1、Chemical disinfectants - Quantitative suspension test for the evaluation of sporicidal activity of chemical disinfectants used in food, industrial, domestic and institutional areas -Test method and requirements (phase 2, step 1) The European Standard EN 13704:2002 has the status of a British Standar
2、d ICs 71.100.35 BS EN 13704:2002 BS EN 13704:2002 Amd. No. National foreword Date Comments This British Standard, having been prepared under the direction of the Health and Environment Sector Policy and Strategy Committee, was published under the authority of the Standards Policy and Strategy Commit
3、tee on 05 March 2002 This British Standard is the official English language version of EN 13704:2002 The UKparticipation in its preparation was entrusted to Technical Committee IH/57, Chemical disinfectants and antiseptics, which has the responsibility to: - - aid enquirers to understand the text; p
4、resent to the responsible European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. - A list of organizations represented on this committee can be obtaine
5、d on request to its secretary. Cr oss-r e fer enc e s The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards Correspondence Index”, or by using the “Find
6、” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal
7、obligations. Summary of pages This document comprises a front cover, an inside front cover, the EN title page, pages 2 to 34, and an inside back cover and a back cover. The BSI copyright date displayed in this document indicates when the document was last issued. O BSI 05 March 2002 ISBN O 580 39201
8、 5 EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 13704 February 2002 ICs 71.100.35 English version Chemical disinfectants - Quantitative suspension test for the evaluation of sporicidal activity of chemical disinfectants used in food, industrial, domestic and institutional areas - Test method
9、 and requirements (phase 2, step 1) Dsinfectants chimiques - Essai quantitatif de suspension pour lvaluation de lactivit sporicide des dsinfectants chimiques utiliss dans le domaine de lagro-alimentaire, dans lindustrie, dans les domaines domestiques et en collectivit - Mthode dessai et prescription
10、s (phase 2, tape 1) Chemische Desinfektionsmittel - Quantitativer Suspensionsversuch zur Bestimmung der sporiziden Wirkung chemischer Desinfektionsmittel in den Bereichen Lebensmittel, Industrie, Haushalt und ffentliche Einrichtungen - Prfverfahren und Anforderungen (Phase 2, Stufe 1) This European
11、Standard was approved by CEN on 11 November 2001 CEN members are bound to comply with the CENKENELEC 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 concern
12、ing such national standards may be obtained on application to the 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 lang
13、uage and notified to the Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Spain, Swe
14、den, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION EUROPISCHES KOMITEE FR NORMUNG COMIT EUROPEN DE NORMALISATION Management Centre: rue de Stassart, 36 B-1050 Brussels O 2002 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Memb
15、ers. Ref. No. EN 13704:2002 E EN 13704:2002 (E) Contents page Foreword 3 Introduction . 4 1 Scope 5 2 Normative references 6 3 Terms and definitions 6 4 Requirements . 7 5 5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8 Test method 7 Principle 7 Materials and reagents 8 Apparatus and glassware . 1 0 Preparation of
16、spore test suspension and test solutions . II Procedure . 12 Calculation and expression of results . 15 Conclusion 17 Test report 18 Annex A (normative) Preparation of Bacillus spore stock suspensions 20 A.l Material and reagents 20 A.2 Preparation of Bacillus spore stock suspensions . 20 Annex B (n
17、ormative) Validation of dilution-neutralization and membrane filtration methods . 21 B.l Principle 21 B.2 Preparation of spore suspension . 21 B.3 Preparation of product test solution 21 B.4 Test for validation 21 B.5 Validation 24 Annex C (informative) Preparation of Clostridium sporogenes spore st
18、ock suspension . 25 C.l Culture media and reagents 25 C.2 C.3 C.4 Preparation of Clostridium spore stock suspension . 26 Annex D (informative) Neutralizers 27 Annex E (informative) Rinsing liquids . 28 Annex F (informative) Example of a typical test report . 29 Annex G (informative) Referenced strai
19、ns in national collections . 32 Apparatus and glassware . 26 Preparation of regenerated media and incubation conditions . 26 G.l Bacillus subtilis 32 G.2 Bacillus cereus . 32 G.3 Clostridium sporogenes 32 Annex H (informative) Information on the application and interpretation of European standards o
20、n chemical disinfectants and antiseptics . 33 H.l General guidelines for the application and interpretation of test methods in accordance with European Standards for chemical disinfectants and antiseptics . 33 H.2 Guide to interpretation of tests for chemical disinfectants and antiseptics . 34 2 EN
21、13704:2002 (E) Foreword This European Standard has been prepared by Technical Committee CEN/TC 21 6 “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
22、 or by endorsement, at the latest by August 2002, and conflicting national standards shall be withdrawn at the latest by August 2002. In this standard the annexes A and B are normative. The annexes C, D, E, F, G and H are informative. According to the CENKENELEC Internal Regulations, the national st
23、andards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. 3
24、 EN 13704:2002 (E) Introduction This European Standard describes a suspension test method for establishing whether a chemical disinfectant has or does not have a sporicidal activity in the fields described in clause 1. The laboratory test closely simulates practical conditions of application. Chosen
25、 conditions (contact time, temperature, in suspension, etc.) reflect parameters which are found in practical situations including conditions which can influence the action of disinfectants. Each utilization concentration found from this test corresponds to defined experimental conditions. The condit
26、ions 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. 4 EN 13704:2002 (E) 1 Scope This European Standard spe
27、cifies a test method (phase 2/step 1) (see annex H) and the minimum requirements for sporicidal activity of chemical disinfectant products that form a homogeneous, physically stable preparation in hard water and that are used in food, industrial, domestic and institutional areas, excluding areas and
28、 situations where disinfection is medically indicated and excluding products used on living tissues except those for hand hygiene in the above considered areas. This European Standard applies at least to the following : a) processing, distribution and retailing of 1) food of animal origin : - milk a
29、nd milk products ; - meat and meat products ; - fish, seafood, and related products ; - eggs and egg products ; - animal feeds ; - etc. ; 2) food of vegetable origin - beverages ; - fruits, vegetables and derivatives (including sugar, distillery, etc.) ; - flour, milling and baking ; - animal feeds
30、; - etc. ; b) institutional and domestic areas : - catering establishments ; - public areas ; - public transports ; - nurseries ; - sports rooms ; - waste containers (bins, etc.) ; - hotels ; 5 EN 13704:2002 (E) - dwellings ; - clinically non sensitive areas of hospitals ; - offices : - etc. ; c) ot
31、her industrial areas - packaging material ; - biotechnology (yeast, proteins, enzymes, etc.) ; - pharmaceutical ; - cosmetics and toiletries ; - textiles ; - space industry, computer industry ; - etc. Using this European Standard, it is not possible to determine the sporicidal activity of undiluted
32、product as some dilution is always produced by adding the inoculum and interfering substance. Products can only be tested at a concentration of 80 % or less. NOTE spores in the conditions in which they are used. The method described is intended to determine the activity of commercial formulations or
33、 active substances on 2 Normative references This European Standard incorporates by dated or undated reference, provisions from other publications. These normative references are cited at the appropriate places in the text, and the publications are listed hereafter. For dated references, subsequent
34、amendments to or revisions of any of these publications apply to this European Standard only when incorporated in it by amendment or revision. For undated references the latest edition of the publication referred to applies (including amendments). EN 1040, Chemical disinfectants and antiseptics - Ba
35、sic bactericidal activity - Test method and requirements (phase I). 3 Terms and definitions For the purposes of this European Standard, the following terms and definitions apply. 3.1 product (for chemical disinfection andlor antisepsis) chemical agent or formulation used as a chemical disinfectant o
36、r antiseptic EN 10401 3.2 sporicide (in this standard) product which kills spores of the genus Bacillus NOTE The adjective derived from a sporicide ) is a sporicidal . 6 EN 13704:2002 (E) 3.3 sporicidal activity (in this standard) capability of the product to produce at least a reduction of IO3 in t
37、he number of bacterial spores belonging to reference strain of Bacillus subtilis under conditions defined by this European Standard 3.4 clean conditions conditions representative of surfaces which have received a satisfactory cleaning programme and/or are known to contain minimal levels of organic a
38、nd/or inorganic materials 4 Requirements The product, when diluted in hard water and tested in accordance with clause 5 under simulated clean conditions (0,3 g/l bovine albumin see 3.4) according to its practical applications and under the required test conditions (20 OC, 60 min, 1 selected referenc
39、e strain), shall demonstrate at least a IO3 reduction in viable counts. The sporicidal activity shall be evaluated using the test organism Bacillus subtilis. The determined sporicidal concentration of the test product is suggested as being suitable for practical situations of use. Where appropriate,
40、 additional specific sporicidal activity shall be determined under other conditions of time, temperature and additional strains (see 5.2.1 and 5.5.1) in accordance with 5.5.1 in order to take into account intended specific use conditions. NOTE initial test conditions of 20 “C, 60 min, 1 selected ref
41、erence strain. For these additional conditions, the concentration defined as a result can be lower than the one obtained under the 5 Test method 5.1 Principle 5.1.1 A test suspension of bacterial spores in a solution of interfering substance, simulating clean conditions, is added to a prepared sampl
42、e of the product under test diluted in hard water. The mixture is maintained at 20 “C f 1 “C for 60 min f 10 s (required test conditions). At this contact time, an aliquot is taken ; the sporicidal action in this portion is immediately neutralized or suppressed by a validated method. The method of c
43、hoice is dilution-neutralization. If a suitable neutralizer cannot be found, membrane filtration is used. The number of surviving bacterial spores in each sample are determined and the reduction in viable counts is calculated. 5.1.2 The test is performed using spores of Bacillus subtilis. Additional
44、 and optional exposure times, temperatures and strains are specified. 7 EN 13704:2002 (E) 5.2 Materials and reagents 5.2.1 Test organisms The sporicidal activity shall be evaluated by using spores of the following strain : - Bacillus subtilis ATCC 6633 I). If required for specific applications, addi
45、tional strains may be chosen from, for example : - Bacillus cereus ATCC 12826 ; - Clostridium sporogenes CIP 7939 I). NOTE 1 See annex G for corresponding strain numbers in some other culture collections. NOTE 2 See annex C for particular culture and handling conditions for Clostridium sporogenes. I
46、f 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 do not correspond to the specified strains, their suitability for supplying inocula of sufficient concentration shal
47、l be verified. If the additional strains tested 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 under a reference for 5 years. 5.2.2 Culture media and reagents 5.2.2.1 General
48、 The reagents shall be of analytical grade and/or appropriate for microbiological purposes. NOTE To improve reproducibility, it is recommended that commercially available dehydrated material is used for the preparation of culture media. The manufacturers instructions relating to the preparation of t
49、hese products should be rigorously fol lowed. 5.2.2.2 Water The water shall be free from substances that are toxic or inhibiting to the bacterial spores or to the bacteria. It shall be freshly glass distilled water and not demineralized water. Sterilize in the autoclave (see 5.3.1). NOTE 1 If the water is sterilized during the sterilization of the reagents, this is not necessary. NOTE 2 be used. If distilled water of adequate quality is not available, water for injectable preparation_(European Pharmacopoeia) can ATCC 6633 and ATCC 12826 are the collection numbers of st
