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GuidanceonValidationandQualificationofProcessesandOperationsInvolvingRadiopharmaceuticals

Abstract ThisdocumentismeantasanAppendixofPartBoftheEANM“GuidelinesonGoodRadiopharmacyPractice (GRPP)” issued by the Radiopharmacy Committee of the EANM(http://www.eanm.org/publications/guidelines/gl_radioph_cgrpp.pdf),coveringthequalificationandvalidationaspectsrelatedtothesmall-scale“inhouse” preparationofradiopharmaceuticals.Theaimistoprovidemoredetailedandpractice-orientedguidancetothosewhoare involved inthesmall-scale preparation of radiopharmaceuticals which are not intended for commercial purposes ordistribution.

Abbreviations ART ActivityReferenceTimeCFU ColonyFormingUnitCLV CleaningValidationCPP CriticalProcessParametersCPV ContinuousProcessVerificationCV CoefficientofvariationDQ DesignQualificationEANM EuropeanAssociationofNuclearMedicineEMA EuropeanMedicineAgencyEtOH EthanolEU EuropeanUnionFAT FactoryAcceptanceTestingFID FlameIonisationDetectorFDA USFoodandDrugAdministrationGAMP GoodAutomatedManufacturingPracticeGC GasChromatographyGMP GoodManufacturingPracticecGRPP CurrentgoodradiopharmacypracticeHEPA HighEfficiencyParticulateAirHPGe HighPurityGermaniumHPLC High-performanceliquidchromatographyHVAC Heating,VentilationandAirConditioningICH International Conference on Harmonization of technical requirements for

pharmaceuticalsforhumanuseIMPD InvestigationalMedicinalProductDossierIQ InstallationQualificationISO InternationalOrganisationforStandardizationLOD LimitofdetectionLOQ LimitofquantitationMDA MinimumDetectableActivityOQ OperationalQualificationsPET PositronEmissionTomography

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PIC/SScheme ThePharmaceuticalInspectionConventionandPharmaceuticalInspectionCo-operationScheme

Ph.Eur EuropeanPharmacopoeiaPLC ProgrammableLogicControllerPQ PerformanceQualificationQC QualitycontrolR2 CoefficientofDeterminationRP(s) Radiopharmaceutical(s)Rs ResolutionFactorRSD RelativeStandardDeviationSAT SiteAcceptanceTestingSD StandardDeviationSOP StandardoperatingprocedureSPC SummaryofproductcharacteristicsSPE SolidphaseextractionSPECT SinglePhotonEmissionComputedTomographySSRP Small-Scale“in-house” RadiopharmaceuticalTLC ThinLayerChromatographyTSB TrypticSoyBrothUPS UninterruptiblePowerSupplyURS UserRequirementsSpecificationUV UltravioletVMP ValidationMasterPlanWHO WorldHealthOrganization

Definitions Goodradiopharmacypractice

Good radiopharmacy practice is described in the “Guidelines on Current Good RadiopharmacyPractice (cGRPP)” issued by the Radiopharmacy Committee of the EANM (seehttp://www.eanm.org/publications/guidelines/gl_radioph_cgrpp.pdf).

Radiopharmaceutical

A radiopharmaceutical (RP) is anymedicinal product which, when ready for use, contains one ormoreradionuclides(radioactiveisotopes)includedforamedicinalpurpose.

Small-scaleradiopharmaceutical

Asmall-scaleradiopharmaceuticalisanyin-houseradiopharmaceuticalnotintendedforcommercialpurposes or distribution prepared on a small scale (for PET, SPECT or therapeutic applications),including extemporaneous preparations based on generators and kits, and excluding simple kitpreparationbasedonSPC.

Small-scaleradiopharmacy

Asmall-scaleradiopharmacyisafacilitywherethesmall-scalepreparationofradiopharmaceuticalsiscarried out in accordance with national regulations. The term small-scale radiopharmacy is not

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related to thephysical sizeof the facility, butonly to thekindof radiopharmaceuticalpreparationperformed.

Preparation

Preparation includesalloperations involved inthepurchaseofmaterialsandproducts,production,QC,releaseandstorageofamedicinalproduct.

Finishedproduct

Afinishedproductisamedicinalproductwhichhasundergoneallstagesofproduction,includingQCandproduct/batchrelease,packaginginitsfinalcontainerandproperlabelling.

Automatedmodule

Anautomatedmoduleisadeviceabletoperformautomaticallyasequenceofoperationsneededinthepreparationofradiopharmaceuticals.Anautomatedmodulecanbecommercialorcustommade.Itconsistsoftwoassembledparts:amechanicalpartandachemistrypart.

The mechanical part consists of an assembly of electric and/or pneumatic, linear and/or circularactuators, power supplies, pumps, coolers, heaters, sensors for monitoring different parameters(suchastemperature,pressure,flow,radioactivity)oranyotherphysicaldevicenotindirectcontactwithchemicals.

Thechemistrypartisaninterconnectednetworkofcontainersinwhichgaseous,liquidand/orsolidreagentsandcomponentscanbemoved,mixedand/ortransformedtoobtainthedesiredproduct.Themechanicalpartandthechemistrypartareconnectedtoeachother.Thechemistrypartcanbepermanent(non-disposabledevice)orsingleuse(disposabledevice,or“cassette”).

SOP

SOP,orStandardOperatingProcedure(s)aredocumentswhichprovide instructions, ina clearandconciseform,toperformaspecifictask.Theydealwithalltheoperationsandstepsinvolvedinthelifecycleofthepreparationofaradiopharmaceutical.

Validation

Validation is the action of proving that any procedure, process, equipment, material, activity orsystemactuallyleadstotheexpectedresults(1).

Qualification

Actionofprovinganddocumentingthatanypremises,systemsandequipmentareproperlyinstalled,and/orworkcorrectlyandleadtotheexpectedresults.Qualificationisoftenapart(theinitialstage)ofvalidation,buttheindividualqualificationstepsalonedonotconstituteprocessvalidation(1).

Validationprotocol

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A documentwhich contains all the information required to perform the validation of an intendedinstrument/method/process.

DesignQualification(DQ)

DQ is aimed to verify that the system / instrument has been designed suitably for the intendedpurpose.Inparticular:

- thedesignmeetstheuserrequirementspecification(URS);- thedesigncomplieswithalltheapplicableguidelinesandstandards- thedesigncomplieswiththevalidationmasterplan(VMP)

InstallationQualification(IQ)

IQisaimedtoverifythatthefacility/system/instrumenthasbeeninstalledcorrectly,basedonthemanufacturer’srecommendationsand/ortheapprovedspecificationsoftheUser.

OperationalQualification(OQ)

OQ is aimed to verify that the facility / system / instrument are operating properly, and that theresponseofcriticalcomponents(e.g.sensors)matchwiththeintendedvaluesandwithinthedesiredrange.

PerformanceQualification(PQ)

ThegoalofPQisverifythatthefacility/system/instrumentperformproperlyandreproduciblyinthe intended routine conditions set for the specific preparation process, and using approvedmethods.

CleaningValidation

Cleaning validation has the purpose to demonstrate that the cleaning of a facility / system /equipment,orthatpartsofitwhichcomeintocontactwiththefinishedproductorwithreagents/solvents during the preparation process, is suitable for the intended purposes, and that residues(chemical,radiochemical,microbiological,cleaningagents)areremovedbelowadefinedlevelbythecleaningprocedure.

ProcessValidation

ProcessValidation (PV)has tobe intendedas amean toestablish that all theprocessparametersthatbring to thepreparationof the intendedRPs and their quality characteristics are consistentlyandreproduciblymet.

ValidationSummaryReport(VSR)VSRisthefinaldocumentthatsummarizesthewholeprotocolresultsandcomments/opinionsabouttheirsuitability.UserRequirementSpecification(URS)

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A set of specifications, thatmay be related to production/QC equipment, aswell as to thewholefacilityorpartsofitsuchasutilitiesorsystems/sub-systems,definedbytheUserandthatrepresentausefulreferenceforthetheirdesignand/orpurchase,andduringthevalidationactivities.ValidationMasterPlan(VMP)VMP is a general document that summarizes validation policy and all the intended validation /qualificationactivities,togetherwithadescriptionofthefacilityandorganisationalstructure.

Generalitiesandscope The present EANM guidance covers the qualification and validation aspects intertwined with thepreparation of small-scale radiopharmaceuticals. It concerns the preparation ofradiopharmaceuticalswhicharenotintendedforcommercialpurposesordistribution.

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Summary

Abstract...............................................................................................................................................1

Abbreviations......................................................................................................................................1

Definitions...........................................................................................................................................2

Generalitiesandscope........................................................................................................................5

1.GeneralPrinciples...............................................................................................................................7

2.ValidationMasterPlan........................................................................................................................7

3.Documentation...................................................................................................................................9

4.Userrequirementspecifications.........................................................................................................9

5.Qualificationofequipment...............................................................................................................11

5.1Qualificationofproductionequipment.......................................................................................12

5.2QualificationofQCinstrumentation...........................................................................................16

6.Validationofanalyticalmethods.......................................................................................................19

7.Computerisedsystems......................................................................................................................21

8.Validationofclassifiedrooms...........................................................................................................22

9.CleaningValidation............................................................................................................................24

9.1Validationoftheprocedureforthecleaningofproductionequipmentinternalsurfaces..........24

9.2Validationoftheprocedureforcleaningexternalsurfaces.........................................................26

10.Processvalidation............................................................................................................................27

11.ValidationofasepticoperationsviaMediafill................................................................................28

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1.GeneralPrinciplesValidation is the action of proving that any procedure, process, equipment, material, activity orsystemactuallyleadstotheexpectedresults,withtheaimtocontributetoguaranteethequalityofa(radio)pharmaceutical.Theconceptofqualificationisverysimilartothatofvalidation,butwhiletheformer ismoregeneral and reliesonabroad rangeofactivities, the latter ismore“practical”andindicates the actions andoperations aimed todemonstrate that a system / equipment is properlyinstalled,workscorrectlyandleadstotheexpectedresults.Qualificationmaybeconsideredasapartofvalidation.GeneralPrinciplesonValidationandQualificationareoutlined indifferent importantreferencedocuments,themostimportantandrelevantofwhich,forprofessionalsoperatingwithintheEuropeanUnion,istheAnnex15(2)ofGoodManufacturingPractice(GMP)guidelines,thatapplyto the manufacturing of medicinal products aimed to obtain a Marketing Authorization, and ingeneraltothosewhoarerequestedtocomplywithGMP.Annex15hasbeenrecentlyrevised,andmost recent version came into operation on 1st October 2015. Other useful guidelines have beenreleased by Institutions such as World Health Organization (WHO)(1) or the US Food and DrugAdministration(FDA)(3),orevenbyinstrumentationsuppliers(4),thelatterbeingusuallyaddressedtospecific proprietary technology, while the former are typically conceived as general guidanceprinciples for industry. Although principles described in the above documents are generallyapplicable to any process, equipment, system or facility, their practical implementation in thepreparation and quality controls of radiopharmaceuticals may require adaptations that meet thepeculiar nature of the RPs themselves and of the equipment used for their preparation. Anotherimportant issue related to the validation concept is the validation of analytical methods, whosegeneral principles are outlined in ICH Q(2) R1 – Note for Guidance on validation of analyticalprocedures:textandmethodology(5),whichdefinethetypeofanalyticalmethodstobevalidatedandsetparametersofconcernandacceptancecriteriatobeconsidered.Thesameconsiderationsstatedabove apply: ICH guidelines are very general and capable to embrace a broad range of analyticalprocedures, including those procedures specifically developed for the quality control ofradiopharmaceuticals; however, the intrinsic nature of radioactivity, which decreases with timefollowingthedecaylaw,andthephysicalcharacteristicsofthedetectionofradioactivity,promptforspecific validation protocols. Only a brief, general description of the principles of validation ofanalyticalmethodswillbegiveninthistext;indeed,duetothecomplexityandvarietyoftheinvolvedprocedures, instrumentation, etc., they will be the subject of a separate, dedicated guidancedocument.Ithastobeunderlinedherethatvalidationmayultimatelybeconsideredasausefulwaytoincreasereliability and prevent deviations and out of specification in the day by day operation in theradiopharmaceuticalpreparationprocess,as it isaimedtoguaranteethatprocesses/procedures/equipmentworkcorrectlyandleadtotheexpectedresults.Asstatedabove,theaimofthisguidelineistoprovidemoredetailedandpractice-orientedguidancetothoseprofessionalswhoareinvolvedinthesmall-scalepreparationofradiopharmaceuticals,notintendedforcommercialpurposesordistribution.

2.ValidationMasterPlanValidation activities should be planned in a validation plan, in an ordered manner. For instance,processvalidationshouldbeperformedafterthevariousproductionandqualitycontrolequipment

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havebeenqualified,andnotviceversa.Moreover,validationactivitiesshouldbeconsideredasanintegralpartofthequalityassurancesystem,andshouldthusbedocumentedinordertoguaranteethe necessary traceability. To this regard, the overall validation activities should be described in ageneraldocument,usuallyknownasValidationMasterPlan(VMP).VMPshouldinclude:i) a general validation policy, with a description of the intended working methodology, and

factorsthatmayaffectthequalityoftheintendedradiopharmaceutical(s);ii) adescriptionofthefacility,withadetaileddescriptionofthecriticalpoints;iii) adescriptionoftheradiopharmaceuticalpreparationprocess(es);iv) thelistofproductionequipmenttobequalified,includingtheextentofqualificationrequired

(e.g. IQ, OQ and PQ vs PQ only), and indications about equipment defined as critical (e.g.dispensingsystemworkinginaclassAenvironment);

v) a list of the quality control equipment to be qualified, including the extent of qualificationrequired;

vi) a list of other ancillary equipment, utilities, system and/or facilities to be qualified (e.g.Heating,VentilationandAirConditioning,orHVAC,or thegasdistributionsystem), includingtheextentofqualificationrequired;

vii) thegeneralpolicyrelatedtoprocessvalidation;viii) analytical methods to be validated; generally only thosemethods which are different from

EuropeanPharmacopoeia(Ph.Eur.)methods, forwhichonlypartialvalidation,performedbytesting the most significant parameters (e.g. in case of a method for evaluation ofradiochemicalpuritytheymaybelinearityandreproducibility),isusuallyrequired;

ix) cleaningvalidationofpremisesandequipment;x) thegeneralpolicyonre-validation,indicatingfrequencyandconditionsforre-validation;xi) thepersonnelinvolvedinthevariousactivities,andadefinitionofresponsibilities;xii) a general change control and deviation policy, to be applied to all the involved protocols,

aimed to specify how and when actions are required in case e.g. of test failures or anacceptancecriteriaisnotmet.

Itisimportanttonotethatvalidation/qualificationmayrepresentasignificant“burden”,intermsoftherequiredtime,personnelandfinancialresources,whichareproportionaltothecomplexityofthepreparation process(es); this means that in case the Facility is dedicated to the preparation ofdifferent radiopharmaceuticals, to be used for different clinical purposes, and multiple hot cells,automated systems and analytical equipment are used, an inadequate planning of validationactivitiesmayleadtoanunnecessaryworkloadandhighcosts.Thus,it isofparamountimportancetoclearlydefineintheVMPwhatithastobevalidated,theextentofvalidationrequiredforeachfacility / system / equipment / analyticalmethod, the actions to be taken in case of a significantchange (e.g. the replacement of a production / quality control instrument with a different one)togetherwiththeconditionsforre-validation/re-qualification.VMPshouldbeperiodicallyreviewed,especially in the light of the need for re-validation, and risk assessment methodology should beappliedtotakescientificallysounddecisions.AusefulreferenceinthepreparationofVMPisthedocumentissuedbyPIC/S“RecommendationonValidationMasterPlan….”(6),butmanyotherresourcesmaybeeasilyfoundontheweb.

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3.DocumentationValidation/qualification activities should be documented. Validation/qualification protocols shouldincludegeneralinformationsuchas:

- A general statement on validation policy, with a description of workingmethodology andwhichvalidationstageistobeperformed;

- adescriptionoftheinstrument/process/step/analyticalmethodtobevalidated;- a list of the key personnel involved in the validation activities, including their individual

trainingprogramandacleardefinitionoftheirresponsibilities;- incaseofqualificationofaninstrument/equipment,acleardescriptionofthetest(s)tobe

performed (e.g. a pressure / temperature / radioactivity measurements), together withacceptancecriteria;

- In caseof validationofanalyticalmethods:amatrixwith theparameters tobeconsidered(e.g. linearity, accuracy, etc.), together with statistical functions (e.g. linear regression,standarddeviation(SD),etc.)usedtomakecalculationsandrelatedacceptancecriteria;

- the reference equipment used during the above test (e.g. calibrated gauge, thermometer,activimeter,etc.),andrelatedcalibrationcertificate,ifapplicable;

- areferencetootherrelevantdocumentation(e.g.SOPs,instructionmanuals,etc.);- therawdataobtainedduringtheexecutionoftheexperimentaltests;- a listof thedeviations (if any)encounteredduring theexecutionof theprotocol, together

with a discussion about their possible impact on the considered instrument / process/operational step, and preventive / corrective actions, if applicable, which may provideuseful suggestions to e.g. modify SOPs and operating protocols in general, prompt forpossibleequipmentfailuresandallowformonitoringrisksinherenttotheintendedsystems/processes.

- a final summary,where results are commented, anda final statement about the intendedprocess/instrument/method(theValidationSummaryReport).

Validation protocols should be numbered following a logical order, using appropriate codesdependingonthevalidationextent.ForinstanceapossiblecodemightbePQ-yyyy-nnn,where:

- PQisacronymofPerformancequalification- yyyyindicatestheyeartheprotocolisperformed- nnnisaprogressivenumber

PossiblytheaboveinformationcouldbecodedinasuitableSOPorstatedintheVMP.At leastthemost significant information, such as test approval or rejection, as well as comments related topossibledeviations,shouldbehandwritten.In order to meet the necessary traceability, general quality assurance policy for documentationapply;forinstance,typeorhandwritingerrorsshouldneverbefullyblurredorcancelled,butrathersimplymarkedwithathickline,andupdatedinformationshouldbehandwritten,datedandsigned.

4.UserrequirementspecificationsUser Requirement Specifications (URS) may be considered as the first step in the validation“flowchart”,whichcanbesummarizedinthefigurebelow:

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Fig.1TheV-shapeddevelopmentmodel

TheaimofURSistosetparametersandrelatedperformanceconsideredbytheUserassuitabletoconsiderthesystem/equipmentacceptable.URSshouldinclude:

- a description of the process /method to be carried outwith the specific equipment (e.g.whichkindofanalyticalproceduresaretobeperformedwithanHPLCsystem,theexpectedimpurities,andtherelatedlimits,etc.);

- a detailed description of the intended instrument / equipment including computerizedsystems,ifapplicable;

- a brief description of the room / environment where the instrument / equipment issupposedtobeinstalled;

- whichutilities(e.g.gassupply,electricity,etc.)arerequiredbythesystem/instrument,andwhichonesareavailableintheconsideredaboveroom/environment;

- required performance of the instrument / equipment (e.g. eluent flow range for an HPLCpump,sensitivityforUV(ultraviolet)detector,etc.);

- documentation to be requested to themanufacturer (e.g.manuals, drawings, schematics,etc.);

- otherusefulinformationsuchasserviceandtechnicalsupport.

A fewexamplesmayhelp tounderlineandclarify theabovestatement.AnURS foranautomatedsystem for the preparation of RPs could include a list of significant parameters to bemet by thesystemitself,suchasradiochemicalyield,preparationtimeandlevelofacceptableknownimpuritiesfor a specificRP (e.g. [18F]FDG); general specifications could include thedesignof the system (e.g.cassettesystemvsre-usablesystems), thepurificationprocess(e.g.semi-preparativeHPLCvsSPE),thepresenceof“built-in”additional features (e.g.asub-systemfortheautomatedperformanceoffilter integrity test, etc.), and software specifications (e.g. the possibility to modify automated

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sequencesthatdrivethepreparationofRPsorthepossibilitytodefinesoftwareaccessandrelatedprivileges).

Followingthesameabovedescribedprinciples,aURS fora radio-HPLCcould includespecificationsforsensitivityofradiochemicaldetectors,and/oraccuracy,sensitivity,etc.,forUVdetectors,aswellasspecificationsforsoftwareuseraccess,audittrailpolicy,andmore.

URS are of the utmost importance in case the intended system / equipment is not commerciallyavailable, and it has to be specifically designed. An example is represented by the Heating,VentilationandAirConditioning (HVAC) system,which isusually tailored to theneedsof theUser(e.g.airtreatmentunits,aswellasthesizeoftheairconduits,willbechosenbasedontherequestedlevelof“GMP”classificationoftheenvironments,thesizeandvolumeoftheclassifiedrooms,etc.),andwhosedesignhastobespecificallyadaptedtothelocalbuildinglayout.Anotherexamplecouldbe the need to have custom made hot cells, specifically designed for non-standard research orproductionpurposes, thatmayrequireadditionalshieldingor larger internalworkingareas. Intheabovesituations,URSareclearlytobeconsideredasthefirststepinthe“V-shaped”diagrams,andtheyarethebasisfordesignqualification.

URSarealsoparticularlyusefulincaseofinvitationtotenderprocedures,wheretheymayrepresentthebasisfortenderofficialdocumentation,buttheyaregenerallyconsideredasausefulreferencedocumenttodefinetheintendeduseoftheinstrumentandrelatedacceptancecriteria.

5.QualificationofequipmentWiththeterm“equipment”,ithastobeintendedalltheinstrumentationwhichisdirectlyinvolvedinthepreparationandqualitycontrolofradiopharmaceuticals.Theirfunctions,andgeneralprinciplestobeaccountedfor,willbedescribedinthefollowingtwoparagraphs,dedicatedtotheequipmentforproductionandqualitycontrol,respectively.Althoughcyclotronsandnuclearreactorsare,strictlyspeaking,directly involved in thepreparationofanessential ingredient, the radionuclide, theywillnotbe coveredby thepresent guidelines,which is also in agreementwithAnnex3 –GMP(7), thatconsider this important step in the preparation of RPs as a “non-GMP” step, and as such it’s notrequested to be described and justified by the radiopharmaceutical manufacturers. There arepractical reasonsbehindtheabovechoice, that take intoaccountthecomplexityandmulti-taskingintrinsic nature of the radionuclide production equipment/infrastructures. More important, thequality of produced radionuclide(s) is carefully controlled, thus indirectly ensuring that theequipmentisworkingproperlyanditisproducingtheintendedradionuclideinproperamountsandquality.Anothergeneralcommentisrelatedtothesoftwaresystems,thatareintegralpartsofmostoftheproductionandQCequipment,todate.Theyoftenplayacriticalrole,performingthefollowingtasks:

- instrumentationcontrol (e.g. runningaRPpreparationoradispendingsequence incaseofautomatedradiosynthesisordispensingsystems,regulatingHPLCpumpflowrateorhandlingthesampleinjectionsincaseofradio-HPLC,etc.);

- acquisitionofdatacomingfromsensors/detectors;- logging/storageoftheacquireddata;

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- processingoftheacquireddataandcreatingreports;- handleandassuretraceabilitythroughaudittrailfunctions;- ensuresafetyandreliabilityofthepreparations.

For theabove reasons,aparagraphwillbespecificallydedicated to thevalidationof softwareandcomputerised systems, although reference will also be given when necessary throughout thediscussiononvalidationofequipment.Finally, it is very important to note that, as said above, qualification protocols are aimed todemonstrate that a system / equipment is properly installed, works correctly and leads to theexpected results. This means that the successful outcome of a qualification protocol allows theequipment to be routinely used for the preparation / QC of radiopharmaceuticals, but do noteliminatetheneedforperiodictestingoftheinstrumentationthroughouttheirlifecycle.Thetypeofperiodic tests, their recommended frequency and responsibilities are specific for each intendedequipment,andtheyareusuallypartofthegeneralqualityassuranceprogrammes,thatshouldbeinplace in every radiopharmacy.Often they include tests alreadyperformedduring theexecutionofqualification protocols, but that need to be periodically repeated to verify and ensure the correctfunctionalityof the intendedequipment.Although theirdetaileddescription isoutof the scopeofthepresentdocument,usefulreferencewillbeprovidedinthefollowingparagraphs,especially(butnot only) for the routine quality control testing of radioactivity detection and measurementinstruments,suchasdosecalibrators,radio-HPLC“flow”detectorsandgammaspectrometers.

5.1QualificationofproductionequipmentEquipmentusedinthepreparationofRPsusuallyinclude:i)radiosynthesissystem,whichareoften,butnotnecessarily,fullyautomated;ii)dispensingsystems,whichareoften,butnotnecessarily,fullyautomated;iii)suitablyshieldedhotcells,whereradiosynthesisanddispensingsystemsarelocated,forradiationprotectionpurposes; telepliersandmanipulatorsaresometimeused inthosesystemsnotequippedwithfullyautomateddevices;iv)hotcells/isolatorsformanualpreparationofRPs(e.g.these are frequently used in the preparation of Tc-99m labelled kits or in cell labelling); v) dosecalibrators.Otherinstrumentsoraccessoriesmaybeused,buttheywillnotbeconsideredindetailby thepresent guidelines.On theotherhand, the sameprinciples andmethodologies thatwill bedescribed for the typical equipment also apply to less frequently used instruments. It has to beconsideredthatproductionequipmentcomplexityrangefromrelativelysimpleinstruments,suchasdose calibrator, to more complicated devices such as automated systems for radiosynthesis ordispensing. Qualification activities should be focused on the most critical components, selectedevaluatingthepossibleeffectoffailureormiscalibrationonthegeneralperformanceofthesystemand,inturn,onthequalityandsafetyofthedesiredRPproducts.Installation of production equipment may be preceded by additional evaluation steps, namelyFactoryAcceptanceTesting(FAT)andSiteAcceptanceTesting(SAT).FAT/SATareparticularlyusefulincaseofcomplexand/orbulkyequipment,orwhentheintendedinstrumenthasbeenspecificallydesigned,basedonURS.Agoodexample is representedbyhot cells,whetherornot they includeautomated systems. Hot cells are indeed bulky, and sometimes require some degree ofcustomization, especially in case they have to be used for non-standard processes. During FAT,functionalityofmajorcomponents (buttons, fan, filters,etc.)maybetested,aswellasparameters

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typicallyverifiedduringOQsuchasairvelocity, leaktightnessorparticlecontamination.Theabovetests might help to reveal possible malfunctions or deviations, that may be fixed directly at theFactory,beforetheshipment.FATareusuallyrepeatedonSite(SAT),andtheycanbeconsideredaspartofthewholequalification.a)Radiosynthesissystem“Initialqualificationandperiodicqualificationshouldbeplannedinthemasterdocumentdescribingeach automatedmodule. Initial qualification should include IQ,OQ and PQ. IQ should include theverification of the designedmodule specifications, the check of installed instrumentation and theintegration ofworking andmaintenance instructions in themaster document of themodule. Thefunctionalities of the automated module without reagents nor chemical components should becheckedduringOQ.Ifthemoduleisacommercialone,theusershouldaskthesuppliertoperformaqualification according to internal procedures or to propose a procedure to be performed by theuser.Ifthemoduleiscustommade,theusershouldcheckthatallfunctionalities,definedintheURSdocument, meet the specifications included in the master document describing the module. Thisshould include themovement of actuators and the calibration status of the probes (temperature,pressure,andradioactivity).PQof themoduleshouldbeconductedbyperforming threecompleteruns of a representative process covering all normal operations for the concerned preparationprocess.Forexample,amoduleincludingapreparativechromatographicsystemshouldbevalidatedselecting a RP preparation process which includes a chromatographic purification. PQ shoulddemonstratethatthemoduleissuitablefortheintendedapplicationinrealconditionsofuse.

Each automated module should follow a programme of periodic qualifications of the probes(temperature,pressure,andradioactivity)inordertore-calibratethemifneeded.Formajorupdatesor repairsof themechanicalpart, or in caseofmajormodificationsof the control software, a riskassessmentshouldbeperformedinordertoevaluatethepotentialimpactontheprocessperformedwith the module. OQ and PQ should eventually be performed as conclusions of the riskassessment”(8).

b)Dispensingsystems

Generalprinciplesoutlinedforradiosynthesisdevicesalsoapplyfordispensingsystems,whichallowsfor vial or syringe dispensing starting from a “mother” solution prepared using the automatedradiosynthesismodule.IQshouldinclude:i)averificationofthedocumentation,suchasinstructionmanuals,drawings,electrical/pneumaticschematics,specifications,certificates; ii)averificationofthemajorinstalledcomponents,suchasvalves,tubing,software,programmablelogiccontroller(PLC)orothersuitablecontrol,pumps,balances,controlpanels,PC,softwareversion,etc.Verification isaimedtocheckthatcomponentsareinstalledinaproperway,bycomparisonwithURSand/orthedocumentationprovidedbythemanufacturer,ifapplicable;iii)interconnectionsbetweenthemajorcomponents (e.g. cables, tubing assemblies, etc.); iv) a general check of the electrical connectionsandgassupplypiping.

OQ should consider: i) a verification of the software user access policy, with reference to thedifferent possible level of privileges (e.g. administrators usually have the right to modify anyparameters,sequences,methods,etc.,whileoperatorsshouldhavethepossibilitytorundispensingprograms only); ii) a verification of the software sequences, if applicable; iii) a verification of the

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possibleeffectsofageneralpowerfailure (e.g. tocheckforthepresenceand/ortheneedforanUPS; iv) a verification of the calibration status of the major components; for instance, in severaldispensing systems, vial filling accuracy is based on balances thatweigh the solution during fillingoperations; balance is in this case a critical component and its performance could be evaluatedduringOQbycomparisonwithacalibratedprecisionbalance,usingcertifiedweights.Certificateofcalibrationofthereferencebalanceandweightsshouldnotbeexpiredandshouldbeincludedinthevalidation documentation. Dispensing systems for individual syringes preparation are preferablybasedondirectradioactivitydeterminationusingdosecalibrators:inthiscasetheactivimeteristhecritical component,whosecalibrationstatusneed tobeverifiedduringOQ (seebelow).Onemoreexampleofcriticalcomponentsindispensingsystemsarethepumpsoftenusedtodraw/pushfluidsthrough tubing assemblies; again, a verification of their calibration (e.g. by measuring dispensedvolumeswithareferenceprecisionbalance)shouldbeperformedduringOQ;v)averificationofdatabackupandrestore.

PQ of dispensing systemsmight be carried out by performing at least three successful dispensingcycles in typical working conditions, i.e. using radioactive solutions of the intended activities andradioactiveconcentrations,dispensedinarepresentativenumberofvials/syringes.

c)ShieldedHotCells

Hotcellsmaybeusedtoaccommodateautomatedorremotelycontrolledradiosynthesisapparatusor,moresimply, toprovide theoperatorsa suitableenvironment toprepareRPs,manuallyorwiththehelpoftele-pliers,theirmajorfunctionsbeingtoprotecttheoperatorsfromradiationburden(auseful calculator to determine the required shielding thickness may be found at the followingreference: http://www.radprocalculator.com/Gamma.aspx(9)), and to guarantee an environmentwith suitable air quality and cleanliness, which is critical for the microbiological quality of theproducts. Generally, working area is tightly sealed, and a negative pressure is operating, to allowpotential radioactive exhaust to be collected to safe containment systems, such as shielded gascylindersorretardationpipes.Validationextentforhotcellsisdependentontheircomplexity,thatmay range from a simple workingsurface surrounded by an adequate lead shielding, to fullyautomateddispensing systemwhichareembeddedand integrated in thehotcellwhole structure.However, there are common characteristics that may allow to set general principles for theirvalidation.

IQfollowsthesamegeneralconceptabovedepictedforautomatedsystems,andbasicallyconsistsofa series of verification of the documentation, the major installed components and theirinterconnections.SpecifictestforOQmightconsider:

i) a leak test, to verify the tightness of the working area with respect for the externalenvironment; the testmaybeperformedby simplymeasuring leak rate afternegativepressure has been brought to its maximum, and ventilation / extraction have beenswitchedoff,thusisolatingthehotcellitself;

ii) an air velocity test, to determine the suitability of ventilation above theworking area,whereRPpreparationanddispensingoperationstakeplace;analternativetestmaybe

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themeasurementofairparticlecontamination,usingportableorstand-alonecalibratedparticlecounterdevices,whichprovideandindirect,butnonethelesseffective,measureofairquality;indeed,classBorclassAenvironment,asdefinedbyEUGMP–Annex1(10),areoftenclaimedbyhot cellmanufacturer. InOQ, these testareperformed“at rest”,withworkingareainnormaloperatingconditions,butwithoutpersonnelintervention.

iii) hotcellsdoorsareusuallyinterlockedforsafetyreasons;forinstance,incaseofhotcellsused for the preparation of PET RPs, radionuclide transfer from the cyclotron is notallowedifhotcelldoorsareopen;othercommonsafetyinterlockslinksradiationlevelsinsidetheworkingareawithhotcelldooropening,whichisnotallowedincasethelevelisaboveadefinedthreshold,orstop.TesttoverifyfunctionalityofinterlocksaretypicaloperationstobeincludedinOQprotocols.

iv) Othertest,suchaslaminarflowpatterndetermination(e.g.usingasmoketrace),maybeperformed,dependingonthehotcellcharacteristics.Ithastobeunderlinedonceagaintheneedtofocusoncriticalparameters.

It is appropriate to consider PQ of hot cells in conjunction with OQ, as there is no significantdifferenceintheirmodeofoperationduringthepreparationoftheRPsoratrest.Ontheotherhand,this is not true in case ofmanual or semi-automated operations, whenmanipulationsmay affectlaminarflowpattern,e.g.duetothemovementoftheoperatingpersonnelarmsthroughthegloves.Thus, theabove test shouldbeexecutedbothat rest (OQ)and“inoperation” (PQ).As forparticlemonitoring,ithastobenotedthatradioactivitymaystronglyinfluencetheinstrumentresponse,asradiationpulsesmaybeerroneously“counted”bytheparticlemonitoringsystem,andthusparticlecontamination may be overestimated. Keeping this in mind, for hot cells used dedicated toautomatedpreparationordispensingoperations,PQtestsshouldbeperformedduringtheirnormalworkcycles(e.g.duringthepreparationorthedispensingoftheintendedradiopharmaceuticals).Incase of hot cells used for manual RP preparations, which are typically performed by personnelthroughsuitable sealedgloves,PQ tests shouldbeperformedduringnormalworkflow (e.g.duringthepreparationofTc-99mlabelledkits).

d)Dosecalibrators

IQ for dose calibrators should be straightforward, considering that these kind of instruments arerelativelysimpleandconsistofasuitabledetectorconnectedtoaPCortoanelectroniccontrolcasewith a display. Thus, a check of documentation and installation conditions (electrical connections,cabling betweendetector andmeasuring / display device) should be sufficient.OQ test should beaimed to verify calibration status of the activimeter. This could be done with accuracy andreproducibility tests, to be performed using suitable calibrated radioactivity sources. Chosenradionuclides should be of adequate activity, energy and half-life (e.g. Cs-137,with γ emission at662keV,whoseT1/2is=30years).OQmightalsoincludeothertests,suchasverificationofcalibrationthrough accurate measurements of the output current in response to increasing activities, butusuallythemonitoringoftheabovecitedparametersisconsideredsufficienttoprovideanadequateoperational qualification of the instrument. The same test should be performed with the aim ofqualifying theperformanceof the instruments, butusing the intended (oroneof the intended, incasemoreradionuclidesaremeasuredwiththesameinstrument)radionuclide(s).Forinstance,ina[18F]FDGpreparationfacility,accuracy,reproducibilityandlinearityshouldbedeterminedusingF-18

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withactivityinthenormalworkingrange.ForPQpurposes,alsoLimitofQuantitation(LOQ)shouldbedetermined.OQandPQtests should take intoaccount thegeometryof thesample (e.g. shapeandsizeofthecontainer,anddistancetothesensitivesurfaceofthedetector).Re-qualificationpolicyofdosecalibratorsshouldaccountthatdailychecks(e.g.constancytests)areusuallyperformed,andalsoverificationoflinearityandreproducibilityarerelativelyfrequent,soastoavoidtheneedofre-qualification,thatshouldbeonlydoneincasetheinstrumentismovedtoadifferentlocationorduetoothersignificantchanges.Thereareanumberofusefulreferencedocumentsthatmayhelpduringthe implementation of the IQ, OQ and PQ validation steps. Table 6 of EANM guidelines on“Acceptancetestingfornuclearmedicineinstrumentation”(11)providealistofteststobeperformedboth at the acceptance of the instrument and to periodically verify its correct functionality.Moreexperimental details related to the above suggested tests are described in EANM guidelines on“Routine quality control recommendations for nuclear medicine instrumentation”(12). Finally,recommendations relevant to assuring the continuing acceptability of the performance ofradionuclide calibrators are set by European Commission Radiation Protection document n° 162“Criteria forAcceptabilityofMedicalRadiologicalEquipmentused inDiagnosticRadiology,NuclearMedicineandRadiotherapy”(13).

5.2QualificationofQCinstrumentationQualitycontrolactivitiesmayrangeincomplexityfromrelativelysimpletestwithTLC,whichisoftenthemainQCtestrequiredincaseofTc-99mlabelledkits,toafullseriesofQCtests,includingHPLC,TLC,GC,gammaspectrometry,etc, incaseofPETradiopharmaceuticalpreparation.QCequipmentmay include instruments normally used in analytical chemistry, such as pHmeters, analyticalbalances,HPLCorGC,andinstrumentsspecificallydesignedfortheanalysisofradioactivesamples,suchasactivitydetectorsconjointlyusedwithHPLCorTLC,andgammaspectrometers.Moreover,the need to control microbiological contamination of injectable radiopharmaceutical preparationsmakedevicesdesignedtomonitorendotoxinlevelsfamiliartotheradiopharmacists.

IQ for QC equipment follows the general rules already depicted for production equipment, andverificationofthedocumentation,drawings,schematics,cablesandpiping,ageneralcheckonSOP,logbook,etc.,andverificationofenvironmentalconditionsandutilitiesherealsoapply.OQandPQaremorespecific forthevarious instruments,andwillbedescribedwithmoredetails. Ithastobeunderlined once again that IQ, and also OQ, may be performed by the instrumentationmanufacturer,exceptfortestsonradiochemicaldetectors.

Radio-HPLC

Aradio-HPLCsystemis typicallycomposedofapump,whichdrivestheeluentthroughthevariousdetectors and columns, the detectors themselves, one ofwhich is always a radioactivity detector,whiletheothersareneededto identifyandquantifynonradioactivespecies,andtheirselection isdependingon the intendedapplication. Themost frequentlyuseddetectors areUVdetectors,butconductivity or electrochemical (or others) detectors are also used for specific applications. Thesedetectorswillbehereinafterdefinedas“massdetectors”.Injectionofthesamplemaybeperformed

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manuallyorautomatically,bymeansofanautosampler.Chromatographiccolumnsmaybekeptatroom temperature or heated, by means of a column oven. Finally, most of the HPLC systemscurrentlyavailablearecontrolledviaasuitablesoftware,which isalsousedtoacquireandprocesssignalscomingfromdetectors.Fromavalidationperspective,HPLCmaybeconsideredasasumofdifferent components that may be tested individually. Thus, OQ and PQ test should be designedspecificallyfore.g.UVdetectors,aswellasforradiochemicaldetectors,whilecontrolandacquisitionsoftware may be evaluated as a whole. OQ on radiochemical detectors may include a linearityverificationof thevoltageoutput, in response todecreasing levelof radioactivity.A sampleof theintended radionuclide/radiopharmaceutical is suitable for this purpose. OQ test on UV detectorsusuallyinclude:i)testonwavelengthaccuracy,usingasuitableknownreferencestandard;ii)noiseanddrifttest,whichcanbeperformedrunningflowforasuitabletime(e.g.60min)andrecordingand allowing software to record the above parameters (some instruments may already havesoftwareroutinesdesignedtorunthetests);iii)averificationofabsorbanceaccuracyusingreferencestandard,whichcanbeeasilypurchasedfromcommercialsupplier, iv)testonsoftwareuseraccessandrelatedprivileges.Similarly,other“massdetectors”suchasconductivitydetectorsmightbeOQcheckedforlinearityandreproducibilityusingstandardionicsolution(e.g.chlorides,sulphates,etc.).HPLCpumpmaybetestedforaccuracyandprecisionbycollectingandweighing,usingacalibratedanalytical balance, a statistically significant number of samples (e.g. 10 samples, collected at aflowrateof1ml/min).Columnoven,ifpresent,shouldbecheckedforitscapabilitytomaintaintheselectedtemperature,bysettingarangeandmeasuring,usingacalibratedthermometer,arangeoftemperatures. Similarly, accuracy, precision and linearity test might be performed on theautosampler,withtheaimtoverifytheircapabilitytoreliablyinjectsamplesofthedesiredvolumes.Irrespectiveofthewaythesamplesareinjected(manualorautomated),theinjectionsystemneedstobecleanedbetweeninjections:carryoverisanothertypicalOQtest,aimedtoprovetheefficacyof the cleaningprocedure.Carryover shouldbe testedby repeatedly analysing samplesofmobilephasefollowingtheinjectionofsamplescontainingsignificantamountsoftheintendedanalytes;toverify carry over of UV or other “mass detectors”, samples should be taken from the higherconcentrationsolutionused in linearitytest; forradiationprotectionpurposes,carryovertests onradiochemicals shouldbeavoided,and the resultsobtainedwith testonmassdetectorsshouldbeconsideredassufficienttodemonstratethecleaningefficacy.

PQ protocols onmass detectors should include tests to verify precision (or reproducibility) of thedetector output, by injecting at least 5 samples of the intended “cold” counterpart of thedesiredradiopharmaceuticals (e.g. [19F]FDG for [18F]FDG, or [12C]methionine for [11C]methionine) at aconcentration included in the typical working range; also linearity test, using at least 5 differentsolutionswithincreasingconcentrationinthenormalworkingrange,shouldbeperformed.PQtestonradiochemicaldetectorsshouldbeaimedtocheckprecisionandlinearityaswell.However,duetoradioactivedecay,asinglesampleofsuitableactivitymightbeused,andareavaluesobtainedfromtherelatedchromatogramsshouldberecalculatedusingthedecaylaw(A=A0e-λt).

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GasChromatography

Inthefieldofradiopharmacy,gaschromatographicisveryoften(althoughnotalways)usedfortheanalysisofresidualsolvents.Theinstrumentismadeofacolumn(oftenacapillarycolumn)placedinanoven,throughwhichacarriergasisswept;themostpopulardetectorisFlameIonizationDetector(FID),whichissuitableforresidualsolventsanalysis,butothertypesmaybeused,dependingontheselectedapplication.SimilarlytoHPLC,sampleinjectionsystemmaybemanualor,morefrequently,automated (e.g. head space injection system). IQ protocols don’t differ significantly from thosealreadydescribed for radio-HPLC.OQ testsonFIDdetectors should include sensitivity.Head spaceinjectionsystem,ifpresent,shouldbetestedforprecisionandaccuracyusingreferencestandard,inorder to verify its capability to reliably inject the selected volumes, and for temperature, using acalibratedthermocouple.A leaktest, tocheckthetightnessof the injectionsystem,hasalsotobeperformed. Finally, test on carry over within the injection system is also recommended. Oventemperature is another critical parameter that should be checked during OQ, by means of acalibratedthermometer;aseriesofmeasurementsallowsforaccuracyandprecisiondetermination.Also carrier gas flowmeter should be checked, by comparisonwith a calibrated flowmeter. PQ, asusual,helpstodemonstratethatthesystemiscapabletoyieldtheexpectedperformanceinnormaloperatingconditions.Precisionandlinearityshouldbecheckedusingareferencesolutionofoneormore of the analytes that are expected to be quantified during normal QC operations (e.g.acetonitrile, ethanol), while for linearity determination, a series of solutions with increasingconcentrationsoftheinterestedanalytesshouldbepreparedandanalysed.Thesamedataobtainedfollowingtheabovetests,couldthenbeusedforthevalidationofanalyticalmethods.

Radio-TLC

Radio-TLC scanners are mainly used to determine radiochemical purity of radiopharmaceuticalpreparations.Radio-TLCareoften scanners thatdriveaTLC sheetorplateundera suitable sensorcapable to detect radioactivity. Autoradiography systemsmay also be used for this purpose, thattake advantage of the capability of a suitable phosphor plate to store the radioactive signal andreleaseitintheformofasuitableluminescence,andthatmaythuscreateakindof“latent”imageofthe spots generated during the TLC run by the separation of the analytes. IQ follows the sameprinciples already depicted for other analytical instruments. OQ and PQ may be consideredconjointly, and usually tests on reproducibility and linearity, using a solution of the desiredradionuclidewithsuitableactivityrangeshouldbeperformed.Reproducibilitymaybeevaluatedbydeposition, using preferably a calibrated micro-pipette, of a few microliters of the radioactivesolutionindifferentpositionoftheTLCplate.Duringdataacquisitionandcalculations,decayshouldbe accounted for, especially in case of very short half-life radionuclides. For linearity purposes, asinglespotcouldbedepositedandacquiredatsuitableuserdefinedintervals.OtherOQtestsmayberelated, as usual, to the software system, by checking software access policy and privileges, andarchiving/backupfunctions.

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Gammaspectrometer

Gamma spectrometers are used both for identification and radionuclidic purity determinationpurposes.MostcommondetectorsareHPGe(highpuritygermanium),whicharetobepreferreddueto higher energy resolution, and thallium activated NaI detectors, that have better sensitivity butmuchlowerenergyresolution.Detectorsaresuitablyshielded,toreducebackground,andconnectedthroughacabletoaPCequippedwithapropersoftware.HPGealsoneedtobecooled,inordertoreducethermallyinducedleakagecurrent,andeffectivenessofthecoolingsystem,irrespectiveifthisisdonevialiquidnitrogenorelectrically,hastobecarefullycontrolledduringnormaloperationsandduringqualificationtestsaswell.Withsuchasimpleconfiguration,IQisalsoquitesimple,anditmaybeperformed,asalreadydescribedforotherinstruments,byageneralcheckofthedocumentation(order, manuals, other documents provided by the supplier, logbook, dedicated SOPs, installedsoftware, versions, etc.).OQmay includeanenergy calibrationof the instrument,with theaim toverifythatdetectedenergiesmatchwithexpectedvalues.Bothmono-andmultinuclidecalibrationsourcesmay be used.Multinuclide or single-nuclidemulti-energy (e.g. Eu-152) sources should bepreferred,soastocheckcalibrationstatusinabroaderenergyrange.Usually,typicalworkingrangeoftheaboveinstrumentsisindeed0-2000KeV.Aminimumof6acquisitionsforeachoftheselectedenergy signals, followed by coefficient of variation (CV%) calculation allow for energy calibrationdetermination. Efficiency is another parameter to be considered in OQ, especially when gammaspectrometryisusedforquantificationpurposes.Herealsomultinuclidesourcesareideallysuited,astheyallowforquantificationofradioactivityamountofthevariousnuclides,providedthattheyaresufficientlylonglived(mediumhalf-liferadionuclidesmightalsobeused,buterrorsarehigher).PQisdepending on the intended use of the instrument, but it generally includes reproducibility andlinearity tests, tobeperformedwith the radionuclidesexpected in theRPpreparationof concern.The sensitivity of an instrument is usuallymeasured, as already described above, using calibratedstandardsattheproperconcentration.Incaseofgammaspectrometer,sensitivitymaybeexpressedbyaparameterknownasMinimumDetectableActivity(MDA),whichmaybeconsideredsimilartotheLimitofDetection(LOD),andwhichisdependentonmanyfactors(background,geometry,etc.)and it may vary from run to run for the same radionuclide. Thus, although MDA might bedetermined, forexample,duringOQ testwith calibrated source(s)orduringPQwith the intendedradionuclide, it would make more sense to evaluate it during validation of the specific analyticalmethod.Itisalsoimportanttoestablishthemaximumdetectableactivityrange,asthesaturationofthedetectormayleadtounderestimationoftheradioactivity.

6.ValidationofanalyticalmethodsQualityControlofradiopharmaceuticals, forwhichdedicatedmonographsofEur.Ph.areavailable,are straightforwardas themonographsdefinewhich controlshave tobeperformed, including therelatedmethodofanalysiswithexperimentaldetails(e.g.stationaryphase,mobilephase,flowrate,wavelength in caseofHPLCanalysiswithUVdetector, etc.) andacceptance criteria. Following theabove specifications, the analytical methods of concern do not need to be validated, but ratherverifiedineachindividuallaboratorythroughtheexecutionoftestsaimedtoverifythatthemethodhasbeenimplementedproperly(e.g.testtoverifylinearityandprecision).IncaseamonographfortheintendedRPisnotpublished,orincasethemonographexistsbutforanyreasonsitispreferredtouseadifferentmethod,itssuitabilityneedtobeassessedanddemonstratedthroughavalidation

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procedure. Guidelines for validation of analytical methods have been released by ICH(5), whichprovide general information and guidance about the parameters to be tested (e.g. accuracy,precision, linearity,etc.),howtotest themandwhen; for instance, theaboveguidelinesstatethatthedeterminationofrepeatabilityshouldbeperformedafteraminimumof9analyses,coveringthespecifiedrangeoftheprocedure,etc.

Duringvalidationofanalyticalprocedurethreecommonanalyticalproceduresmayberecognized:

a) Identificationtest,which isaimedtocontributetothe identificationofthedesiredproductor other analytes in the sample. In case of RPs, identification of the intended RP is oftencarriedoutexploitingthetwodistinctcharacteristicsofanyRP:i)the“pharmaceutical”partis identified through the chromatographic comparison of the retention time of the mainradioactive peak with retention time of the “cold” standard (e.g. [18F]FDG is identifiedthroughtheanalysisofa[19F]FDGsample);ii)theradionuclidepartcontributestotheoverallidentificationoftheRPthankstothegammaspectrometryanalysisoftheemittedenergies(e.g.incaseTc-99mistheintendedradionuclide,anemissionat140KeVisexpected)

b) Testing for impurities,whichcanbequantitativeor limit tests.TheseanalyticalproceduresareoftennotofconcernincaseofRPs,wherevalidationofcharacteristicssuchasaccuracy,precision,etc.ispracticallyimpossibleforradiochemicalimpurities,astheyarenotavailableassuitablestandards,andtheymaybedifficultalsofor“cold”impurities.However,whentheimpurities are, for instance, the chemical precursor of the radiolabelling, or they arewellcharacterized,testingmaybefeasible,andrulessetbyICHguidelinesapply.

c) Assaytest,thatareperformedtoquantitativelymeasuretheanalyteofinterest(usually,thedesiredRPs)

Ananalyticalmethodshouldbere-validatedincaseof:i)changesintheRPpreparationprocessthatmay affect the quality of the final products; examples of such changes are represented by themodificationoftheselectedprecursor,orbychangesinreactionparameters(e.g.temperatureandreactiontime),whenpurificationcomponentsarereplacedbydifferentones(e.g.aluminacartridgesarereplacedbyionexchangecartridges)orthepurificationmethodischanged(e.g.HPLCvsSPE);ii)changes in the composition of the final product; an example of such changes is a variation inradioactive concentration, that could potentially increase radiolysis and related growth ofradiolabelledimpurities;iii)significantchangesinanalyticalprocedure;examplesofsuchchangesarethe replacementofexistingHPLCcolumnwithanewonewithadifferentstationaryphase,or thereplacementofadetector.

Analytical methods used for the QC and characterization of RPs are sometimes typical analyticalmethods(forexample,analysisofresidualsolventsusingGC); inthesecases, ICHguidelinescanbeappliedwithoutsignificantadaptations.Ontheotherhand,specificadjustmentsarerequiredincaseofradioanalyticalmethods,suchasradio-HPLC,radio-TLCandgammaspectrometry,andtheywouldneedtobeconsideredwithmoredetails.Forthisreason,andinconsiderationofthewidevarietyofpossibleapplicationinthefieldofradiopharmaceuticalpreparations,validationofanalyticalmethodswillbethesubjectofadedicateddocument.Moreover,practicalexamplesofvalidationofanalyticalmethods of routinely used RPs may be found in the recently published EANM guidelines on thepreparationofIMPD(14).

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7.ComputerisedsystemsAs already mentioned earlier, computerized systems are ubiquitously used and most of theinstrumentation of concern in the field of radiopharmacy are controlled by a wide variety ofhardware/softwaresystems.Thus,validationofsoftwareshouldbeconsideredasanintegralpartofthe general validation policy(15). Two different general approaches are possible: i) validation /qualification of a production / QC instrument as a whole (holistic approach), in which thecomputerised system is considered as a part, although significant, of the whole instrument, andvalidation of hardware / software is thus performed consistently; ii) validation of computerisedsystemasanindependententity.Whateveristhechosenroute,thefollowingprinciplesapply:

- requestedcharacteristicsofthesoftware/hardwareshouldbedescribedinURS;- datasafetyshouldbeensured,soastominimizetheriskoflossofdataorwrongdataentry

bytheoperators;- initialconfigurationofthesystemsshouldbeperformedbyqualifiedpersonnel;- access to the software should be allowed for authorized persons only, and it should be

regulatedbymeansofappropriate login/password,andtheallowedoperationsshouldbedifferent, depending on the various functions (e.g. in case of a QC software, differentprivilegesmaybeattributedtotheanalystsandtotheresponsibleoftheQCdepartment);

- backupfunctionsshouldalwaysbeenabled,andtheway,thechosenstoragemedium(e.g.stand-alonedevicessuchasUSBpendrives,CDrom,externalharddisk,ornetworkbackupdisks)andbackupfrequencyshouldbedefinedthroughappropriateSOPs;restorefunctionsofthebackedupdatashouldbeverified,aswell;

- printoutfunctionofthestoreddatashouldbechecked;- audit trails functions should be considered mandatory, in order to have the necessary

traceability of the operations performed using the interested software; for instance,information about access (who and when accessed the software), changes (e.g. aboutdeleted files, change to operating methods, new methods, etc.), software / hardwareupdates should be automatically recorded by the software; in case the audit trail is notenabled,alternativeprocedurestoensureoperationtraceabilityshouldbeputinplace(e.g.printingand/orrecordinginformationaboutperformedoperationsondedicatedlogbooks);

- the risk related to possible accidental loss of data or software functionality should becarefullyevaluated,andexecutablecopyoftheinterestedsoftwareshouldbeavailableandfullycompatiblewiththehardwareequipment;

Ausefulreferencewhilevalidatingcomputerisedsystemsarethesocalled“GAMP”(goodautomatedmanufacturingpractice(16)),whosemaingoalistohelpUsersinunderstandingrequirementsandthelevel of validation to be performed and, which is even more important, to help suppliers indevelopingtheirsystemscomplyingwithgeneralrulesofgoodpractice.Itmeansthatwheneverthepurchased systems have been developed complyingwithGAMP, validation extent required to theenduserisminimized.

IQforacomputerisedsystemmightincludethefollowingcontrols:

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- checkpurchaseorder,shipmentdocumentation,packaging,etc.;- thesoftwarehasbeenloadedcorrectly;- powersupplyandelectricalprotectionaresuitablefortheintendedapplications;- PCandhardwarehavebeenproperlyconnected;- thesystemisworkingproperlyafterpowerup;- collectinformationaboutsoftware/hardware/operatingsystemversions,dateandplaceof

installation;- includeabriefdescriptionoftheroom/sitewherethesystemshavebeeninstalled;- software/hardwarecertificationareavailable(ifany).

OQ would be more focused on a functional verification of the software / hardware, and mightconsiderthefollowingverifications:

- checkondataintegrity,accuracy,securityandreliability;- verification of operating sequence, such as automated synthesis module routines used to

perform part of the preparation process (e.g. a purification operation, or fluid transfersthrough different parts of the modules, etc.) or acquisition sequences for an analyticalinstrument;

- a verification that different login/password credentials for access areworking and lead todifferentoperatingprivileges;

- verifysoftware/hardwaresafety,e.g.throughbreakinguppfpowersupply;- verifydatasafety,bymeansofattemptstochange,delete,copy/paste,etc.

Finally,PQoncomputerisedsystemsmightincludethefollowingtests:

- test specific SOPs, dedicated to the intended RP preparation process, for use andmaintenanceofthecomputerisedsystem;

- asPQistypicallyaimedtoverifythatthesystemiscapabletoproperlyperformthetasksforwhichithasbeenpurchased/built,PQforcomputerisedsystemstestscouldbemergedwithgeneral PQ of the intended instrument / system / utility. Thus, please refer to theinformation provided in the relevant section for e.g. PQ on automated synthesis systems,dispensingsystemsorforanalyticalinstrumentation

8.ValidationofclassifiedroomsPreparationofparenteralinjectablesolutionsrequiresspecialcareinthemanipulationofthestartingmaterials /intermediates / finished products, that may potentially be subject to microbiologicalcontamination in the form of bacterial endotoxins and vital microorganisms such as bacteria andfungi. To this regard, Annex 1 – GMP(10) set general guidance about technical characteristics ofclassified environment, as well as of the tests to be performed together with related acceptancecriteria for particle andmicrobiological contaminations. Thepossibility to establish andmaintain aclassified environment depends on several factors, such as the technical specification of HVACsystem,constructiondetailsofthepremises,characteristicsofequipment,dressingandbehaviouralrulesfortheoperatingpersonnel,cleaningandsanitizationprocedures,sterilization,etc.Validation

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of classified environments is challenging for typical radiopharmacies, as it requires skills andinstrumentation which are often not available. Moreover, differently than the above describedproductionandQCinstrumentation,whichareusuallycommerciallyavailable,evenDQplayshereacrucial role, as rooms and HVAC are specifically designed for the intended use, and theircharacteristics may significantly affect day-by-day operations and general compliance with EUguidelines.DQwillhavetobeperformedintightconnectionwithURSrequirements,andwillhavethegoaltoverifythate.g.requestedutilityservicesareavailableandsuitedfortheintendedpurposeorthatthesystemswillbeeasytobecalibratedandmaintainedandmayoperateinamannersafefortheproductsandfortheoperatingpersonnel.IQofHVACincludeacarefulverificationofalltheinstalled components, to check that e.g. valves, pipes, shutters, ventilationmachines are properlyinstalled compared with project layout, and that they are properly labelled. Of course a generalcheckondocumentation(drawings, layout,componentspecification,listofthesuppliers,operatingmanuals, etc.) is here of paramount importance. OQ of HVAC, which plays a critical role indeterminingthequalityofair,usuallyforeseetestsonairflowrate,HEPAfiltersintegrity,thenumberof air exchange / hour, particle and microbiological contamination. For these reasons, fullqualification of classified environments is usually sub-contracted to suitable specialized servicecompanies. However, the following tests, that can be considered as representative of the generalclassificationstatusoftheintendedrooms,couldbeperformed,providedthatatleastanairparticlecounterandanincubatorareavailable.

1)theeffectoflackofpowersupplyonHVACefficiency;thistestmaybeeasilyperformedbyturningoff and on the general power supply, and checking whether the main functions are correctlyrecoveredornot;

2) particle contamination test, that has to be carried out using a suitable calibrated instrument,following generalmethodology setby ISO standard(17).Numberof air samplesmaybedeterminedusingthefollowingformula:

ANL =

Where “NL” is the number of samples to be taken, and “A” is the surface of the classified area(expressed inm2); aminimum of two samples should be considered, notwithstanding the surfacearea. In case of class “A”, it is necessary to collect at least 1m3 of air / point (17),while for otherclasses,1m3shouldbethetotalvolumefortheconsideredwholeroom/environment.Thefollowinggeneralformulamaybeusedtocalculatetheminimumsampledvolume:

𝑉𝑠 =20

𝐶𝑛,𝑚𝑥1000

Where:

- Vsistheminimumsinglesamplevolumeperlocation,expressedinlitres- Cn,m is the class limit (number of particles / m3) for the largest considered particle size

specifiedfortherelevantclass- 20isthedefinednumberofsamplesthatcouldbecountediftheparticleconcentrationwere

attheclasslimit

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Testshouldbedonein“atrest”conditions,asdefinedbyAnnex1–GMP(10).

3)decay / recovery test,which is intended todetermine the timeneeded to recover thespecifiedclassaftere.g.HVACisintentionallyswitchedoffforadefinedtime.

4)clean-uptest; inprinciple, this test isaimedtodetermine the timerequired toswitch fromoneconditiontoanother;incaseofcleanroom,thatmayberepresentedbythetimeittakesto“clean-upfrom “in operation” to “at rest” conditions, and can be experimentally measured monitoringappropriateparameters,suchasairbornecontamination.

PQmaybeperformedby: i) repeatingtheparticlecontaminationtest in“inoperationconditions”,which means with personnel normally operating in the lab; ii) verification of the microbiologicalcontaminationoftheairandsurfaces,thelatterbeingcheckedbymeansofagarcontactplatesfilledwithasuitablemedia,andtheformerusingagarsettleplates;numberofplatesandtheirpositionhavetobechosenwitharationalebasedontheexpectedmicrobiologicalrisk;tothisregard,contactplatesshouldbescratchedonrepresentativepositionsonthefloor,wallsandmajorinstrumentation(inside/outsidehotcells,externalsurfaceofautomatedsystem,workbench,etc.),whilesettleplatesshouldbelocatedmoreorlessinthesamepointsabovecited.Agarplatesshouldthenincubatedat25±2°Cforthreedays,andat35±2°Cforthenexttwodays.Acceptancecriteriaaredependingofthedesiredclassificationlevelandareset,asusual,byAnnex1–GMP(10).

9.CleaningValidationCleaningvalidationisaimedtoverifytheeffectivenessofacleaningprocedure.TwogeneralcleaningproceduresareofconcerninthepreparationofRPs:i)cleaningofproduction/dispensingapparatus,with special emphasis for those parts of the equipment which come into contact with reagents/solvents /intermediates / finishedproducts; ii) cleaningof theexternal surfacesof theequipment(e.g. hot cells, laminar flow cabinet,workbench, furniture, etc.). Considered that aim, procedures,andcriticalstepsaredifferent,theywillbedescribedseparately.

9.1ValidationoftheprocedureforthecleaningofproductionequipmentinternalsurfacesProduction of RPs is often performed using automated or at least remotely controlled devices. Auseful guidance, edited under the umbrella of EANM Radiopharmacy Committee, for the use,installation, cleaning, and validation of automated systems has been recently published(8), andgeneralprinciplesofcleaningvalidationmaybefound.Ingeneral,automatedsystemsmaybeoftwodistinct types, depending on the nature of the so called “chemistry part” of the system,which isdefinedas“an interconnectednetworkofcontainers inwhichgaseous, liquidand/orsolid reagentsand components can bemoved,mixed and/or transformed to obtain the desired final product”(8).With “cassette” systems, the chemistry part is disposable, and replaced every time a newpreparationbegins,while innon-disposablesystemsthechemistrypartmaypotentiallybere-usedfor an undefined number of times. In the latter case cleaning operations and, in turn, cleaningvalidationareclearlymorecriticalthanintheformer.“Validationofthecleaningprocessesshouldbe

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performedprior to theuseof theautomatedmodule, todemonstrate that cleaningoperationsareefficient to fulfil the established specifications in the area of effective operation”(8). A thoroughknowledge of the chemistry involved in the preparation process is required, so as to identify thepossible impurities leftover insidethechemistrypartsurfaces,selectproper limitsandacceptancecriteria of carry over and, which is of the utmost importance, design a suitable cleaning process.Cleaningvalidationneedtobeperformedboth incasetheautomatedsystemisusedtoproduceasingleradiopharmaceutical(e.g.[18F]FDG)andincaseitisusedtopreparedifferentRPs,whichmaypose additional problemsof cross contamination.Cleaning validation should includeat least threeproductionsof thedesired radiopharmaceutical, followedby three cleaningprocedures. The lattershouldbedesignedwiththeaimtokeepcarryoverataminimumextent.Forvalidationpurposes,cleaningstepsshouldbefollowedbyacarefulsweepingoftheinnersurfacesofthechemistrypartwith a suitable (aqueousororganic, orboth)media, capable to solubilizemostof the residualsofimpurities.Theaboveoperations shouldbedesigned soas toensure thatall thepossible surfacesthat get in contactwith reagents / intermediates / final product are suitably swept by the abovemedia. Washing solutions should then be collected, and samples submitted to quality controlprocedures.Analyticalmethodsshouldbesufficientlysensitivetodetecttheestablishedacceptablelevel of the residue or contaminant. The above “sweeping” step should keep out multiple usechromatographic support, such as liquid chromatography columns, due to their inherentcharacteristicsandcapabilitytoretainimpurities. Incasetheautomatedsystemisusedtoproducedifferent RPs, cleaning validation protocols should demonstrate that cleaning procedures areeffectiveirrespectiveoftheorderthatthevariousRPsareproduced.Asalreadystatedabove,cleaningvalidationprotocolsare lesscritical incasesingle-use,disposablesystems are used. This general consideration apply to both “cassette” automatedmodules for RPproduction, and to dispensing systems used to prepare syringes with individual patient doses ormulti-dosevials.However,shouldtheabovesystemsincludenon-disposableparts(e.g.typicallytheradionuclidetransfer line fromthecyclotron), thesameprinciplespreviouslydepictedapply to theinterestedparts.Thesameconsiderationsapplyincaseofmicrobiologicalcontamination,whichislesscriticalincaseof“cassette”systems,duetotheirsingle-usecharacteristics.Moreover,somecommerciallyavailablekits are sterile. In case of non-disposable system, bioburden is the method of choice to validatecleaningprocedures.Typically,threepreparationrunsareperformedusingthesameconditionssetfornormalroutinepreparations,butwithoutusingradioactivityandavoidingfinalsterilization(e.g.in case the RP solution has to be sterilized by filtration, filter is not included in the preparationsdedicated to bioburden testing). As ionizing radiations, depending on the amount and radiationpatternof the starting radionuclide,mayplaya role in keeping themicrobialpopulations low, thelack of radioactivity during the simulation of the preparation procedure may be considered as aworstcasescenario.Thethreesimulatedpreparationrunsyieldsolutions,whicharethenanalysedfollowing routine procedures for bioburden test. Typical acceptance criteria is 10 Colony FormingUnit(CFU)/100ml.Thechoiceofthecleaningmedia,whichmaybeasolvent(e.g.acetone,ethanol,isopropanol,etc.),agas (e.g. ozone, ethylene oxide), or a combination of different solvents/gases, is clearly veryimportant, as they should be: i) effective in the removal of both chemical and microbiologicalimpurities,ii)easytoberemovedfromthesystemaftertheendofthecleaningprocedures,iii)non-toxic, iv) inert with respect for the interested internal surfaces, and v) they should not yield

26

additional impurities.For instance,acetone issuitable insolubilizingchemical impurities,dueto itspolar characteristics, and it’s easy to be removed, due to its low boiling point, but it is not veryeffectivewithmicrobiological impurities, and ethanol, isopropyl alcohol or amixture of the abovesolventsmightbepreferable.

9.2ValidationoftheprocedureforcleaningexternalsurfacesExternal surfacesmay include laboratory floors andwalls orworkbenches, aswell as internal andexternal surfaces of the hot cells / laminar flow cabinets, and external surface of the preparationequipment. They should be cleaned following written procedures, that set which detergents /disinfectants/biocideshavetobeused,thecleaningoperationfrequencies(e.g.daily,weekly,etc.),andturnoverpolicyonbiocides.Chemical,radiochemicalandradionuclidicresiduesareusuallynotof concern for contact surfaces, as their presence should be considered as a consequence ofinadvertent contamination. Further, RPs are generally prepared in small scale, and low amount ofreagents / solvents are used,which further decrease the risk of “chemical” contamination e.g. onworkbenchesoraroundtheautomatedsystemssurface.Thesmall scale“size”ofRPspreparationshasalsotobeconsideredinviewofariskevaluationduetotheoperatingpersonnel,whichisusuallylowinnumberandoccupancyfactor.Thus,validationofcleaningofcontactsurfacesismostlyaimedtodemonstratethatmicrobiologicalcontaminationiskeptwithintheproperlimits,dependingonthedesiredclassificationlevel(10).Suchacleaningvalidationprotocolshouldinclude:

- adescriptionofthefacility,includingtherequestedlevelofroomclassification;- adescriptionoftheinstrumentation/equipmentinstalledintheclassifiedrooms,andtheir

locations;- thecleaningprocedures(alsothroughareferencetothededicatedSOPs);- adescriptionofthecleaningproducts(detergents,disinfectants,biocides);- adescriptionoftheprocess(es)carriedoutintheinterestedrooms,withspecialcareincase

of“multitracer”productioninthesameenvironments;- the sampling procedures (e.g. direct surface sampling using swabs or contact plated petri

dishes,etc.);- thesamplingcampaignandcriteria;samplingpointsconsideredtobemostcritical(e.g.hot

cell working surface, the laboratory area where personnel spend most of the time, etc.)shouldbediscussedandjustified;

- test methodology, including the media used to collect samples (e.g. contact plates, airsamplinginstrumentation,typeofmicroorganismgrowingmedia,etc.);

- acceptancecriteria;- protocoldiscussionandconclusion.

The design of a cleaning validation protocols might take advantage of risk analysis based on theknowledge of the intended RP preparation processes and of the established cleaning procedures,which may provide information related to the hazard associated with the use of both startingmaterialsandcleaningagents,andthewaytheresiduesareeffectivelyremovedanddetected.Cleaningvalidationprotocolsshouldalsotakeaccountofthepersonnelaccessingtheworkingrooms,includingcleaningservicepersonnel,andsamplingandtestingshouldberepeatedforareasonablenumber of times, considering theworst case in terms of number of persons entering the labs, of

27

operations performed and of “hot spots” where cleaning may be more difficult for accessibilityreasons(recesses,hiddenpartsofequipment/labs).Worstcaseapproachmightallowto“bracket”the different cleaning products and procedures, thus reducing the need for multiple validationprotocols.Cleaning validation protocol should be considered as amean to validate cleaning procedures andcleaningmediaatthesametime.

10.ProcessvalidationAs already stated above, Process Validation (PV) should be viewed as the final step of validation,aimed to verify that the preparation process of a RP is capable to prepare the product with therequestedcharacteristicsofyield,quality,reliability,safetyandefficacy,andthattheRPispreparedwithin a suitable environment, with the necessary safety for the operating personnel and for theproduct. For the above reasons, it is expected that process validation is being performed whenprocessdesign,andallthedetailsoftheprocessareadequatelyknown.PreparationoftestbatchesisusuallyofhelpandincreasetheprobabilityofasuccessfulPV.PVshouldbecompletedpriortotheuse of the intended RP in routine clinical activity, while this is not strictly required in case ofinvestigationalRPs,where it isconsideredthepossible lackofwellestablishedroutineprocedures.Objectives of PV should be clearly stated. In general, traditional approach to PV consists of thepreparation of three consecutive batches of the intended RP, in different days and in the sameconditionssetfortypicalroutinepreparations.Thismeansthatstartingactivityoftheradionuclide,reagents / solvents and their quantities, automated system parameters (if applicable), reaction,purification and formulation conditions should be the same as those intended for the routinepreparation of the desired radiopharmaceutical. Moreover, the three productions should yieldbatcheswiththeintendedsize(e.g.numberofvials),andwithappropriatelabellingandpackaging.EverybatchofRPpreparedaccordinglywiththeabovesetofconditionsshouldbefullycharacterizedfromtheanalyticalpointofview,withtheaimtoverifythattheproductmeettheacceptancecriteriaas for all the established quality parameters (e.g. radiochemical purity, pH, sterility, radioactiveconcentration,etc.).

APVprotocolshouldinclude,butnotnecessarilylimited,to:

- a description of the process to be validated, including the instrumentation used (e.g.,automated radiosynthesis system, automated dispensing system, the hot cell where theautomateddevicesare installedorwherethepreparation ismanuallyperformed,thedosecalibrator,etc.),apreparationflowchart,includingalistofreagents/solvents/excipientsandrelatedquantities,theproposedpurificationprocess(e.g.HPLCvsSPE,orboth),ifapplicable,togetherwithmajorcomponents (e.g.detectorsandseparationconditions incaseofHPLCpurification,or thecartridgesandmethod incaseofSPEpurification), the formulationandfinalsterilizationmethodsofconcern;

- a list of key personnel involved in validation activities, their functions and their trainingstatus;

- alistofthequalificationprotocolscodenumbersrelatedtothevariousinstrumentwhichareusedinthepreparationprocess,togetherwiththerelatedqualificationdates,withtheaim

28

todemonstrate that theabove instruments status is compliantwith thegeneral validationpolicy;

- the listof the intendedanalytical testsand the relateddocumentation, includinganalyticalmethod validation protocols code numbers, if applicable, which are expected to beperformedduringtheprotocolexecution;

- finishedproductreleasespecifications;- indicationofthetesttobeperformedaftertherelease(e.g.sterility,radionuclidicpurity);- samplingplan,withindicationsofwhich,howmanyandwhensamplestobesenttoQuality

Control(QC)arecollected,andselectioncriteria;- a listofthedeviationsactuallyoccurred(ifany)duringtheexecutionofthetests,together

with a discussion about their potential impact on the quality of the final product and therequestedcorrectiveaction;

- afinaldiscussionabouttheresults(theValidationSummaryReport).

ResultsobtainedfromPVhelptoidentifycriticalprocessparametersandtheiracceptancecriteria/limits.Inparticular,radioactiveconcentrationhastobeconsideredasabetterindicator/criteriathanthe amount of radioactivity as such. In case of RPs labelled with short or very short half-liferadionuclides (e.g. C-11 or Ga-68), it might be difficult to comply with European Union (EU)guidelines, thatoftenclaim for radioactivityatActivityReferenceTime (ART) tobedefined for thefinal radiopharmaceutical product, and process validation is then used to establish a suitableradioactivity concentration range.Process validation is alsoaimed todefine volume (or a rangeofvolumes),which isanotherparameter thatmaybedifficult,duetotechnical reasons, tounivocallysetincaseRPsarepreparedwithanautomatedsystem,andnodispensingsystemsareavailable.

Retrospective validation, that’s the process validation based on historical data ofwell known andestablishedpreparationprocesses,isnolongerconsideredandacceptableapproach(8).

An alternative approach, set by the European Medicine Agency (EMA) Guidelines on ProcessValidation(18),andbyAnnex15–GMP(2) is thesocalled“ContinuousProcessVerification” (CPV), inwhichthecontinuousmonitoringandevaluationofsuitableparametersmayreplacethetraditionalprocess validation approach above described. CPV makes sense in case of well known and fullydevelopedpreparationprocesses,andrequiresthemonitoringofprocessperformanceandproductqualityoneachbatchoftheintended(radio)pharmaceuticals.AsthesecriteriaareoftenmetbythepreparationofRPs,whicharefullycharacterizedbeforetheirrelease,thisapproachseemstobewellsuitedanditmayreplacetheneedforre-validation,providedthatthepreparationprocessdoesnotundergosignificantchanges.

11.ValidationofasepticoperationsviaMediafillTheobjectiveofasepticprocessing is tomaintain the sterilityofaproduct that is assembled fromcomponents,eachofwhichhasbeensterilizedbyoneofthemethodsdescribedinPh.Eur(19).Thisisachievedbyusingconditionsandfacilitiesdesignedtopreventmicrobialcontamination.Inordertomaintainthesterilityofthecomponentsandtheproductduringprocessing,carefulattentionneedstobegivento:environment,personnel,criticalsurfaces,container/closuresterilizationandtransferprocedures,maximumholdingperiodoftheproductbeforefillingintothefinalcontainer.

29

Aseptic operationsmay be validated bymeans of process simulation tests usingmicrobial growthmedia,whicharethenincubatedandexaminedformicrobialcontamination(mediafilltests).

Mostradiopharmaceuticalsaredesignedforparenteralapplicationandthusforeseeoperationstobeperformed under aseptic conditions. A media fill is the performance of an aseptic proceduremimicking theconditionsof the realprocedure,butusingasterilemicrobiologicalgrowthmediuminsteadofthesolutionsotherwiseusedinthepreparationoftheradiopharmaceutical.Thepurposeof media fill procedure is to test whether the aseptic procedures are adequate to preventcontamination during actual RP production. Media fill may thus be considered as a part of theprocessvalidationoftheRPpreparation.

Themediafillshouldevaluatetheasepticassemblyandoperationofthecritical(sterile)equipment,qualifytheoperatorsandassesstheirtechnique,anddemonstratethattheenvironmentalcontrolsare adequate to meet the basic requirements necessary to produce a sterile RP by asepticprocessing(20).

The media fill should include positive control, which may be represented by a sealed productcontainerofthegrowthmediuminoculatedwithasmallnumberofmicroorganisms,andanegativecontrol,toensuretheabsenceoffalsepositiveresults.Anegativecontrolmaybepreparedbypre-incubating the medium, or by aseptically transferring medium into a separate suitable sterilecontainerandincubatingthecontrolsimultaneouslywiththemediafilltestcontainers.Thecontrolsshouldbeincubatedunderthesameconditionsasthemediafillcontainers(17).

All steps in a media fill should be done in the same locations as those typical for theradiopharmaceutical production. To initially qualify an aseptic process at a specific facility, threemedia fills should be conducted on three separate days, following the procedures of the specificproductionprocess that is beingqualified.Additionally,media fills shouldbe conductedwheneversignificant changes are made to the aseptic process (e.g. changes in personnel, components, orequipment)andwheneverthereisevidenceofafailuretomaintainproductsterility.Mediafilltestsperformedtovalidateanasepticprocessataspecificfacilityshouldbedonebyoperatorswhohavepreviously been trained and qualified in aseptic techniques (e.g., proper gowning, disinfectionpractices,handlingsterilematerials).

Media fills arean importantelementofoperatorqualification.Tobecomeaqualifiedoperator forradiopharmaceutical product production, an operator should perform three media fills on threeseparatedays.Aqualifiedoperatorshouldperformamediafillatleastannually(20).

Duringmediafilltestmonitoringoftheair,personnelandcriticalsurfacesshouldbedone.

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“This guideline summarizes the viewsof theRadiopharmacyCommittee of the EANMand reflectsrecommendationsforwhichtheEANMcannotbeheldresponsible.Therecommendationsshouldbetaken into context of good practice of nuclear medicine and do not substitute for national andinternationallegalorregulatoryprovisions”.“Theguidelineswerebrought to theattentionof all other EANMCommittees and to theNationalSocietiesofNuclearMedicine.ThecommentsandsuggestionsfromtheDrugDevelopment,Physics,Oncology and Bone & Joint Committees and the xxx and the xxx National Society are highlyappreciatedandhavebeenconsideredforthisGuideline.”

31

References

(1)http://apps.who.int/prequal/info_general/documents/TRS937/WHO_TRS_937-Annex4.pdf

(2)http://ec.europa.eu/health/files/eudralex/vol-4/2015-10_annex15.pdf

(3)http://www.fda.gov/downloads/Drugs/Guidances/UCM070336.pdf

(4)http://www.chem.agilent.com/Library/primers/Public/5990-3288EN.pdf

(5)http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf

(6)http://www.picscheme.org/pdf/25_pi-006-3-recommendation-on-validation-master-plan.pdf

(7)http://ec.europa.eu/health/files/eudralex/vol-4/2008_09_annex3_en.pdf

(8)AertsJ,BallingerJR,BeheM,DecristoforoC,ElsingaPH,Faivre-ChauvetA,MindtTL,KolencPeitlP,ToddeSandKoziorowskiJ.Guidanceoncurrentgoodradiopharmacypracticeforthesmall-scalepreparationof radiopharmaceuticalsusingautomatedmodules:aEuropeanperspective. J.LabelledCompd.Radiopharm.2014;57:615-620

(9)http://www.radprocalculator.com/Gamma.aspx

(10)http://ec.europa.eu/health/files/eudralex/vol-4/2008_11_25_gmp-an1_en.pdf

(11)http://www.eanm.org/publications/guidelines/index.php?navId=37

(12)http://www.eanm.org/publications/guidelines/index.php?navId=37

(13)http://ec.europa.eu/energy/sites/ener/files/documents/162.pdf

(14)ToddeS,WindhorstAD,BeheM,BormansG,DecristoforoC, Faivre-ChauvetA, FerrariV,GeeAD,GulyasB,HalldinC,KolencPeitlP,Koziorowski J,MindtTL,SolliniM,Vercouillie J,Ballinger JRandElsingaPH.EANMguidelineforthepreparationofanInvestigationalMedicinalProductDossier(IMPD). Eur. J.Nucl. Med. Mol. Imaging. 2014; 41: 2175-2185. Appendix 1: IMPD for [11C]choline;Appendix 2: IMPD for [68Ga]DOTA.NOC. Appendix 3: IMPD for [177Lu]DOTA-peptide(http://link.springer.com/article/10.1007/s00259-014-2866-8).

(15)http://ec.europa.eu/health/files/eudralex/vol-4/annex11_01-2011_en.pdf

(16)https://www.google.it/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0ahUKEwi_v9WmhOvLAhUEFCwKHU4YDToQFggfMAA&url=http%3A%2F%2Fwww.picscheme.org%2Fpdf%2F27_pi-011-3-

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recommendation-on-computerised-systems.pdf&usg=AFQjCNF_BpZN91VAEzsmLTE6GlvgWZC5WQ&bvm=bv.118353311,d.bGg&cad=rja

(17)EN/ISO14644-1Cleanroomandassociatedcontrolledenvironments–Part1:classificationofaircleanliness

(18)(http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2014/02/WC500162136.pdf)

(19) European Pharmacopoeia, current version, chapter 5.1.1 “methods of preparations of sterileproducts”

(20)http://www.fda.gov/downloads/Drugs/.../Guidances/UCM273766.pdf

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Section2–PracticalExamples

Example 1: Qualification of a production instrument: automated system for thepreparationofF-18labelledradiopharmaceuticals

InstallationQualification

Aim

• Verifyinstallationstatusandutilities;

• Identifythecriticalcomponents

• CheckfordedicatedSOPsaimedtouseandmaintenanceofthesystem;

• Checkfordocumentation(manuals,administrativedocumentation)

• Checkforlistofspareparts

Description

Thesystemincludesthefollowingcomponents:

- Theradiosynthesismodule;

- AUVdetector;

- Asemi-preparativeHPLCpump;

- AcontrolPC;

- APLC.

Test

Test1:documentation

Pleaseusethefollowingformtocollectdata:

34

Document(e.g.manual,SOP)andtitle Code Location

YES NO

CONFORM: r r

Test2–Systemdescription

RadiosynthesisModule

Components

Description Identification(ID#) Installed

Radiosynthesismodule qyes qNo

PC qyes qNo

PLC qyes qNo

HPLCpump qyes qNo

UVdetector qyes qNo

Foreachcomponentpleasecollectthefollowinginformation:

35

Radiosynthesismodule

Identification

Supplier

Model

Serialnumber

Location(room)

Roomtemperature

Roomrelativehumidity

OperationalQualification

Test1–heliumflowverificationAim:toverifytheefficiencyofheliumflowcontrollerMethodology:- connectthecalibratedflowmeterinaproperpositiondownstreamwithrespectforthehelium

flowcontroller;- usingthecontrolsoftware,enterthedesiredheliumflowratevalue- report theautomated synthesis and calibrated flowmeteroutput and recordon the following

datasheet.

Acceptancecriteria:thedifferencebetweenthetwoaboveoutputshouldnotbe>2%

Recordedflowvalues

Test#1 Test#2

Actualflowvalues(mL/Min)

Setvalues Measuredvalues Setvalues Measuredvalues

Setvalues Measuredvalues

36

Test2:reactortemperatureverificationAim:toverifythatthetemperatureinsidethereactorareconsistentwiththesetvaluesMethodology:- installthereactorfollowingthenormalprocedure;- placeacalibratedthermocoupleclosetothefollowingcriticalpoint:a)insidethereactorb)close

totheautomatedsystemsensor;- startasimulationoftheintendedRPpreparationprocedure,avoidingtheuseofradioactivity;- waituntiltheautomatedprocedurereachthecriticalreactionstep(e.g.nucleophilicsubstitution

reaction),whichisusuallycarriedoutat80°Cfor10minutes;- recordatdefinedtimeintervals(e.g.every30seconds)theactualtemperaturevalues;- takeanoteofthemeasurementtime;- reportexperimentaldatainthefollowingtable.

Acceptancecriteria:theaverageexperimentaltemperaturesshouldfallwithinthefollowingrange:78-82°C.

NucleophilicsubstitutionReactiontemperatures

Startreactiontime(pointa):

Endreactiontime(pointa):

Startreactiontime(pointb):

Endreactiontime(pointb):

# Time Thermocouplemeasured

temperaturesatpointa

Automatedsystem

measuredtemperature

# Time Measuredtemperaturesat

pointb

Automatedsystemmeasuredtemperature

37

PerformanceQualification

Processverificationtest

Aim: to verify that the preparation process of the intended radiopharmaceutical is performedcorrectly.

Methodology:

- preparetheautomatedsynthesissystemfollowingthenormalSOPdedicatedtothepreparationof the intended radiopharmaceutical, loading reagents / solvents / components in the properquantitiesandfollowingthedesiredorder;

- performafullpreparationprocessuntilthefinalproductiscollectedinthepropercontainer;- recordandreportthedataobtainedduringthepreparationinthefollowingdatasheet;- repeatthepreparationprocessthreetimes.

Acceptancecriteria:

The preparation of the desired radiopharmaceutical has to be performed correctly, meetingspecificationandacceptancecriteriaforthevariousparameters,withoutsignificantdeviations.Theprocessmaybesplitinthefollowingsteps:

- 18F-fluoridetransferfromthetargettotheautomatedsynthesissystem;transferredradioactivityismeasured;

- purification/trappingof18F-fluorideusingQMAcartridge;- nucleophilicsubstitutionreaction;- hydrolysis;- SPEpurification;- sterilizationusingmembranefilters;- finalformulation;- activityoffinalproductshouldfallwithinthefollowinginterval:2-8GBq;- radioactiveconcentrationshouldmeetthefollowingcriteria:100–500MBq/mL;- thevolumeofthefinalproductsolutionshouldbe15-20mL.

38

Semi-preparativeHPLCpurification

Retentiontimeofthedesiredproductis:

Specification:15-17min

Sterilizationandfinalformulation

Thesolutioncorrectlypassesthroughthefilters rYesrNO

Thevialiscorrectlytransferredtothedosecalibrator rYesrNO

Detectedfinalproductactivity ______MBq

Specification:2-8GBq rYesrNO

PERFORMANCEQUALIFICATIONTEST

Automatedsynthesissystemmodel…..

Batchnumber Date:

18F-fluoridetransfer

Measuredtransfertime: ___________min Expectedtransfertime: max.5min

TheF-18transferstepwasperformedasexpected rYesrNO

DetectedF-18activity: _______MBq

Purification/trappingof18F-fluorideusingQMAcartridge

DetectedactivityafterpurificationusingQMA: _______MBq

Specification:≥80%ofstartingtheactivityhasbeen

recoveredConform: rYesrNO

Nucleophilicsubstitutionreaction

Desiredreactortemperature 100°C Actualtemperature: ______°C

Detectedactivityattheendofthenucleophilicsubstitutionreaction: _______MBq

Hydrolysis

Desiredreactortemperature: 110°C Actualtemperature ______°C

Detectedactivityattheendofhydrolysisstep: ____________MBq

39

Example2:ProcessValidation

Aim: toensure that thepreparationprocessof the radiopharmaceutical [18F]123yield theintendedproductwithcharacteristicssuitableforitsuseinclinicaldiagnosticroutine.

Process validation requires the preparation and quality control of three batches of theradiopharmaceutical[18F]123,followingthesameprocedures,usingthesameinstruments,andusingthesamereagents/solvents/componentsinthesameamountsandcharacteristicsintendedfortheroutinepreparationoft[18F]123.

Descriptionofthepreparationprocess:[18F]123ispreparedviathefollowingsteps:

1. Alltherequiredstartingmaterialsneedtobeready;

2. Aseptic assembly, in a laminar flow cabinet, of the final container (sterile,pyrogenfree,15mlvial)withsterilizingfilter,ventfilterandsterileneedles;

3. radiosynthesis, purification and sterilization of the desiredradiopharmaceutical, fromtheproductionof theradionuclideF-18,until thefinal sterile, pyrogen free solution of [18F]123 is formulated in the finalcontainer;

4. labellingofthecontainer(vial)andofthesecondarycontainer(shieldedleadcontainer);

5. samplingandQCtests;

Here the process has been described in brief; in the real validation protocol it should bemoredetailed.

Tests

Preparationof[18F]123

Prepare threebatchesof [18F]123, following the indicationsdescribed in theproper SOPs.Reporttheexperimentaldatainthefollowingdatasheet.

40

Step2:preparingtheautomatedradiosynthesissystem

2.1Reactorleaktest(leakrate≤5kPa/min)ConformqYesqNO

2.3Listofstartingmaterials: Amount BatchN. Expirydate

[18O]H2O 2g

Kryptofix 15mg/1mlacetonitrile

K2CO3 3.5mg/0.5mlwaterforinjection(wfi)acquap.p.i.

Precursor 10mg

Acetonitrile 1ml

HCl1M 0.3ml

WFI 5ml

HPLCmobilephase(wfi/acetonitrile;95/5) 1L

QMA-lightcartridge 1

2.4Shieldedleadcontainerisready:qYesqNO

2.5Labelforshieldedcontainerisready:qYesqNO

Radiopharmaceutical:[18F]123

Nameandaddressofthepreparationfacility Module#002

Pharmaceuticalform:Injectablesolution

Dateofpreparation: Preparationtime: Batchn°:

Step1:cyclotronproductionofF-18

1.1.startingbeamtime: 1.2.endofirradiationtime(EOB):

1.3.Targetn°: 1.4.targetaveragecurrent:

1.5.Targetintegratedcurrent: 1.6.DetectedF-18radioactivity*

1.7Annexes:irradiationdatasheet

1.8.Comments:*F-18activityisdeterminedbyabuilt-inprobeoftheautomatedsystem

1.9.Operator: 1.10.Operator’ssignature:

41

2.6Comments

2.7Operator: 2.8Operator’ssignature:

Step3:assemblyofsterilevial+filters

3.1.Laminarflow(m/s): Limit(m/s):0.36÷0.54

Conform:Yes�NO�

3.2Materials: Amount BatchN. Expiry

Sterilevials 1

Sterilizingfilter 1

Ventingfilter 1

Needles22Gx1/4"(0.70x30mm) 2

3.3labelforvialisready:Yes�NO�

3.4Operator: 3.5Operator’ssignature:

Step4:radiosynthesisof[18F]123

4.1.Activityatstartofsynthesis(afterelutionofQMA),inMBq:

4.2.Startsynthesistime:

4.3.Activityattheendofsynthesis(EOS)inMBq:

4.4.EOStime:

4.5.Radiochemicalyield%(notcorrectedfordecay):

4.6.Expirytime:

4.7.Annexes:automatedsynthesismodulereportprintout

4.8.Operator 4.9.Operator’ssignature

Acceptancecriteria

Radioactiveconcentration:200-400MBq/MlConform:Yes�NO�

Finalvolume:8-12mLConform:Yes�NO�

Resp.ofthepreparation: Resp.ofthepreparation’ssignature:

42

Qualitycontrolofthepreparedbatchesof[18F]123

Batchesof[18F]123arecontrolledfollowingapprovedandvalidated(ifapplicable)analyticalmethodsimplementedduringradiopharmaceuticaldevelopment.

CERTIFICATEOFANALYSIS MOD.#003

Radiopharmaceutical [18F]123

Date Batchnumber

Analysis results Acceptancecriteria date Conform

pH 4.5–8.5 Yes NO

Appearance Clear,colourlesssolution Yes NO

Identification:gammaspectrometry 511±10KeV Yes NO

Identification:HPLC

Mainradioactivepeakhasretentiontimesimilartothatofthecoldstandardof[19F]123

Yes NO

Residualsolvents:acetonitrile ≤4.1mg/V* Yes NO

Residualsolvents:acetone ≤50mg/V* Yes NO

Chemicalpurity:123 ≤20µg/V* Yes NO

Chemicalpurity:K222 ≤2.2mg/V* Yes NO

Otherimpurities 0.5mg/V*isthesumof«other»impurities Yes NO

Radiochemicalpurity:HPLC ≥95%di[18F]123 Yes NO

Radionuclidicpurity:gamma F-18>99.9% Yes NO

Half-life 105-115min Yes NO

Sterility Sterile(Ph.Eur.) Yes NO

Bacterialendotoxins <175IU/V(Ph.Eur.) Yes NO

*V=maximumrecommendeddoseinmillilitres

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Conclusions

Validation protocol of the radiopharmaceutical [18F]123 has been carried out satisfactorily.Radiochemicalyield,radioactiveconcentrationandqualityofthedesiredproductalwaysmetalltheacceptancecriteria,andnodeviationsoroutofspecificationswereobservedduringtheexecutionofthetests.Thus,thepreparationprocess[18F]123showedtobereliableandeffective.Inconclusion,preparation process of [18F]123 is suitable for its intended use in routine diagnostic activity usingPET/CT.

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Example3:ValidationofoperatingpersonnelwithMediafilltest

1. Purpose

This operating procedure applies for validation and periodic revalidation of the process of asepticpreparationanddispensingofradiopharmaceuticalsobtainedfromradionuclideprecursorandkitforradiopharmaceutical preparation. It is also intended for qualification and requalification of thepersonnelworkingintheasepticpreparationofradiopharmaceuticals.

2. Generalrequirements

Allstepsinamediafillshouldbedoneinthesamelocationsastheradiopharmaceuticalproductionsteps. To initially qualify an aseptic process at a specific facility, three media fills should beconducted on three separate days. The manipulation of materials such as vials and disposablesshould be done in a samemanner as usually used in the hot-lab (i.e. sanitization of thematerialbefore the introduction in theworkarea; sanitizationof the rubberstopperof thebottlesprior tosampling, using syringe and vials shielding, taking into account measuring steps, incubation step,etc.).

Althoughduring themedia fill simulation there isno radioactivity, it is essential toact accordinglywiththerulesadoptedindailyroutineradioprotection,sinceyoumustreproducethemethodsusedinthedailyroutine.

Duringmediafilltestmonitoringoftheair,personnelandcriticalsurfacesshouldbedone(accordingtoSOP:Microbiologicalmonitoring).

Tovalidateorrevalidatetheprocessexecutionofthreesimulationsisnecessary.

The culture media should support growth of a wide range of microorganisms (fungi, yeasts andaerobicbacteria) suchas theTSB (Tryptic soybroth)mediumdistributed inbottlesneeded for themediafilltest.

Afterthetest,atleast2mLofthemediashouldbeleftineachvial,whichenablessterilitytesting.

Checkattheendofthesimulationtheabsenceofradioactivecontaminationofthevials,beforeshipmenttothelaboratoryforincubation.

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3. Materials:

TSBsolution20mLin25mLvial 1vial

TSBsolution10mLin12mLvial 1vial

SterileVacuumvials10mL 1+4=5vials

Syringe10mL 1syringe

Syringe2.5mL 4syringes

Syringe1mL 1syringe

Needles23G qs

Agarplates asprescribedinSOP(Microbiologicalmonitoring)

Rodacplates asprescribedinSOP(Microbiologicalmonitoring)

Swabs for microbiologicalmonitoring

asprescribedinSOP(Microbiologicalmonitoring)

TSB(TrypticSoyBroth)isobtainedfromMicrobiologydepartment,wherethegrowthpromotiontestaccordingtoEur.Ph.isperformedandpositiveandnegativecontrolsolutionsprepared.

4. OperatingProcedure

TSBsolution20mLin25mLvial S1

TSBsolution10mLin12mLvial R1

SterileVacuumvials10mL A1;

P1-1,P1-2,P1-3,P1-4

Syringe10mL 1syringe

Syringe2.5mL 4syringes

Syringe1mL 1syringe

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1. Prepareallmaterialneeded forperforming themedia fill test, andcarefully sanitize it andputitintothematerialair-lockasrequiredbyspecificSOP(Introductionofthematerialintothecleanrooms);

2. enter into the hot-lab (laboratory for the preparation of radiopharmaceuticals). Wearappropriateclothing,cap,maskandglovesasrequiredbyspecificSOP(Entryofpersonnelincleanrooms);

3. transfer all neededmaterials into the laminar-flow cell, use appropriate shielding aswhenworkingwithradioactivity;

4. makeanoteofthebatchrecordalllotnumbersofexpirationdates;5. labelthevialsaccordingtotheschemeabove:

S1: Bottle containing 20 mL of TSB media - Simulates the saline used to dilute theradionuclide to the required concentration for the reconstitution of the kit forradiopharmaceuticalpreparation(lyophilized)-usedinthesimulation1

S2: Bottle containing 20 mL of TSB media - Simulates the saline used to dilute theradionuclide to the required concentration for the reconstitution of the kit forradiopharmaceuticalpreparation(lyophilized)-usedinthesimulation2,secondday

S3: Bottle containing 20 mL of TSB media - Simulates the saline used to dilute theradionuclide to the required concentration for the reconstitution of the kit forradiopharmaceuticalpreparation(lyophilized)-usedinthesimulation3,thirdday

R1: Bottle containing 10mL of TSBmedia - Simulates the solution of radiopharmaceuticalprecursor used to radio-label the kit for radiopharmaceutical preparation - used in thesimulation1

R2: Bottle containing 10mL of TSBmedia - Simulates the solution of radiopharmaceuticalprecursor used to radio-label the kit for radiopharmaceutical preparation - used in thesimulation2,secondday

R3: Bottle containing 10mL of TSBmedia - Simulates the solution of radiopharmaceuticalprecursor used to radio-label the kit for radiopharmaceutical preparation - used in thesimulation3,thirdday

A1: 10mL bottle - Simulates the kit for radiopharmaceutical preparation (containing thelyophilizeddrugtoberadiolabeled)inthesimulation1

A2: 10mL bottle - Simulates the kit for radiopharmaceutical preparation (containing thelyophilizeddrugtoberadiolabeled)inthesimulation2,secondday

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A3: 10mL bottle - Simulates the kit for radiopharmaceutical preparation (containing thelyophilizeddrugtoberadiolabeled)inthesimulation3,thirdday

P1-1:10mLbottle-Simulatesthedispensingstepforpatient1inthesimulation1

P1-2:10mLbottle-Simulatesthedispensingstepforpatient2inthesimulation1

P1-3:10mLbottle-Simulatesthedispensingstepforpatient3inthesimulation1

P1-4:10mLbottle-Simulatesthedispensingstepforpatient4inthesimulation1

Secondday

P2-1:10mLbottle-Simulatesthedispensingstepforpatient1inthesimulation2

P2-2:10mLbottle-Simulatesthedispensingstepforpatient2inthesimulation2

P2-3:10mLbottle-Simulatesthedispensingstepforpatient3inthesimulation2

P2-4:10mLbottle-Simulatesthedispensingstepforpatient4inthesimulation2

Thirdday

P3-1:10mLbottle-Simulatesthedispensingstepforpatient1inthesimulation3

P3-2:10mLbottle-Simulatesthedispensingstepforpatient2inthesimulation3

P3-3:10mLbottle-Simulatesthedispensingstepforpatient3inthesimulation3

P3-4:10mLbottle-Simulatesthedispensingstepforpatient4inthesimulation3

6. IntroducewithintheLAFallthematerialneededtoperformthefirstsimulation:S1,R1,A1,P1-1,P1-2,P1-3,P1-4

7. Insert thebottleR1 in the radionuclide calibrator to simulate the activitymeasurementoftheeluate.Putitbackintotheledvialshield

8. Takea10mLsyringe,usesyringeshieldwithdraw2mLofthesolutionfromtheR1vialanddiluteto10mLwiththesolutionfromtheS1vial

9. Insertthesyringecontaining2mLofTSBfromactivityvialR1and8mLofTSBfromthesalinevialS1intheradionuclidecalibratortosimulateactivitymeasurementinthesyringe.

10. Usesyringeshieldtotransferthevolumefromthesyringeinsidethekitvial-A1(shielded).11. Shake the vial, insert the kit vial - A1 in the radionuclide calibrator to simulate activity

measurementofthefinalproduct.12. Simulatetheincubationstepfor10minutesonambienttemperature(20-25°C).13. Simulatingthecoolingphasefor5minutes.

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14. Take1mLsyringeandwithdraw0,3mLofsolution–thisistosimulatetheasepticsamplingintendedtoperformQC.

15. DispensethecontentofthevialA1to4patientdosesasfollows:P1-1:

- Witha2.5sterilesyringeWithdraw2mLofthesolutionfromthevialA1- Measuretheactivityintheradionuclidecalibrator- TransferthecontentsofthesyringeintothevialP1-1P1-2:

- Witha2.5sterilesyringeWithdraw2mLofthesolutionfromthevialA1- Measuretheactivityintheradionuclidecalibrator- TransferthecontentsofthesyringeintothevialP1-2P1-3:

- Witha2.5sterilesyringeWithdraw2mLofthesolutionfromthevialA1- Measuretheactivityintheradionuclidecalibrator- TransferthecontentsofthesyringeintothevialP1-3P1-4:

- Witha2.5sterilesyringeWithdraw2mLofthesolutionfromthevialA1- Measuretheactivityintheradionuclidecalibrator- TransferthecontentsofthesyringeintothevialP1-4

16. Removeall thematerialusedduringthefirstsimulation.Fill intheformforsterilitytesting

andsendvialsS1,R1,A1,P1-1,P1-2,P1-3andP1-4totheMicrobiologydepartment

17. Forsecondandthirdsimulationrepeatstepsfrom5-17.

Conformity

SterilitytestingisperformedatMicrobiologydepartment.Saveresultsfromthesterilitytestingtogetherwiththebatchrecord.Themedia-filltestconformstotherequirementsifallsamplesinallthreerunsaremarkedasSterile.