EN 14204-2012 en Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of mycobactericidal activity of chemical disinfectants and antiseptics use.pdf

1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN 14204:2012Chemical disinfectants andantiseptics Quantitativesuspension test for theevaluation of mycobactericidalactivity of chemicaldisinfectants and antisepticsused in th

2、e veterinary area Test method and requirements(phase 2, step 1)BS EN 14204:2012 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 14204:2012. Itsupersedes BS EN 14204:2004 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommitt

3、ee CH/216, Chemical disinfectants and antiseptics.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 correctapplication. The British Sta

4、ndards Institution 2013. Published by BSI StandardsLimited 2013ISBN 978 0 580 74073 2ICS 71.100.35Compliance with 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.

5、Amendments issued since publicationDate Text affectedBS EN 14204:2012EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 14204 November 2012 ICS 71.100.35 Supersedes EN 14204:2004English Version Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of mycobacteric

6、idal activity of chemical disinfectants and antiseptics used in the veterinary area - Test method and requirements (phase 2, step 1) Antiseptiques et dsinfectants chimiques - Essai quantitatif de suspension pour lvaluation de lactivit mycobactricide des antiseptiques et des dsinfectants chimiques ut

7、iliss dans le domaine vtrinaire - Mthode dessai et prescriptions (Phase 2, tape 1) Chemische Desinfektionsmittel und Antiseptika - Quantitativer Suspensionsversuch zur Bestimmung der mykobakteriziden Wirkung chemischer Desinfektionsmittel und Antiseptika fr den Veterinrbereich - Prfverfahren und Anf

8、orderungen (Phase 2, Stufe 1) This European Standard was approved by CEN on 22 September 2012. CEN members are 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-da

9、te 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 in three official versions (English, French, German). A version in any other language made by translation

10、 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 standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, For

11、mer Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT

12、 EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2012 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 14204:2012: EBS EN 14204:2012EN 14204:2012 (E) 2 Contents Page Forew

13、ord 3Introduction .41 Scope 52 Normative references 53 Terms and definitions .54 Requirements .55 Test method 65.1 Principle 65.2 Materials and reagents 75.2.1 Test organisms 75.2.2 Culture media and reagents .75.3 Apparatus and glassware 105.3.1 General . 105.3.2 Usual microbiological laboratory eq

14、uipment and, in particular, the following: . 105.4 Preparation of mycobacterial test suspension and product test solutions . 115.4.1 Mycobacterial test suspensions (test and validation suspension) . 115.4.2 Product test solutions 135.5 Procedure for assessing the mycobactericidal activity of the pro

15、duct 145.5.1 General . 145.5.2 Test procedure - Dilution-neutralization method 155.5.3 Test procedure - Membrane filtration method . 175.6 Experimental data and calculation 195.6.1 Explanation of terms and abbreviations 195.6.2 Calculation . 195.7 Verification of methodology 235.7.1 General . 235.7.

16、2 Control of weighted mean counts . 235.7.3 Basic limits 235.8 Expression of results . 245.8.1 Reduction 245.8.2 Control of active and non-active product test solution (5.4.2) 245.8.3 Mycobactericidal concentration 245.9 Interpretation of results - conclusion . 245.9.1 General . 245.9.2 Mycobacteric

17、idal activity for general purposes 245.9.3 Qualification for certain fields of application 245.10 Test report . 25Annex A (informative) Referenced strain in national collections 27Annex B (informative) Examples of neutralizers of the residual antimicrobial activity of chemical disinfectants and anti

18、septics and rinsing liquids . 28Annex C (informative) Graphical representations of dilution-neutralization method . 30Annex D (informative) Example of a typical test report 32Bibliography . 36BS EN 14204:2012EN 14204:2012 (E) 3 Foreword This document (EN 14204:2012) has been prepared by Technical Co

19、mmittee 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 May 2013, and conflicting national standards

20、 shall be withdrawn at the latest by May 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 document supersedes EN 14204:2004.

21、 Data obtained using the former version of EN 14204 may still be used. This document was revised to correct obvious errors and ambiguities, to harmonize the structure and wording with other quantitative suspension tests of CEN/TC 216 (existing or in preparation), and to improve the readability of th

22、e standard and thereby make it more understandable. The following technical changes have been made: Membrane filtration (5.5.3) method and associated media and equipment have been added. According to the CEN/CENELEC Internal Regulations, the national standards organisations of the following countrie

23、s 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, Netherlands, Norway, Poland

24、, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 14204:2012EN 14204:2012 (E) 4 Introduction This European Standard specifies a suspension test for establishing whether a chemical disinfectant or antiseptic has mycobactericidal activity in the

25、area and fields described in the scope. This laboratory test takes into account practical conditions 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 ar

26、e intended to cover general purposes and to allow reference between laboratories and product 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

27、of a product may differ and therefore additional test conditions need to be used. BS EN 14204:2012EN 14204:2012 (E) 5 1 Scope This European Standard specifies a test method and the minimum requirements for mycobactericidal activity of chemical disinfectant and antiseptic products that form a homogen

28、eous, physically stable preparation when diluted with hard water or in the case of ready-to-use-products with water. Products can only be tested at a concentration of 80 % or less, as some dilution is always produced by adding the test organisms and interfering substance. This European Standard appl

29、ies to products that are used in the veterinary area 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 anothe

30、r and to “use recommendations”. 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 1 test. 2 Normative references The following documents, in w

31、hole 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 amendments) applies. EN 12353, Chemical disinfectants and

32、 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 disinfectants and antiseptics 3 Terms and definitions For the p

33、urposes 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 when tested in accordance with Table 1 and Clause 5 under simulated low level (3,0 g/l bovine albumin) or high level soiling (10 g/l yeast

34、 extract and 10 g/l bovine albumin). BS EN 14204:2012EN 14204:2012 (E) 6 Table 1 - Obligatory and additional test conditions Test conditions Mycobactericidal activity Test organism a) obligatory Mycobacterium avium b) additional any relevant test organism Test temperature a) obligatory 10C 1C b) add

35、itional 4C 1C; 20C 1C; 40C 1C Contact time a) obligatory 60 min 10 sab) additional 1 min 5 s; 5 min 10 s; 10 min 10 s, 15 min 10 s, 30 min 10 s, 120 min 10 s Interfering substance a) obligatory low level soiling high level soiling 3,0 g/l bovine albumin 10 g/l yeast extract plus 10 g/l bovine albumi

36、n b) additional any relevant substance NOTE For the additional conditions, the concentration defined as a result can be lower than the one obtained under the obligatory test conditions.a The obligatory contact time for disinfectants stated in Table 1 was chosen to enable comparison of standard condi

37、tions. The referenced test conditions are by no means intended as requirements for the use of a product. The recommended contact time for the use of the product is within the responsibility of the manufacturer. Any additional specific mycobactericidal activity shall be determined in accordance with

38、5.2.1 and 5.5.1.1 in order to take into account intended specific use conditions. 5 Test method 5.1 Principle A sample of the product as delivered and/or diluted in hard water (or water for ready to use products) is added to a test suspension of mycobacteria in a solution of an interfering substance

39、. The mixture is maintained at (10 1) C for 60 min 10 s. At the end of this contact time, an aliquot is taken and the mycobactericidal action in this portion is immediately neutralized or suppressed by a validated method. The number of surviving mycobacteria in each sample is determined and the redu

40、ction in viable counts calculated. BS EN 14204:2012EN 14204:2012 (E) 7 The test is performed using Mycobacterium avium as the test organism (Clause 4, Table 1). Additional and optional contact times and temperatures are specified (Clause 4, Table 1). Additional interfering substances and test organi

41、sms may be used. 5.2 Materials and reagents 5.2.1 Test organisms 5.2.1.1 The mycobactericidal activity shall be evaluated using the following strain : Mycobacterium avium ATCC 157691)NOTE See Annex A for corresponding strain numbers in some other culture collections. The required incubation temperat

42、ure for this test organism is (36 1) C or (37 1) C. The same temperature (either 36 C or 37 C) shall 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,

43、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 are not classified at a reference centre, their identification

44、 characteristics shall be stated. In addition, they shall be held by the testing laboratory or national culture collection under a reference for five years. 5.2.2 Culture media and reagents 5.2.2.1 General All weights of chemical substances given in this European Standard refer to the anhydrous salt

45、s. Hydrated forms may be used as an alternative, but the weights required shall be adjusted to allow for consequent molecular weight differences. The reagents shall be of analytical grade and/or appropriate for microbiological purposes. They shall be free from substances that are toxic or inhibitory

46、 to the test organisms. If additional strains do not grow on the media (5.2.2.3) or cannot be used with diluent (5.2.2.4) additional media shall be used and shall be reported as well as additional incubation conditions. To improve the reproducibility, it is recommended that commercially available de

47、hydrated material is used for the preparation of culture media. The manufacturers instructions relating to the preparation of these products should be rigorously followed. Ready-to-use media may be used if it complies with the required specification. For each culture medium and reagent, a limitation

48、 for use should be fixed. 5.2.2.2 Water The water shall be freshly glass distilled and not demineralised water. If distilled water of adequate quality is not available, water for injectable preparations (see bibliographic reference 1 Pharmacopoeia) can be used. 1) ATCC 15769 is the collection number

49、 of strain supplied by the American Type Culture Collections. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of the product named. BS EN 14204:2012EN 14204:2012 (E) 8 Sterilize in the autoclave 5.3.2.1a). Sterilization is not necessary if the water is used e. g. for