EN ISO 14729:2001

Ophthalmic optics - Contact lens care products - Microbiological requirements and test methods for products and regimens for hygienic management of contact lenses (ISO 14729:2001)

Specification of uniform test methods in order to achieve comparable test results.

Augenoptik - Kontaktlinsenpflegemittel - Mikrobiologische Anforderungen und Prüfverfahren für Produkte und Systeme zum Hygienemanagement von Kontaktlinsen (ISO 14729:2001)

Diese Internationale Norm beschreibt zwei Testverfahren sowie Anforderungen, die bei der Bewertung der antimikrobiellen Wirkung von Produkten anzuwenden sind, die zur chemischen Desinfektion von Kontaktlinsen vermarktet werden sollen. Der "Stand Alone Test" (Pflegemittel Direktprüfung) soll einzelne Lösungen mit eindeutig antimikrobieller Wirkung als Desinfektionsmittel für Kontaktlinsen qualifizieren. Der "Regimen Test" (Prüfung gemäß Pflegeanweisung) soll einzelne Lösungen als Teil einer umfassenden Pflegeanweisung zur Desinfektion von Kontaktlinsen qualifizieren.

Optique ophtalmique - Produits d'entretien des lentilles de contact - Exigences microbiologiques et méthodes d'essai des produits et protocoles d'entretien des lentilles de contact (ISO 14729:2001)

La présente Norme internationale décrit deux méthodes d'essai et les exigences devant être utilisées pour l'évaluation de l'activité antimicrobienne des produits destinés à être commercialisés pour la décontamination des lentilles de contact par des moyens chimiques, ainsi que des produits faisant partie d'un protocole d'entretien des lentilles de contact.  La présente Norme internationale ne concerne pas l'entretien des lentilles expérimentales.  
NOTE Les normes générales relatives aux produits de décontamination (par exemple l'EN 1040:1997 et l'EN 1275:1997) ne sont pas applicables aux produits d'entretien des lentilles de contact

Očesna optika - Kontaktne leče in izdelki za vzdrževanje kontaktnih leč – Mikrobiološke zahteve in preskusne metode za izdelke in postopki higienskega ravnanja s kontaktnimi lečami (ISO 14729:2001)

Ta mednarodni standard določa dve preskusni metodi za vrednotenje antimikrobne dejavnosti izdelkov, ki se tržijo za razkuževanje kontaktnih leč s kemičnimi sredstvi in izdelkov, ki so del postopkov nege kontaktnih leč. Ta mednarodni standard ne velja za higiensko ravnanje s poskusnimi lečami.

General Information

• Status: Published
• Publication Date: 14-Apr-2001
• ICS: 11.040.70 - Ophthalmic equipment
• Technical Committee: CEN/TC 170 - Ophthalmic optics
• Drafting Committee: CEN/TC 170 - Ophthalmic optics
• Current Stage: 9093 - Decision to confirm - Review Enquiry
• Start Date: 20-May-2008
• Completion Date: 20-May-2008
• Ref Project: SIST EN ISO 14729:2002 - Ophthalmic optics - Contact lens care products - Microbiological requirements and test methods for products and regimens for hygienic management of contact lenses (ISO 14729:2001)

Relations

• Amended By: EN ISO 14729:2001/A1:2010 - Ophthalmic optics - Contact lens care products - Microbiological requirements and test methods for products and regimens for hygienic management of contact lenses - Amendment 1 (ISO 14729:2001/Amd 1:2010)
• Effective Date: 22-Dec-2008

Overview

EN ISO 14729:2001 (ISO 14729:2001) specifies microbiological requirements and test methods for contact lens care products and care regimens used for the hygienic management of contact lenses. Its purpose is to provide uniform test methods so manufacturers, test laboratories and regulators can achieve comparable results when evaluating antimicrobial performance of chemical disinfecting products and multi‑step regimens for contact lens disinfection.

Key topics and technical requirements

• Scope and intent
  • Applies to liquid contact lens care products and disinfecting regimens (not trial lenses).
  • Emphasises that multidose products must be sterile until opened and adequately preserved or packaged to limit in‑use contamination.
• Two principal test methods
  • Stand‑alone test (inoculum challenge) - evaluates an individual solution’s antimicrobial activity against a standard panel of microorganisms over predetermined time points.
  • Regimen test - evaluates a multifunctional care regimen (cleaning, rinsing, soaking) by applying the microbial challenge to contact lenses and carrying them through the labelled regimen.
• Performance criteria
  • Primary criteria (stand‑alone):
    • Bacteria: average reduction ≥ 99.9% (3.0 log) within the minimum recommended soaking period.
    • Moulds and yeasts: average reduction ≥ 90% (1.0 log) within minimum soaking period, with no increase at ≥ 4× that period (±0.5 log experimental error).
  • Secondary criteria (for products failing primary criteria):
    • Sum of averages ≥ 5.0 log reduction for three bacterial species within the recommended soaking period, with a minimum average of 1.0 log for any single bacterium; stasis observed for yeast/moulds (±0.5 log).
  • Regimen criteria: average recovery after the regimen must be ≤ 10 cfu for each lens type/storage solution combination.
• Validation and controls
  • Methods require validated neutralization or removal of residual antimicrobial agents during culturing and appropriate controls to demonstrate method reliability.
• Annexes and scope notes
  • Informative annexes discuss virus and Acanthamoeba testing rationale (Annex C and D) and organic soil/artificial tears considerations (Annex E).

Applications and users

EN ISO 14729 is used by:

• Contact lens care product manufacturers for product development, labeling (disinfecting product vs. regimen), and shelf‑life claims.
• Microbiological test laboratories performing stand‑alone and regimen tests to regulatory standards.
• Regulatory bodies and conformity assessment organizations evaluating safety and performance of lens care products.
• Ophthalmic device engineers and quality assurance teams responsible for formulation, preservation and in‑use contamination risk management.

Practical uses include qualifying disinfecting solutions, validating multi‑step care systems, and supporting regulatory submissions and product labeling related to contact lens disinfection and microbiological efficacy.

Related standards

• ISO 8320 (vocabulary for contact lenses and care products) is referenced.
• General disinfection standards such as EN 1040:1997 and EN 1275:1997 are not applicable to contact lens care products; EN ISO 14729 provides the specific microbiological framework for that product class.

EN ISO 14729:2001 delivers standardized, comparable test methods and microbiological requirements essential for ensuring safe, effective contact lens disinfection in manufacturing and regulatory practice.

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.KLJLHQVNHJDAugenoptik - Kontaktlinsenpflegemittel - Mikrobiologische Anforderungen und Prüfverfahren für Produkte und Systeme zum Hygienemanagement von Kontaktlinsen (ISO 14729:2001)Optique ophtalmique - Produits d'entretien des lentilles de contact - Exigences microbiologiques et méthodes d'essai des produits et protocoles d'entretien des lentilles de contact (ISO 14729:2001)Ophthalmic optics - Contact lens care products - Microbiological requirements and test methods for products and regimens for hygienic management of contact lenses (ISO 14729:2001)11.040.70Oftalmološka opremaOphthalmic equipmentICS:Ta slovenski standard je istoveten z:EN ISO 14729:2001SIST EN ISO 14729:2002en01-maj-2002SIST EN ISO 14729:2002SLOVENSKI
STANDARD
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN ISO 14729April 2001ICS 11.040.70English versionOphthalmic optics - Contact lens care products - Microbiologicalrequirements and test methods for products and regimens forhygienic management of contact lenses (ISO 14729:2001)Optique ophtalmique - Produits d'entretien des lentilles decontact - Exigences microbiologiques et méthodes d'essaides produits et protocoles d'entretien des lentilles decontact (ISO 14729:2001)Augenoptik - Kontaktlinsenpflegemittel - MikrobiologischeAnforderungen und Prüfverfahren für Produkte undSysteme zum Hygienemanagement von Kontaktlinsen (ISO14729:2001)This European Standard was approved by CEN on 15 April 2001.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards 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 translationunder the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2001 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 14729:2001 ESIST EN ISO 14729:2002

Page 2EN ISO 14729:2001ForewordThe text of the International Standard ISO 14729:2001 has been prepared by TechnicalCommittee ISO/TC 172 “Optics and optical instruments” in collaboration with TechnicalCommittee CEN/TC 170 “Ophthalmic optics”, the secretariat of which is held by DIN.This European Standard shall be given the status of a national standard, either by publicationof an identical text or by endorsement, at the latest by October 2001, and conflicting nationalstandards shall be withdrawn at the latest by October 2001.According to the CEN/CENELEC Internal Regulations, the national standards organizationsof the following countries are bound to implement this European Standard: Austria, Belgium,Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy,Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the UnitedKingdom.Endorsement noticeThe text of the International Standard ISO 14729:2001 was approved by CEN as a EuropeanStandard without any modification.SIST EN ISO 14729:2002

ReferencenumberISO14729:2001(E)©ISO2001INTERNATIONALSTANDARDISO14729Firstedition2001-04-15Ophthalmicoptics—Contactlenscareproducts—MicrobiologicalrequirementsandtestmethodsforproductsandregimensforhygienicmanagementofcontactlensesOptiqueophtalmique—Produitsd'entretiendeslentillesdecontact—Exigencesmicrobiologiquesetméthodesd'essaidesproduitsetprotocolesd'entretiendeslentillesdecontactSIST EN ISO 14729:2002

ISO14729:2001(E)PDFdisclaimerThisPDFfilemaycontainembeddedtypefaces.InaccordancewithAdobe'slicensingpolicy,thisfilemaybeprintedorviewedbutshallnotbeeditedunlessthetypefaceswhichareembeddedarelicensedtoandinstalledonthecomputerperformingtheediting.Indownloadingthisfile,partiesacceptthereintheresponsibilityofnotinfringingAdobe'slicensingpolicy.TheISOCentralSecretariatacceptsnoliabilityinthisarea.AdobeisatrademarkofAdobeSystemsIncorporated.DetailsofthesoftwareproductsusedtocreatethisPDFfilecanbefoundintheGeneralInforelativetothefile;thePDF-creationparameterswereoptimizedforprinting.EverycarehasbeentakentoensurethatthefileissuitableforusebyISOmemberbodies.Intheunlikelyeventthataproblemrelatingtoitisfound,pleaseinformtheCentralSecretariatattheaddressgivenbelow.©ISO2001Allrightsreserved.Unlessotherwisespecified,nopartofthispublicationmaybereproducedorutilizedinanyformorbyanymeans,electronicormechanical,includingphotocopyingandmicrofilm,withoutpermissioninwritingfromeitherISOattheaddressbeloworISO'smemberbodyinthecountryoftherequester.ISOcopyrightofficeCasepostal[e-mail]©ISO2001–AllrightsreservedSIST EN ISO 14729:2002

ISO14729:2001(E)©ISO2001–AllrightsreservediiiContentsPageForeword.ivIntroduction.v1Scope.12Normativereferences.13Termsanddefinitions.14Principle.24.1General.24.2Stand-alonetest(Inoculumchallengetest).24.3Regimentest.35Performancerequirements.35.1Stand-alonetest:Primarycriteria(seealsoTable1).35.2Stand-alonetest:Secondarycriteria(seealsoTable1).45.3Regimentest:Regimencriteria(seealsoTable1).46Testmethods.56.1Materialsandreagents.56.2Preparationofmicrobialchallenge(Inoculum).66.3Stand-aloneprocedure.76.4Regimenprocedure.10AnnexA(informative)Testorganismsfromotherculturecollections.13AnnexB(informative)Exampleofamembranefiltrationprocedure.14AnnexC(informative)Technicalreport:Virustesting.16AnnexD(informative)Technicalreport:Acanthamoebatesting.17AnnexE(informative)Technicalreport:Artificialtears(organicsoil)inlaboratorytesting.18Bibliography.19SIST EN ISO 14729:2002

ISO14729:2001(E)iv©ISO2001–AllrightsreservedForewordISO(theInternationalOrganizationforStandardization)isaworldwidefederationofnationalstandardsbodies(ISOmemberbodies).TheworkofpreparingInternationalStandardsisnormallycarriedoutthroughISOtechnicalcommittees.Eachmemberbodyinterestedinasubjectforwhichatechnicalcommitteehasbeenestablishedhastherighttoberepresentedonthatcommittee.Internationalorganizations,governmentalandnon-governmental,inliaisonwithISO,alsotakepartinthework.ISOcollaboratescloselywiththeInternationalElectrotechnicalCommission(IEC)onallmattersofelectrotechnicalstandardization.InternationalStandardsaredraftedinaccordancewiththerulesgivenintheISO/IECDirectives,Part3.DraftInternationalStandardsadoptedbythetechnicalcommitteesarecirculatedtothememberbodiesforvoting.PublicationasanInternationalStandardrequiresapprovalbyatleast75%ofthememberbodiescastingavote.AttentionisdrawntothepossibilitythatsomeoftheelementsofthisInternationalStandardmaybethesubjectofpatentrights.ISOshallnotbeheldresponsibleforidentifyinganyorallsuchpatentrights.InternationalStandardISO14729waspreparedbyTechnicalCommitteeISO/TC172,Opticsandopticalinstruments,SubcommitteeSC7,Ophthalmicopticsandinstruments.AnnexesAtoEofthisInternationalStandardareforinformationonly.SIST EN ISO 14729:2002

ISO14729:2001(E)©ISO2001–AllrightsreservedvIntroductionProductsforcontactlensdisinfectionbychemicalmeansareintendedtoreducemicrobialcontaminationintroducedduringlenswearandremoval,cleaningandstorageandarerequiredtocontainantimicrobialagentscapableofachievingthis.Itisessentialthatallliquidcontactlenscareproductsaresterileuntilopened.Dryproducts(tablets,granules,etc.)shouldbesubjecttocontrolofmicrobialcontaminationandshouldbedissolvedinasuitablediluentimmediatelypriortouse.Multidosecontactlenscareproductsmustbeadequatelypreservedorbepackagedinacontainerdesignedandlabelledtominimizetheriskofinjuryresultingfromin-usecontamination.Contactlensesarenormallysubjecttoaregimenofcleaningandcontactlensdisinfectionbetweenperiodsofwear.Aqueoussolutionscontainingcleaningand/ordisinfectingagentsarecommonlyusedforthispurpose.Theseproductsmaybemarketedassolutionsorastabletsfordissolutionimmediatelypriortouseinasuitablediluentsuchassaline.Thepast20yearsofexperienceintheuseandregulationofcontactlensdisinfectingproductshasshowndistinctdisinfectingantimicrobialcriteriaforthisclassofmedicaldevices.Oculartoxicologyconcerns,processconvenienceandproductcomfortontheeye,havemeantanevolutionofproductswhichmaintainalowincidenceofcontactlensassociatedocularinfectionwhenusedasinstructedbythemanufacturer.ThisInternationalStandardgivesthesedistinctcontactlensdisinfectingantimicrobialcriteriaalongwithannexestoexplainwhyviruses(annexC)andAcanthamoeba(annexD)arenotincludedaschallenges.Organicsoilisnotrequiredforevaluationofcontactlenscaredisinfectingproductsbutmaybeused;aninformativeannex(annexE)isincludedtodiscussorganicsoilinthecontextofcontactlensesandcontactlenscareproducts.SIST EN ISO 14729:2002

INTERNATIONALSTANDARDISO14729:2001(E)©ISO2001–Allrightsreserved1Ophthalmicoptics—Contactlenscareproducts—Microbiologicalrequirementsandtestmethodsforproductsandregimensforhygienicmanagementofcontactlenses1ScopeThisInternationalStandardspecifiestwotestmethodsforevaluatingtheantimicrobialactivityofproductstobemarketedforcontactlensdisinfectionbychemicalmeansandforproductsthatarepartofacontactlenscareregimen.ThisInternationalStandardisnotapplicabletothehygienicmanagementoftriallenses.NOTEGeneraldisinfectionproductstandardsarenotapplicabletocontactlenscareproducts,e.g.EN1040:1997andEN1275:1997.2NormativereferencesThefollowingnormativedocumentscontainprovisionswhich,throughreferenceinthistext,constituteprovisionsofthisInternationalStandard.Fordatedreferences,subsequentamendmentsto,orrevisionsof,anyofthesepublicationsdonotapply.However,partiestoagreementsbasedonthisInternationalStandardareencouragedtoinvestigatethepossibilityofapplyingthemostrecenteditionsofthenormativedocumentsindicatedbelow.Forundatedreferences,thelatesteditionofthenormativedocumentreferredtoapplies.MembersofISOandIECmaintainregistersofcurrentlyvalidInternationalStandards.ISO8320-1:—1),Contactlensesandcontactlenscareproducts—Vocabulary—Part1:Contactlenses.ISO8320-2:—1),Contactlensesandcontactlenscareproducts—Vocabulary—Part2:Contactlenscareproducts.3TermsanddefinitionsForthepurposesofthisInternationalStandard,thedefinitionsgiveninISO8320applytogetherwiththefollowing.3.1contactlensdisinfectingproductproductthatpossessescidalactivity(kills,destroysand/orinactivates)meetingtheprimarycriteriaofthestand-alonetestspecifiedinthisInternationalStandard3.2contactlensdisinfectingregimencontactlenscareregimendesignedtomeetboththesecondarycriteriaofthestand-alonetestandtheregimentestasspecifiedinthisInternationalStandard1)Tobepublished.(RevisionofISO8320:1986)SIST EN ISO 14729:2002

ISO14729:2001(E)2©ISO2001–Allrightsreserved3.3contactlensdisinfectionchemicalorphysicalprocesstoreducethenumberofviablemicroorganismsasspecifiedintheperformancerequirementsectionsofthisInternationalStandard4Principle4.1GeneralThestand-alonetestisdesignedtoqualifyindividualsolutionswithasuitablelevelofantimicrobialactivityascontactlensdisinfectionproducts.Theregimentestisdesignedtoqualifyindividualsolutionsaspartofacontactlensdisinfectingregimen.Productsmeetingtheregimentestcriteriashallalsomeettheminimumperformancerequirementsofthestand-alonetest.Itisfundamentalthatsuchproducts(unopenedcontainers)arecapableofmeetingtherequirementsofthetestthroughouttheirlabelledshelflife.AsdescribedinFigure1,contactlenscaresolutionswhicharedesignedtopossessdisinfectingpropertiesshallbetestedinthestand-alonetestfirst.Iftherespectiveprimarycriteriaaremet(see5.1),theproductmaybelabelledasacontactlensdisinfectingproduct.Iftheproductfailstheprimarycriteriaofthestand-alonetest,theproductmustexhibitsufficientantimicrobialactivitytomeetthesecondarycriteriaofthestand-alonetestaslistedin5.2.Ifthesesecondarycriteriaaremet,theregimentestshallbeperformedinordertoqualifytheproductaspartofacontactlensdisinfectingregimenbymeetingtheregimencriteria(see5.3).Iftheproductmeetsboththesecondarycriteriaofthestand-alonetestandtheregimentestbutfailstheprimarycriteriaofthestand-alonetest,itshallbelabelledaspartofacontactlensdisinfectingregimen.Thedesignofcontactlenscareproductsforcleaningandcontactlensdisinfectionshalltakeintoconsiderationtheneedsofpatientcomplianceandtheprobabilityofnon-compliance.Forexample,disinfectingtimemustbeappropriateforcontactlenswear.NOTEUseofmultipleormixedmicrobialchallengescaninfluencetheapparentdisinfectingactivityofaparticularproduct.TheevaluationofthesevariablestogetherwithtestingagainstalargerpanelofmicroorganismsandtestingofsamplesfrompartiallyusedcontainersmaybeofvalueindevelopingacontactlenscareproductbutareexcludedfromthescopeofthisInternationalStandard.(SeeannexesCandD).4.2Stand-alonetest(Inoculumchallengetest)Thestand-alonetestchallengesadisinfectingproductwithastandardinoculumofarepresentativerangeofmicroorganismsandestablishestheextentoftheirviabilitylossatpre-determinedtimeintervalscomparablewiththoseduringwhichtheproductmaybeused.Thesizeofthemicrobialchallengechoseninthistestisnotintendedtoberepresentativeofthelikelychallengeinpracticebuttoprovidecountablenumbersfromwhichestimationoftherateandextentofviabilitylosscanbedetermined.Incarryingoutthetestforantimicrobialactivitythequalitativeandquantitativecompositionsoftheproducthavetobeknownatthetimeoftestingbyeitheranalyticaltestingorextrapolation.Appropriatemeasuresshallbetakentoinactivateorremoveresidualantimicrobialagentsduringculturingandcountingofchallengeorganismsurvivors,andtheeffectivenessofthesemeasuresshallbevalidated.Theactionofthisprocessduringthetestshallbedemonstratedbytheconstructionofsuitablecontrols.NOTEForinformationaboutvirustesting,seeannexC,andforAcanthamoebatesting,seeannexD.SIST EN ISO 14729:2002

ISO14729:2001(E)©ISO2001–Allrightsreserved3aProductmaybelabelledasacontactlensdisinfectingproduct.bProductshallbelabelledaspartofacontactlensdisinfectingregimen.Figure1—Flowchartforstand-alonetestandregimentest4.3RegimentestThisisatestwhichchallengesamultifunctionaldisinfectingregimenwithastandardinoculumofarepresentativerangeofmicro-organismsandwhichmeasurestheviabilitylossafterapredeterminedtimeinterval;theinoculumiscarriedthroughtheregimenbybeingappliedtoacontactlens.Thisprocedureisapplicabletomultifunctionaldisinfectingregimenswhichmayincludethestepsofcleaning,rinsingandsoaking.Incarryingouttheregimentestprocedure,theproductsareusedinthemannerandquantityrecommendedinproductlabellingand/orpatientinstructions.Thedisinfectingstageofanyproposedcontactlensdisinfectingregimenevaluatedbythistestshallsatisfytheminimumrequirementsofthestand-alonetestasdescribedinFigure1.Onlythoseproductsthatsatisfytheminimumperformancerequirementsforthestand-alonetestmaybeincorporatedintoadisinfectingregimen.Incarryingoutthetest,qualitativeandquantitativecompositionofallproductsusedinthetestregimenhavetobeknownatthetimeoftesting,eitherbyanalyticaltestingorextrapolation.Appropriatemeasuresshallbetakentoinactivateorremoveresidualantimicrobialactivityduringculturingandcountingofchallengeorganismsurvivors,andtheeffectivenessofthesemeasuresshallbevalidated.Theactionofthisprocessduringthetestshallbedemonstratedbytheconstructionofsuitablecontrols.NOTEForproblemsassociatedwiththeuseofhuman-wornlenses,seeannexE.5Performancerequirements5.1Stand-alonetest:Primarycriteria(seealsoTable1)5.1.1BacteriaThenumberofeachchallengeorganismrecoveredpermillilitreshallbereducedbyanaveragevalueofnotlessthan99,9%(3,0logs)withintheminimumrecommendedsoakingperiod.SIST EN ISO 14729:2002

ISO14729:2001(E)4©ISO2001–AllrightsreservedNOTEThevalueisdeterminedbytakingtheaverageofthelogreductionsforeachchallengeorganismfortheindividuallotstested.5.1.2MouldsandyeastsThenumberofeachchallengeorganismrecoveredpermillilitreshallbereducedbyanaveragevalueofnotlessthan90%(1,0log)withintheminimumrecommendedsoakingperiodwithnoincreaseatnotlessthanfourtimestheminimumrecommendedsoakingperiodwithinanexperimentalerrorof0,5logs.NOTEThevalueisdeterminedbytakingtheaverageofthelogreductionsforeachchallengeorganismfortheindividuallotstested.5.2Stand-alonetest:Secondarycriteria(seealsoTable1)Productsfailingtomeetthecriteriain5.1.1or5.1.2shallbeevaluatedbytheregimentestproceduredescribedin6.4,providedthesumoftheaveragesisaminimumof5,0logunitsreductionforthethreespeciesofbacteriawithintherecommendedsoakingperiodwithaminimumaverageof1,0logunitreductionforanysinglebacteria.Stasisfortheyeastandmouldshallbeobservedfortherecommendedsoakingperiodwithinanexperimentalerrorof0,5logs.5.3Regimentest:Regimencriteria(seealsoTable1)Foreachmicrobialspecies,theaverageregimenrecoverycount(foralllotstested)shallbenomorethan10cfuforeachlenstype/storagesolutioncombination.Datafrommorethanonelenstypeshouldnotbecombinedtocalculatetheaverage.NOTEWhenqualifyingalenscareproductregimenforusewithonelenstype,theaveragecountforeachspeciesrequiresaveragingthedatafromthe24inoculatedandtreatedlensesoftheonelenstype.Whenqualifyingalenscareproductregimenforusewithmorethanonelenstype,theaveragecountforeachspeciesbylenstypewouldrequireaveragingthedatafromthe12inoculatedandtreatedlensesforeachlenstype.SeeTable4fornumbersoflensestobeused.SIST EN ISO 14729:2002

ISO14729:2001(E)©ISO2001–Allrightsreserved5Table1—SummaryofperformancerequirementscriteriaforcontactlensdisinfectionproceduresAveragelogreductionatsoakingtimeFungiBacteriaTestFSaCAaSMaPAaSAaStand-alonetest:PrimaryCriteria11333Stand-alonetest:SecondaryCriteriabbcccRegimentest:Regimencriteriad4to54to54to54to54to5aPA=P.aeruginosaATCC9027;SA=S.aureusATCC6538;SM=S.marcescensATCC13880;CA=C.albicansATCC10231;FS=F.solaniATCC36031.bStasisatthesoakingtime.cTheminimumacceptablelogreductionforallthreebacteriacombinedis5.Theminimumacceptablelogreductionforanysinglebacterialtypeis1.dEquivalenttoanaverageofnotmorethan10cfuperlenstype/storagesolutioncombination.6Testmethods6.1MaterialsandreagentsThematerialsandreagents(i.e.thetestorganisms,mediaandreagents,equipmentandsamples)arecommontoboththestand-aloneprocedurefordisinfectingproductsandtheregimenprocedureforcontactlensdisinfection.6.1.1TestorganismsThestrainslistedinTable2shallbeused.NOTETestorganismsfromotherculturecollectionsthatmaybeusedarelistedinannexA.Table2—TestorganismsPseudomonasaeruginosaATCC9027StaphylococcusaureusATCC6538SerratiamarcescensATCC13880CandidaalbicansATCC10231FusariumsolaniATCC360316.1.2Culturemediaandreagents6.1.2.1Potatodextroseagar(PDA).SIST EN ISO 14729:2002

ISO14729:2001(E)6©ISO2001–Allrightsreserved6.1.2.2Tryptonesoyaagar(TSA).6.1.2.3SabouraudDextroseAgar(SDA).6.1.2.4Dulbecco'sPhosphateBufferedSaline,withoutcalciumchlorideandmagnesiumchloride(DPBS):200mg/lKCl,200mg/lKH2PO4,8000mg/lNaCl,and2160mg/lNa2HPO47H2Oorsuitablediluent.6.1.2.5Dulbecco'sPhosphateBufferedSaline,plus0,05%(mass/volume)polysorbate80(DPBST)orsuitablediluent.6.1.2.6Validatedneutralizingagents/mediaasrequired,forexample,Dey-EngleyNeutralisingBroth(DEB)andLetheenBroth2).6.1.3TestequipmentThefollowingcommonlaboratoryequipmentisrequired.6.1.3.1Sterilepipettes.6.1.3.2Swabs.6.1.3.3Tubes.6.1.3.4Petridishes(90mmto100mm20mm).6.1.3.5Incubator.6.1.3.6Spectrometer,fordeterminationofcelldensity.6.1.3.7Instrumentforcolonycounting.6.1.3.8Centrifuge.6.1.4TestsamplesTheproducttobetestedshallberepresentativeoftheproducttobemarketed.Aliquotsshouldbetakendirectlyfromthefinalproductcontainerimmediatelypriortotesting.Threelotsofproductshallbetested.Eachlotofproductshallbetestedwithaseparateinoculumpreparationforeachchallengeorganism.6.1.5CulturemaintenanceMaintainthetestculturesasrecommendedbythecuratoroftheappropriateculturecollection.Culturesshouldbenogreaterthan5passesremovedfromthedepositorystock(ATCC,NCIB,NCTC,NCPForotherrecognizedculturedepository;seeannexA).Eachpassisasubcultureofthepreviouspass.6.2Preparationofmicrobialchallenge(Inoculum)Thepreparationofthemicrobialchallengeorganisms(inoculum)iscommontoboththestand-aloneprocedurefordisinfectingproductsandtheregimenprocedureforcontactlensdisinfection.2)Dey-EngleyNeutralisingBroth(DEB)andLetheenBrothareexamplesofsuitableproductsavailablecommercially.ThisinformationisgivenfortheconvenienceofusersofthisInternationalStandardanddoesnotconstituteanendorsementbyISOofthisproduct.SIST EN ISO 14729:2002

ISO14729:2001(E)©ISO2001–Allrightsreserved7Fortheregimenprocedureforcontactlensdisinfection,organicsoilmaybeincludedaspartoftheinoculum.SeeannexEforanexample.CultureeachtestorganismonagarslopesundertheconditionsgiveninTable3.Table3—MediaandincubationconditionsforgrowthofchallengeorganismsOrganismMediumTemperature°CIncubationtimeP.aeruginosaTSA30to3518hto24hS.aureusTSA30to3518hto24hS.marcescensTSA30to3518hto24hC.albicanxSDA20to2542hto48horC.albicansSDA30to3518hto24hF.solaniPDA20to2510dto14dUsesterileDPBSTorsuitablediluenttoharvesteachculture;washthesurfacegrowth,transferittoasuitablevesselandvortex.FiltertheF.solanisuspensionsthroughsterileglasswool,cheeseclothorgauzetoremovehyphalfragments.Afterharvesting,theculturedorganismsmaybewashedusingcentrifugation.Thebacterialsuspensionsmaybefiltered(e.g.3µmto5µmporesize)toproduceasinglecelldispersion.ThenadjustallchallengecellsuspensionswithDPBSTorothersuitablediluenttoaconcentrationofbetween1107cfu/mland1108cfu/ml.Estimatetheapproximatecellconcentrationofeachsuspensionbymeasuringtheturbidityofthesuspensionoradilutionofthesuspensionusingaspectrophotometer.Theactualconcentrationofcolonyformingunitspermillilitreshallbedeterminedforeachsuspension,e.g.bytheplatecountmethod,atthetimeofthetest.Ifacentrifugeisused,eachcentrifugationshouldbeconductedat20°Cto25°Cfornolongerthantheequivalentof10minat4000gorless.Longercentrifugationtimesmayberequiredatlowerspeeds.Usebacterialandyeastcellsuspensionsonthedayofpreparation.Sporesuspensionsmaybeuseduptosevendaysfollowingpreparationifstoredunderrefrigeration(2°Cto8°C).6.3Stand-aloneprocedure6.3.1Inoculumchallengetestprocedure6.3.1.1Prepareoneormoretubes(foreachlottested)containingaminimumof10mloftestproductsolutionperchallengeorganism.NOTESampletubesareusedratherthanlenscasestoalloweffectivetechnicalexecutionofthetest.Sinceincompatibilitiescanexistbetweensolutioningredientsandtubematerials,tubesofanappropriatematerial,whicharecompatiblewiththeingredients,shouldbeused.Inoculatethesampletubeoftheproducttobetestedwithasuspensionoftestorganismssufficienttoprovideafinalcountofbetween1,0105and1,0106cfu/ml.Ensurethatthevolumeofinoculumdoesnotexceed1%ofthesamplevolume.Ensurecompletedispersionoftheinoculumbyadequatemixing.6.3.1.2Storetheinoculatedproductat20°Cto25°C.Thetemperatureshallbemonitoredusingacalibrateddeviceandthetemperaturedocumented.Iftheproductissensitivetolightitshouldbeprotectedduringtheperiodofthetest.SIST EN ISO 14729:2002

ISO14729:2001(E)8©ISO2001–Allrightsreserved6.3.1.3Take1,0mlaliquotsoftheinoculatedproductfordeterminationofviablecountat25%,50%,75%and100%oftheminimumrecommendeddisinfectingtimeforallorganisms,and,inaddition,notlessthan400%oftheminimumrecommendeddisinfectingtimeforyeastandmould.Ifovernightcontactlensdisinfectionisrecommended,useasoakingtimeof8h.6.3.1.4Subjecteachofthe1,0mlaliquots,removedatthespecifiedtimeintervals,toasuitableseriesofdecimaldilutionsinvalidatedneutralizingmedia.Mixthesuspensionwellbyvortexingvigorouslyandletstandtoallowneutralizationtobecompleted.Neutralizationconditionsshallbebasedonrecoverymediumcontroltesting(see6.3.2.2).Ifanantimicrobialagentintheformulationcannotbeadequatelyinactivatedorneutralized,eliminateitusingavalidatedmembranefiltrationprocedure(seeannexB).6.3.1.5Determinetheviablecountoforganismsinappropriatedilutionsbypreparationoftriplicateplates(unlessotherwisejustified)ofasuitablerecoverymedium(e.g.TSAforbacteriaandSDAformouldandyeast).Ifmembranefiltrationhasbeenemployedtoremoveorneutralizeantimicrobialagents,culturethemembranesonthesemediaasappropriate.Ifthepourplatemethodisutilized,keeptheagarforpourplatesbelow50Cpriortopouring.Theagarmediausedfordeterminationofviablecountsmayalsocontainantimicrobialinactivatorsorneutralizers,ifrequired.6.3.1.6Incubatebacterialrecoveryplatesat30Cto35C.Incubateyeastrecoveryplatesat20Cto25Cor30Cto35C.Incubatemouldrecoveryplatesat20Cto25C.Incubationtimesforoptimalrecoveryofbacteria,yeastandmouldsshallbedetermined.Minimumincubationtimesshallbebasedonrecoverymediumcontroltesting(see6.3.2).Recordthenumberofcfuobservedoncountableplates.NOTEPlatesshouldbeobservedperiodicallyduringincubationtopreventtheoccurrenceofuncountableplatesduetoovergrowth.6.3.1.7Determinetheaveragenumberofcolonyformingunitsoncountableplates.Calculatethemicrobialreductionatthespecifiedtimepoints.NOTECountableplatesreferto30cfu/plateto300cfu/plateforbacteriaandyeast,and8cfu/plateto80cfu/plateformoulds,exceptwhencoloniesareobservedonlyforthe100or10-1dilutionplates.6.3.1.8Theabsenceofmicroorganismsshallbedocumented,e.g.byrecordingan"0”or"NR”(norecovery),whenplatesforalldilutionsofasampleatasingletimepointhavezerocolonies.6.3.2Controls6.3.2.1InoculumControlPrepareaninoculumcountbydispersinganidenticalaliquotoftheinoculumintothesamevolumeasusedin6.3.1.1ofasuitablediluent(e.g.DPBST)toachieveafinalconcentrationof1,0105cfu/mlto1,0106cfu/ml.Ensurethatthevolumeofinoculumdoesnotexceed1%ofthesamp
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