1、BRITISH STANDARD BS EN 13704:2002 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
2、the status of a British Standard ICS 71.100.35 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBS EN 13704:2002 This British Standard, having been prepared under the direction of the Health and Environment Sector Policy and Strategy Committee, was published under the authority
3、of the Standards Policy and Strategy Committee on 05 March 2002 BSI 05 March 2002 ISBN 0 580 39201 5 National foreword This British Standard is the official English language version of EN 13704:2002 The UK participation in its preparation was entrusted to Technical Committee IH/57, Chemical disinfec
4、tants and antiseptics, which has the responsibility to: A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BS
5、I Standards Catalogue under the section entitled “International Standards Correspondence Index”, or by using the “Find” 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 respon
6、sible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible European committee any enquiries on the interpretation, or proposals for change, and keep the UK interes
7、ts informed; monitor related international and European developments and promulgate them in the UK. 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 doc
8、ument indicates when the document was last issued. Amendments issued since publication Amd. No. Date CommentsEUROPEANSTANDARD NORMEEUROPENNE EUROPISCHENORM EN13704 February2002 ICS71.100.35 Englishversion ChemicaldisinfectantsQuantitativesuspensiontestforthe evaluationofsporicidalactivityofchemicald
9、isinfectantsusedin food,industrial,domesticandinstitutionalareasTestmethod andrequirements(phase2,step1) DsinfectantschimiquesEssaiquantitatifdesuspension pourlvaluationdelactivitsporicidedesdsinfectants chimiquesutilissdansledomainedelagroalimentaire, danslindustrie,danslesdomainesdomestiqueseten c
10、ollectivitMthodedessaietprescriptions(phase2, tape1) ChemischeDesinfektionsmittelQuantitativer SuspensionsversuchzurBestimmungdersporiziden WirkungchemischerDesinfektionsmittelindenBereichen Lebensmittel,Industrie,Haushaltundffentliche EinrichtungenPrfverfahrenundAnforderungen(Phase2, Stufe1) ThisEu
11、ropeanStandardwasapprovedbyCENon11November2001. CENmembersareboundtocomplywiththeCEN/CENELECInternalRegulationswhichstipulatetheconditionsforgivingthisEurope an Standardthestatusofanationalstandardwithoutanyalteration.Uptodatelistsandbibliographicalreferencesconcernings uchnational standardsmaybeobt
12、ainedonapplicationtotheManagementCentreortoanyCENmember. ThisEuropeanStandardexistsinthreeofficialversions(English,French,German).Aversioninanyotherlanguagemadebytra nslation undertheresponsibilityofaCENmemberintoitsownlanguageandnotifiedtotheManagementCentrehasthesamestatusasthe official versions.
13、CENmembersarethenationalstandardsbodiesofAustria,Belgium,CzechRepublic,Denmark,Finland,France,Germany,Greece, Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal,Spain,Sweden,SwitzerlandandUnitedKingdom. EUROPEANCOMMITTEEFORSTANDARDIZATION COMITEUROPENDENORMALISATION EUROPISCHESKOMITE
14、EFRNORMUNG ManagementCentre:ruedeStassart,36B1050Brussels 2002CEN Allrightsofexploitationinanyformandbyanymeansreserved worldwideforCENnationalMembers. Ref.No.EN13704:2002EEN13704:2002(E) 2 Contents page Foreword3 Introduction .4 1 Scope 5 2 Normativereferences 6 3 Termsanddefinitions. .6 4 Requirem
15、ents .7 5 Testmethod. .7 5.1 Principle7 5.2 Materialsandreagents 8 5.3 Apparatusandglassware . 10 5.4 Preparationofsporetestsuspensionandtestsolutions . 11 5.5 Procedure .12 5.6 Calculationandexpressionofresults. 15 5.7 Conclusion17 5.8 Testreport . .18 AnnexA (normative) Preparationof Bacillus spor
16、estocksuspensions 20 A.1 Materialandreagents . .20 A.2 Preparationof Bacillus sporestocksuspensions .20 AnnexB (normative) Validationofdilutionneutralizationandmembranefiltrationmethods .21 B.1 Principle21 B.2 Preparationofsporesuspension. 21 B.3 Preparationofproducttestsolution 21 B.4 Testforvalida
17、tion . .21 B.5 Validation24 AnnexC (informative) Preparationof Clostridiumsporogenes sporestocksuspension.25 C.1 Culturemediaandreagents 25 C.2 Apparatusandglassware . 26 C.3 Preparationofregeneratedmediaandincubationconditions .26 C.4 Preparationof Clostridium sporestocksuspension .26 AnnexD (infor
18、mative) Neutralizers 27 AnnexE (informative) Rinsingliquids. 28 AnnexF (informative) Exampleofatypicaltestreport .29 AnnexG (informative) Referencedstrainsinnationalcollections.32 G.1 Bacillussubtilis 32 G.2 Bacilluscereus .32 G.3 Clostridiumsporogenes 32 AnnexH (informative) Informationontheapplica
19、tionandinterpretationofEuropeanstandardson chemicaldisinfectantsandantiseptics . 33 H.1 Generalguidelinesfortheapplicationandinterpretationoftestmethodsinaccordancewith EuropeanStandardsforchemicaldisinfectantsandantiseptics.33 H.2 Guidetointerpretationoftestsforchemicaldisinfectantsandantiseptics .
20、34EN13704:2002(E) 3 Foreword ThisEuropeanStandardhasbeenpreparedbyTechnicalCommitteeCEN/TC216“Chemicaldisinfectantsand antiseptics“,thesecretariatofwhichisheldbyAFNOR. ThisEuropeanStandardshallbegiventhestatusofanationalstandard,eitherbypublicationofanidenticaltextor byendorsement,atthelatestbyAugus
21、t2002,andconflictingnationalstandardsshallbewithdrawnatthelatestby August2002. InthisstandardtheannexesAandBarenormative.TheannexesC,D,E,F,GandHareinformative. AccordingtotheCEN/CENELECInternalRegulations,thenationalstandardsorganizationsofthefollowing countriesareboundtoimplementthisEuropeanStandar
22、d:Austria,Belgium,CzechRepublic,Denmark,Finland, France,Germany,Greece,Iceland,Ireland,Italy,Luxembourg,Malta,Netherlands,Norway,Portugal,Spain, Sweden,SwitzerlandandtheUnitedKingdom.EN13704:2002(E) 4 Introduction ThisEuropeanStandarddescribesasuspensiontestmethodforestablishingwhetherachemicaldisin
23、fectanthas ordoesnothaveasporicidalactivityinthefieldsdescribedinclause1. Thelaboratorytestcloselysimulatespracticalconditionsofapplication.Chosenconditions(contacttime, temperature,insuspension,etc.)reflectparameterswhicharefoundinpracticalsituationsincludingconditions whichcaninfluencetheactionofd
24、isinfectants.Eachutilizationconcentrationfoundfromthistestcorrespondsto definedexperimentalconditions. Theconditionsareintendedtocovergeneralpurposesandtoallowreferencebetweenlaboratoriesandproduct types. Howeverforsomeapplicationstherecommendationsofuseofaproductcandifferandthereforeadditionaltest
25、conditionsneedtobeused.EN13704:2002(E) 5 1Scope ThisEuropeanStandardspecifiesatestmethod(phase2/step1)(seeannexH)andtheminimumrequirementsfor sporicidalactivityofchemicaldisinfectantproductsthatformahomogeneous,physicallystablepreparationinhard waterandthatareusedinfood,industrial,domesticandinstitu
26、tionalareas,excludingareasandsituationswhere disinfectionismedicallyindicatedandexcludingproductsusedonlivingtissuesexceptthoseforhandhygienein theaboveconsideredareas. ThisEuropeanStandardappliesatleasttothefollowing: a) processing,distributionandretailingof: 1) foodofanimalorigin: milkandmilkprodu
27、cts; meatandmeatproducts; fish,seafood,andrelatedproducts; eggsandeggproducts; animalfeeds; etc. ; 2) foodofvegetableorigin: beverages ; fruits,vegetablesandderivatives(includingsugar,distillery,etc.); flour,millingandbaking; animalfeeds; etc. ; b) institutionalanddomesticareas: cateringestablishmen
28、ts; publicareas; publictransports; schools ; nurseries ; shops ; sportsrooms; waste containers (bins, etc.) ; hotels ;EN13704:2002(E) 6 dwellings ; clinicallynonsensitiveareasofhospitals; offices ; etc. ; c) otherindustrialareas: packagingmaterial; biotechnology(yeast,proteins,enzymes,etc.); pharmac
29、eutical ; cosmeticsandtoiletries; textiles; spaceindustry,computerindustry; etc. UsingthisEuropeanStandard,itisnotpossibletodeterminethesporicidalactivityofundilutedproductassome dilutionisalwaysproducedbyaddingtheinoculumandinterferingsubstance.Productscanonlybetestedata concentrationof80%orless. N
30、OTE Themethoddescribedisintendedtodeterminetheactivityofcommercialformulationsoractivesubstanceson sporesintheconditionsinwhichtheyareused. 2 Normativereferences ThisEuropeanStandardincorporatesbydatedorundatedreference,provisionsfromotherpublications.These normativereferencesarecitedattheappropriat
31、eplacesinthetext,andthepublicationsarelistedhereafter.For datedreferences,subsequentamendmentstoorrevisionsofanyofthesepublicationsapplytothisEuropean Standardonlywhenincorporatedinitbyamendmentorrevision.Forundatedreferencesthelatesteditionofthe publicationreferredtoapplies(includingamendments). EN
32、1040,ChemicaldisinfectantsandantisepticsBasicbactericidalactivityTestmethodandrequirements (phase1). 3 Termsanddefinitions ForthepurposesofthisEuropeanStandard,thefollowingtermsanddefinitionsapply. 3.1 product(forchemicaldisinfectionand/orantisepsis) chemicalagentorformulationusedasachemicaldisinfec
33、tantorantisepticEN1040 3.2 sporicide (inthisstandard) productwhichkillssporesofthegenus Bacillus NOTE Theadjectivederivedfromsporicideissporicidal.EN13704:2002(E) 7 3.3 sporicidalactivity (inthisstandard ) capabilityoftheproducttoproduceatleastareductionof10 3 inthenumberofbacterialsporesbelongingto
34、 referencestrainof Bacillussubtilis underconditionsdefinedbythisEuropeanStandard 3.4 cleanconditions conditionsrepresentativeofsurfaceswhichhavereceivedasatisfactorycleaningprogrammeand/orareknownto containminimallevelsoforganicand/orinorganicmaterials 4Requirements Theproduct,whendilutedinhardwater
35、andtestedinaccordancewithclause5undersimulatedcleanconditions (0,3g/lbovinealbuminsee3.4)accordingtoitspracticalapplicationsandundertherequiredtestconditions(20C, 60min,1selectedreferencestrain),shalldemonstrateatleasta10 3 reductioninviablecounts. Thesporicidalactivityshallbeevaluatedusingthetestor
36、ganism Bacillussubtilis . Thedeterminedsporicidalconcentrationofthetestproductissuggestedasbeingsuitableforpracticalsituationsof use. Whereappropriate,additionalspecificsporicidalactivityshallbedeterminedunderotherconditionsoftime, temperatureandadditionalstrains(see5.2.1and5.5.1)inaccordancewith5.5
37、.1inordertotakeintoaccount intendedspecificuseconditions. NOTE Fortheseadditionalconditions,theconcentrationdefinedasaresultcanbelowerthantheoneobtainedunderthe initialtestconditionsof20C,60min,1selectedreferencestrain. 5 Testmethod 5.1Principle 5.1.1 Atestsuspensionofbacterialsporesinasolutionofint
38、erferingsubstance, simulatingcleanconditions,is addedtoapreparedsampleoftheproductundertestdilutedinhardwater.Themixtureismaintainedat 20C1Cfor60min10s(requiredtestconditions). At this contact time, an aliquot is taken; the sporicidal action in this portion is immediately neutralized or suppressedby
39、avalidatedmethod.Themethodofchoiceisdilutionneutralization.Ifasuitableneutralizercannot befound,membranefiltrationisused.Thenumberofsurvivingbacterialsporesineachsamplearedeterminedand thereductioninviablecountsiscalculated. 5.1.2 Thetestisperformedusingsporesof Bacillussubtilis.Additionalandoptiona
40、lexposuretimes, temperaturesandstrainsarespecified.EN13704:2002(E) 8 5.2 Materialsandreagents 5.2.1 Testorganisms Thesporicidalactivityshallbeevaluatedbyusingsporesofthefollowingstrain: Bacillussubtilis ATCC6633 1) . Ifrequiredforspecificapplications,additionalstrainsmaybechosenfrom,forexample: Baci
41、lluscereus ATCC12826 1) ; Clostridiumsporogenes CIP7939 1) . NOTE1 SeeannexGforcorrespondingstrainnumbersinsomeotherculturecollections. NOTE2 SeeannexCforparticularcultureandhandlingconditionsfor Clostridiumsporogenes . Ifadditionalstrainsareused,theyshallbeincubatedunderoptimumgrowthconditions(temp
42、erature,time, atmosphere)andnotedinthetestreport. Iftheadditionalstrainsselecteddonotcorrespondtothespecifiedstrains,theirsuitabilityforsupplyinginoculaof sufficientconcentrationshallbeverified.Iftheadditionalstrainstestedarenotclassifiedatareferencecentretheir identificationcharacteristicsshallbest
43、ated.Inaddition,theyshallbeheldbythetestinglaboratoryornational cultureunderareferencefor5years. 5.2.2 Culturemediaandreagents 5.2.2.1General Thereagentsshallbeofanalyticalgradeand/orappropriateformicrobiologicalpurposes. NOTE Toimprovereproducibility,itisrecommendedthatcommerciallyavailabledehydrat
44、edmaterialisusedforthe preparationofculturemedia.Themanufacturersinstructionsrelatingtothepreparationoftheseproductsshouldberigorousl y followed. 5.2.2.2Water Thewatershallbefreefromsubstancesthataretoxicorinhibitingtothebacterialsporesortothebacteria.Itshall befreshlyglassdistilledwaterandnotdemineralizedwater. Sterilizeintheautoclave(see5.3.
