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1229.4STERILIZINGFILTRATIONOFLIQUIDS
INTRODUCTION
Sterilizationprocessesaredividedbroadlyintotwocategories:destructionofmicroorganismsand
theirphysicalremovalfromthematerialtobesterilized.Autoclavingisanexampleoftheformer,and
sterilizingfiltrationisanexampleofthelatter.Thephysicalremovalofmicroorganismsdependson
thebioburdenofthesolutiontobefiltered,thepropertiesofthesolution,thefiltrationconditions,and
thefilteritself.
Sterilizingfiltrationisaprocessthatcanbevalidatedtoconsistentlyyieldfiltratesthataresterile,
asdefinedinSterilizationofCompendialArticles1229.Thischapterprovidesanoverviewof(1)
variousfactorsthataffectthefiltrationprocess,(2)thefilterintegritytestandwhentoperformit,(3)
prefiltrationbioburdencontrol,(4)responsibilitiesofthefiltermanufactureranduser,and(5)
troubleshootingthefiltrationprocess.
Multiplefactorscontributetotheeffectivenessofanysterilizingfiltrationprocess.Theseincludethe
typeandnumberofmicroorganisms,thepropertiesoftheliquid,thefilterdesignandmembrane
polymer,andthefiltrationprocessparameters.Propertiesoftheliquidthatinfluencefiltration
effectivenessincludeitschemistry,viscosity,surfacetension,pH,osmolarity,ionicstrength,and
temperature,aswellasthepresenceofinsolublematerials.Aspectsofthefilterthataffectthe
filtrationincludeeffectivefilterarea,nominalporesize,poresizedistribution,membranethickness,
porosity,membranepolymer,filterpleatdensity,nonwovensupportlayers,electrostaticcharge,and
thehydrophilicnatureofthefiltermembrane.Thefiltrationprocessparametersthatinfluence
microbialretentionincludetemperature,flowrate,volume,filtrationtime,differentialpressure,and
pressurepulsations.
Additionally,effectivesterilizingfiltrationdependson(1)theproductioncontrolsandquality
assurancesystemsusedbythefiltermanufacturertoensurethequalityanduniformityofthefilter
membranesandfabricatedfilters,(2)thequalificationandvalidationstudiesconductedby,orfor,the
filterusertodemonstratethatthechosensterilizingfiltrationprocessachievesasterilefiltrate,(3)
effectivecontrolstoensurethatprefiltrationbioburdenandoperatingparametersremainwithinthe
validatedranges,and(4)filterintegrity.Filterusersshouldensurethatthefiltrateremainssterileby
usingvalidatedsterilizationprocessesforthefiltrationassemblyandalldownstreammanufacturing
equipmentandeffectiveaseptichandlingofthesterilizedmaterials.Filterusersshouldcarefully
considerplacementofthefilter(e.g.,proximitytothefillinglineorholdtank)tominimizethe
possibilityofpostfiltrationcontamination.
SterilizingGradeFilters
Asterilizinggradefilterisonethatiscapableofretainingaminimum1107cfuofB.diminuta
(ATCC19146)persquarecentimeterofeffectivefilterareawhentestedinaccordancewithASTM
F83805(2013),StandardTestMethodforDeterminingBacterialRetentionofMembraneFiltersUtilized
forLiquidFiltration(2).
Thedesignationsterilizinggradeimpliesasterilizingactiononlyifotherconditionsaremet,
includingtheintegritytestspecificationestablishedbythefiltermanufacturerandvalidatedbythe
user(seeValidation,below).
Sterilizinggradefilterstypicallyaremicroporousmembranesthathavenominalporesizeratingsof
about0.2m.Thesemembranesarefabricatedwithvariousmaterials,haverelativelynarrowpore
sizedistributions,andcanbeintegritytested.Theintegritytestresultscanbecorrelatedwith
microbialretention.Membranefiltersthathaveanominalporesizeof0.45mcanbevalidatedto
producesterilefiltratesundersomeconditionsforexample,someliquidsrequirehighdifferential
pressurestoachieveusefulflowrates,andthesepressuresarenotsuitableforusewith0.2mrated

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filters.Whenmanufacturersuse0.45mfilters,theyshouldensureparticularlystringentcontrolof
presterilizationbioburden.
RetentionMechanisms
Microporousmembranesremovemicroorganismsfromtheliquidbytwoprimarymechanisms:sieve
retention,whichreliesonphysicalblockageofparticlesthatarelargerthantheporestheyencounter,
andadsorption,whichisachargerelatedphenomenonwherebyparticlesareboundtothemembrane
surfaces.Bothmechanismsshouldbeconsideredduringthedevelopment,qualification,andvalidation
ofsterilizingfiltrationprocesses.
Sieveretention,forthemostpart,isindependentoffiltrationconditionsandthemicrobialchallenge
level,butthecompositionoftheliquid,temperature,andfiltrationconditionscanaffectsieveretention
undercertainconditions.
Adsorptionisachargerelatedphenomenonthatisinfluencedbythecompositionofthemembrane,
thepropertiesofthefilteredliquid,thefiltrationconditions,andthenumberandtypeof
microorganismspresentintheliquid.
Thenumberandtypeofmicroorganismsandotherparticlespresentinthematerialtobefilteredcan
affectretention.Sieveretentionimpliesthateverymicroorganismislargerthanthelargestporein
thefilter.Microorganismsandfilterporesarenotuniforminsize,andsomemicroorganismsmaybe
smallerthanthelargestpores.Evenifamicroorganismissmallerthantheporeitencounters,itmay
beretainedifasiteisavailableandthefiltrationconditionsareconducivetoadsorption.Retention
probabilityisrelatedtothenumberofmicroorganismsintheupstreambioburden(4,5).
FACTORSTHATAFFECTRETENTION
NatureofPores
Microporousmembranesconsistofapolymermatrixthatispenetratedbyintersticescommonly
referredtoaspores.Comparedwiththoseindepthfilters,theporesinmicroporousmembraneshave
arelativelynarrowsizedistribution.Thesize,number,andshapeoftheporesdeterminethefilters
retentioncapabilities.Withtheexceptionoftheporesintracketchedfiltermembranes,poresarenot
cylindricaltheyaremadeupofaseriesofpseudopolyhedralstructureswithvaryinginternal
diameters(6).
Theporesize,poresizedistribution,andmembraneporosityareafunctionofthemanufacturing
process.Carefuldesignandcontrolofthatprocessarenecessarytoensurethattheresulting
membranehasthedesiredintegritytestvalue,microbialretentioncapability,anduniformity.
NatureofMicroorganisms
Microorganismshaveavarietyofshapesandsizes.Ifthemicroorganismencountersamembrane
porethatissmallerthanitssmallestdiameter,themicroorganismlikelywillbecapturedbysieve
retention.If,however,themicroorganismissmallerthantheporeitencounters,itmayberetainedby
adsorptioniftheresidencetime,electrostaticcharge,pH,fluidchemistry,andmembranematerialare
conducivetoadsorption.
Somegeneraofmicroorganismsaredeformable,sothatathighpressuredifferentialsorflowrates
amicroorganismmaybeforcedthroughafilterporethatisonlyslightlysmallerthantheorganism
(7).Mollicutebacterialackcellwallsandthusaresmallandpliableenoughtopassthroughfilter
membranesundercertainconditions.
Growthrough(i.e.,passageofmicroorganismsthroughafilterasafunctionoftime)mayresult
fromoneormorescenarios.Themicroorganismmaypenetrateasitmultiplies:aparentcelldivides
intotwosmallerdaughtercellsthatnegotiateporepassageways.Penetrationcanoccurwithtime,
becausetheincreasingnumberofmicroorganismsoverwhelmsthefewlargerporesthatare
encountered.However,becauseofthelimitedtimeperiodstypicallyinvolved,thecontrolson
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bioburden,andtheoftenlimitedavailabilityofnutrients,growthroughisconsideredarare
phenomenoninpharmaceuticalprocesses(seeMonitoringofBioburden1229.3)(8).
CompositionandStructureoftheFilterMatrix
Severalfactorsrelatedtothefiltermatrixcanaffectmicrobialretention.Theseincludethematerial
fromwhichthefilterismade,theporesizeandporesizedistribution,whetherthemembraneis
isotropicoranisotropic(i.e.,whethermembraneporestructureisuniformfromfacetofaceor
tapersfromonefacetotheother),membranethickness,andwhetherthefilterconsistsofsingleor
multiplelayers.
Themembranematerialisespeciallyimportantifadsorptionisasignificantmechanismina
particularfiltrationscheme.Forexample,polyamideexhibitsstrongermicrobialadsorptionthandoes
celluloseester(3).
Microporousmembranesthathavearelativelywideporesizedistributionarelesslikelytoretain
microorganisms,particularlyathighchallengelevels,thanmembranesofcomparableporesize
ratingswithnarrowerporesizedistributions.Thisrelatestotheprobabilityofamicroorganisms
encounteringaporelargerthanitself.
Thickermembranesgenerallyaremoreretentivethanthinnermembranesofthesametypeand
poresizerating,owingtothehigherprobabilityofentrapmentoradsorptionwithintheporestructure
becauseoftheincreaseddistancethatbacteriamusttravelinthickermembranes.Thisdistancefavors
entrapment,asdoesincreasedresidencetimewithinthepore,whichfavorsadsorptiveretention(3).
Multilayeredmembranefiltersexhibitahigherprobabilityofretentionthandosinglelayered
membranefiltersofthesamethicknessbecausethesmallnumberoflargestporesisthefactor
affectingretentionandtheprobabilityofalargeporeinonelayerbeingcongruenttoalargeporein
theadjoininglayerofadoublelayerfilterisnegligible(3).
CompositionoftheFilteredSolution
Thecompositionofthefilteredsolutioncanadverselyaffectthemembranematerialifan
incompatibilityexists,causingdamagetothemembraneandaffectingbothretentionandphysical
integrity,unlessdetectedbeforethefilterisselectedforuse.Inaddition,ifadsorptionisasignificant
retentionmechanism,thensolutionpropertiessuchaspHandthepresenceofsurfactantsbecome
important.
Surfacechargeandionicstrengthareimportantvariables.Bacterialandmembranesurfacesin
aqueousmediaarenegativelycharged,resultinginrepulsion.Therepulsiveforceisbalancedby
attractiveforces,whichincludehydrophobicsurfaceenergyminimizingforces,operativeonlyover
shortdistances,andhydrogenbonding.Highionicstrengthallowsthesurfacestoclosebecauseof
dischargethroughtheelectrolytetothepointwherehydrophobicadsorptioncanoccur(3).Also,high
ionicstrengthcandrawwateroutofthecell,reducingitssize,whichmaylowertheprobabilityof
retention,dependinguponthecompositionoftheprefiltrationbioburden.Surfacetensionandthe
presenceofsurfactantsinfluenceretention.Adsorptionofsurfactantbythefilterandthe
microorganismcreatesrepulsion,leadingtoadecreasedprobabilityofretention.Surfactantsin
concentrationsaslowas0.05%havebeenshowntoinhibitadsorptionanddecreasetheretentionof
latexspheres(9).
FiltrationConditions
Differentialpressure,flowrate,andtemperatureareamongthefactorsthatcanaffectmicrobial
retention.Hydraulicshockshouldbeavoided,notonlybecauseitaffectspressuredifferentialandflow
ratebutalsobecauseitcandamagethefilter.Flowrateisproportionaltodifferentialpressure,and
higherflowratesreduceadsorptionbecausethecontacttimeisreduced(10).
Microbialretentionmaybereducedathighertemperatureswhentheseresultinhigherflowrates
duetodecreasedsolutionviscosity.Temperatureeffectsarenotsignificantinfilterswheresieve
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retentionistheprimaryremovalmechanism.
FilterEfficacy:LogReductionValue
Filtrationefficacycanbedefinedintermsofalogreductionvalue,whichisthelogarithmofthe
quotientproducedbydividingtheupstreamchallengepopulationbytherecovereddownstream
population.
Thelogreductionvalueisinfluencedbythenumberandsizeofthechallengemicroorganisms,the
filterdesignandmembranepolymer,thefiltrationprocessparameters,andthepropertiesofthe
solution.
Todeterminethespecificlogreductionvalueofafilter,thechallengetestshouldpermitsome
passageofthetestmicroorganismthroughthefilterinordertoproduceadenominator.Sterilizing
grademembranefiltersshouldnotpermitpassageofthespecifiedchallengemicroorganismsforthat
filterrating.Forthisreasonthelogreductionvalueofsterilizinggradefiltersisdescribedasequalto
orgreaterthanthelogofthechallengepopulation.
Validation
Asnoted,microbialretentioninsterilizingfiltrationreliesonacombinationofsieveretentionand
adsorption.Validationofsterilizingfiltrationthereforerequiresdeterminationoftheeffectoftheliquid
onthefilter,determinationoftheeffectofthefilterontheliquid,anddemonstrationthatthefiltercan
consistentlyyieldsterilesolutionsundertheintendedconditionsofuse.Theliquidtobefilteredcan
affecttheporestructureofthemembrane,canhaveelectrostaticpropertiesdifferentfromthe
standardchallengesuspensionusedtoestablishintegritytestspecifications,andcanchangethesize
andshapeofthechallengemicroorganisms.Factorsthatshouldbeconsideredwhendevelopinga
sterilizingfiltrationvalidationprotocolincludethesurfacetension,pH,temperature,andosmolarityof
theliquidtobefilteredthecompatibilityofthematerialorsolutioncomponentswiththefilteritself
thepressures,flowrates,andhydraulicshocklikelytobeencounteredandthemaximumfiltration
timeandvolumetobefiltered.Theeffectofsterilization(steam,radiation,orgas)onthefilters
retentioncapabilityalsoshouldbeconsidered.
B.diminuta(ATCC19146)isusedasthechallengeorganismunlessitisnotviableintheliquidtobe
filtered.Viabilitystudiesshouldbeusedtoconfirmthattheliquidhasnoadverseeffectsonthe
challengeorganism.Ifthechallengeorganismisviableintheliquidtobefiltered,theliquidshouldbe
inoculatedtoachieveachallengelevelof1107cfu/cm2,andthefiltrateshouldbeevaluatedforthe
presenceofthechallengeorganism.IfB.diminutaisnotviableintheliquid,severaloptionsare
available,andanalystscan(1)modifytheliquidtoensuretheviabilityofthechallengeorganism
(e.g.,adjustpHorremovethebactericidalcomponent),(2)reducetheexposuretimetoensurethat
thechallengeorganismremainsviable,or(3)changethechallengeorganismfromB.diminutatoone
thathasbeenisolatedfromtheliquidtobefiltered.Thesestudiesshouldemployproductionprocess
pressuredifferentialsorprocessfluxvaluesasappropriate.
Ifpossible,theliquidtobefilteredshouldbeusedbecauseinsomeinstancesthechallengeorganism
haspenetratedafilterincontactwiththeliquidbuthasbeenretainedbythesamefilterwhen
inoculatedintoasurrogatefluid(11).
Threedifferentlotsoffiltermembranesshouldbeusedforthemicrobialretentionstudies.The
membranesshouldhavepreuseintegritytestvaluesthatarenearthefiltermanufacturer's
specificationinordertominimizethepossibilitythatproductionfilterswillfailtomeettheintegrity
testvalueestablishedduringthevalidationexercise.Successfulmicrobialchallengestudiesresultin
nomicroorganismsdetectableinthefiltrate.Thesterilizationprocessforthefilter,itshousing,and
associatedequipmentshouldbevalidated.Thefiltershouldbesterilizedwithinitshousingratherthan
relyingonasepticassemblyfollowingsterilizationofthefiltrationsystemcomponents.Theassembled
filtrationapparatuscanbesteamsterilizedinanautoclave,usingavacuumcycle,withparticular

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attentiontotheorientationandwrappingofthehousingandanyassociatedtubingtoallowcondensate
drainageandsteampenetration.Validatedsteamsterilizationinplacecyclesalsocanbeused.
Sterilizationmethodsandcyclesshouldbecarefullychosenanddesignedtoprecludedamagetothe
filter.Preandpoststerilizationintegritytestingcanbeusedtoconfirmthatthesterilizationprocedure
doesnotchangetheintegritytestvalueandtodemonstratethatthesterilizationprocessdoesnot
damagethefilter.
Finally,thescaleofthevalidationapproachshouldbeconsidered.Twoapproachesarecommon.One
usesasmallsectionofmembranematerial,typicallya47mmdisk,torepresentthefilter.This
approachvalidatesthemicrobialretentioncapabilityofthemembrane.Asecondapproachusesthe
intendedfilterconfiguration,typicallya10inchcartridgeinitshousing.Thisapproachvalidatesthe
filtersystemperformanceinadditiontotheretentioncapabilityofthemembranematerial.Analysts
shouldconsiderthevolumeofthetestliquidandoperationalconsiderationswhentheychoosewhich
approachtouse.
IntegrityTestingPrinciplesandMethods
Integritytestingcanbeusedtoshowthatafilterhasthecorrectporesizerating,isinstalled
properlyinitshousing,andhasnotbeendamagedbytheprocessusedtosterilizeit.Theseintegrity
testmethodsgenerallyrelyondetectinggasflowcausedbypressuredifferentialacrossawetted
membrane.Figure1showstherelationshipbetweengasflowandpressuredifferentialforhigh
surfaceareaandlowsurfaceareamembranes(a47mmmembranediskcanbeconsideredlow
surfacearea,andamulticartridgearrayof10inchcartridgefilterscanbeconsideredhighsurface
area).

Figure1.Relationshipbetweengasflowandpressuredifferentialforhighsurfaceareaandlow
surfaceareamembranes(seethesectionIntegrityTestingPrinciplesandMethodsfordefinitions).
Afilterthatsuccessfullypassesanintegritytestbasedonspecificationsdevelopedduringan
effectivesterilizingfiltrationvalidationstudyiscapableofproducingasterilefiltrate.

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Integritytestmethodsformicroporousmembranefiltersinclude bubble point,diffusiveflow,and

pressurehold.Membranetypeandsizeandtheparticularfiltrationprocessinfluencethechoiceofan
appropriateintegritytestmethod.Thesensitivityofthe bubble pointtestdecreaseswithincreasing
filterareabecauseofincreasingdiffusiveflow. Bubble point,diffusiveflow,andpressurehold
integritytestsareusedforhydrophilicmicroporousmembranefilters.AsshowninFigure1,diffusive
flowoccursatpressuresbelowthe bubble point,andbulkflowtakesplaceabovethe bubble point.
The bubble pointmarksthetransitionbetweendiffusiveflowandbulkflow.Theexact bubble point
isdifficulttodetectinhighsurfaceareafilters,becausethehighmembranesurfaceareagenerates
significantdiffusion.Integritytestingmaybeperformedmanuallyorbyautomatedequipment
designedspecificallyforthatpurpose.
BUBBLE

POINT

The bubble pointoccurswhenagasdisplacesawettingliquidfromthelargestmembranepores,

resultinginbulkgasflowthroughthosepores.Theflowisevidencedbyasteadystreamofbubbles
throughacolumnofwateronthedownstreamsideofthemembrane.
The bubble pointrelatestotheporesizeofthemembraneandthecontactanglethatthewetting
liquidmakeswiththeporewall. Bubble pointisindirectlyproportionaltomembraneporesizeand
isdirectlyproportionaltothesurfacetensionofthewettingliquidthatis,filterswithsmallerpores
havehigher bubble pointsthanthosewithlargerpores,andliquidsthateasilywetthemembrane
exhibithigher bubble pointsthanthosethatdonot. Bubble pointisdefinedas:
P=4cos/D
P=pressuretoevacuatethepore
=surfacetension
=angleofwetting
D=porediameter
Microbialretentionand bubble pointcorrelate:numerousstudieshavedemonstratedthatwhen
themicrobialreductionratioisplottedagainstthe bubble point,alineofconstantsloperesults
(12).
Theactual bubble pointisindependentofthemembranesurfacearea,butdiffusiveflowthrough
highsurfaceareafilterscanmaskthetrue bubble point.The bubble pointtestiseasytoperform
onsmalltomediumscalefilters,thetesttimeisshort,andtemperatureeffectsareminor.Manual
bubble pointtestingrequiresmanipulationofthedownstreamsideofthefilterandissubjective.
DIFFUSIVEFLOW

Indiffusiveflowtestingawettedfilterprovidesaliquidlayeracrosswhichgascanflowbymeans
ofdiffusion.Diffusiveflowismeasureddirectlyatconstantpressure.
Diffusiveflowisproportionaltothedifferentialpressureofthetestgas,thediffusivityofthetest
gasinthewettingliquid,thethickness(depth)ofthewettingliquid,theporosity(i.e.,voidvolume)
ofthemembrane,andtheeffectivefiltrationarea.DiffusiveflowisdefinedbyFick'sLawof
Diffusion,shownas:
N=[DH(p1p2)]/(L)
N=permeationrate(molesofgasperunittime)
D=diffusivityofthegasintheliquid
H=solubilitycoefficientofthegas
p1p2=transmembranepressure(differentialpressure)
L=thicknessofliquidinthemembrane
=voidvolume(porosity)ofthemembrane
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Unlikethe bubble pointtest,diffusiveflowtestingmeasuresgasflowthroughallthewetted

pores,andthusdiffusiveflowdoesnotcorrelatedirectlywithmicrobialretention.However,bacterial
challengetestswithaseriesoffiltersthathavedecreasingdiffusionratesshowthatagasdiffusion
rateexistsbelowwhichsterilefiltratesareobtained.
Diffusiveflowtestingishighlysensitive,especiallyforhighersurfaceareamembranes.Larger
poresorflawscanbedetectedbyathinningoftheliquidlayerandcorrespondinglyhigherdiffusive
flowrates.Diffusiveflowtestingisusefulformembraneswithsmallporesizes(e.g.,0.1mand
smaller)becauseofthehighpressuresrequiredfor bubble pointtesting.Diffusiveflowtesting
measuresflowacrossthetotalporevolume,whichmaymaskaflaw,especiallyinhighsurfacearea
multiplecartridgearrays(13).Thistestalsoishighlysensitivetotemperature.
PRESSUREHOLD

Thepressureholdintegritytestisavariationofthediffusiveflowtest.Thewettedmembrane
providesaliquidlayeracrosswhichgascanflowbymeansofdiffusion.Thegasflowisproportional
todifferentialpressureandismeasuredbypressuredecayontheupstreamsideofthemembrane.
Therateofpressuredecayisinfluencedbyupstreamvolumeoftheparticularholderfilter
combination,valveplacement,andtubingvolume.Temperatureshouldremainconstantbecauseof
therelationshipbetweentemperatureandpressuredefinedbytheidealgaslaw.
Conversionofpressuredecaytestresultstodiffusiveflowvaluesallowscorrelationwithmicrobial
retention.Analystsestablishtherelationshipbetweenpressuredecayanddiffusiveflowby
calculatingdiffusiveflowonthebasisofpressuredropperunittime,withaknownupstreamvolume
andreferencepressure.Diffusiveflowasitrelatestopressuredecayisshownas
D=[(p1V1)/(p0t)]ln[p1/(p2Dp)]
D=diffusion
p1=startingtestpressure
V1=upstreamvolumeoffiltersystem
p0=atmosphericpressure
t=testtime
p2=endingtestpressure
Dp=p1p2
Theadvantagesanddisadvantagesofthepressureholdtestaresimilartothoseofthediffusive
flowtest.Thepressureholdtesthastheadditionalabilitiesofrevealingimperfectionsinthe
assemblyandsealingofthehousingandfilterseatingandavoidingdownstreammanipulation.Its
disadvantagesarethatitisstronglyinfluencedbytemperatureandthataccuratemeasurementof
theupstreamvolumeisrequired(14).
WhentoTestIntegrity
Thedecisionnottoperformpreuse,presterilizationintegritytestingshouldbebasedonaformalrisk
assessment.
Preuse,poststerilizationintegritytestingmaycreateanunnecessaryriskformicrobialcontamination
ofthefilterandassociateddownstreamtubingandequipment.Preuse,poststerilizationintegrity
testingisunnecessaryifeffectivevalidationstudieshavedemonstratedthattheprocessforsterilizing
thefilterdoesnotaffecttheintegritytestvalueofthefilter.
Postfiltrationintegritytestingshouldbeconductedtoensurethatthefilterwasnotdamagedduring
thefiltrationprocess.
PREFILTRATIONBIOBURDENCONTROL

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Thebioburdenremovalcapabilitydependsontheavailablefilterretentioncapacity,whichisa
functionoftheinherentbioburdenloadpresentintheentirevolumeoftheliquidtobefilteredandthe
effectivefiltersurfacearea.Studieshavedemonstratedthatmicrobialretentioninsterilizingfiltration
isafunctionoftheupstreambioburden(3,15,16).Also,nonviableparticulatematterthatmaybe
presentinthesolutioncaninfluencetheretentivecapacityofthefilter(17).Therefore,the
prefiltrationbioburdenandparticulatelevelsofthesolutionshouldbeminimizedandcontrolledbefore
thefinalsterilizingfiltrationstep.
Variousfilterconfigurationsandprocessescanbeusedtocontrolthebioburdenandnonviable
particulatelevelspresentedtothefinal,sterilizinggradefilter.Oneconfigurationisamultifilter
arrangementthatconsistsoftwosterilizinggradefilters(orabioburdenreductionfilterfollowedbya
sterilizinggradefilter)connectedinseries(Figure2).

Figure2.Multifilterconfigurationtocontrolbioburdenandnonviableparticulates.
Anotherconfigurationappropriateforprefiltrationbioburdencontrolusestwofiltrationsteps
separatedintime:theliquidissterilefilteredintoasterilizedtank,whereitisthenheldbeforea
final,sterilizingfiltration(Figure3).

Figure3.Bioburdencontrolusingtwofiltrationsteps.
Ineachofthesescenarios,thebioburdenandparticulatelevelspresentedtothefinal,sterilizing
gradefilterarelowandarecontrolledbyprefiltration.
Whentheprocessresultsinaconsistentlylowandcontrolledprefiltrationbioburden,useofasingle
sterilizinggradefilterisappropriate.
Irrespectiveofthestrategyemployed,validationstudiesshoulddemonstratethecapabilityto
consistentlyachievetherequisitelevelsofprefiltrationbioburdenandparticulatelevelreductionand
control.
RESPONSIBILITIES
FilterManufacturer
Thefiltermanufacturerisresponsibleforensuringthatthefilterproductionprocesshasbeen
validatedandiswellcontrolledandthatthesterilizinggradefiltersmeettherequirementsofASTM
F83805(2013).Thefiltermanufacturerdeterminestheintegritytestspecificationforthefilters,
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usuallyaddingasafetyfactortoensurethateachfilterwillmeetthatspecification.Thefilter
manufacturerconductsextractableandleachablestudiestoensurethatthefilterdoesnotrelease
objectionablelevelsofthesematerialsintothesolventsystemstypicallyemployedinpharmaceutical
manufacturing.Thefiltermanufacturerconductscleanlinessteststoassurethatthefilterdoesnot
adverselyaffecttheUSPparticulaterequirementsoftheproduct.Thefiltermanufacturerprovides
technicalsupportandtroubleshootingadviceifthefilteruserencountersaproblem.
FilterUser
Thefilteruserisresponsibleforestablishingmicrobialretentionatthevalidatedintegritytestvalue,
establishingmicrobialretentionintheliquidtobefiltered,andvalidatingtheuseandsterilizationof
thefilterandhousing.Thefilteruserisresponsiblefordeterminingthatthefilterisnotadditiveor
extractivetotheextentthatthefilteredliquidisadverselyaffected.
TROUBLESHOOTING
Failureofafiltertopassanintegritytestmaymeanthatthefilterisdamaged,isimproperlysealed
inthehousing,isincompletelywetted,orisnonintegral.Italsocouldmeanthatthefilterisincorrectly
labeled(e.g.,hasthewrongporesize)orthattheintegritytestapparatushasbeenimproperlysetup
orcalibrated.
Thecauseofanintegritytestfailurecanbedeterminedbyevaluatingthetestsetup,test
parameters,wettingfluid,andwettingprocedureensuringthatthesystemisleakfreeandthe
temperatureisconstantandensuringthatthetestequipmenthasbeenproperlycalibrated.
Ifthecauseofthefailurecannotbedetermined,analystscanrewetthefilterandrepeatthe
integritytest,increasingflushtime,flushvolume,andpressuredifferential.Itmaybebeneficialto
usealowersurfacetensionreferencefluid(e.g.,70%isopropylalcohol).Ifthefilterfailstheintegrity
testusingthereferencefluid,itshouldbeconsiderednonintegral.Inaddition,thefiltercanbe
returnedtothemanufacturerforafullanalysistofurtherelucidatethecauseoftheintegritytest
failure.
REFERENCES
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