Home>RuggednessofVisibleResidueLimitsforCleaningValidation

RuggednessofVisibleResidueLimitsforCleaning
Validation
Visibleresiduelimitshavebeenshowntobeavaluabletoolinvalidatedcleaning
validationprogram.
Apr02,2016
ByRichardJ.Forsyth[1]
PharmaceuticalTechnology
Volume40,Issue4,pg5057
Peerreviewed:
Submitted:July31,2015.Accepted:September11,2015.
Abstract
Visibleresiduelimits(VRLs)havebeenshowntobeavaluabletoolinavalidated
cleaningvalidationprogram.Asanorganizationgainsexperienceinestablishing
VRLs,theestablishedlevelsgenerallygoloweraspersonnelbecomemore
familiarwiththeappearanceofvisibleresiduesandspikeincreasinglylower
levelsofresiduetoobserve.Theselowerlevelshaveledtoaconundrumthat
requiresanadjustmentinthewayVRLsaredefined.
Visibleresiduelimits(VRLs)canbequantitativelyestablishedforpharmaceutical
products,APIs,excipients,anddetergents(13).TheVRListhelowestlevelof
residuethatcanbeseenbyapanelofobserversunderadefinedsetofobservation
parameters.IfaVRLislowerthanthecalculatedacceptablecleaningresiduelimit
forapieceofequipmentoranequipmenttrain,thentheVRLcouldbeconsidered
thecleaninglimitforthatequipment.Theregulatoryagencieshaveacceptedtheuse
ofVRLsasavaluabletooltohelpdevelop,validate,andmaintainarobustcleaning
validationprogram(4,5).
TheruggednessofVRLshasbeenestablishedfordifferentresidues,personneland
viewingconditions(6),presentationofcouponsforpersonnelobservation(7),andfor
differentmaterialsofconstruction(MOCs)(8).Throughallofthesestudies,the
methodofpreparationoftheVRLsampleswasheldconstant.Asolutionor
suspensionwaspreparedinavolatilesolvent,typicallymethanol,serialdilutions
weremadetodecreasingconcentrationlevelsandcouponswerespikedbypipetting
100LontotheMOCcoupon.Themethanolspreadouttoacircleofabout5cm
diameterandrapidlyevaporated,leavingaringofvisibleresidue.

Figure1:Measuringfull
area.Therehavebeentwoprimaryoutstandingconcernswiththisapproachto
establishingVRLs.ThefirstconcernwasthataVRLestablishedunderdefined
viewingconditionsmightnotbereplicatedinthefieldandthatapracticalVRLmight
behigher.Althoughthisissuehasbeenexplored(9),ithasnotbeendefinitively
addressedandtakingspikedcouponsintothefieldorreplicatingVRLspikelevelon
actualequipmentisnotpracticalonanongoingbasis,butitdoeswarrantadditional
work.
Thesecondconcernwasthattheresiduesformedaringandwerenotevenly
distributedoverthecoupons.Whileitwasacknowledgedthattheresidueswerenot
evenlydistributedoverthecoupons,inthefield,residuesoncleanedequipment
weremorelikelytoshowupasringsthanasanevencoverageontheequipment,
especiallyatlowresiduelevels.Theentireareaoftheresultingcirclewasusedin
thecalculationoftheVRL(seeFigure1).ThistechniqueofdeterminingVRLswas
notanissueuntilexperienceandimprovedtechniqueloweredtheexperimentally
determinedVRLstolevelsthatmadethemimpractical.Alternativeapproachesto
determiningVRLsareproposed,whichbringVRLlevelsbacktoamorepractical
level.Theadvantagesanddisadvantagesofthevariousapproacheswerealso
comparedtodeterminethemostexpeditiouspathforward.

Background

[2] CLICKTABLETOENLARGE

TableI:Spikingpreparationandtargetconcentrations.
TheinitialworkonVRLs(1)startedwiththe100glevelwithatargetof4g/cm2 to
correlatewiththelevelcitedintheFourmanandMullinarticle(10).Additionallevels
atdecreasingvalueswerealsodetermined,butatruevisuallimitwasnot
determinedifthelowestleveltestedwasvisibleanditwassufficientlylowerthanthe
acceptablecleaningresiduelimit(ARL)(i.e.,atleasttwotimesloweror50%ofthe
ARL).TheinitialworksurveyedawiderangeofAPIs,excipients,andformulations
overarelativelyshortperiodoftimetoestablishacompletedatabaseforacleaning
validationprograminaclinicalmanufacturingfacility.AsapplicationsofVRLs
expanded(2)andexperiencegrew,thespikinglevelsoftheresidueswerelowered
untilafinalsetofspikelevelswerefinalized,asshowninTableI.
TheaverageofseveralhundredVRLdeterminationswas1.1g/cm2 (6),which
madeitapracticalandachievablecorrelationtousetheVRLastheARL,althoughit
wasnotedthatroughlytwothirdsoftheVRLswereabletobeseenatthelowest
levelspikedontothecoupons.Therewasaclearindicationofthecurrentsituation.In
addition,theaveragenumberswerealittlemisleadinginthatasexperience
increasedindeterminingtheVRLlevels,thenumbersgeneratedbegantoskewto
lowerlevels(6).Now,when1gofmaterialisspikedontoastainlesssteelcoupon
andallowedtospreadouttoa5cmdiametercircle,itisoftenstillvisibleunder
definedconditionsofviewingdistance,viewingangle,andlightlevel.Theareaofthe
samplewitha5cmdiameteris19.6cm2 (x(2.5cm)2 ),makingtheconcentrationof
thesample1g/20cm2 or0.05g/cm2 .Thiswouldequatetoaswabsampleof1.2
gusinga25cm2 swabarea.Extractingthe1.2gsampleintosolventforanalysis
resultsinasolutionconcentrationwellbelow1.0g/mL.

[3] CLICKTABLETOENLARGE

TableII:Spikingpreparationandconcentrations.
Anattemptwasmade(10)toincorporateVRLsalongwithswabandrinserecoveries
andrelativecleanabilityintoanefficient,cohesiveefforttogeneraterelevantworst
casesoils,ARL,andrecoveryfactorsintoashortseriesofstudies.Itwasunderstood
thatanARLwascalculatedusinganacceptabledailyexposure(ADE)toassure
patientsafetyandiftheVRLwaslowerthanthecalculatedARL,thentheVRLlevel
wasthedefactocleaninglimit.Allcleaningvalidationtestsamplesshouldcome
backlowerthanthecleaninglimittovalidateacleaningprocedure.

Usinga1g/20cm2 or0.05g/cm2 VRLasacleaninglimitisnotpractical.VRLs

establishedusingthisapproachatthislevelcouldresultinapracticalquantitative
contradictionbetweentheVRLandthelimitofquantitation/limitofdetection
(LOQ/LOD)oftheanalyticaltestmethodiftheVRLislowerthantheLOQ/LOD.Per
regulatoryguidance,thesensitivityoftheanalyticalmethodshouldextendbelowthe
cleaninglimittoassesstheeffectivenessofthecleaningprocedure.Thisapproach,
however,isnotpossibleifthevisuallimitislowerthantheanalyticallimit.Evenwith
aVRLatornearthequantitationlimitsoftheanalyticalmethod,therewouldbea
verynarrowmarginbetweentheVRLlevelforapieceofequipmentandtheabilityof
theanalyticaltestmethodtodeterminetheresidualanalyteofinterest.

Figure2:Measuring'donut'areaAslongas
theVRLisabovetheLOQ/LODoftheanalyticalmethod,thisapproachto
determiningVRLsisacceptable.However,aspersonnelgainexperience,theVRLs
starttrendingtolowervaluesandtheanalyticalsensitivitywillbecomeanissue.
Eitheranalternative,morepracticalquantitativeapproachisnecessary,orVRLs
generatedusingthisapproachcanbetreatedasmorequalitativewiththeassurance
thattheequipmentwillbevisuallysoiledlongbeforethereisaconcernforpatient
safety.
Methodology
ThereareseveralalternativestotheVRLmethodologydescribedinthefirstpartof
thisarticle,bothqualitativeandquantitative.Asecond,qualitativealternativewould
betospikeresidueattheARLandatsomelevelbelowtheARL(50%or25%)using
thesamespikingprocedureasdescribedpreviously.Iftheresiduesarevisually
detected,thentherewouldbeconfidencethattheresiduewouldbevisibleon
equipmentatlevelslowerthantheARLand,therefore,visualinspectionwouldbe
appropriatetodetermineequipmentcleanliness.TheVRLwouldbelistedasless
thanthelowestresidueleveltested.
Forthisstudy,twoadditionalalternativemethodsofdeterminingpractical,
quantitativeVRLswereexplored.Athirdapproachistocontinuetopreparethe
residuesamplesinthesamemannerasdescribedandmeasuretheareaofonlythe
residueringbysubtractingtheareaoftheinnercirclefromthelargercircleleaving
onlytheareacoveredbytheresidueasshowninFigures2and3usingEquation1:
[Eq.1]RingArea=r2r2 2=(r1 2r2 2 )=(r1 r2 )(r1 r2 )

Figure3:Measuring'donut'area.For
example,spiking100Lofa10g/mLspikingsolutionresultsinaresiduewithan
outerdiameterof5cmandaninnerdiameterof4.8cm,theareaoftheresidueis1.5
cm2 comparedtotheareaofthe5cmcirclealone,whichis19.6cm2 .Fora1g
residuesample,thischangestheconcentrationfrom0.05g/cm2 to0.67g/cm2 .
Thiswouldresultinaswabsampleof17gusinga25cm2swabarea.
Afourthapproach,whichshouldalsobemorequantitative,istogeneratearesidue
withauniformcoverage.Becauselowspikelevelssuchas1gin100Lresultina
ringresidueatlargerareas,minimizingtheareaoftheresiduecanbeaccomplished
byspikingaminimumvolumeofahigherconcentrationsample.Spiking20Lofa
75g/mLspikingsolutionresultedinamoreuniform1.5gresidue.Theresulting
uniformresiduespreadoveracirclewithadiameterof0.8cmandanareaof0.5cm2
andaconcentrationof3g/cm2 or75g/25cm2 .

Figure4:Measuringuniformarea.
Serialdilutionsweremadeandsimilarlyspiked.Theadditionalsamplesareshown
inTableIIandFigure4.ItcanbeseeninFigure4thatthefirstthreespotsare
uniformwhilespotsfourandfivearenotuniformlydistributed.TheVRLcanbe
determinedusingthelowestuniformlydistributedresiduefromspotthreeforaVRL
of0.5g/cm2 .Althoughspotsfourandfivearevisible,theyarenotuniformly
distributedandattemptingtodetermineaVRLwouldresultinthesameissueaswith
theFigure1determination(i.e.,overestimatingthearearesultinginalowerVRL).
ThissituationoftheuniformandnonuniformresiduesisanalogoustotheLOQand
LODofananalyticaltestmethod.Thethirdspotcanbequantitativelydetermined,but
quantitationofspotsfourandfiveisnotpossibleeventhoughtheyarevisible.

Discussion

[4] CLICKTABLETOENLARGE

TableIII:Optionsforestablishingvisibleresiduelimits(VRLs).MeOHismethanol,
ARLisacceptablecleaningresiduelimit,LOQislimitofquantitation.Eachofthefour
approachesforVRLshasitsadvantagesaswellasdrawbacks.TableIIIprovidesa
summaryforthedifferentapproachesincludingpreparation,levelofVRL,and
primaryissue.Eachisareasonableapproachaslongasthecompanydefinestheir
processandisawareoftheintentandlimitationsfortheapproachtheyhavechosen.
Theinitialapproachdescribedinthebackgroundsectionofspikingaknownquantity
ofresidueindecreasingamountsontomultiplecouponsandthenmeasuringthe
areaoftheentirecirculardriedresidueprovidesassurancethatvisibleresidue
wouldbedetectedontheequipmentsurfaceswellbeforetheresiduebecomesa
safetyriskinthesubsequentmanufacturedbatch.ThisdeterminationofaVRL
providesaquantitativeassessmentofthemarginofsafetybetweentheVRLlevel
andtheARLofthespikedAPIorformulation.Thisapproachservesasamuchbetter
trainingtoolinthatpersonnelwillgetabetterimpressionoftheappearanceofthe
residuesastheyneartheVRL,whichisavaluableperspectivetohave,particularly
foroldercleanedequipmentthatissubjecttootherpermanentmarksonthe
surfaces.Itisalsopracticalinthatresiduewilloftenappearasringsorwaterspotson
cleaned,driedequipment.
ThemajorpotentialdrawbacktothisapproachcanbeseeninTableI.Ifa1g
spikedsampleisvisible,whichisoftenthecase,itwillhavearesultingconcentration
of0.05g/cm2 .Swabbingthissampleandextractinginto10mLofsolventwould
resultinasampleconcentrationof0.1g/mL.IfnotalreadybelowtheLOQofthe
analyticalmethod,itwouldbeveryneartoitformostanalyticaltestingtechnologies,
includingthecommonlyusedtechnologiesfortestingcleaningsamples,suchas
highperformanceliquidchromatography(HPLC)andtotalorganiccarbon(TOC)
analysis.BecausetheVRLshouldbeconsideredequivalenttotheARL,itis
expectedthatthetestmethodforcleaningsamplesshouldhaveanLOQlowerthan
theARLfortheresidue.Theanalyticaltestmethodmightnotbeadequately
sensitive,orifitissensitiveenough,sampleswouldbeindangeroffailingifanything
isdetectedinthetest.
ThisdangerofapotentialcleaningfailureusingthisVRLapproachisunjustified
becausethemajorityofthespiked,driedresidueareaisnotoccupiedwithresidue.
Attheselowspikedconcentrations,aringisformedandtheresidueoccupiesonlya
smallareaaroundtheperimeterofthedriedcircle.Althoughthisapproachto
determinationofVRLsisefficientandprovidesanaccuratedeterminationofthe
amountofresiduedividedbytheareaoftheresidue,theresidueisnotauniform
residueandresultsinaVRLlowerthanispractical.AVRLdeterminedthiswaycan
bemisleadingifanattemptismadetorelateittotheLOQoftheanalyticalmethod.
VRLsestablishedusingthisapproach,however,cancertainlybecomparedtothe
ADEcalculatedcleaninglimittodeterminethatvisualinspectionisadequatefor
cleaningconfirmationandtodeterminehowwidethemarginofsafetyisbetween
thevisuallimitandthecleaninglimit.
Thesimplestapproachisthesecondonedescribedpreviously,whichistospike
couponsatasinglelevelthatislower(e.g.,50%)thantheARL.Ifthespikedresidue
isvisible,itprovidesassurancethatvisibleresiduewouldbedetectedonthe
equipmentsurfacesbeforetheresiduebecameasafetyriskinthesubsequent
manufacturedbatch.Thisapproachavoidsthelowlevelpracticalityissueandalso
hastheadvantageofspeedinthatsamplescanbepreparedrelativelyquicklyand
calculationsarenotnecessary.Fewercouponswouldbeusedperresidue,
therefore,moreresiduescouldbequalifiedatthesametime.Thisapproachmightbe
recommendedifacompanywishedtoestablishaVRLprograminanefficienttime
frameorquicklydeterminethelimitationsofdetectingthevisibleresidueusing
multipledistancesandviewingangles.Thisapproachwouldalsobeusefulfor
trainingorqualifyingalargegroupofpersonnel,suchasallequipmentwashersand
inspectionpersonnel.Thisapproachisalsopracticalinthatresiduewillmostoften
appearasringsorwaterspotsoncleaned,driedequipment.

Thedrawbackswiththisapproacharethatitprovidesaqualitativedetermination
only,whilenotwelldefiningthemarginofsafetybetweentheVRLandtheARL.A
minordrawbackisifthespikedresidueisnotvisible,onecannotbecertainthat
somelevelbetweenthespikedlevelandtheARLmightstillbevisibleandadditional
couponswouldhavetobespikeduntilavisualresidueisobserved.Thebiggest
drawbacktoapurelyqualitativeapproachisthatthevisuallimitshouldactuallybe
thecleaningARLifresidueisvisibleatlevelslowerthanthecalculatedcleaning
ARL.However,ifacompanystatesintheirstrategythattheyarelookingatVRLs,
solelyfromaqualitativeperspectiveandthatthelowerspikelevelprovidesalevelof
confidencetoensurethatvisiblysoiledequipmentwillnotbedeemedclean,and
usesthecalculatedARLsastheircleaninglimit,thisshouldbeaviableapproach
anddefensiblewiththeagencies.
Thethirdapproachistousethesamespikingtechniqueasthefirsttwoapproaches,
butthenmeasureonlytheringareathatiscoveredbytheresidue.Thereareseveral
advantageswiththisstrategy.Thespikingofthecouponsisstraightforward,the
residuesappearastheywouldoncleanedanddriedequipment,andmost
importantly,thisapproachresultsinamorerealisticdeterminationofaVRLbecause
itincludesonlytheareacoveredbytheresidue.ThehigherVRLdeterminationsalso
shouldbewellaboveanachievableLOQandLODforananalyticaltestmethod.
Theprimarydrawbackofthisapproachisthepotentialinconsistencyofthering
thicknessandthesubsequentsubjectivityfordeterminationoftheringthickness.The
insideedgeoftheringmightnotbewelldefinedandasomewhatsubjective
assessmentwillneedtodeterminethemeasurementinneredgeofthering.The
widerthering,thelowertheresultingconcentrationofthedriedresidue.Thisissue
wouldneedtobeaddressedinaprotocolpriortogeneratingofficialVRLdatausing
thisapproach.
Theobjectiveofthefourthapproachwastohaveauniformresidueratherthanaring
typeresidue.AdvantagestothisapproachincludethemorerealisticVRLbasedonly
onareacoveredbytheresidueaswellasthefactthatbecausethesmallerresidues
takeuplessspaceandthereforefewercoupons,moreresidueVRLscanbe
generatedusingfewercoupons.
Potentialdisadvantageswiththisapproachinclude:pipettingverysmallvolumes(20
L)accuratelyandtheconceptofLOQandLODofvisibleresidue.Atsomepoint,
residuewouldstillbevisiblebuttheconcentrationnotabletobeaccurately
determined.
Conclusion
DifferentapproachestoestablishingVRLswereproposedanddiscussed.Eachof
thefourapproachesisviableaslongasitisspelledoutintheoverallcleaning
validationmasterplanstrategyandappliedconsistently.Twomoreaccurately
quantitativeapproachesweredefined,whichseemtoberoughlyequivalentinthe
resultingVRLs,butthepracticalityofeachcanonlybebetterassessedthrough
generationofdataandasidebysidecomparison.
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ArticleDetails
PharmaceuticalTechnology
Vol.40,No.4
Page:5057
Citation
Whenreferringtothisarticle,pleaseciteitasJ.Forsyth,RuggednessofVisible
ResidueLimitsforCleaningValidation,"PharmaceuticalTechnology40(4)2016.

2016AdvanstarCommunications,Inc.Allrightsreserved.Reproductioninwholeorinpartisprohibited.Pleasesendanytechnicalcommentsorquestionstoourwebmasters.

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[2]http://images.alfresco.advanstar.com/alfresco_images/pharma/2016/04/04/1eff1d581bb244a4b08bfeeee0cb8361/PT0416VRL_TableI.jpg
[3]http://images.alfresco.advanstar.com/alfresco_images/pharma/2016/04/04/1eff1d581bb244a4b08bfeeee0cb8361/PT0416VRL_TableII.jpg
[4]http://images.alfresco.advanstar.com/alfresco_images/pharma/2016/04/04/1eff1d581bb244a4b08bfeeee0cb8361/PT0416_TableIII.jpg