1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN 14349:2012Chemical disinfectants andantiseptics Quantitativesurface test for the evaluationof bactericidal activity ofchemical disinfectants andantiseptics used in theveter
2、inary area on non-poroussurfaces without mechanicalaction Test method andrequireCopyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14349:2012 BRITISH STANDARDNational foreword
3、This British Standard is the UK implementation of EN 14349:2012. Itsupersedes BS EN 14349:2007 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee CH/216, Chemical disinfectants and antiseptics.A list of organizations represented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 beobt
4、ained on request 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 StandardsLimited 2013ISBN 978 0 580 74074 9ICS 71.100.35Compliance with
5、 a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 28 February 2013.Amendments issued since publicationDate Text affectedCopyright European Committee for Standardization Provided by
6、 IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14349:2012EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 14349 November 2012 ICS 71.100.35 Supersedes EN 14349:2007English Version Chemical disinfectants and antiseptics - Quanti
7、tative surface test for the evaluation of bactericidal activity of chemical disinfectants and antiseptics used in the veterinary area on non-porous surfaces without mechanical action - Test method and requirements (phase 2, step 2) Antiseptiques et dsinfectants chimiques - Essai quantitatif de surfa
8、ce pour lvaluation de lactivit bactricide des antiseptiques et des dsinfectants chimiques utiliss dans le domaine vtrinaire sur des surfaces non poreuses sans action mcanique - Mthode dessai et prescriptions (phase 2, tape 2) Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflchenve
9、rsuch zur Bestimmung der bakteriziden Wirkung chemischer Desinfektionsmittel und Antiseptika fr den Veterinrbereich auf nicht-porsen Oberflchen ohne mechanische Wirkung - Prfverfahren und Anforderungen (Phase 2, Stufe 2) This European Standard was approved by CEN on 22 September 2012. CEN members ar
10、e bound to comply with the CEN/CENELEC Internal 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
11、the CEN-CENELEC Management Centre or to 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 Ce
12、ntre has the same status as the official 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, Lithua
13、nia, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, 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
14、 2012 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 14349:2012: ECopyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IH
15、S-,-,-BS EN 14349:2012EN 14349:2012 (E) 2 Contents Page Foreword 3Introduction .41 Scope 52 Normative references 53 Terms and definitions .54 Requirements .55 Test method 75.1 Principle 75.2 Materials and reagents 75.2.1 Test organisms 75.2.2 Culture media and reagents .85.2.3 Test surface . 105.3 A
16、pparatus and glassware 105.3.1 General . 105.3.2 Usual microbiological laboratory equipment 115.4 Preparation of test organism suspensions and product test solutions . 125.4.1 Test organism suspensions (test and validation suspension) 125.4.2 Product test solutions 145.5 Procedure for assessing the
17、bactericidal activity of the product . 145.5.1 General . 145.5.2 Test procedure (Dilution-neutralization method). 155.5.3 Observation of test surface agar . 185.6 Experimental data and calculation 185.6.1 Explanation of terms and abbreviations 185.6.2 Calculation . 195.7 Verification of methodology
18、225.7.1 General . 225.7.2 Control of weighted mean counts . 225.7.3 Basic limits 225.8 Expression of results and precision . 225.8.1 Reduction 225.8.2 Control of active and non-active product test solution (5.4.2) 235.8.3 Limiting test organism and bactericidal concentration 235.8.4 Precision, repet
19、itions . 235.9 Interpretation of results conclusion 235.9.1 General . 235.9.2 Bactericidal activity for general purposes . 235.9.3 Qualification for certain fields of application 245.10 Test report . 24Annex A (informative) Referenced strains in national collections 26Annex B (informative) Examples
20、of neutralizers of the residual antimicrobial activity of chemical disinfectants and antiseptics 27Annex C (informative) Graphical representation of the method . 29Annex D (informative) Example of a typical test report 33Bibliography . 36Copyright European Committee for Standardization Provided by I
21、HS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14349:2012EN 14349:2012 (E) 3 Foreword This document (EN 14349:2012) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the secretariat of whic
22、h 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 May 2013, and conflicting national standards shall be withdrawn at the latest by May 2013. Attention is drawn to the possibility
23、 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 document supersedes EN 14349:2007. Data obtained using the former version of EN 14349 may still be used. It was revise
24、d to correct obvious errors and ambiguities, to harmonize 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 more understandable. According to the CEN/CENELEC Internal Regulations, the national sta
25、ndards organisations 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,
26、Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without licen
27、se from IHS-,-,-BS EN 14349:2012EN 14349:2012 (E) 4 Introduction This European Standard specifies a surface test for establishing whether a chemical disinfectant or antiseptic has bactericidal activity in the area and fields described in the scope. This laboratory test takes into account practical c
28、onditions of application of the product including contact time, temperature, test organisms and interfering substances, i.e. conditions which may influence its action in practical situations. The conditions are intended to cover general purposes and to allow reference between laboratories and produc
29、t types. Each utilization concentration of the chemical disinfectant or antiseptic, found by this test corresponds to the chosen experimental conditions. However, for some applications, the instructions of use of a product may differ and therefore additional test conditions need to be used. Copyrigh
30、t European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14349:2012EN 14349:2012 (E) 5 1 Scope This European Standard specifies a test method and the minimum requirements for bactericidal a
31、ctivity of chemical disinfectant and antiseptic products that form a homogeneous physically stable preparation when diluted with hard water, or in the case of ready-to-use-products with water. This European Standard applies to products that are used in the veterinary area on non-porous surfaces with
32、out mechanical action i.e. in the breeding, husbandry, production, transport and disposal of all animals except when in the food chain following death and entry to the processing industry. EN 14885 specifies in detail the relationship of the various tests to one another and to “use recommendations”.
33、 NOTE 1 The method described is intended to determine the activity of commercial formulations or active substances under the conditions in which they are used. NOTE 2 This method corresponds to a Phase 2 Step 2 test. This method cannot be used to evaluate the activity of products against mycobacteri
34、a. 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 (including any am
35、endments) applies. EN 12353, Chemical disinfectants and antiseptics Preservation of test organisms used for the determination of bactericidal, mycobactericidal, sporicidal and fungicidal activity EN 14885, Chemical disinfectants and antiseptics Application of European Standards for chemical disinfec
36、tants and antiseptics 3 Terms and definitions For the purposes of this document, the terms and definitions given in EN 14885 apply. 4 Requirements The product shall demonstrate at least a 4 decimal log (lg) reduction from a water control, when tested in accordance with Table 1 and Clause 5 under sim
37、ulated low level (3,0 g/l bovine albumin) or high level soiling (10 g/l yeast extract and 10 g/l bovine albumin) on a surface. Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS
38、EN 14349:2012EN 14349:2012 (E) 6 Table 1 - Obligatory and additional test conditions Test conditions Bactericidal activity on non-porous surfaces without mechanical action in the veterinary area Test organism a) obligatory Enterococcus hirae Proteus vulgaris Pseudomonas aeruginosa Staphylococcus aur
39、eus b) additional any relevant test organism Test temperature a) obligatory 10 C + 1 C b) additional 4 C + 1 C; 20 C + 1 C; 40 C + 1 C Contact time a) obligatory 30 min + 10 s b) additional 1 min + 5 s; 5 min + 10 s; 60 min + 10 s Interfering substance a) obligatory low level soiling high level soil
40、ing 3,0 g/l bovine albumin 10 g/l yeast extract plus 10 g/l bovine albumin b) additional any relevant substance The obligatory contact time for surface disinfectants stated in Table 1 were chosen to enable comparison of standard conditions. The recommended contact time for the use of the product is
41、within the responsibility of the manufacturer. NOTE For the additional conditions, the concentration defined as a result can be lower than the one obtained under the obligatory test conditions. Any additional specific bactericidal activity shall be determined in accordance with 5.2.1 and 5.5.1.1 in
42、order to take into account intended specific use conditions. Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14349:2012EN 14349:2012 (E) 7 5 Test method 5.1 Principle A tes
43、t suspension of bacteria mixed with interfering substance is inoculated onto the test surface and dried. After a drying time, 0,1 ml of the product is transferred to the surface, in a manner which covers the dried film. The surface is maintained at a specified temperature for a defined period of tim
44、e specified in Clause 4 and 5.5.1.1. At the end of that contact time the surface is transferred to a neutralizer so that the action of the disinfectant is immediately neutralized. The numbers of surviving organisms which can be recovered from the surface is determined quantitatively. The number of b
45、acteria on a surface treated with water in place of the disinfectant is also determined and the reduction is calculated. The test is performed using Enterococcus hirae, Proteus vulgaris, Pseudomonas aeruginosa and Staphylococcus aureus as test organisms (Clause 4, Table 1). Additional and optional c
46、ontact times and temperatures are specified (Clause 4, Table 1). Additional interfering substances and test organisms may be used. 5.2 Materials and reagents 5.2.1 Test organisms The bactericidal activity shall be evaluated using the following strains1): Enterococcus hirae ATCC 10541; Proteus vulgar
47、is ATCC 13315; Pseudomonas aeruginosa ATCC 15442; Staphylococcus aureus ATCC 6538. NOTE See Annex A for strain references in some other culture collections. The required incubation temperature for these test organisms is 36 C 1 C or 37 C 1 C (5.3.2.3). The same temperature (either 36 C or 37 C) shal
48、l be used for all incubations performed during a test and its control and validation. If additional test organisms are used, they shall be incubated under optimum growth conditions (temperature, time, atmosphere, media) noted in the test report. If the additional test organisms selected do not correspond to the specified strains, their suitability for supplying the required inocula shall be verified. If these additional test organisms ar
