EN 1499-2013 en Chemical disinfectants and antiseptics - Hygienic handwash - Test method and requirements (phase 2 step 2)《化学消毒剂和防腐剂 手卫生消毒法 试验方法和要求(2阶段 2级)》.pdf

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1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN 1499:2013Chemical disinfectants andantiseptics Hygienichandwash Test method andrequirements (phase 2/step 2)BS EN 1499:2013 BRITISH STANDARDNational forewordThis British St

2、andard is the UK implementation of EN 1499:2013. Itsupersedes BS EN 1499:1997 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee CH/216, Chemical disinfectants and antiseptics.A list of organizations represented on this committee can beobtained on request

3、to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2013.Published by BSI Standards Limited 2013 ISBN 978 0 580 74827 1 ICS 11.080.20; 71.100.35 Compliance with a

4、British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 31 May 2013. Amendments issued since publicationDate T e x t a f f e c t e dBS EN 1499:2013EUROPEAN STANDARD NORME EUROPENNE EUROPISC

5、HE NORM EN 1499 April 2013 ICS 11.080.20; 71.100.35 Supersedes EN 1499:1997English Version Chemical disinfectants and antiseptics - Hygienic handwash - Test method and requirements (phase 2/step 2) Antiseptiques et dsinfectants chimiques - Lavage hyginique des mains - Mthode dessai et prescriptions

6、(phase 2/tape 2) Chemische Desinfektionsmittel und Antiseptika - Hygienische Hndewaschung - Prfverfahren und Anforderungen (Phase 2/Stufe 2) This European Standard was approved by CEN on 1 March 2013. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the condi

7、tions 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 any CEN member. This European Standard exists

8、 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 versions. CEN members are the national stand

9、ards 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, Norway, Poland, Portugal, Romania, Slovakia, Slo

10、venia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of exploitation in any form and by any means reserved worldwide

11、for CEN national Members. Ref. No. EN 1499:2013: EBS EN 1499:2013EN 1499:2013 (E) 2 Contents Page Foreword 3 1 Scope 4 2 Normative references 4 3 Terms and definitions .4 4 Requirements .4 5 Test methods 5 5.1 Principle 5 5.2 Materials and reagents 5 5.2.1 Test organisms 5 5.2.2 Culture media and re

12、agents .5 5.3 Apparatus and glassware .7 5.3.1 General 7 5.3.2 Usual microbiological laboratory equipment .7 5.4 Preparation of test organism suspensions and product test solutions 9 5.4.1 Test organism suspensions (test and validation suspension) .9 5.4.2 Product test solutions 10 5.5 Procedure for

13、 assessing the bactericidal activity of the product on volunteers hands 11 5.5.1 General . 11 5.5.2 Neutralization control and validation 12 5.5.3 Test procedure with volunteers 13 5.5.4 Incubation and counting of the test mixture and the control and validation mixtures . 15 5.6 Experimental data an

14、d calculation 15 5.6.1 Explanation of terms and abbreviations 15 5.6.2 Calculation . 15 5.7 Verification of the methodology - Test validation . 18 5.7.1 Acceptance criteria for test results. 18 5.7.2 Control of weighted mean counts . 19 5.7.3 Basic limits 19 5.8 Statistical evaluation (significance

15、testing), expression of results and precision 19 5.9 Conclusion 19 5.10 Test report . 19 Annex A (normative) Standard handwash procedure 22 Annex B (informative) Neutralizers and rinsing liquids . 23 Annex C (informative) Graphical representation of neutralizer control and method validation 25 Annex

16、 D (informative) Quality control of soft soap 26 Annex E (informative) Examples of reporting of results and significance testing 27 Annex F (normative) WILCOXONS matched-pairs signed-ranks test 31 Bibliography . 32 BS EN 1499:2013EN 1499:2013 (E) 3 Foreword This document (EN 1499:2013) has been prep

17、ared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the secretariat of which is held by AFNOR. 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 October 2013, and confli

18、cting national standards shall be withdrawn at the latest by October 2013. 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. This docume

19、nt supersedes EN 1499:1997. This document was revised to adapt it to the latest state of science, to correct errors and ambiguities, to harmonise the structure and wording with other tests of CEN/TC 216 existing or in preparation, and to improve the readability of the standard and thereby make it mo

20、re understandable. The following technical changes have been made: Neutralization (5.5.1.2). The procedure (Annex A). The annexes have been completely revised. Data obtained using the former version of EN 1499 may still be used, if it is supplemented by data on neutralization. If the neutralizer use

21、d in the test using the former version is not sufficiently neutralizing, a complete new test will be run. The changed procedure in Annex A is regarded as having no (or negligible) influence on the results. According to the CEN-CENELEC Internal Regulations, the national standards organisations of the

22、 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, Luxembourg, Malta, Netherla

23、nds, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 1499:2013EN 1499:2013 (E) 4 1 Scope This European Standard specifies a test method simulating practical conditions for establishing whether a product for hygienic handwash red

24、uces the release of transient microbial flora on hands when used to wash the artificially contaminated hands of volunteers. NOTE 1 Attention is drawn to the fact that tests on human volunteers are the subject of legal provisions in certain European countries/regions. This European Standard applies t

25、o products for hygienic handwash for use in areas and situations where disinfection is medically indicated. Such indications occur in patient care, for example: in hospitals, in community medical facilities and in dental institutions, in clinics of schools, of kindergartens and of nursing homes; and

26、 may occur in the workplace and in the home. It may also include services such as laundries and kitchens supplying products directly for the patient. EN 14885 specifies in detail the relationship of the various tests to one another and to “use recommendations”. NOTE 2 This method corresponds to a ph

27、ase 2, step 2 test. 2 Normative references The 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 applies. For undated references, the latest edition of the referenced document

28、(including any amendments) applies. EN 12353, Chemical disinfectants and antiseptics Preservation of test organisms used for the determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal (including bacteriophages) activity EN 14885, Chemical disinfe

29、ctants and antiseptics Application of European Standards for chemical disinfectants and antiseptics 3 Terms and definitions For the purposes of this document, the terms and definitions given in EN 14885 apply. 4 Requirements When tested in accordance with Clause 5, the mean reduction of the release

30、of the test organism Escherichia coli K12 achieved by the hygienic handwash with the product under test shall be larger than that achieved by a specified reference hygienic handwash (unmedicated liquid soap). BS EN 1499:2013EN 1499:2013 (E) 5 5 Test methods 5.1 Principle Hands of volunteers are arti

31、ficially contaminated with test organisms. The number of test organisms released from their fingertips into sampling fluids is assessed before and after the hygienic handwash. The ratio of the two resulting values represents a measure for the antimicrobial activity of the product tested. The necessa

32、ry precision is achieved by repeating the test on 12 to 15 volunteers. To compensate for extraneous influences, it is compared with the reduction obtained by a reference handwash which is performed with the same volunteers, on the same day and under comparable environmental conditions. Prior to the

33、test, a suitable neutralizer is validated. The neutralizer is used as a sampling fluid for recovering the test organisms after the hygienic handwash to ensure that the bactericidal and/or bacteriostatic activity in the sampling fluids is neutralized or suppressed. 5.2 Materials and reagents 5.2.1 Te

34、st organisms E. coli , Escherichia coli K12 NCTC 10538; CIP 54.117; NCIMB 100831)NOTE This test organism has been specifically chosen to meet health and safety guidance and ethical committee considerations. It is a K12 strain of E. coli of normal flora origin internationally recognised as being non-

35、pathogenic. According to the UK catalogue of the National Collections of Industrial prepare solution B: dissolve 35,02 g sodium bicarbonate (NaHCO3) in water (5.2.2.2) and dilute to 1000 ml. Sterilise by membrane filtration (5.3.2.7). Store the solution in the refrigerator (5.3.2.8) for no longer th

36、an one week; place 600 ml to 700 ml of water (5.2.2.2) in a 1 000 ml volumetric flask (5.3.2.12) and add 6,0 ml (5.3.2.9) of solution A, then 8,0 ml of solution B. Mix and dilute to 1 000 ml with water (5.2.2.2). The pH of the hard water shall be 7,0 0,2, when measured at 20 C 1C (5.3.2.4). If neces

37、sary, adjust the pH by using a solution of approximately 40 g/l (about 1 mol/l) of sodium hydroxide (NaOH) or approximately 36,5 g/l (about 1 mol/l) of hydrochloric acid (HCl). The hard water shall be freshly prepared under aseptic conditions and used within 12 h. NOTE When preparing the product tes

38、t solutions (5.4.2), the addition of the product to the hard water produces a different final water hardness in each test tube. In any case, the final hardness, expressed as calcium carbonate (CaCO3) in the test tube, is lower than 375 mg/l. 5.3 Apparatus and glassware 5.3.1 General Sterilise all gl

39、assware and parts of the apparatus that will come into contact with the culture media and reagents or the sample, except those that are supplied sterile by one of the following methods: a) by moist heat, in the autoclave 5.3.2.1 a); b) by dry heat, in the hot air oven 5.3.2.1 b). 5.3.2 Usual microbi

40、ological laboratory equipment2)In particular, the following: 5.3.2.1 Apparatus for sterilisation 2) Disposable equipment is an acceptable alternative to reusable glassware. BS EN 1499:2013EN 1499:2013 (E) 8 a) for moist heat sterilisation, an autoclave capable of being maintained at (12130+) C for a

41、 minimum holding time of 15 min; b) for dry heat sterilisation, a hot air oven capable of being maintained at (18050+) C for a minimum holding time of 30 min, at (170 50+) C for a minimum holding time of 1 h or at (16050+) C for a minimum holding time of 2 h. 5.3.2.2 Water baths, capable of being co

42、ntrolled at 20 C 1 C, at 45 C 1 C (to maintain melted TSA and TSSA in case of pour plate technique) and at additional test temperatures 1 C (5.5.1) 5.3.2.3 Incubator, capable of being controlled at 36 C 1 C or 37 C 1 C (5.5.2). The same temperature shall be used for incubations performed during a te

43、st and its control and validation 5.3.2.4 pH-meter, having an inaccuracy of calibration of no more than 0,1 pH units at 20 C 1 C. A puncture electrode or a flat membrane electrode should be used for measuring the pH of the agar media (5.2.2.3) 5.3.2.5 Stopwatch 5.3.2.6 Shakers a) electromechanical a

44、gitator, e.g. Vortexmixer3); b) mechanical shaker. 5.3.2.7 Membrane filtration apparatus, constructed of a material compatible with the substances to be filtered The apparatus shall have a filter holder of at least 50 ml volume. It shall be suitable for use with filters of diameter 47 mm to 50 mm an

45、d 0,45 m pore size for sterilisation of hard water (5.2.2.7). The vacuum source used shall give an even filtration flow rate. To prevent overlong filtration, the device shall be set so as to obtain the filtration of 100 ml of rinsing liquid in 20 s to 40 s. 5.3.2.8 Refrigerator, capable of being con

46、trolled at 2 C to 8 C 5.3.2.9 Graduated pipettes, of nominal capacities 10 ml, 1 ml and 0,1 ml, or calibrated automatic pipettes 5.3.2.10 Petri dishes (plates), of size 90 mm to 100 mm 5.3.2.11 Glass beads (diameter 3 mm to 4 mm) 5.3.2.12 Volumetric flasks 5.3.2.13 Spreader, made of glass or other m

47、aterial 5.3.2.14 Container of sufficient capacity to immerse two hands vertically up to the mid-metacarpals simultaneously in 2 l of contamination fluid 5.3.2.15 Two bottles of at least 1 l capacity 3) Vortex in an example of a suitable product available commercially. This information is given for t

48、he convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. BS EN 1499:2013EN 1499:2013 (E) 9 5.4 Preparation of test organism suspensions and product test solutions 5.4.1 Test organism suspensions (test and validation suspension) 5.4.1.1 General

49、For the test organism, two different suspensions have to be prepared: the “test suspension”, i.e. contamination fluid to perform the test, and the “validation suspension” to perform the controls and method validation. 5.4.1.2 Preservation and stock cultures of test organisms The test organism and its stock cultures shall be prepared and kept in accordance with EN 12353. 5.4.1.3 Working culture of test organisms In order to prepare the working culture of the test organism (5.2.1), prepare a first subcult