BRITISH STANDARD BS EN 1276:2009 Incorporating corrigendum August 2010 Chemical disinfectants and antiseptics — 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) ICS 11.080.20; 67.020; 71.100.35 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW BS EN 1276:2009 National foreword This British Standard is the UK implementation of EN 1276:2009, incorporating corrigendum August 2010 It supersedes BS EN 1276:1997 which is withdrawn The UK participation in its preparation was entrusted to Technical Committee CH/216, Chemical disinfectants and antiseptics A list of organizations represented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 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 May 2010 © BSI 2010 ISBN 978 580 72821 Amendments/corrigenda issued since publication Date Comments 30 November 2010 Implementation of CEN corrigendum August 2010: 5.7.3 corrected EN 1276 EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM October 2009 ICS 71.100.35 Supersedes EN 1276:1997 Incorporating corrigendum August 2010 English Version Chemical disinfectants and antiseptics - 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 désinfectants chimiques - Essai quantitatif de suspension pour l'évaluation de l'activité bactéricide des antiseptiques et des désinfectants chimiques utilisés dans le domaine de l'agro-alimentaire, dans l'industrie, dans les domaines domestiques et en collectivité - Méthode d'essai et prescriptions (Phase 2, étape 1) Chemische Desinfektionsmittel und Antiseptika Quantitativer Suspensionsversuch zur Bestimmung der bakteriziden Wirkung chemischer Desinfektionsmittel und Antiseptika in den Bereichen Lebensmittel, Industrie, Haushalt und öffentliche Einrichtungen - Prüfverfahren 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 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 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 Management 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, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom EUROPEAN COMMITTEE FOR STANDARDIZATION COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR 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 1276:2009: E BS EN 1276:2009 EN 1276:2009 (E) Contents Page Foreword Introduction 5 1 Scope 2 Normative references 3 Terms and definitions 7 4 Requirements 5 Test method 5.1 Principle 5.2 Materials and reagents 5.2.1 Test organisms 5.2.2 Culture media and reagents 5.3 Apparatus and glassware 13 5.4 Preparation of test organism suspensions and product test solutions 14 5.4.1 Test organism suspensions (test and validation suspension) 14 5.4.2 Product test solutions 15 5.5 Procedure for assessing the bactericidal activity of the product 17 5.5.1 General 17 5.5.2 Dilution-neutralization method 18 5.5.3 Membrane filtration method 20 5.6 Experimental data and calculation 22 5.6.1 Explanation of terms and abbreviations 22 5.6.2 Calculation 23 5.7 Verification of methodology 26 5.7.1 General 26 5.7.2 Control of weighted mean counts 26 5.7.3 Basic limits 26 5.8 Expression of results and precision 26 5.8.1 Reduction 26 5.8.2 Control of active and non-active product test solution (5.4.2) 27 5.8.3 Limiting test organism and bactericidal concentration 27 5.8.4 Precision, replicates 27 5.9 Interpretation of results - conclusion 27 5.9.1 General 27 5.9.2 Bactericidal activity for general purposes 27 BS EN 1276:2009 EN 1276:2009 (E) 5.9.3 Bactericidal activity for specific purposes 27 5.10 Test report 28 Annex A (informative) Referenced strains in national collections 30 Annex B (informative) Neutralizers and rinsing liquids 31 Annex C (informative) Graphical representations of dilution neutralization method and membrane filtration method 33 C.1 Dilution-neutralization method 33 C.2 Membrane filtration method 35 Annex D (informative) Example of a typical test report 37 Test results (bactericidal suspension test) 39 Annex E (informative) Precision of the test result 41 Bibliography 44 BS EN 1276:2009 EN 1276:2009 (E) Foreword This document (EN 1276:2009) 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 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 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 1276:1997 It 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 (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, the 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, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom BS EN 1276:2009 EN 1276:2009 (E) Introduction This European Standard specifies a suspension test for establishing whether a chemical disinfectant or antiseptic has or does not have bactericidal 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 conditions 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 of use of a product may differ and therefore additional test conditions need to be used BS EN 1276:2009 EN 1276:2009 (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, as some dilution is always produced by adding the test organisms and interfering substance This document applies 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 included: a) processing, distribution and retailing of: 1) food of animal origin: food of vegetable origin: milk and milk products; beverages; meat and meat products; fruits, vegetables and derivatives (including sugar, distillery, etc.); fish, seafood, and related products; flour, milling and baking; eggs and egg products; animal feeds; animal feeds; etc etc b) institutional and domestic areas: catering establishments; public areas; public transports; schools; nurseries; shops; sports rooms; waste containers (bins, etc.); hotels; dwellings; 2) clinically non sensitive areas of hospitals; offices; etc BS EN 1276:2009 EN 1276:2009 (E) 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 another and to “use recommendations” 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 2 This method corresponds to a phase step test 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 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, mycobactericidal, sporicidal and fungicidal activity EN 14885:2006, 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 14885:2006 apply Requirements The product shall demonstrate at least a 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 under simulated clean 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, or (for hands disinfection)) The bactericidal activity shall be evaluated using the following four test organisms: Pseudomonas aeruginosa; Escherichia coli; BS EN 1276:2009 EN 1276:2009 (E) Staphylococcus aureus; Enterococcus hirae 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 additional conditions, the concentration defined as a result can be lower than the one obtained under the obligatory test conditions 5.1 Test method 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 ± 10 s (obligatory test conditions) during (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 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 aureus 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 1276:2009 EN 1276:2009 (E) Phenolic and related compounds: orthophenylphenol, phenoxyethanol, triclosan, phenylethanol, etc… Lecithin - Polysorbate 80, 30 g/l + lecithin, g/l Polysorbate 80 Anilides Ethylene oxide condensate of fatty alcohol b) - Ethylene oxide condensate of fatty alcohol, g/l + lecithin, 20 g/l, + polysorbate 80, g/l Rinsing liquid : tryptone, g/l + NaCl, g/l; polysorbate 80, g/l Alcohols Lecithin, Saponin, Polysorbate 80 e) - Polysorbate 80, 30 g/l + saponin, 30 g/l + lecithin, g/l Rinsing liquid : tryptone, g/l + NaCl, g/l; polysorbate 80, g/l a) According to the pH of the tested product, the pH of the neutralizer or the rinsing liquid may be adjusted at a suitable value or prepared in phosphate buffer [ex: phosphate buffer 0,25 mol/l: potassium dihydrogen phosphate (KH2PO4) 34 g; distilled water (500 ml); adjusted to pH 7,2 ± 0,2 with sodium hydroxide (NaOH) mol/l; distilled water up to 000 ml] b) The carbon chain-length varies from C12 to C18 carbon atoms c) Egg and soya; egg is preferable d) The toxic effect of sodium thiosulphate differs from one test organism to another e) For the neutralization of short chain alcohols (less than C5), simple dilution may be appropriate Care should be taken if the alcohol-based -products contain additional antimicrobial agents NOTE Other neutralizer mixtures may be required for products containing more than one antimicrobial agent NOTE The concentrations of the various neutralizing compounds or of the neutralizer as such may not be adequate to neutralize high concentrations of the products 32 BS EN 1276:2009 EN 1276:2009 (E) Annex C (informative) Graphical representations of dilution neutralization method and membrane filtration method C.1 Dilution-neutralization method Key Figure C.1 33 BS EN 1276:2009 EN 1276:2009 (E) Key Figure C.2 34 BS EN 1276:2009 EN 1276:2009 (E) C.2 Membrane filtration method Key Figure C.3 35 BS EN 1276:2009 EN 1276:2009 (E) Key Figure C.4 36 BS EN 1276:2009 EN 1276:2009 (E) Annex D (informative) Example of a typical test report NOTE All names and examples in Annex D are fictitious apart from those used in this standard NOTE Only the test results of one replicate for Pseudomonas aeruginosa are given as an example NOTE Test reports for bactericidal activity should be entitled “EN 1276, BACTERICIDAL ACTIVITY”, and be presented in the same format - 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 1276, BACTERICIDAL ACTIVITY (obligatory and additional conditions) Client: Mult Formulations Inc., Mannheim / Euroland Disinfectant-sample Name of the product: W Batch number: 26-01-48 Manufacturer or - if not known - supplier: Centipede 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: 2006-05-01 Dates of tests: see “Test results” (attached) 37 BS EN 1276:2009 EN 1276:2009 (E) Experimental conditions Product diluent: hard water Concentrations of the product tested: see “Test results” (attached) Obligatory conditions: test organisms: Pseudomononas aeruginosa ATCC 15442, Escherichia coli ATCC 10536, Staphylococcus aureus ATCC 6538, Enterococcus hirae ATCC 10541 test temperature: 20 °C contact time: interfering substance: 0,3 g/l bovine albumin = clean conditions Incubation temperature: 36 °C Additional conditions: test organism: Pseudomononas aeruginosa ATCC 15442 Test temperature: 40 °C contact time:60 Interfering substance: 3,0 g/l bovine albumin = dirty conditions Incubation temperature: 36 °C Test results: see attached sheets test organism: Salmonella Typhimurium ATCC 13311 Test temperature: °C contact time: 15 Interfering substance: 10,0 g/l reconstituted milk 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 an lg reduction of less than lg No precipitate during the test procedure (test mixtures were homogeneous) 38 BS EN 1276:2009 EN 1276:2009 (E) Conclusion: For the product W (batch 26-01-48), the bactericidal concentration for general purposes determined according to the EN 1276 standard (obligatory conditions) under clean conditions is: 1,0 % (v/v) (the mean reduction of six replicates with the limiting test organism Pseudomononas aeruginosa was 1,2 x 10 Staphylococcus aureus, Escherichia coli and Enterococcus hirae were tested once and showed a lg reduction or more at a lower concentration than Pseudomononas aeruginosa) For the product W (batch 26-01-48), the bactericidal concentration for specific purposes determined according to the EN 1276 standard at 40 °C, with 60 contact time under dirty conditions, using Pseudomononas aeruginosa ATCC 15442 as test organism is: 0,25 % (v/v) For the product W (batch 26-01-48) the bactericidal concentration for specific purposes determined according to the EN 1276 standard at °C, with 15 contact time with milk as interfering substance, using Salmonella Typhimurium ATCC 13311 as test organism is: 0,25 % (v/v) Antiseptville, 2001-11-11 Alexandra May, MD, PhD, Scientific Director Test results (bactericidal suspension test) EN 1276… .(Phase 2, step 1) Product-name: W Batch No.: 26-01-48 Remarks: Dilution neutralization method X Pour plate Spread plate x Number of plates .2 / ml Neutralizer: Lecithin 3,0 g/l in diluent Membrane filtration method Test temperature: 20 °C Rinsing liquid: interfering substance:….bovine albumin: 0,3 g/l ……………………………………… Test organism: Pseudomonas aeruginosa ATCC 15442 Incubation temperature: 36 °C Internal lab No : QS 68/00 Date of test:2006-06-06 Responsible person: Fang Signature: Fang 39 BS EN 1276:2009 EN 1276:2009 (E) Diluent used for product test solutions: hard.water Appearance of the product test solutions: clear Validation and controls Validation Experimental suspension (Nv0) Conditions control (A) Vc1 86 x= Vc1 92 86 Vc1 x= (43 + 36) (40 + 46) Vc2 79 Neutralizer or filtration control (B) 89 Vc2 (47 + 45) 30 ≤ x of Nv0 ≤ 160 ? yes no Method validation (C) Product conc.: 10 ml/l 75 Vc1 x= (42 + 44) 84 81,5 (35 + 40) 91 Vc2 (39 + 45) 88,5 87 Vc2 (43 + 48) yes no yes Test-suspension (41 + 46) Conc of the product % no N (N and N0): Vc1 Vc1 yes no Vc2 x wm = 193,64 x 106 ; lgN = 8,29 10 -6 168 213 No = N/10 ; lg No = 7,29 10 -7 20 25 7,17 ≤ lgN0 ≤ 7,70? Vc2 Na = x x10 lgNa yes no lgR Contact- (N0 = 7,29) time (min) 0,50 > 660 > 630 > 450 > 3,81 < 3,48 0,75 122 154 380 3,14 4,15 1,00 < 140 < 2,15 > 5,14 Remarks: Counting per plate for; –6 10 : 80 + 88; 105 + 108 (Na) –7 10 : + 11; 15 + 10 0,75 % : 66 + 56; 71 + 83 1,00 % Vc1: + Explanations: Vc = count per ml (one plate or more) x wm = weighted mean of x x = average of Vc1 and Vc2 (1 + duplicate) R = reduction (lg R = lgN0 – lgNa) 40 81 x of A is ≥ 0,5 x x of Nv0 ? x of B is ≥ 0,5 x x of Nv0 ? x of C is ≥ 0,5 x x of Nv0 ? Test suspension and Test (N) x= BS EN 1276:2009 EN 1276:2009 (E) Annex E (informative) Precision of the test result A collaborative study (ANDISTAND 1997 - 1999) was carried out to determine the precision of the test method within and between different laboratories The study involved 16 laboratories from different European countries Each laboratory replicated the test conditions three times The tests were performed using the dilution neutralization or membrane filtration methods with sodium dichloroisocyanurate, benzalkonium chloride and phenol on Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and Enterococcus hirae (as test organisms) The complete results and the statistical evaluation of this study is described in documents CEN/TC 216 HWG N 121 + N 121 Annexes + N 121 Corrigendum The agreement between laboratories, expressed in terms of bactericidal effect (reduction ≥ log), is very good at low and high concentrations but less good at intermediate levels Uncountable data were replaced by theoretical fake values of reduction factor When counts were below the lower counting limit (15 viable colonies), the reduction factor could vary between 3,33 x 10 (the maximum value when data were countable) and 5,00 x 10 (no surviving colonies), corresponding respectively to 5,52 and 7,70 log reduction In order to simulate a possible variability, three theoretical reduction factors (one for each replicate) were chosen in this range: 5,5, 6,6 and 7,7 (in logarithmic terms) When counts were above the upper counting limit (300 viable colonies), the reduction factor could vary between 1,00 x10 (no reduction) and 5,00 x10 (the minimum value when data were countable), giving the log reduction values of, respectively, 0,00 and 3,70 In this case, the three chosen fake theoretical reduction factors were 0,0; 1,8 and 3,6 (instead of 3,7 for calculation simplifying reason) All these theoretical values were collected in the following table (Table E.1) Table E.1 — Theoretical reduction factors used for bacteria (in logarithmic terms) All the replicates are Only replicates are Only replicate is below the lower limit (15) above the upper limit (150) 5,5 – 6,6 – 7,7 0,0 – 1,8 – 3,6 5,5 – 7,7 0,0 – 3,6 6,6 1,8 Variances analyses were performed on the reduction factors in order to point out the tested conditions with significant obtained results and to estimate the two types of variability: "within" and "between" labs The differences in reduction factors across the lab were essentially obtained with medium and high dilutions of the tested products Some of those significant results were due to outliers, certain laboratories giving completely different reduction factor from most of the others In other cases, it was more difficult to point out the outliers but the significant differences were the consequences of the important "within lab" variability 41 BS EN 1276:2009 EN 1276:2009 (E) In most of the cases, the "between" estimated standard deviation, which varied (in logarithmic terms) from 1,5 to 3,0, was higher than the "within" value, which oscillated between 1,0 and 2,0 Nevertheless, in some cases, this ratio was inverted ("between" dispersion lower than "within" one), due probably to the techniques used for replacing uncountable data, which overestimated the "within" variability and reduced the "between" variation The "inside" variability was also used to estimate the precision of the obtained reduction factors Two different hypotheses were taken into account: the first one, which could be qualified of "worst" case, used the maximum of the calculated "inside" variability (σ = 2,20) while the second o~e was based on an average "inside" dispersion (σ = 1,62) The estimated precision depended also on the sample size (number of replicates) and the confidence level (90 % probability of used in the calculation) With three replicates, the "worst" case lead to a reduction factor precision ± 3,71 while the "mean" case was about ± 2,73 So, if the precision target is ± log reduction, 15 replicates were needed in the "worst" case and in the "mean" one (Figure E.1 or Table E.2) Key X Y Precision (in log terms) Sample size (number of replicates) Worst case Mean case Figure E.1 — Precision of the reduction factor obtained with bacteria (in logarithmic terms) 42 BS EN 1276:2009 EN 1276:2009 (E) Table E.2 — Precision of the reduction factors obtained for bacteria (in logarithmic terms) Standard Deviation (σ) Repli cates 1,0 1,1 1,2 1,3 1,4 1,5 1,6 1,7 1,8 ± 6,70 ± 7,14 ± 7,59 ± 8,04 ± 8,48 ± 8,93 ± 9,38 ± 9,82 ± 10,27 ± 2,53 ± 2,70 ± 2,87 ± 3,03 ± 3,20 ± 3,37 ± 3,54 ± 3,71 ± 3,88 ± 1,77 ± 1,88 ± 2,00 ± 2,12 ± 2,24 ± 2,35 ± 2,47 ± 2,59 ± 2,71 ± 1,43 ± 1,53 ± 1,62 ± 1,72 ± 1,81 ± 1,91 ± 2,00 ± 2,10 ± 2,19 ± 1,23 ± 1,32 ± 1,40 ± 1,48 ± 1,56 ± 1,65 ± 1,73 ± 1,81 ± 1,89 ± 1,10 ± 1,18 ± 1,25 ± 1,32 ± 1,40 ± 1,47 ± 1,54 ± 1,62 ± 1,69 ± 1,00 ± 1,07 ± 1,14 ± 1,21 ± 1,27 ± 1,34 ± 1,41 ± 1,47 ± 1,54 ± 0,93 ± 0,99 ± 1,05 ± 1,12 ± 1,18 ± 1,24 ± 1,30 ± 1,36 ± 1,43 10 ± 0,87 ± 0,93 ± 0,99 ± 1,04 ± 1,10 ± 1,16 ± 1,22 ± 1,28 ± 1,33 11 ± 0,82 ± 0,87 ± 0,93 ± 0,98 ± 1,04 ± 1,09 ± 1,15 ± 1,20 ± 1,26 12 ± 0,78 ± 0,83 ± 0,88 ± 0,93 ± 0,99 ± 1,04 ± 1,09 ± 1,14 ± 1,19 13 ± 0,74 ± 0,79 ± 0,84 ± 0,89 ± 0,94 ± 0,99 ± 1,04 ± 1,09 ± 1,14 14 ± 0,71 ± 0,76 ± 0,80 ± 0,85 ± 0,90 ± 0,95 ± 0,99 ± 1,04 ± 1,09 15 ± 0,68 ± 0,73 ± 0,77 ± 0,82 ± 0,86 ± 0,91 ± 0,96 ± 1,00 ± 1,05 16 ± 0,66 ± 0,70 ± 0,75 ± 0,79 ± 0,83 ± 0,88 ± 0,92 ± 0,96 ± 1,01 17 ± 0,64 ± 0,68 ± 0,72 ± 0,76 ± 0,80 ± 0,85 ± 0,89 ± 0,93 ± 0,97 18 ± 0,62 ± 0,66 ± 0,70 ± 0,74 ± 0,78 ± 0,82 ± 0,86 ± 0,90 ± 0,94 19 ± 0,60 ± 0,64 ± 0,68 ± 0,72 ± 0,76 ± 0,80 ± 0,84 ± 0,88 ± 0,91 20 ± 0,58 ± 0,62 ± 0,66 ± 0,70 ± 0,73 ± 0,77 ± 0,81 ± 0,85 ± 0,89 NOTE It cannot be excluded that the precision may be better or worse when other test organism, products, and/or interfering substances are tested However, it seems likely that the precision in these cases will be in the same range 43 BS EN 1276:2009 EN 1276:2009 (E) Bibliography [1] European Pharmacopoeia (EP), Edition 1997 supplement 2000, Water for injections 44 This page has been intentionally left blank BS EN 1276:2009 BSI - British Standards Institution BSI is the independent national body responsible for preparing British Standards It presents the UK view on standards in Europe and at the international level It is incorporated by Royal Charter Revisions British Standards are updated by amendment or revision Users of British Standards should make sure that they possess the latest amendments or editions It is the constant aim of BSI to improve the quality of our products and services We would be grateful if anyone finding an inaccuracy or ambiguity while using this British Standard would inform the Secretary of the technical committee responsible, the identity of which can be found on the inside front cover Tel: +44 (0)20 8996 9000 Fax: +44 (0)20 8996 7400 BSI offers members an individual updating service called PLUS 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