EN 13623-2010 en Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of bactericidal activity against Legionella of chemical disinfectants for .pdf

1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN 13623:2010Chemical disinfectants andantiseptics Quantitativesuspension test for theevaluation of bactericidalactivity against Legionellaof chemical disinfectantsfor aqueous

2、 systems Testmethod and requirements(phase 2, step 1)BS EN 13623:2010 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 13623:2010.The UK participation in its preparation was entrusted to TechnicalCommittee CH/216, Chemical disinfectants and antiseptics.A list of

3、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. BSI 2010ISBN 978 0 580 53071 5ICS 11.080.20; 71.100.35Compliance with a Bri

4、tish Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 31 October 2010.Amendments issued since publicationDate Text affectedBS EN 13623:2010EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM E

5、N 13623 September 2010 ICS 71.100.35 English Version Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of bactericidal activity against Legionella of chemical disinfectants for aqueous systems - Test method and requirements (phase 2, step 1) Antiseptiques et ds

6、infectants chimiques - Essai quantitatif de suspension pour lvaluation de lactivit bactricide contre des lgionelles des dsinfectants chimiques pour les systmes aqueux - Mthode dessai et prescriptions (phase 2, tape 1) Chemische Desinfektionsmittel und Antiseptika - Quantitativer Suspensionsversuch z

7、ur Bestimmung der bakteriziden Wirkung gegen Legionella von chemischen Desinfektionsmitteln fr wasserfhrende Systeme - Prfverfahren und Anforderungen (Phase 2, Stufe 1) This European Standard was approved by CEN on 15 April 2010. CEN members are bound to comply with the CEN/CENELEC Internal Regulati

8、ons 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 the CEN Management Centre or to any CEN member. This Eur

9、opean 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 Management Centre has the same status as the official versions. CEN members are the n

10、ational standards bodies of Austria, Belgium, Bulgaria, Croatia, 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, Swit

11、zerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2010 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No.

12、 EN 13623:2010: EBS EN 13623:2010EN 13623:2010 (E) 2 Contents Page Foreword 3Introduction .41 Scope 52 Normative references 53 Terms and definitions .54 Requirements .55 Test methods 65.1 Principle 65.2 Materials and reagents 65.2.1 Test organism 65.2.2 Culture media and reagents .65.3 Apparatus and

13、 glassware 105.3.1 General . 105.3.2 Usual microbiological laboratory equipment 105.4 Preparation of test organism suspensions and test solutions 115.4.1 Test organism suspension (test and validation suspension) 115.4.2 Product test solution 135.5 Procedure for assessing the bactericidal activity of

14、 the product . 135.5.1 General . 135.5.2 Dilution-neutralization method 155.5.3 Membrane filtration method 175.6 Experimental data and calculations 185.6.1 Explanation of terms and abbreviations 185.6.2 Calculation . 195.7 Verification of methodology 225.7.1 General . 225.7.2 Control of weighted mea

15、n counts . 235.7.3 Basic limits 235.8 Expression of results and precision . 235.8.1 Reduction 235.8.2 Control of active and non-active product test solution (5.4.2) 235.8.3 Bactericidal concentration . 235.8.4 Precision, repetition . 245.9 Interpretation of results conclusion 245.9.1 General . 245.9

16、.2 Bactericidal activity for general purposes . 245.9.3 Bactericidal activity for specific purposes 245.10 Test report . 24Annex A (informative) Determination of the bactericidal activity against Legionella pneumophila . 26Annex B (informative) Neutralizer . 28Annex C (informative) Graphical represe

17、ntation of test procedures 30C.1 Dilution-neutralization method 30C.2 Membrane filtration method 32Bibliography . 34BS EN 13623:2010EN 13623:2010 (E) 3 Foreword This document (EN 13623:2010) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the secretariat

18、 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 March 2011, and conflicting national standards shall be withdrawn at the latest by March 2011. Attention is drawn to the

19、 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. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countr

20、ies are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Sloven

21、ia, Spain, Sweden, Switzerland and the United Kingdom. BS EN 13623:2010EN 13623:2010 (E) 4 Introduction This European Standard specifies a suspension test for establishing whether a chemical disinfectant has a bactericidal activity against Legionella pneumophila in the fields described in the scope.

22、 This standard is specifically prepared for water treatment products, but it may also be possible to use it for other products. Proliferation of Legionella only occurs in waters under certain conditions, and predominantly poses a risk when aerosolised. Many systems containing water do not require tr

23、eatment. A decision to add chemical disinfectants to any water should be based on a risk assessment. If the product complies with the requirements of this standard, it can be considered bactericidal against Legionella pneumophila, but it should not necessarily be inferred that the product is accepta

24、ble for a specific site of application without consideration of other relevant factors such as the pH, water, chemistry, temperature and degree of biological fouling at that site of application. It does not take into account the protective effect conveyed by biofilm on the organisms. The conditions

25、are intended to cover general purposes and to allow reference between laboratories and product types. Each concentration of the chemical disinfectant found by this test corresponds to defined experimental conditions. However, for some applications the recommendations of use of a product may differ a

26、nd therefore additional test conditions need to be used. BS EN 13623:2010EN 13623:2010 (E) 5 1 Scope This European Standard specifies a test method and the minimum requirements for bactericidal activity of chemical disinfectant products intended to be used for treatment in aqueous systems against Le

27、gionella pneumophila that form a homogeneous, physically stable preparation when diluted with buffered ferrous hard water or hard water. Whenever Legionella pneumophila poses a risk to human health, this method is suitable for water used in cooling towers and water for general purposes, like spas, p

28、ools, showers and other uses. The method is not suitable for electro-chemical disinfection. The European Standard applies to products used to treat water in order to kill Legionella pneumophila. NOTE 1 The method described is intended to determine the activity of commercial formulations or active su

29、bstances under the conditions in which they are used. NOTE 2 This method corresponds to a phase 2 step 1 test . NOTE 3 This method does not take into account the fact that Legionella pneumophila is often found in cells of amoebae and/or biofilms and that thereby a products activity against the bacte

30、ria may be reduced. EN 14885 specifies in detail the relationship of the various tests to one another and to “use recommendation“. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. F

31、or undated references, the latest edition of the referenced document (including any amendments) applies. EN 14885, Chemical disinfectants and antiseptics Application of European Standards for chemical disinfectants and antiseptics 3 Terms and definitions For the purposes of this document, the terms

32、and definitions given in EN 14885 and the following apply. 3.1 cooling water water used to remove heat from a process or environment 3.2 water for general purposes water used in premises other than water used as cooling water 4 Requirements The product shall demonstrate al least a four decimal log (

33、lg) reduction, when diluted with buffered ferrous hard water (5.2.2.10) or hard water (5.2.2.7), and tested in accordance with Clause 5 under the obligatory test conditions (one selected test organism, at either 20 C or 30 C) within 60 min for rapid acting products or 15 h for slower acting products

34、. The bactericidal activity shall be evaluated using the following test organism: Legionella pneumophila. Where indicated, additional specific bactericidal activity shall be determined applying other contact times and test organisms (in accordance with 5.2.1 and 5.5.1.1) in order to take into accoun

35、t intended specific use conditions. BS EN 13623:2010EN 13623:2010 (E) 6 NOTE For these additional conditions, the concentration defined as a result can be lower than the one obtained under the obligatory test conditions. 5 Test methods 5.1 Principle 5.1.1 A sample of the product diluted with hard wa

36、ter (5.2.2.7 or 5.2.2.10) is added to a test suspension of bacteria in a solution of an interfering substance. The mixture is maintained at either (20 1) C or (30 1) C for 60 min 10 s or (15 1) h (obligatory test conditions). At the end of the chosen contact time, an aliquot is taken, and the bacter

37、icidal and/or the bacteriostatic 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 deter

38、mined and the reduction is calculated. 5.1.2 The test is performed using Legionella pneumophila as test organism (obligatory test conditions). 5.1.3 Additional and optional contact times are specified. Additional test organisms may be used. 5.2 Materials and reagents 5.2.1 Test organism The bacteric

39、idal activity shall be evaluated using the following strain as test organism1): Legionella pneumophila: serogroup 1, Philadelphia (NCTC 11192; ATCC 33152). If required for specific applications, additional test organisms may be used, e.g. Legionella pneumophila serogroup 1 Benidorm (NCTC 12006, ATCC

40、 43108). The required incubation temperature for this test organism 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 incubated u

41、nder 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 are not cla

42、ssified 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 Unless specifically stated, all weights of

43、chemical substances 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 pur

44、poses. They shall be free from substances that are toxic or inhibitory to the test organisms. 1) The NCTC and ATCC numbers are the collection numbers of strains supplied by the National Type Culture Collection (NCTC) and American Type Culture Collection (ATCC). This information is given for the conv

45、enience of users of this standard and does not constitute an endorsement by CEN of the product named. BS EN 13623:2010EN 13623:2010 (E) 7 NOTE 1 To improve reproducibility, it is recommended that commercially available dehydrated material is used for the preparation of culture media. The manufacture

46、rs instructions relating to the preparation of these products should be rigorously followed. NOTE 2 For each culture medium and reagent, a limitation for use should be fixed. 5.2.2.2 Water The water shall be freshly glass-distilled water and not demineralized water. If distilled water of adequate qu

47、ality is not available, water for injections (bibliographic reference 1) can be used. Sterilize in the autoclave (5.3.2.1, a). Sterilization is not necessary if the water is used e.g. for preparation of culture media and subsequently sterilized. NOTE See 5.2.2.10 for the procedure to prepare buffere

48、d ferrous hard water. 5.2.2.3 Buffered Charcoal Yeast Extract (BCYE) Agar BCYE agar, consisting of yeast extract (bacteriological grade) 10,0 g; agar 12,0 g; activated charcoal 2,0 g; alpha-ketoglutarate, monopotassium salt 1,0 g; ACES buffer (N-2-acetamido-2-aminoethanesulfonic acid) 10,0 g; potass

49、ium hydroxide (KOH) (pellets) 2,8 g; L-cysteine hydrochloride monohydrate 0,4 g; iron(III) pyrophosphate Fe4(P207)3 0,25 g; distilled water to 1 000,0 ml. Preparation a) Cysteine and iron solutions Prepare fresh solutions of L-cysteine hydrochloride and iron(III) pyrophosphate by adding 0,4 g and 0,25 g respectively to 10-ml-volumes of water (5.2.2.2). Sterilize each solution by membrane filtration (