EN 16438-2014 en Chemical disinfectants and antiseptics - Quantitative surface test for the evaluation of fungicidal or yeasticidal activity of chemical disinfectants and antisepti.pdf

1、BSI Standards PublicationBS EN 16438:2014Chemical disinfectants and antiseptics Quantitative surface test for the evaluation of fungicidal or yeasticidal activity of chemical disinfect-ants and antiseptics used in the veterinary area on non-porous surfaces without mechanical action Test method andre

2、quirements (phase 2, step 2)BS EN 16438:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 16438:2014. The UK participation in its preparation was entrusted to TechnicalCommittee CH/216, Chemical disinfectants and antiseptics.A list of organizations represente

3、d on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2014.Published by BSI Standards Limited 2014ISBN 978 0 580 767

4、81 4ICS 71.100.35Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 28 February 2014.Amendments/corrigenda issued since publicationDate T e x t a f f e c t e dBS EN

5、 16438:2014EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16438 February 2014 ICS 71.100.35 English Version Chemical disinfectants and antiseptics - Quantitative surface test for the evaluation of fungicidal or yeasticidal activity of chemical disinfectants and antiseptics used in the veterina

6、ry area on non-porous surfaces without mechanical action - Test method and requirements (phase 2, step 2) Antiseptiques et dsinfectants chimiques - Essai quantitatif de surface pour lvaluation de lactivit fongicide ou levuricide des antiseptiques et des dsinfectants chimiques utiliss dans le domaine

7、 vtrinaire sur des surfaces non poreuses sans action mcanique - Mthode dessai et prescriptions (phase 2, tape 2) Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflchenversuch zur Bestimmung der fungiziden oder levuroziden Wirkung chemischer Desinfektionsmittel und Antiseptika fr de

8、n Veterinrbereich auf nicht-porsen Oberflchen ohne mechanische Wirkung - Prfverfahren und Anforderungen (Phase 2, Stufe 2) This European Standard was approved by CEN on 30 November 2013. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for givi

9、ng 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 in three offi

10、cial 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 standards bodies of

11、 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, Slovenia, Spain,

12、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 2014 CEN All rights of exploitation in any form and by any means reserved worldwide fo

13、r CEN national Members. Ref. No. EN 16438:2014 EBS EN 16438:2014EN 16438:2014 (E) 2 Contents Page Foreword 3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Requirements .5 5 Test method 6 5.1 Principle 6 5.2 Materials and reagents 7 5.2.1 Test organism 7 5.2.2 Cultur

14、e media and reagents .7 5.2.3 Test surface . 10 5.3 Apparatus and glassware 10 5.3.1 General . 10 5.3.2 Usual microbiological laboratory equipment . 10 5.4 Preparation of test organism suspensions and product test solutions . 11 5.4.1 Test organism suspension (test and validation suspension) 11 5.4.

15、2 Product test solutions 15 5.5 Procedure for assessing the fungicidal or yeasticidal activity of the product 16 5.5.1 General . 16 5.5.2 Test procedure (Dilution-neutralisation method) 17 5.5.3 Observation of the test surface agar 20 5.6 Experimental data and calculation 20 5.6.1 Explanation of ter

16、ms and abbreviations 20 5.6.2 Calculation . 21 5.7 Verification of methodology 24 5.7.1 General . 24 5.7.2 Control of weighted mean counts . 24 5.7.3 Basic limits 24 5.7.4 Microscopic observation . 24 5.8 Expression of results and precision . 24 5.8.1 Reduction 24 5.8.2 Control of active and non-act

17、ive product test solution (5.4.2) 25 5.8.3 Fungicidal or yeasticidal concentration . 25 5.8.4 Precision, repetitions . 25 5.9 Interpretation of results - conclusion . 25 5.9.1 General . 25 5.9.2 Fungicidal or yeasticidal activity for general purposes . 26 5.9.3 Qualification for certain fields of ap

18、plication 26 5.10 Test report . 26 Annex A (informative) Referenced strains of national collections . 28 Annex B (informative) Examples of neutralisers of the residual antimicrobial activity of chemical disinfectants and antiseptics 29 Annex C (informative) Graphical representations of dilution-neut

19、ralisation method . 31 Annex D (informative) Example of a typical test report 35 Bibliography . 39 BS EN 16438:2014EN 16438:2014 (E) 3 Foreword This document (EN 16438:2014) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the secretariat of which is held

20、 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 August 2014 and conflicting national standards shall be withdrawn at the latest by August 2014. Attention is drawn to the possibility tha

21、t 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 standards organisations of the following countries are bound to

22、 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, Portugal, Ro

23、mania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 16438:2014EN 16438:2014 (E) 4 Introduction This European Standard specifies a surface test for establishing whether a chemical disinfectant or antiseptic has or does not have fungicidal or yeasticidal activit

24、y in the 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

25、e intended to cover general purposes and to allow reference between laboratories and product types. Each utilisation 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

26、of a product may differ and therefore additional test conditions need to be used. BS EN 16438:2014EN 16438:2014 (E) 5 1 Scope This European Standard specifies a test method and the minimum requirements for fungicidal or yeasticidal activity of chemical disinfectant and antiseptic products that form

27、a homogeneous physically stable preparation in hard water or in the case of ready-to-use products with water. This European Standard applies to products for use in the veterinary area i.e. in the breeding, husbandry, production, transport and disposal of all animals except when in the food chain fol

28、lowing 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”. NOTE 1 The method described is intended to determine the activity of commercial formulations or active substances under the conditions in

29、 which they are used. NOTE 2 This method corresponds to a Phase 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

30、 references, the latest edition of the referenced document (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 (

31、including bacteriophages) activity EN 14885, Chemical disinfectants and antiseptics - Application of European Standards for chemical disinfectants and antiseptics ISO 4793, Laboratory sintered (fritted) filters Porosity grading, classification and designation 3 Terms and definitions For the purposes

32、 of this document, the terms and definitions given in EN 14885 apply. 4 Requirements The product shall demonstrate at least a 3 decimal log (lg) reduction from a water control, when tested in accordance with Table 1 and Clause 5 under simulated low level (3,0 g/l bovine albumin) or high level soilin

33、g (10 g/l yeast extract and 10 g/l bovine albumin) on a surface. BS EN 16438:2014EN 16438:2014 (E) 6 Table 1 Obligatory and additional test conditions Test Conditions Fungicidal or yeasticidal activity on non-porous surfaces without mechanical action in the veterinary area Test organism Fungicidal a

34、ctivity a) obligatory Aspergillus brasiliensis (conidiospores) and Candida albicans (vegetative cells) Test organism Yeasticidal activity a) obligatory Candida albicans (vegetative cells) b) additional any relevant test organism Test temperature a) obligatory 10 C 1 C b) additional 4 C 1 C; 20 C 1 C

35、; 40 C 1 C Contact time a) obligatory 60 min 10 s b) additional* 5 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 albumin b) additional any relevant substance The obligatory c

36、ontact times 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 within the responsibility of the manufacturer. NOTE For the additional conditions, the concentration defined as a result can b

37、e lower than the one obtained under the obligatory test conditions. Any additional specific fungicidal activity shall be determined in accordance with 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 test suspension of fungal conidiospor

38、es or yeast and interfering substance is inoculated onto the test surface and dried. After a drying time, an aliquot of the product under test 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 time spe

39、cified in Clause 4 and 5.5.1.1. At the end of that contact time the surface is transferred to a neutraliser so that the action of the disinfectant is immediately neutralised. The numbers of surviving organisms which can be recovered from the surface is determined quantitatively. The number of fungi

40、or yeast on a surface treated with water in place of the disinfectant is also determined and the reduction is calculated. The test is performed using the conidiospores of Aspergillus brasiliensis (formerly niger) and the vegetative cells of Candida albicans (fungicidal activity) or only the vegetati

41、ve cells of Candida albicans (yeasticidal BS EN 16438:2014EN 16438:2014 (E) 7 activity) as test organisms (Clause 4, Table 1). Additional and optional contact times and temperatures are specified (Clause 4, Table 1). Additional interfering substances and test organisms may be used. 5.2 Materials and

42、 reagents 5.2.1 Test organism The fungicidal activity shall be evaluated using the following strains as the test organisms1): Aspergillus brasiliensis (formerly Aspergillus niger) ATCC 16404 Candida albicans ATCC 10231 The yeasticidal activity shall be evaluated using only Candida albicans. NOTE See

43、 Annex A for strain references in some other culture collections. The required incubation temperature for these organisms is 30 C 1 C (5.3.2.3). The same temperature shall be used for all incubations performed during a test and its control and validation. If additional test organisms are used, they

44、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 o

45、rganisms are not classified at a reference centre, their identification 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

46、 substances given in this European Standard refer to the anhydrous salts. 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 pu

47、rposes. They shall be free from substances that are toxic or inhibitory to the test organism. To improve reproducibility, it is recommended that commercially available dehydrated material is used for the preparation of culture media. The manufacturers instructions relating to the preparation of thes

48、e products should be rigorously followed. Ready-to-use medium may be used if it complies with the required specification. For each culture medium and reagent a limitation for use should be fixed. 1) The ATCC numbers are the collection numbers of strains supplied by the American Type Culture Collecti

49、on (ATCC). 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 16438:2014EN 16438:2014 (E) 8 5.2.2.2 Water The water shall be freshly glass-distilled and not demineralised water. Sterilise in the autoclave 5.3.2.1 a). NOTE 1 Sterilisation is not necessary if the water is used e.g. for preparation of culture media and subsequently sterilised. NOTE 2 If distilled water of adequate quality is not available, water for injecti