BS EN 13697:2015 BSI Standards Publication Chemical disinfectants and antiseptics — Quantitative non-porous surface test for the evaluation of bactericidal and/or fungicidal activity of chemical disinfectants used in food, industrial, domestic and institutional areas — Test method and requirements without mechanical action (phase 2, step 2) BS EN 13697:2015 BRITISH STANDARD National foreword This British Standard is the UK implementation of EN 13697:2015 It supersedes BS EN 13697:2001 which is withdrawn The UK participation in its preparation was entrusted to Technical Committee CH/216, Chemical disinfectants and antiseptics A list of organizations represented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 78888 ICS 11.080.20; 71.100.35 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 30 April 2015 Amendments/corrigenda issued since publication Date Text affected BS EN 13697:2015 EN 13697 EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM April 2015 ICS 11.080.20; 71.100.35 Supersedes EN 13697:2001 English Version Chemical disinfectants and antiseptics - Quantitative non-porous surface test for the evaluation of bactericidal and/or fungicidal activity of chemical disinfectants used in food, industrial, domestic and institutional areas - Test method and requirements without mechanical action (phase 2, step 2) Antiseptiques et désinfectants chimiques - Essai quantitatif de surface non-poreuse pour l'évaluation de l'activité bactéricide et/ou fongicide des désinfectants chimiques utilisés dans le domaine de l'agro-alimentaire, dans l'industrie, dans les domaines domestiques et en collectivité - Méthode d'essai sans action mécanique et prescriptions (phase 2/étape 2) Chemische Desinfektionsmittel und Antiseptika Quantitativer Oberflächen-Versuch nicht poröser Oberflächen zur Bestimmung der bakteriziden und/oder fungiziden Wirkung chemischer Desinfektionsmittel in den Bereichen Lebensmittel, Industrie, Haushalt und öffentliche Einrichtungen - Prüfverfahren und Anforderungen ohne mechanische Behandlung (Phase 2, Stufe 2) This European Standard was approved by CEN on 20 January 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 13697:2015 E BS EN 13697:2015 EN 13697:2015 (E) Contents Page Foreword Introduction Scope Normative references Terms and definitions Requirements 5.1 5.2 5.2.1 5.2.2 5.2.3 5.3 5.4 5.4.1 5.4.2 5.5 5.5.1 5.5.2 5.5.3 5.6 5.6.1 5.6.2 5.6.3 5.6.4 5.7 Test methods Principle Materials and reagents Test organisms Culture media and reagents Test surface 12 Apparatus and glassware 13 Preparation of test organism suspensions and product test solutions 14 Test organism suspensions 14 Product test solutions 17 Procedure 17 Choice of experimental conditions 17 Test procedure 18 Counting of the test mixtures 20 Calculation and expression of results 21 Elaboration of data: counting of weighed average values 21 Verification of methodology 22 Expression of results 23 Conclusion 23 Test report 24 Annex A (informative) Corresponding referenced strains 26 Annex B (informative) Neutralizers 28 Annex C (informative) Expression of results with the dilution-neutralization method 30 Annex D (informative) Bactericidal activity in surfaces in general use conditions (for clean conditions) 32 Annex E (informative) Precision of the test result 33 Bibliography 34 BS EN 13697:2015 EN 13697:2015 (E) Foreword This document (EN 13697:2015) 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 October 2015 and conflicting national standards shall be withdrawn at the latest by October 2015 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 supersedes EN 13697:2001 The changes between this edition and EN 13697:2001 are the following: — interfering substance has been changed from 0,03 % bovine albumin to 0,85 % skimmed milk (see Clause 4, Table 1) for Pseudomonas aeruginosa under clean conditions only; — A brasiliensis (ex A niger) spore preparation has been updated in order to harmonize this step with the QST fungicidal test method amendment issued in 2012 (see 5.4.1.3 b)); — Calculations of the weighed means and of the results have been modified in order to be harmonized with new CEN TC 216 standards (see 5.4.1.5, 5.5.2, 5.5.3 and 5.6); — Other paragraphs have been harmonized to new CEN TC 216 standards (e.g preparation of hard water, 5.2.2.7) Results obtained from the previous standard for Aspergillus niger need to be repeated to take into account the new spore morphology requirement and the change in interfering substance 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 13697:2015 EN 13697:2015 (E) Introduction This European Standard describes a surface test method for establishing whether a product proposed as a disinfectant in the fields described in Clause has or does not have bactericidal and/or fungicidal or yeasticidal activity on non-porous surfaces This European Standard has been revised in order to modify the interfering substance under “clean conditions” adopted for P aeruginosa; in order to modify the calculation of N, NC, NT, Nc, Na and consequently the final results and to harmonize the standard with the other recent CEN TC 216 standards The laboratory test closely simulates practical conditions of application Chosen conditions (contact time, temperature, organisms on surfaces ) reflect parameters which are found in practical situations including conditions which may influence the action of disinfectants Each use concentration found from this test corresponds to defined experimental conditions The conditions are intended to cover general purposes and to allow reference between laboratories and product types However, for some applications the recommendations of use of a product can differ and therefore additional test conditions need to be used BS EN 13697:2015 EN 13697:2015 (E) Scope This European Standard specifies a test method (phase 2/step 2) and the minimum requirements for bactericidal and/or fungicidal or yeasticidal activity of chemical disinfectants that form a homogeneous physically stable preparation in hard water or – in the case of ready-to-use products – with water in food, industrial, domestic and institutional areas, excluding areas and situations where disinfection is medically indicated and excluding products used on living tissues The scope of this European Standard applies at least to the following: a) Processing, distribution and retailing of: 1) Food of animal origin: i) milk and milk products; ii) meat and meat products; iii) fish, seafood and products; iv) eggs and egg products; v) animal feeds; vi) etc 2) Food of vegetable origin: i) beverages; ii) fruits, vegetables and derivatives (including sugar distillery); iii) flour, milling and backing; iv) animal feeds; v) b) etc Institutional and domestic areas: 1) catering establishments; 2) public areas; 3) public transports; 4) schools; 5) nurseries; 6) shops; 7) sports rooms; 8) waste container (bins); BS EN 13697:2015 EN 13697:2015 (E) 9) hotels; 10) dwellings; 11) clinically non sensitive areas of hospitals; 12) offices; 13) etc c) Other industrial areas: 1) packaging material; 2) biotechnology (yeast, proteins, enzymes ); 3) pharmaceutical; 4) cosmetics and toiletries; 5) textiles; 6) space industry, computer industry; 7) etc Using this European Standard, it is possible to determine the bactericidal or fungicidal or yeasticidal activity of the undiluted product As three concentrations are tested, in the active to non active range, dilution of the product is required and, therefore, the product forms a homogeneous stable preparation in hard water EN 14885 specifies in detail the relationship of the various tests to one another and to use recommendations NOTE The method described is intended to determine the activity of commercial formulations or active substances on bacteria and/or fungi in the conditions in which they are used NOTE 2 This method cannot be used to evaluate the activity of products against mycobacteria 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 ISO 4793, Laboratory sintered (fritted) filters - Porosity grading, classification and designation Terms and definitions For the purposes of this document, the terms and definitions given in EN 14885 apply BS EN 13697:2015 EN 13697:2015 (E) Requirements The product shall demonstrate at least a decimal log (lg) reduction for bacteria and at least a decimal log (lg) reduction for fungi, when tested in accordance with Table and 5.5.1 Table — Obligatory and additional conditions Test Conditions Bactericidal activity on non-porous surfaces Yeasticidal activity on non-porous surfaces without mechanical action without mechanical action Enterococcus hirae Fungicidal activity on non-porous surfaces without mechanical action Candida albicans Candida albicans Aspergillus brasiliensis (ex A niger) any relevant test organism Enterobacter cloacae Saccharomyces cerevisiae (for breweries) Saccharomyces cerevisiae var diastaticus (for breweries) Test temperature obligatory Between 18 °C ± °C and 25 °C ± °C Between 18 °C ± °C and 25 °C ± °C Between 18 °C ± °C and 25 °C ± °C additional °C ± °C; 10 °C ± °C; 40 °C ± °C °C ± °C; 10 °C ± °C; 40 °C ± °C °C ± °C; 10 °C ± °C; 40 °C ± °C Contact time obligatory ± 10 s 15 ± 10 s 15 ± 10 s additional ± s; 15 ± 10 s; 30 ± 10 s; 60 ± 10 s ± s; ± 10 s; 30 ± 10 s; 60 ± 10 s ± s; ± 10 s; 30 ± 10 s; 60 ± 10 s 0,3 g/l Bovine Albumin for Staphylococcus aureus, Enterococcus hirae and Escherichia coli ; 0,3 g/l Bovine Albumin for C albicans 0,3 g/l Bovine Albumin for C albicans and A brasiliensis dirty conditions 3,0 g/l Bovine Albumin for Staphylococcus aureus, Enterococcus hirae, Pseudomonas aeruginosa and Escherichia coli 3,0 g/l Bovine Albumin for C albicans 3,0 g/l Bovine Albumin for C albicans and A brasiliensis additional any relevant substance any relevant substance any relevant substance Log reduction from a water control (decimal log) ≥ 4Log ≥ 3Log ≥ 3Log Test organism (see 5.2.1) obligatory example Interfering substance obligatory clean conditions Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus Salmonella typhimurium Lactobacillus brevis 8,5 g/l skim milk for Pseudomonas aeruginosa The obligatory contact times for surface disinfectants stated in Table were chosen to enable comparison of standard conditions The referenced test conditions are by no means intended as requirements for the use of a product, nor as requirements for the evaluation and acceptance of products by regulatory authorities The recommended contact time for the use of the product is within the responsibility of the manufacturer Where appropriate (specific purposes), additional specific bactericidal/yeasticidal/fungicidal activity shall be determined under other conditions of time, temperature, additional strains and interfering substances in order to take into account intended specific use conditions NOTE For the additional conditions, the concentration defined as a result can be lower than the one obtained under the obligatory test conditions BS EN 13697:2015 EN 13697:2015 (E) Test methods 5.1 Principle A test suspension of bacteria or fungi in a solution of interfering substances is inoculated onto a test stainless steel surface and dried A prepared sample of the product under test is applied in a manner which covers the dried film The surface is maintained at a specified temperature for a defined period of time The surface is transferred to a previously validated neutralization medium so that the action of the disinfectant is immediately neutralized The number of surviving organisms which can be recovered from the surface is determined quantitatively The number of bacteria or fungi on a surface treated with hard water in place of the disinfectant is also determined and the reduction in viable counts attributed to the product is calculated by difference 5.2 5.2.1 Materials and reagents Test organisms The bactericidal activity shall be evaluated using the following four strains: 1) — Pseudomonas aeruginosa ATCC 15 442 ; — Staphylococcus aureus ATCC 538; — Enterococcus hirae ATCC 10 541; — Escherichia coli ATCC 10 536 The fungicidal or yeasticidal activity shall be evaluated using the following two strains: — Candida albicans ATCC 10 231; — Aspergillus brasiliensis (ex A niger) ATCC 16 404 If required for specific applications, additional strains may be chosen from, for example: — Salmonella typhimurium ATCC 13 311; — Lactobacillus brevis DSM 235; — Enterobacter cloacae DSM 234; — Saccharomyces cerevisiae (for breweries) or ATCC 763 or DSM 333; — Saccharomyces cerevisiae var diastaticus (for breweries) DSM 70 487 NOTE See Annex A for corresponding strain numbers in some other culture collections If additional strains are used, they shall be incubated under optimum growth conditions (temperature, time, atmosphere) and noted in the test report If the additional strains selected not correspond to the specified strains, their suitability for supplying inocula of sufficient concentration shall be verified If the additional strains tested are not classified at a 1) ATCC 15 442, ATCC 538, ATCC 10 541, ATCC 10 536, ATCC 10 231, ATCC 16 404 and ATCC 13311 are the collection numbers of strains supplied by the American Type Culture Collections This information is given for the convenience of users of this standard and does not constitute an endorsement by CEN of the product named Equivalent products can be used if they can be shown to lead to the same results BS EN 13697:2015 EN 13697:2015 (E) If observed counts from two consecutive dilutions fall within the 14-330 or 14-165 range, then a weighted average value is calculated as follows: (m+m’+n+n’)/2.2xVxd (4) where m, m’ are the two replicas at the lower dilution expressed as cfu; n, n’ are the two replicas at the higher dilution expressed as cfu; V is the volume of the inoculated into the plate expressed in ml; d is the lower dilution factor In case not all the replicas at each dilution fall within the 14-330 or 14-165 range, the weighted average will take this into account as follows: Case 1: only m and n fall within the 14-330 or 14-165 range, the formula will change as follows: (m+n)/1.1xVxd (5) Case 2: only m, m’ and n fall within the 14-330 or 14-165 range, the formula will change as follows: (m+m’+n)/2.1xVxd (6) Case 3: only m, n and n’ fall within the 14-330 or 14-165 range, the formula will change as follows: (m+n+n’)/1.2xVxd (7) Case 4: only m falls within the 14-330 or 14-165 range, the formula will change as follows: m/Vxd 5.6.2 (8) Verification of methodology For each test, check that: a) the mean counts form duplicate plate used for calculation of N, Nc, Nd, NC, NT are between 14 and 330 for bacteria and yeast strains and 14 and 165 for mould strains; b) 6,57 ≤ N ≤ 7,10 Logs for bacteria; c) 5,57 ≤ N ≤ 6,10 Logs for fungi; d) Nc ≥ 6,27 Logs for bacteria; e) Nc ≥ 5,27 Logs for fungi; f) NC > 0,5 Nc; g) NT > 0,5 Nc is not greater than ± 0,3; h) Nts is less than 100 cfu/ml for active concentrations If not, the recovery of microorganisms has not been sufficient For non active concentrations, Nts may be not countable; 22 BS EN 13697:2015 EN 13697:2015 (E) i) control of weighted mean counts: quotient is not lower than and not higher than 15 It applies only to N calculation 5.6.3 5.6.3.1 Expression of results Reduction The reduction (R) is expressed in logarithm For each test organism record the number of cfu/ml in the test procedure for microbicidal activity of the product (5.5.2.2) and the control procedure (5.5.2.3).” For each product concentration and each experimental condition, calculate and record the decimal log (lg) reduction separately using the equation: R = Nc – Nd 5.6.4 5.6.4.1 (9) Conclusion Activity on non-porous surfaces for general purposes Bactericidal activity on surfaces for general purposes is characterized by the concentration of the tested product for which criteria 5.6.1 and 5.6.2 are met and for which a log or more reduction in viability is demonstrated under the required test conditions: ± 10 s, 18 °C and 25 °C and clean or dirty conditions (see Clause 4), when the test organisms are Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus hirae and Escherichia coli Fungicidal activity on surfaces for general purposes is characterized by the concentration of the tested product for which criteria 5.6.1 and 5.6.2 are met and for which a log or more reduction in viability is demonstrated under the required test conditions: 15 ± 10 s, 20 °C ± °C and clean or dirty conditions (see Clause 4), when the test organisms are Candida albicans and Aspergillus brasiliensis (ex A niger) Yeasticidal activity on surfaces for general purposes is characterized by the concentration of the tested product for which criteria 5.6.1 and 5.6.2 are met and for which a log or more reduction in viability is demonstrated under the required test conditions: 15 ± 10 s, 20 °C ± °C and clean or dirty conditions (see Clause 4), when the test organisms are Candida albicans Bactericidal and fungicidal or yeasticidal activity on surfaces for general purposes is characterized by the concentration of the tested product for which criteria 5.6.1 and 5.6.2 are met and: — for which a log or more reduction in viability is demonstrated under the required test conditions: ± 10 s, 20 °C ± °C and clean or dirty conditions (see Clause 4), when the test organisms are Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus hirae and Escherichia coli; — for which a log or more reduction in viability is demonstrated under the required test conditions: 15 ± 10 s, 20 °C ± °C and clean or dirty conditions (see Clause 4), when the test organisms are Candida albicans and Aspergillus brasiliensis (ex A niger); — for which a log or more reduction in viability is demonstrated under the required test conditions: 15 ± 10 s, and 20 °C ± °C and clean or dirty conditions (see Clause 4), when the test organisms are Candida albicans 5.6.4.2 Activity on non-porous surfaces for specific purposes Bactericidal and/or fungicidal or yeasticidal activity on surfaces for specific purposes is characterized by the concentration of the tested product for which criteria 5.6.1 and 5.6.2 are met and for which: 23 BS EN 13697:2015 EN 13697:2015 (E) — a log or more reduction in viability is demonstrated, when the test organisms are Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus hirae and Escherichia coli, and if required additional test organisms; and/or for which: — a log or more reduction in viability is demonstrated, when the test organisms are Candida albicans and Aspergillus brasiliensis (ex A niger), and if required additional test organisms; — a log or more reduction in viability is demonstrated, when the test organisms are Candida albicans, and if required additional test organisms under additional conditions: t in minutes, θ in degrees °C, clean or dirty conditions and additional interfering substances (see 5.5.1) 5.7 Test report The test report shall refer to this European Standard The test report shall state, at least, the following information: a) identification of the laboratory; b) identification of the sample: c) 24 1) name of the product; 2) batch number; 3) manufacturer; 4) date of delivery: 5) storage conditions; 6) active substance(s) and its/their concentration(s) (optional); experimental conditions: 1) period of analysis; 2) product diluent used during the test; 3) product test concentrations; 4) appearance product dilutions; 5) contact time(s); 6) test temperature(s); 7) interfering substance; 8) reaction between the inoculum in the presence of interfering substances and product; 9) temperature of incubation; BS EN 13697:2015 EN 13697:2015 (E) 10) neutralizer of the bacterial or fungal strains used; 11) identification of the bacterial and/or fungal strains used; 12) identification of the test surface; d) operating procedure: 1) e) full details for the test for validation of the neutralization medium shall be given; test results: 1) validation tests; 2) evaluation of bactericidal and/or fungicidal and/or yeasticidal activity; f) conclusion; g) locality, date and identified signature(s) NOTE An example of a typical test report is given in Table C.1 25 BS EN 13697:2015 EN 13697:2015 (E) Annex A (informative) Corresponding referenced strains Pseudomonas aeruginosa : ATCC CIP Staphylococcus aureus: Escherichia coli: Salmonella typhimurium: 939 NCIB 10 421 ATCC 538 CIP 4.83 DSM 799 NCTC 10 788 NCIB 518 ATCC 10 536 CIP 54 127 10 418 NCIMB 879 ATCC 10 541 CIP 855 DSM 3320 NCIMB 192 ATCC 13 311 NCTC DSM CIP Enterobacter cloacae: DSM CIP Saccharomyces cerevisiae: 26 682 NCTC CIP Lactobacillus brevis: 103 467 DSM DSM Enterococcus hirae: 15 442 ATCC 858 74 235 103 474 234 104 674 763 BS EN 13697:2015 EN 13697:2015 (E) IP Candida albicans: Aspergillus brasiliensis (ex A niger): 143 283 DSM 333 CBS 900 ATCC 10 231 IP 872 DSM 1386 CBS 431 NCTC 179 ATCC 16 404 DSM 988 CBS 733.88 IP NCTC CMI 431.83 275 149 007 27 BS EN 13697:2015 EN 13697:2015 (E) Annex B (informative) Neutralizers Any of the following neutralizers can be used: 7) — lecithin g/l; polysorbate 80 30 g/l; sodium thiosulphate g/l; L-histidine g/l; saponine 30 g/l in diluent (see 5.2.2.4) or in phosphate buffer 0,25 mol/l at % (V/V); — phosphate buffer 0,25 mol/l: — KH2 PO4 34 g; — water (see 5.2.2.2) 500 ml; — adjusted to pH 7,2 ± 0,2 with mol/l NaOH; — water (see 5.2.2.2) up to 000 ml; — sterilized in an autoclave (see 5.3.1); — fresh egg yolk diluted to % (V/V) or 0,5 % (V/V); — 30 g/l polysorbate 80 ; g/l sodium lauryl sulphate ; lecithin 3g/l; — % (V/V) fresh egg yolk; 40 g/l polysorbate 80 ; — % (V/V) ethylene oxide condensate of fatty alcohol ; 20 g/l lecithin; % (V/V) polysorbate 80 ; — % (V/V) ethylene oxide condensate of fatty alcohol; g/l lecithin; — 30 g/l polysorbate 80 ; lecithin g/l; L-histidine g/l; — glycine as a function of concentration of product; — 30 g/l polysorbate 80 ; lecithin g/l; — phospholipid emulsion (commercial) at 50 mg/ml (diluted to 10); — sodium thioglycollate at 0,5 g/l or g/l; — L cysteine at 0,8 g/l or 1,5 g/l; — thiomalic acid at 0,075 % (V/V) (adjusted to pH with NaOH); — sodium thiosulphate at g/l; 7) 7) 7) 7) 7) 7) Analytical quality, non-hydrolysed in accordance with European Pharmacopoeia volume TWEEN 80 is an example of a suitable product available commercially This information is given for the convenience of users of this standard and does not constitute an endorsement by CEN of this product 28 BS EN 13697:2015 EN 13697:2015 (E) — catalase or peroxidase: One unit for of these enzymes catalyses the decomposition of µmol of hydrogen peroxide per minutes at 25 °C and at pH 7; — polysorbate 80 30 g/l ; saponin 30 g/l ; L-histidine g/l ; L-cysteine 1g/l 7) The above list is not exhaustive and other neutralizers may be tried 29 EN 13697:2015 (E) Annex C (informative) Expression of results with the dilution-neutralization method Table C.1 — Test results Test organisms Validation test : Bacterial or fungal test suspension : N (See 5.4.1.3) (See 5.5.2.1) NT NC (See Annex A) (See Annex A) Nc : > 330 ; > 33 10 −4 : 123 ;118 10 : 229 ;216 10 −7 : 20 ;17 10 aeruginosa 10 ATCC 15442 −3 −6 10 N : 6,74 −5 10 : ;13 NT : 7,08 : > 330 ; > 33 10 −4 : 153 ;123 10 N : 6,74 −5 10 : ;2 NT : 7,09 30 10 −5 : 14 ;9 10 −2 : 13 ;17 10 10 −0 : ;0 −1 : ;0 10 −1 : ;0 −2 : ;0 10 −2 : ;0 −5 : 11 ;4 Nc : 7,14 Nd : 4,18 Nd : < 0,10 Nd : < 0,10 Nts : > 100 Nts : > 100 Nts : 12 Nts : R : 2,96 R: > 6,04 R: > 7,04 10 NC : 7,14 10 10 : 160 ;138 : ;0 10 : 132 ;113 : 23 ;19 10 −1 −0 10 : 137 ;142 −4 −7 ATCC 10536 : > 330 ; > 33 −4 10 10 −0 : > 330 ; > 33 : > 330 ; > 33 : 230 ;210 1.00 −3 −3 −6 10 0.75 (See 5.5.3) 10 −3 : > 330 ; > 33 10 −4 : 155 ;121 10 −5 : 18 ;23 10 10 Escherichia coli 0.5 −3 10 Pseudomonas Test procedure at concentrations % (V/V) Water control : −0 : > 330 ; > 33 −1 : 166 ;144 10 −2 : 22 ;18 10 −0 : 210 ;198 10 −1 : 27 ;19 10 −2 : ;2 10 10 −0 : ;0 −1 : ;0 −2 : ;0 −3 : > 330 ; > 33 −4 : 143 ;122 10 −5 : 22 ;11 Nc : 7,15 Nd : 4,20 Nd : 3,65 Nd : < 0,10 Nts : > 100 Nts : > 100 Nts : Nts : R: 2,95 R: 3,50 R: > 7,05 10 NC : 7,14 10 EN 13697:2015 (E) −3 : > 330 ; > 33 10 −4 : 162 ;146 10 −5 : 12 ;16 10 10 : > 330 ; > 33 10 −4 : 166 ;134 10 : 227 ;202 10 −7 : 18 ;23 10 10 aureus 10 ATCC 6538 −3 −6 Staphylococcus N : 6,72 −5 10 : 15 ;11 NT : 7,18 10 ATCC 10541 10 N : 7,09 −5 10 : 27 ;18 NT : 7,28 10 −1 : 63 ;52 10 −2 : 33 ;28 10 −2 : ;7 10 −1 : ;0 −2 : ;0 : 13 ;8 Nc : 7,19 Nd : 4,41 Nd : 3,76 Nd : < 0,10 Nts : > 100 Nts : > 100 Nts : 23 Nts : R: 2,78 R: 3,43 R: > 7,09 −0 : > 330 ; > 33 −0 : > 330 ; > 33 −1 : > 300 ; > 30 −1 : 128 ;145 10 −2 : 127 ;132 −2 : ;13 10 10 NC : 7,18 10 10 : 265 ;240 : ;0 −5 : 198 ;178 : 29 ;31 −1 −0 10 10 −4 −7 : > 330 ; > 33 : 158 ;144 10 10 −0 −4 : > 330 ; > 33 : 235 ;255 10 : > 330 ; > 33 −3 −6 : > 330 ; > 33 −3 10 −3 : > 330 ; > 33 −4 : 194 ;179 10 −5 : 23 ;19 10 10 Enterococcus hirae −0 10 10 −0 : ;0 −1 : ;0 −2 : ;0 10 −3 : > 330 ; > 33 −4 : 201 ;187 10 −5 : 17 ;24 Nc : 7,28 Nd : 5,11 Nd : 4,14 Nd : < 0,10 Nts : > 100 Nts : > 100 Nts : Nts : R: … R: … R: > … 10 NC : 7,29 10 10 10 31 BS EN 13697:2015 EN 13697:2015 (E) Annex D (informative) Bactericidal activity in surfaces in general use conditions (for clean conditions) a) Identification of the test laboratory b) Identification of the sample: c) d) Besson test house; Name of the product Z Batch number 94–71–51 Manufacturer Centipede Formulations Inc Date of delivery 1994–02–11 Storage conditions room temperature and darkness Product diluent recommended by the manufacturer for use Potable water Active substance(s) and its (their) concentration(s) (optional) Not indicated Test method and its validation: Method Dilution neutralization; Neutralizer 30 g/l, lecithin, sterilized in the autoclave ; Experimental conditions: Period of analysis 1994–02–20 to 1994–03–12 Product diluent used during the test sterile hard water 300 mg/kg CaCO3 Product test concentrations g/l, g/l and g/l Appearance product dilutions colourless, clear solution Interfering substances 0.3 g/l of bovine albumin (0.85 % skimmed milk fo Pseudomonas aeruginosa) Test temperature θ = between 18 °C and 25 °C Contact time t = ± 10 s Temperature of incubation 37 °C ± °C Identification of the bacterial strains used Pseudomonas aeruginosa ATCC 15 442 Escherichia coli ATCC 10 536 Staphylococcus aureus ATCC 538 Enterococcus hirae ATCC 10 541 e) Test results: See Table C.1 f) Conclusion: In accordance with this European Standard the batch 94–71–51 of the product Z, when diluted at % (V/V), in hard water, possesses bactericidal activity on surfaces in at 20 °C under clean conditions 0,3 g/l bovine albumin for referenced strains Escherichia coli, Staphylococcus aureus a Enterococcus hirae and 0,85 % skimmed milk for reference strain Pseudomonas aeruginosa g) Locality, date and identified signature NOTE 32 The test product, batch N° and manufacturer are given as imaginary examples only BS EN 13697:2015 EN 13697:2015 (E) Annex E (informative) Precision of the test result The formula used for the calculation of the precision of the test results can be found in EN 1040:2005, Annex E The number of repetitions which give a precision of the reduction factor need to be established by appropriate collaborative studies A guidance to the interpretation of the test results concerning their precision and the number of test repetitions is given in the EN 14885 33 BS EN 13697:2015 EN 13697:2015 (E) Bibliography [1] EN 1040:2005, Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of basic bactericidal activity of chemical disinfectants and antiseptics - Test method and requirements (phase 1) [2] EN 10088-1, Stainless steels - Part 1: List of stainless steels [3] EN 10088-2, Stainless steels - Part 2: Technical delivery conditions for sheet/plate and strip of corrosion resisting steels for general purposes 34 This page deliberately left blank NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW British Standards Institution (BSI) BSI is the national body responsible for preparing British Standards and other standards-related publications, information and services BSI is incorporated by Royal Charter British Standards and other standardization products are published by BSI Standards Limited About us Revisions We bring together business, industry, government, consumers, innovators and others to shape their combined experience and expertise into standards -based solutions Our British Standards and other publications are updated by amendment or revision The knowledge embodied in our standards has been carefully assembled in a dependable format and refined through our 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