EN 1656-2009 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 1656:2009ICS 11.080.20; 11.220; 71.100.35NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBRITISH STANDARDChemical disinfectantsand antiseptics Quantitativesuspension test forthe evaluation ofbactericidal activity ofchemical disinfectantsand antiseptics usedin the veterina

2、ry area Test method andrequirements (phase 2,step 1)IncorporatingcorrigendumAugust 2010 National forewordThis British Standard is the UK implementation of EN 1656:2009, incorporating corrigendum August 2010. It supersedes BS EN 1656:2000 which is withdrawn.The UK participation in its preparation was

3、 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 contract. Users are responsible for its co

4、rrect application.Compliance with a British Standard cannot confer immunity from legal obligations.BS EN 1656:2009This British Standardwas published under theauthority of the StandardsPolicy and StrategyCommittee on 31 March2010 BSI 2010Amendments/corrigenda issued since publicationDate Comments 30

5、November 2010 Implementation of CEN corrigendum August 2010. Correction to Annex A, b) Tent disinfectants: ATCC reference for Escherichia coli correctedISBN 978 0 580 73008 5EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 1656 November 2009 ICS 71.100.35 Supersedes EN 1656:2000English Version C

6、hemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of bactericidal 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 suspe

7、nsion pour lvaluation de lactivit bactricide des antiseptiques et des dsinfectants chimiques utiliss dans le domaine vtrinaire - Mthode dessai et prescriptions (phase 2, tape 1) Chemische Desinfektionsmittel und Antiseptika - Quantitativer Suspensionsversuch zur Bestimmung der bakteriziden Wirkung c

8、hemischer Desinfektionsmittel und Antiseptika fr den Veterinrbereich - Prfverfahren und Anforderungen (Phase 2, Stufe 1) This European Standard was approved by CEN on 20 September 2009. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for givin

9、g 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 Management Centre or to any CEN member. This European Standard exists in three official vers

10、ions (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 Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium

11、, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR ST

12、ANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2009 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 1656:2009: EIncorporating corrigendum August 2010

13、EN 1656:2009 (E) 2 Contents Page Foreword . 3Introduction 41 Scope . 52 Normative references . 53 Terms and definitions . 54 Requirements 55 Test method . 6Annex A (informative) Referenced strains in national collections 25Annex B (informative) Examples of neutralizers of the residual antimicrobial

14、activity of chemical disinfectants and antiseptics and rinsing liquids . 27Annex C (informative) Graphical representations of dilution-neutralization method and membrane filtration method 28Annex D (informative) Example of a typical test report 32Bibliography . 37BS EN 1656:2009EN 1656:2009 (E) 3 Fo

15、reword This document (EN 1656:2009) has been prepared 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 endor

16、sement, at the latest by May 2010, and conflicting national standards shall be withdrawn at the latest by May 2010. 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 a

17、ny or all such patent rights. This document supersedes EN 1656:2000. This document was revised to include the results of a collaborative trial (ANDISTAND), to correct obvious errors and ambiguities, to harmonize the structure and wording with other quantitative suspension tests of CEN/TC 216 (existi

18、ng 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 standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,

19、 Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. BS EN 1656:2009EN 1656:2009 (E)

20、4 Introduction This European Standard specifies a suspension test for establishing whether a chemical disinfectant or antiseptic has or does not have a bactericidal activity in the fields described in the scope. This laboratory test takes into account practical conditions of application of the produ

21、ct, including contact time, temperature, test organisms and interfering substance, 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 product types. Each utilization concentrati

22、on of the chemical disinfectant or antiseptic found by this test corresponds to defined experimental conditions. However, for some applications the recommendations of use of a product may differ and therefore additional test conditions need to be used. BS EN 1656:2009EN 1656:2009 (E) 5 1 Scope This

23、European Standard specifies a test method and the minimum requirements for bactericidal activity 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. Products can only

24、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 applies to products that are used in the veterinary area e.g. in the breeding, husbandry, transport and disposal of all animals except when

25、 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”. NOTE 1 The method described is intended to determine the activity of commercial formulations or active substances u

26、nder the conditions in which they are used. NOTE 2 This method corresponds to a phase 2 step 1 test. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the lat

27、est 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, mycobactericidal, sporicidal and fungicidal activity EN 14885:2006, Chemical disinfectants and antise

28、ptics 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 14885:2006 apply. 4 Requirements The product shall demonstrate at least a five-decimal log (lg) reduction when diluted w

29、ith 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 low-level soiling (3,0 g/l bovine albumin solution 5.2.2.8.2) or simulated high-level soiling (10 g/l bovine albumin solution and 10 g/l yeast extract 5.2.2.8.

30、3) (or 10 g/l skimmed milk for teat disinfectants 5.2.2.8.4) according to its practical applications and under the other obligatory test conditions four or three for teat disinfectants selected test organisms, 10 C 30 C for teat disinfectants, 30 min 5 min for teat disinfectants). The bactericidal a

31、ctivity shall be evaluated using the following organisms: a) Products for general disinfection: b) Teat disinfectants: Enterococcus hirae; Escherichia coli; Proteus vulgaris; Staphylococcus aureus; Pseudomonas aeruginosa; Streptococcus uberis. BS EN 1656:2009EN 1656:2009 (E) 6 Staphylococcus aureus.

32、 Where indicated, additional specific bactericidal activity shall be determined applying other contact 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 additio

33、nal conditions, the concentration defined as a result can be lower than the one obtained under the obligatory 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 b

34、acteria in a solution of an interfering substance. The mixture is maintained at (10 1) C (or (30 1) C for teat disinfectants) for 30 min 10 s (5 min 10 s for teat disinfectants) (obligatory test conditions). At the end of this contact time, an aliquot is taken, and the bactericidal and/or the bacter

35、iostatic activity in this portion is immediately neutralized or suppressed by a validated method. The method 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

36、 is calculated. 5.1.2 For general disinfectant products, the test is performed using Enterococcus hirae, Proteus vulgaris, Pseudomonas aeruginosa and Staphylococcus aureus as test organisms. For teat disinfectants the test is performed using Escherichia coli, Staphylococcus aureus and Streptococcus

37、uberis as test organisms. 5.1.3 Additional and optional contact times and temperatures are specified. Additional test organisms can be used. 5.2 Materials and reagents 5.2.1 Test organisms The bactericidal activity shall be evaluated using the following strains as test organisms1): a) General disinf

38、ection products: Enterococcus hirae ATCC 10541 Proteus vulgaris ATCC 13315 Pseudomonas aeruginosa ATCC 15442 Staphylococcus aureus ATCC 6538 b) Teat disinfectants: Escherichia coli ATCC 10536 Staphylococcus aureus ATCC 6538 Streptococcus uberis ATCC 19436 1) The ATCC numbers are the collection numbe

39、rs of strains supplied by the American Type Culture Collection (ATCC). This information is given for the convenience of users of this standard and does not constitute an endorsement by CEN of the product named. BS EN 1656:2009EN 1656:2009 (E) 7 NOTE See Annex A for strain references in some other cu

40、lture 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 performed during a test and its control and validation. If additional test organisms are used, they shall be

41、 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

42、 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 substan

43、ces given in this 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 purposes. They shal

44、l 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 used for the preparation of culture media. The manufacturers instructions relating to the preparation of these product

45、s should be rigorously followed. NOTE 2 For each culture medium and reagent, a shelf life should be fixed (see ISO/IEC 17025). 5.2.2.2 Water The water shall be freshly glass-distilled water and not demineralized water. Sterilize in the autoclave (5.3.2.1 a). NOTE 1 Sterilization is not necessary if

46、the water is used e.g. for preparation of culture media and subsequently sterilized. NOTE 2 If distilled water of adequate quality is not available, water for injections (see 1 in the bibliography) can be used. NOTE 3 See 5.2.2.7 for the procedure to prepare hard water. 5.2.2.3 Tryptone Soya Agar (T

47、SA) Tryptone soya agar, consisting of: Tryptone, pancreatic digest of casein 15,0 g Soya peptone, papaic digest of soybean meal 5,0 g Sodium chloride (NaCl) 5,0 g Agar 15,0 gWater (5.2.2.2) to 1 000,0 ml Sterilize in the autoclave (5.3.2.1 a). After sterilization the pH of the medium shall be equiva

48、lent to 7,2 0,2 when measured at (20 1) C. NOTE In case of encountering problems with neutralization (5.5.1.2 and 5.5.1.3) it may be necessary to add neutralizer to the TSA. Annex B gives guidance on the neutralizers that may be used. BS EN 1656:2009EN 1656:2009 (E) 8 5.2.2.4 Diluent Tryptone sodium

49、 chloride solution, consisting of: Tryptone, pancreatic digest of casein 1,0 g Sodium chloride (NaCl) 8,5 g Water (5.2.2.2) to 1 000,0 ml Sterilize in the autoclave (5.3.2.1 a). After sterilization, the pH of the diluent shall be equivalent to 7,0 0,2 when measured at (20 1) C. 5.2.2.5 Neutralizer The neutralizer shall be validated for the product being tested in accordance with 5.5.1.2, 5.5.1.3 and 5.5.2. It shall be sterile. NOTE Information on