.6.2.2 PreparationStir and adjust pH to (7,2 0,2) (at room temperature). Heat with stirring on a hotplate or in a boiling-w

35、ater bath until the components are completely dissolved. Sterilize with autoclave (5.4) at (121 2) C for 15 min.6.3 Nutrient agar (NA)2 ISO 2013 All rights reservedBS EN ISO 16187:2013ISO 16187:2013(E)6.3.1 CompositionBeef extract, 5,0 g.Peptone, 10,0 g.Sodium chloride (NaCl), 5,0 g.Agar, 15,0 g.Wat

36、er, 1 000 ml.NOTE If solidification is insufficient, 15 g to 18 g of agar can be used.6.3.2 PreparationStir and adjust pH to (7,2 0,2) (at room temperature). Heat with stirring on a hotplate or in a boiling-water bath until the components are completely dissolved. Sterilize with autoclave (5.4) at (

37、121 2) C for 15 min. Cool and shake solution well, then pour into the Petri dishes.6.4 Soybean casein digest broth with lecithin and polyoxyethylene medium (SCDLP)6.4.1 CompositionPeptone, digest of casein, 17,0 g.Peptone, digest of soybean, 3,0 g.Sodium chloride (NaCl), 5,0 g.Potassium dihydrogen p

38、hosphate, 2,5 g.Glucose, 2,5 g.Lecithin, 1,0 g.Polysorbate 80, 7,0 g.Water, 1 000 ml.If the neutralizing power is insufficient, the content of polysorbate 80 or lecithin may be adjusted or another neutralizing agent may be added. The use of any unspecified neutralizer shall be recorded along with th

39、e name and concentration.NOTE Information about selection and evaluation of alternative antibacterial neutralizing agents can be found in ASTM E 1054 and EN 1040.6.4.2 PreparationAfter mixing well, adjust pH to (7,2 0,2) (at room temperature) and sterilize with autoclave (5.4) at (121 2) C for 15 mi

40、n.6.5 Sodium chloride solution (physiological saline)6.5.1 CompositionSodium chloride (NaCl), 8,5 g.Water, 1 000 ml. ISO 2013 All rights reserved 3BS EN ISO 16187:2013ISO 16187:2013(E)6.5.2 PreparationAfter mixing well, adjust pH to (6,9 0,2) (at room temperature) and sterilize at (121 2) C for 15 m

41、in.7 Test microorganisms7.1 Test strainsThe following species shall be used in all antibacterial activity tests.a) Staphylococcus aureus AS 1.89 or ATCC 6538.b) Klebsiella pneumoniae AS 1.1736 or ATCC 4352.NOTE 1 If required, other species or other strains can be used. However, the selected organism

42、s should contain at least one gram-positive and one gram-negative organism as the antibacterial agents may have different activities.Test strains shall be obtained from agencies of the World Federation of Culture Collection (WFCC).The bacteria species and their supply sources shall be included in th

43、e test report.NOTE 2 AS refers to the China General Microbiological Culture Collection Centre (CGMCC), ATCC is the American Type Culture Collection.7.2 Storage of strainsInoculate the strains to the nutrient agar (NA) (6.3), and incubate at (37 2) C for 24 h. Store at (5 3) C (maximum one month) and

44、 keep it as stock culture of the strains. Transfer and incubate one time each month.Strains can be preserved in accordance with the suppliers direction or EN 12353.8 Preparation of test inoculumsUsing a sterile inoculating loop, transfer one colony (7.2) into 20 ml of nutrient broth (NB) (6.2) and i

45、ncubate in the shaking incubator (5.11) at (37 2) C for about 16 h (overnight culture). Estimate the number of bacteria with microscopic observation or other methods. Prepare physiological saline (6.5) with 1 % nutrient broth (NB) (6.2). Use this media to prepare a suspension with a bacterial concen

46、tration of (2,5 approximately 10) 105CFU/ml as test inoculum.If necessary, store the test inoculum on ice and use it within 4 h.9 Preparation of test samples9.1 GeneralTest only the components or material which are claimed to be antibacterial. If the whole footwear is claimed as antibacterial, major

47、 components, including upper, lining, insole, insock, outsole shall be tested separately.In the case where only one material of a component is claimed to be antibacterial, it shall be tested separately, if possible. Otherwise, the whole component shall be tested.Each test sample shall be at least 80

48、 % of the surface area of the component or material. If single material accounts for less than 80 %, take two main materials used in the composition of the component.The test samples can be obtained directly from the footwear raw materials.4 ISO 2013 All rights reservedBS EN ISO 16187:2013ISO 16187:

49、2013(E)9.2 Test specimenThe area of test specimen should be about 500 mm2. For test method A (see Annex A), the area of test specimen shall have a thickness of less than 2,0 mm. The area and the weight shall be reported in the test report. If a larger test specimen is used then the volume of bacterial suspension should be increased proportionally.If it is impossible to lower the thickness of the test specimen (for example, components are thicker and cannot be separated or cut without changing