Bsi bs en 13727 2012 + a2 2015

BS EN 13727:2012+A1:2013 13727:2012+A2:2015 BSI Standards Publication Chemical disinfectants and antiseptics — Quantitative suspension test for the evaluation of bactericidal activity in the medical area — Test method and requirements (phase 2, step 1) BS EN 13727:2012+A2:2015 BRITISH STANDARD National foreword This British Standard is the UK implementation of EN 13727:2012+A2:2015 It supersedes BS EN 13727:2012+A1:2013 which is withdrawn The start and finish of text introduced or altered by amendment is indicated in the text by tags Tags indicating changes to CEN text carry the number of the CEN amendment For example, text altered by CEN amendment A1 is indicated by  The 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 contract Users are responsible for its correct application © The British Standards Institution 2015 Published by BSI Standards Limited 2015 ISBN 978 580 89232 ICS 11.080.20 Compliance 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 31 October 2012 Amendments/corrigenda issued since publication Date Text affected 28 February 2014 Implementation of CEN amendment A1:2013 30 November 2015 Implementation of CEN amendment A2:2015 EN 13727:2012+A2 EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM October 2015 ICS 11.080.20 Supersedes EN 13727:2012+A1:2013 English Version Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of bactericidal activity in the medical area - Test method and requirements (phase 2, step 1) Antiseptiques et désinfectants chimiques - Essai quantitatif de suspension pour l'évaluation de l'activité bactéricide en médecine - Méthode d'essai et prescriptions (Phase 2, Étape 1) Chemische Desinfektionsmittel und Antiseptika Quantitativer Suspensionsversuch zur Bestimmung der bakteriziden Wirkung im humanmedizinischen Bereich - Prüfverfahren und Anforderungen (Phase 2, Stufe 1) This European Standard was approved by CEN on 14 October 2013 and includes Amendment approved by CEN on August 2015 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 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 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, Former 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 COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels © 2015 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members Ref No EN 13727:2012+A2:2015 E BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Contents Page European foreword Introduction Scope Normative references Terms and definitions Requirements 5.1 5.2 5.2.1 5.2.2 5.3 5.3.1 5.3.2 5.4 5.4.1 5.4.2 5.5 5.5.1 5.5.2 5.5.3 5.5.4 5.6 5.6.1 5.6.2 5.7 5.7.1 5.7.2 5.7.3 5.8 5.8.1 5.8.2 5.8.3 5.8.4 5.9 5.9.1 5.9.2 5.9.3 5.9.4 5.9.5 5.10 Test method Principle Materials and reagents Test organisms Culture media and reagents Apparatus and glassware 11 General 11 Usual microbiological laboratory equipment 12 Preparation of test organism suspensions and product test solutions 13 Test organism suspensions (test and validation suspension) 13 Product test solutions 15 Procedure for assessing the bactericidal activity of the product 15 General 15 Dilution-neutralization method 17 Membrane filtration method 19 Modified method for ready-to-use products 21 Experimental data and calculation 23 Explanation of terms and abbreviations 23 Calculation 23 Verification of methodology 28 General 28 Control of weighted mean counts 28 Basic limits 29 Expression of results and precision 29 Reduction 29 Control of active and non-active product test solution (5.4.2) 29 Limiting test organism and bactericidal concentration 30 Precision, repetitions 30 Interpretation of results - conclusion 30 General 30 Bactericidal activity for handrub and handwash products 30 Bactericidal activity for instrument disinfection products 30 Bactericidal activity for surface disinfection products 31 Qualification for certain fields of application 31 Test report 31 Annex A (informative) Referenced strains in national collections 33 Annex B (informative) Neutralizers and rinsing liquids 34 BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Annex C (informative) Graphical representation of test procedures 36 Annex D (informative) Example of a typical test report 44 Annex E (informative) Precision of the test result 48 Annex ZA (informative) Relationship between this European Standard and the Essential Requirements of EU Directive 93/42/EEC 51 Bibliography 52 BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) European foreword This document (EN 13727:2012+A2:2015) 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 endorsement, at the latest by April 2016, and conflicting national standards shall be withdrawn at the latest by April 2016 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 includes Amendment approved by CEN on 2013-10-14 and Amendment approved by CEN on 2015-08-03 This document supersedes #EN 13727:2012+A1:2013$ The start and finish of text introduced or altered by amendment is indicated in the text by tags !" and #$ This document has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association, and supports essential requirements of EU Directive(s) For relationship with EU Directive(s), see informative Annex ZA, which is an integral part of this document #deleted text$ !Data obtained using the former version of EN 13727 may still be used, if a neutralization time of 10 s for all products with contact times of 10 or shorter has been demonstrated to be sufficient Data obtained by using the prEN 12054 should not be used as this project was abandoned in 2001." 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, 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, Sweden, Switzerland, Turkey and the United Kingdom BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Introduction This European Standard specifies a suspension test for establishing whether a chemical disinfectant or an antiseptic has a bactericidal activity in the 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 Each utilization concentration of the chemical disinfectant or antiseptic found by this test corresponds to the chosen experimental conditions BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Scope This 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 be tested at a concentration of 80 % or less (97 % with a modified method for special cases) 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 medical area in the fields of hygienic handrub, hygienic handwash, surgical handrub, surgical handwash, instrument disinfection by immersion, and surface disinfection by wiping, spraying, flooding or other means This European Standard applies to areas and situations where disinfection or antisepsis is medically indicated Such indications occur in patient care, for example: — in hospitals, in community medical facilities and in dental institutions; — in clinics of schools, of kindergartens and of nursing homes; and may occur in the workplace and in the home It may also include services such as laundries and kitchens supplying products directly for the patients NOTE The method described is intended to determine the activity of commercial formulations or active substances under the conditions in which they are used NOTE This method corresponds to a phase step test NOTE This method cannot be used to evaluate the activity of products against Legionella in watersystems against mycobacteria and against bacterial spores EN 14885 specifies in detail the relationship of the various tests to one another and to “use recommendations” 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 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 (including bacteriophages) activity EN 14885, Chemical disinfectants and antiseptics - Application of European Standards for chemical disinfectants and antiseptics Terms and definitions For the purposes of this document, the terms and definitions given in EN 14885 apply Requirements The product shall demonstrate at least a decimal log (lg) reduction (for hygienic hand wash at least a lg reduction), when tested in accordance with Table and Clause BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Table — Minimum and additional test conditions Test Conditions Hygienic handrub and handwash Minimum P aeruginosa, spectrum of S aureus, test organisms E hirae, E coli K12 additional Test temperature Contact time Surgical handrub and handwash P aeruginosa, P aeruginosa, S aureus, S aureus, S aureus, E hirae, E hirae, E hirae E coli K12 when temperature is 40 °C or higher: only E faecium Any relevant test organism according to the manufacturer’s recommendation, but between 20 °C and 20 °C 20 °C and 20 °C Interfering substance additional 20 °C and 70 °C according to the manufacturer’s recommendation 30 s and 60 s dirty conditions Surface disinfection P aeruginosa, but between clean conditions Instrument disinfection and 0,3 g/l bovine 0,3 g/l bovine albumin solution albumin solution (hygienic handrub) b (surgical handrub) b 3,0 g/l bovine 3,0 g/l bovine albumin solution albumin solution plus 3,0 ml/l plus 3,0 ml/l erythrocytes erythrocytes (hygienic handwash) (surgical handwash) c clean or dirty; any relevant substance c clean or dirty; any relevant substance 4°C and 30 °C but no longer than 60 min or 60 a 0,3 g/l bovine albumin solution 0,3 g/l bovine albumin solution 3,0 g/l bovine albumin solution plus 3,0 ml/l erythrocytes 3,0 g/l bovine albumin solution plus 3,0 ml/l erythrocytes any relevant substance any relevant substance and/or and/or NOTE For the additional conditions, the concentration defined as a result can be lower than the one obtained under the minimum test conditions The contact times for surface disinfectants stated in this table are chosen on the basis of the practical conditions of the product The recommended contact time for the use of the product is within the responsibility of the manufacturer Products intended to disinfect surfaces that are likely to come into contact with the patient and / or the medical staff and surfaces, which are frequently touched by different people, leading to the transmission of microorganisms to the patient, shall be tested with a contact time of maximum The same applies where the contact time of the product shall be limited for practical reasons Products for other surfaces than stated above may be tested with a contact time of maximum 60 a b c hygienic and surgical handrub shall be tested as a minimum under clean conditions hygienic and surgical handwash shall be tested as a minimum under dirty conditions BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) 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 one of the temperatures and the contact times specified in Clause and 5.5.1.1 At the end of this contact time, an aliquot is taken; the bactericidal and/or the bacteriostatic action 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 is calculated NOTE Handwash products are always prediluted with hard water (5.2.2.7) The resulting solution is regarded as a ready-to-use product (5.4.2) 5.1.2 The test is performed using Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus hirae and for certain product types Escherichia coli K12 as test-organisms (Clause 4, Table 1) 5.1.3 Other contact times and temperatures within the limits specified in Clause 4, Table may be used Additional interfering substances and test organisms may be used 5.2 Materials and reagents 5.2.1 Test organisms The bactericidal activity shall be evaluated using the following strains as test organisms selected according to Clause (Table 1) 1) : a) Escherichia coli K12, NCTC 10538 b) Pseudomonas aeruginosa, ATCC 15442 c) Staphylococcus aureus, ATCC 6538 d) Enterococcus hirae, ATCC 10541 e) Enterococcus faecium, ATCC 6057 NOTE See Annex A for strain reference in some other culture collections The required incubation temperature for these test organisms is 36 °C ± °C or 37 °C ± °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 under optimum growth conditions (temperature, time, atmosphere, media) noted in the test report If the additional test organisms selected 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 characteristics shall be stated In addition, they shall be held by the testing laboratory or national culture collection under a reference for five years 1) The NCTC and ATCC numbers are the collection numbers of strains supplied by these 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 40 Figure C.5 — Dilution-neutralization method (modified method for ready-to-use products) – Test procedure (Na) BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Figure C.6 — Dilution-neutralization method (modified method for ready-to-use products) – Validation 41 BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) 42 Figure C.7 — Membrane filtration method (modified method for ready-to-use products) – Test procedure (Na) BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Figure C.8 — Membrane filtration method (modified method for ready-to-use products) – Validation 43 BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Annex D (informative) Example of a typical test report NOTE NOTE All names and examples in Annex D are fictitious apart from those used in this European Standard Only the test results of one replicate for Pseudomonas aeruginosa is given as an example NOTE The results of a hygienic handrub product tested according to the modified method 5.5.4 is given without a complete test report as an additional example HHQ Laboratories Antiseptville/Euroland Tel ++011.57 83 62-0 Fax ++011-57 83 62-19 e-mail: h.h.Q.lab@net.com TEST REPORT EN 13727, BACTERICIDAL ACTIVITY (minimum and additional conditions) Client: Cult Formulations Inc., Mannheim/Euroland Disinfectant-sample Name of the product: JN (instruments and surfaces) Batch number: 10-10-76 Manufacturer or – if not known – supplier: MDI Formulations Inc (manufacturer) Storage conditions (temp and other): Room temperature, darkness Appearance of the product: Liquid, clear, yellowish Active substance(s) and their concentration(s): Not indicated Product diluent recommended by the manufacturer for use: Potable water Period of testing Date of delivery of the product: 2010-05-01 Experimental conditions Dates of tests: see "Test results" (attached) Product diluent: hard water; concentrations of the product tested: see "Test results" (attached) test-organisms: Pseudomonas aeruginosa ATCC 15442, Staphylococcus aureus ATCC 6538, Enterococcus hirae ATCC 10541, Enterococcus faecium ATCC 6057 test temperature: 25 °C; contact time: 45 min; (+ 50 °C; 60 min) interfering substance: 0,3 g/l bovine albumin = clean conditions; (+ 3,0 g/l bovine albumin plus 3,0 ml/l erythrocytes = dirty conditions) 44 BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) incubation temperature: 36 °C test results: see attached sheets Special remarks regarding the results: All controls and validation were within the basic limits At least one concentration of the product demonstrated a lg reduction of less than lg No precipitate during the test procedure (test mixtures were homogeneous) Conclusion: For the product JN (batch 10-10-76), the bactericidal concentration for instrument and surface disinfection determined according to the EN 13727 standard under clean conditions at 25 °C within 45 is: 1,0 % (v/v) (the mean reduction of six repetitions with the limiting test organism Pseudomonas aeruginosa was 1,2 x 105 Staphylococcus aureus and Enterococcus hirae were tested once and showed a lg reduction or more at a lower concentration than Pseudomonas aeruginosa) For the product JN (batch 10-10-76), the bactericidal concentration for specific instrument disinfection determined according to the EN 13727 standard at 40 °C, with 60 contact time under dirty conditions, using Enterococcus faecium ATCC 6057 as test organism is: Antiseptville, 2010-11-11 0,50 % (v/v) Alexander May, MD, PhD, Scientific Director 45 BS EN 13727:2012+A2:2015 EN 13727:2012+A2:2015 (E) Test results (bactericidal quantitative suspension test) EN 13727….(Phase 2, step 1)Product-name: JN (Instruments and surfaces) Batch No.: 10-10-76 Remarks: Dilution neutralization X Pour plate Spread plate x Number of plates .2 / ml method Neutralizer: Lecithin 3,0 g/l in diluent Membrane filtration method Rinsing liquid: Test temperature: 25 °C interfering substance:….bovine albumin: 0,3 g/l ……………………………… Test organism: Pseudomonas aeruginosa ATCC 15442 Incubation temperature: 36°C Internal lab No : QS Date of test:.2010-06-06 Responsible person: Fang Signature: Fang 68/00 Diluent used for product test solutions: water Appearance of the product test solutions: clear Validation and controls Validation suspension (Nvo) VC1 86 x= (40 + 46) 89 VC2 92 (47 + 45) 30 ≤ x of Nvo ≤ 160 ? yes no Experimental conditions control (A) VC1 79 x= (43 + 36) 81,5 VC2 84 (39 + 45) x of A is ≥ 0,5x x of Nvo ? yes no Validation suspension (NVB) Test suspension and Test VC1 99 (49 + 50) Test-suspension (N and No): Real conc of the product % 0,50 0,75 1,00 Neutralizer or filtration control (B) x= VC1 86 (42 + 44) 88,5 VC2 91 (43 + 48) x of B is ≥ 0,5x x of Nvo (or NVB/1000) yes no VC2 71 (39 + 32) N x= VC1 168 20 10-6 10-7 VC1 VC2 100 >660 >660 10-1 14 17 Dilution step 10-1 100 100 10-1 77 122 85 154 0 85 30 ≤ VC2 213 25 x Method validation (C) Product conc.: 10 ml/l VC1 75 x= (35 + 40) VC2 81 87 (41 + 46) x of C is ≥ 0,5x x of Nvo? yes no of NVB/1000 ≤ 160 ? Na (= x or lg Na 8100 3,91 1395,5 660 >660 10-1 0 100 77 N 10-7: 80+88; 105+108 10-8: 9+11; 15+10 A 30s: 43+36; 39+45 1min: 45+30; 43+39 Explanations: VC = count per ml (one plate or more) x = average of VC1 and VC2 (1 + duplicate) x of NVB/1000 ≤ 160 ? x wm = 193,64 wm x 10) 8100 85