EN 1276-2009 en Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of bactericidal activity of chemical disinfectants and antiseptics used in .pdf

1、BS EN 1276:2009ICS 11.080.20; 67.020; 71.100.35NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBRITISH STANDARDChemical disinfectants and antiseptics Quantitative suspension test for the evaluation of bactericidal activity of chemical disinfectants and antiseptics used in food,

2、 industrial, domestic and institutional areas Test method and requirements (phase 2,step 1)Incorporating corrigendum August 2010National forewordThis British Standard is the UK implementation of EN 1276:2009, incorporating corrigendum August 2010. It supersedes BS EN 1276:1997 which is withdrawn.The

3、 UK participation in its preparation was entrusted to Technical Committee CH/216, Chemical disinfectants and antiseptics.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a co

4、ntract. Users are responsible for its correct application.Compliance with a British Standard cannot confer immunity from legal obligations.BS EN 1276:2009This British Standardwas published under theauthority of the StandardsPolicy and StrategyCommittee on 31 May 2010 BSI 2010Amendments/corrigenda is

5、sued since publicationDate Comments 30 November 2010 Implementation of CEN corrigendum August 2010: 5.7.3 correctedISBN 978 0 580 72821 1EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 1276 October 2009 ICS 71.100.35 Supersedes EN 1276:1997English Version Chemical disinfectants and antiseptics

6、- Quantitative suspension test for the evaluation of bactericidal activity of chemical disinfectants and antiseptics used in food, industrial, domestic and institutional areas - Test method and requirements (phase 2, step 1) Antiseptiques et dsinfectants chimiques - Essai quantitatif de suspension p

7、our lvaluation de lactivit bactricide des antiseptiques et des dsinfectants chimiques utiliss dans le domaine de lagro-alimentaire, dans lindustrie, dans les domaines domestiques et en collectivit - Mthode dessai et prescriptions (Phase 2, tape 1) Chemische Desinfektionsmittel und Antiseptika - Quan

8、titativer Suspensionsversuch zur Bestimmung der bakteriziden Wirkung chemischer Desinfektionsmittel und Antiseptika in den Bereichen Lebensmittel, Industrie, Haushalt und ffentliche Einrichtungen - Prfverfahren und Anforderungen (Phase 2, Stufe 1) This European Standard was approved by CEN on 20 Sep

9、tember 2009. 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-date lists and bibliographical references concerning such national standards may be

10、obtained on application to the CEN 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 Ma

11、nagement Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands,

12、Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2009 CEN All rights of exploitation in any f

13、orm and by any means reserved worldwide for CEN national Members. Ref. No. EN 1276:2009: EIncorporating corrigendum August 2010EN 1276:2009 (E) 2 Contents Page Foreword 4Introduction .51 Scope 62 Normative references 73 Terms and definitions .74 Requirements .75 Test method 85.1 Principle 85.2 Mater

14、ials and reagents 95.2.1 Test organisms 95.2.2 Culture media and reagents .95.3 Apparatus and glassware 135.4 Preparation of test organism suspensions and product test solutions . 145.4.1 Test organism suspensions (test and validation suspension) 145.4.2 Product test solutions 155.5 Procedure for as

15、sessing the bactericidal activity of the product . 175.5.1 General . 175.5.2 Dilution-neutralization method 185.5.3 Membrane filtration method 205.6 Experimental data and calculation 225.6.1 Explanation of terms and abbreviations 225.6.2 Calculation . 235.7 Verification of methodology 265.7.1 Genera

16、l . 265.7.2 Control of weighted mean counts . 265.7.3 Basic limits 265.8 Expression of results and precision . 265.8.1 Reduction 265.8.2 Control of active and non-active product test solution (5.4.2) 275.8.3 Limiting test organism and bactericidal concentration 275.8.4 Precision, replicates . 275.9

17、Interpretation of results - conclusion . 275.9.1 General . 275.9.2 Bactericidal activity for general purposes . 27BS EN 1276:2009EN 1276:2009 (E) 3 5.9.3 Bactericidal activity for specific purposes . 275.10 Test report 28Annex A (informative) Referenced strains in national collections 30Annex B (inf

18、ormative) Neutralizers and rinsing liquids 31Annex C (informative) Graphical representations of dilution neutralization method and membrane filtration method . 33C.1 Dilution-neutralization method 33C.2 Membrane filtration method . 35Annex D (informative) Example of a typical test report 37Test resu

19、lts (bactericidal suspension test) . 39Annex E (informative) Precision of the test result . 41Bibliography 44BS EN 1276:2009EN 1276:2009 (E) 4 Foreword This document (EN 1276:2009) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the secretariat of which

20、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 April 2010, and conflicting national standards shall be withdrawn at the latest by April 2010. Attention is drawn to the possibili

21、ty 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 1276:1997. It was revised to include the results of a collaborative trial (ANDISTAND), to cor

22、rect 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 the standard and thereby make it more understandable. According to the CEN/CENELEC Internal Regulations, t

23、he national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, N

24、orway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. BS EN 1276:2009EN 1276:2009 (E) 5 Introduction This European Standard specifies a suspension test for establishing whether a chemical disinfectant or antiseptic has or does not have bactericidal

25、activity 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 substance, i.e. conditions which may influence its action in practical situations. The condit

26、ions are 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 defined experimental conditions. However, for some applications, the recommendations

27、of use of a product may differ and therefore additional test conditions need to be used. BS EN 1276:2009EN 1276:2009 (E) 6 1 Scope This European Standard specifies a test method and the minimum requirements for bactericidal activity of chemical disinfectant and antiseptic products that form a homoge

28、neous, 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 document applies

29、to products that are used in food, industrial, domestic and institutional areas excluding areas and situations where disinfection is medically indicated and excluding products used on living tissues except those for hand hygiene in the above considered areas. The following areas are at least include

30、d: a) processing, distribution and retailing of: 1) food of animal origin: milk and milk products; meat and meat products; fish, seafood, and related products; eggs and egg products; animal feeds; etc. 2) food of vegetable origin: beverages; fruits, vegetables and derivatives (including sugar, disti

31、llery, etc.); flour, milling and baking; animal feeds; etc. b) institutional and domestic areas: catering establishments; public areas; public transports; schools; nurseries; shops; sports rooms; waste containers (bins, etc.); hotels; dwellings; clinically non sensitive areas of hospitals; offices;

32、etc. BS EN 1276:2009EN 1276:2009 (E) 7 c) other industrial areas: packaging material; biotechnology (yeast, proteins, enzymes, etc.); pharmaceutical; cosmetics and toiletries; textiles; space industry, computer industry; etc. EN 14885 specifies in detail the relationship of the various tests to one

33、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 which they are used. NOTE 2 This method corresponds to a phase 2 step 1 test. 2 Normative references The following reference

34、d documents are indispensable for the application of this document. 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 antiseptics Preservation of test or

35、ganisms used for the determination of bactericidal, mycobactericidal, sporicidal and fungicidal activity EN 14885:2006, Chemical disinfectants and antiseptics Application of European Standards for chemical disinfectants and antiseptics 3 Terms and definitions For the purposes of this document, the t

36、erms and definitions given in EN 14885:2006 apply. 4 Requirements The product shall demonstrate at least a 5 decimal log (lg) reduction when diluted with hard water (5.2.2.7) or - in the case of ready-to-use products - with water (5.2.2.2) and tested in accordance with Clause 5 under simulated clean

37、 conditions (0,3 g/l bovine albumin solution- 5.2.2.8.2) or simulated dirty conditions (3 g/l bovine albumin solution - 5.2.2.8.3) according to its practical applications and under the other obligatory test conditions (four selected test organisms, 20 C, 5 min or 1 min (for hands disinfection). The

38、bactericidal activity shall be evaluated using the following four test organisms: Pseudomonas aeruginosa; Escherichia coli; BS EN 1276:2009EN 1276:2009 (E) 8 Staphylococcus aureus; Enterococcus hirae. Where indicated, additional specific bactericidal activity shall be determined applying other conta

39、ct times, temperatures, interfering substances and test organisms (in accordance with 5.2.1, 5.2.2.8 and 5.5.1.1) in order to take into account intended specific use conditions. NOTE For these additional conditions, the concentration defined as a result can be lower than the one obtained under the o

40、bligatory test conditions. 5 Test method 5.1 Principle 5.1.1 A sample of the product as delivered and/or diluted with hard water (or water for ready-to-use products) is added to a test suspension of bacteria in a solution of an interfering substance. The mixture is maintained at (20 1) C for 5 min 1

41、0 s (obligatory test conditions) during 1 min (obligatory test conditions for hands disinfection). At the end of this contact time, an aliquot is taken, and the bactericidal and/or the bacteriostatic activity in this portion is immediately neutralized or suppressed by a validated method. The method

42、of choice is dilution-neutralization. If a suitable neutralizer cannot be found, membrane filtration is used. The numbers of surviving bacteria in each sample are determined and the reduction is calculated. 5.1.2 The test is performed using Pseudomonas aeruginosa, Escherichia coli, Staphylococcus au

43、reus and Enterococcus hirae as test organisms. 5.1.3 Additional and optional contact times and temperatures are specified. Additional test organisms can be used. BS EN 1276:2009EN 1276:2009 (E) 9 5.2 Materials and reagents 5.2.1 Test organisms The bactericidal activity shall be evaluated using the f

44、ollowing strains as test organisms1): Pseudomonas aeruginosa ATCC 15442; Escherichia coli ATCC 10536; Staphylococcus aureus ATCC 6538; Enterococcus hirae ATCC 10541. If required for specific applications, additional strains may be chosen, for example from: Salmonella Typhimurium ATCC 13311; Lactobac

45、illus brevis DSM 6235; Enterobacter cloacae DSM 6234. NOTE See Annex A for strain references in some other culture collections. The required incubation temperature for these test organisms is (36 1) C or (37 1) C (5.3.2.3). The same temperature (either 36 C or 37 C) shall be used for all incubations

46、 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 strain

47、s, their suitability for supplying the required inocula shall be verified. If these additional test organisms 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

48、 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 salts. Hydrated forms may be used as an alternative, but the weights required shall be adjusted to allow for consequent molecular we

49、ight 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 to the test organisms. NOTE 1 To improve reproducibility, it is recommended that commercially available dehydrated material is