Manufacturing of sterile preparations

Sterile preparations, parenterals, Zones by gazette of India, Aseptic techniques

1. MANUFACTURING OF
STERILE PREPARATION
Prepared by: SHRUTI TYAGI (LECTURER IN PHARMACY)

2. INTRODUCTION
Parenteralpreparationsaresterile,pyrogen-freeliquids(solutions,
emulsions,orsuspensions)orsoliddosageformscontainingoneor
moreactiveingredients,packagedineithersingle-doseormultidose
containers.Theyareintendedforadministrationbyinjection,infusion,
orimplantationintothebody.
Thedosageformforconveyingadrugbymeansofinjectionthrough
theskinormucousmembranes.Parenteraldrugsareadministered
directlyintotheveins,musclesorundertheskinormorespecialized
tissuessuchasthespinalcord.Circumventedthehighlyefficientfirst
linebodydefensethatisskinandmucusmembrane.Thustheyshould
befreefrommicrobialcontaminationandshouldhavehighpurity
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3. TYPES
•Therearefourmainformsofparenteralpreparations:
•Injections,
•Intravenousinfusions(largevolumeparenterals),
•Powdersforinjections,and
•Implants.
Certaininjectionsandintravenousinfusionsmaybepresentedintheformofsterile
concentratedsolutions,whichmustbesuitablydilutedbeforeuse.
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4. FACILITIES REQUIRED FOR PARENTERAL PRODUCTION
Parenteralpreparationsmaycontainexcipientssuchassolvents,suspendingagents,bufferingagents,
substancestomakethepreparationisotonicwithblood,stabilizers,orantimicrobialpreservatives.
•Theadditionofexcipientsshouldbekepttoaminimum.Whenexcipientsareused,theyshouldnot
adverselyaffectthestability,bioavailability,safety,orefficacyoftheactiveingredient(s),orcausetoxicity
orunduelocalirritation.Theremustbenoincompatibilitybetweenanyofthecomponentsofthedosage
form.
•Waterforinjectionsisusedasthevehicleforaqueousinjections.Itshouldbefreshlydistilledbythe
processdescribedunder"AquaproInjection",befreefromcarbondioxide,andcomplywithTestfor
bacterialendotoxins.Sterilizationatthisstagemaybeomitted,providedthatthesolutionorpreparationis
immediatelysterilizeduponfinalization.Fornon-aqueousinjections,fixedoilsofvegetableoriginareused
asvehicles.
•Unlessotherwisespecifiedintheindividualmonograph,sodiumchlorideorothersuitablesubstance(s),
maybeaddedtoanaqueoussolutionforinjectioninordertorenderthepreparationisotonic.
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5. 5

6. 6

7. TERMINALLY
STERILIZED/
STERILIZATION BY
FILTERATION
MANUFACTURING OF STERILE PREPARATIONS: KEY REQUIREMENTS
CHANGE
ROOMS
PERSONNEL
FLOW
ASEPTIC
FACILITIES-
ZONES
WALLS AND FLOOR
TREATMENTS
LIGHTNING
FIXATURES
As per Gazette of
India: Black, Grey
and White
As per CGMP:
ZONES 7-1
STERILITY TESTING:
a)Direct Transfer
b) Membrane Filtration
c) Rabbit test
d)LAL test
e) Leaker test
f) Particulate matter testing
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8. MANUFACTURE OF STERILE PREPARATIONS
1.Terminallysterilized:usuallyinvolvesfillingandsealingproduct
containersunderhigh-qualityenvironmentalconditions.Productsare
filledandsealedinthistypeofenvironmenttominimizethemicrobial
andparticulatecontentofthein-processproductandtohelpensurethat
thesubsequentsterilizationprocessissuccessful.Inmostcases,the
product,container,andclosurehavelowbio-burden,buttheyarenot
sterile.Theproductinitsfinalcontaineristhensubjectedtoa
sterilizationprocesssuchasheatorirradiation.
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9. TERMINAL
STERILIZATION
OF PRODUCTS
STERILIZATION OF THE
PRODUCT IS DONE
USING HEAT OR
RADIATIONS
PRODUCT IS FILLED IN
SUITABLE CONTAINERS
AND CONTAINER IS
SEALED
PRODUCT IS PREPARED
UNDER CONTROLLED
ENVIRONMENT
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10. 2.SterilizationbyFiltration:-
1. Previously
sterilized container
are taken
2. Filters having
nominal pore size
0.22 μm or less are
used for filtration
5. No fiber
shedding or
asbestos filters
4. Double filter
layer or second
filtration
3. Remove bacteria
and moulds but
Not viruses &
Mycoplasmas
6. Filter integrity
testing
META FILTERS OR MEMBRANE FILTERS ARE USED
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11. 3. Aseptic Preparation: -
1.Thedrugproduct,container,
andclosurearefirstsubjected
tosterilizationmethods
separately
2.Containersarefilledand
sealedinanextremelyhigh-
qualityenvironment
5.Inareaoccupiedbypersonal,theairmustbeexchangedwiththefrequentintervals.Freshoutsideorrecycledair
mustbefirstfilteredtoremoveparticulatematterandthenHEPAfiltersareusedtogetCLASS-100airsystems.
3.Beforeasepticassemblyintoafinal
product,theindividualpartsofthe
finalproductaregenerallysubjected
tovarioussterilizationprocesses
4.Anymanualormechanicalmanipulationofthesterilizeddrug,components,containers,orclosures
priortoorduringasepticassemblyposestheriskofcontaminationandthusnecessitatescarefulcontrol.
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12. GMPREQUIREMENTS FORSTERILEPRODUCTS
MINIMISING THE RISK
OF MICROBIOLOGICAL
CONTAMINATION AND
PARTICULATE MATTER
OR PYROGEN
PRODUCTION IN CLEAN AREAS,
AIRLOCKS FOR ENTRY (PERSONAL
AS WELL AS MATERIAL), SEPARATE
AREAS FOR OPERATIONS SUCH AS
PRODUCT PREPARATION, FILLING
AND SEALING ETC.
LEVEL OF
CLEANLINESS SUCH
AS FILTERED AIR, AIR
CLASSIFICATION-
GRADE A, B, C AND D
LAMINAR AIR FLOW: AIR
SPEED (HORIZONTAL
VERSUS VERTICAL
FLOW), NUMBER OF AIR
CHANGES ANDAIR
SAMPLES
CONFORMITY TO STANDARDS,
WORK STATION AND
ENVIRONMENT AND BARRIER
TECHNOLOGY AND AUTOMATED
SYSTEMS
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13. ZONES AS PER THE C GMP
•ZONE 7:-FILLING LINE
•ZONE 6:-FILLING AREA
•ZONE 5:-WEIGHING, MIXING & TRANSFER AREA.
•ZONE 4:-CLEAN AREA
•ZONE 3:-GENERAL PRODUCTION
•ZONE 2:-WAREHOUSE
•ZONE 1:-EXTERIOR
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14. Zone-7:-fillingline:-
•Thewallsofthefillingareaarethelastphysicalbarriertotheingress
ofcontamination,butwithinthefillingareaatechniqueof
contaminationcontrolknownaslaminarflowmaybeconsideredas
thebarriertocontamination.
Zone-6:-fillingarea:-
•Zone6isadistinctzoneofthecontrolledenvironmentareaforan
asepticfillingprocessbutmaynotbedistinctzonefornon-aseptic
fillingprocesses.
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15. Zone-5:-weighing,mixing,andtransferarea:-
•Zone5encompassesthoseactivitiesof“weighing,mixing,fillingortransfer
operations”addressedbycGMPsection212.81whicharenothandledaszone6but
whichrequireacontrolledenvironment.
Zone-4:-cleanarea:-
•Activitiesinthesemayincludewashingandpreparationsofequipmentor
accumulationandsamplingoffilledproduct.
Zone-3:-generalproductionandadministrationarea:-
•Thethirdzoneofenvironmentalcontrolisformedbytheperipheryofthegeneral
productionarea.Openingsintotheareaareusuallywellsealedandlargeenoughfor
onlyessentialmaterial-handlingequipmentandpersonnel.
Zone-2:-warehouse:-
•Basicwarehousingfunctionsincludereceiving,shipping,andin-processstorage.
Receivingareasincludeunpacking,samplingandincomingquarantine.
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16. Zone-1:-plantexterior:-
•Theenvironmentalwithinwhichaplantlocatedisfirstenvironmental
controlzone.Itisabasepointfromwhichtoworkindeterminingthe
requirementsforthevariouscontrolbarriers.Managementactionstocontrol
zone1mightincludethemaintenanceofsterileareasaroundthefacility
whereweeds,insectsandrodentsarecontrolledoreliminated.
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17. ZONES AS PER GAZZETE OF INDIA
•Whitezone:-Finalstep(fillingofparenteral)
•Greyzone:-weighing,Dissolution&filtration.
•Blackzone:-Storage,Worstareafrom
contaminationviewpoint
•a)Haveaper-cubic-particlecountofnotmore
than100inasizerangeof0.5micronand
largerthroughtheentireworkareaupstreamof
theworkpiece.
•b)Besuppliedatthepointofuseasspecified
insection212.77.
•Thelayoutoftheplantmustbecarefully
developedincoordinationwiththeneedsof
theHVAC(HeatingVentilationandAir
Conditioning)system
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18. 2.WALL&FLOORTREATMENT :
Thedesignoffillingareasormoregenerally,controlledenvironment
areasinvolvesattentiontomanyseeminglyminordetails.Thebasic
clean-abilityrequirementincludessmooth,cleanablewalls,floors,
ceilings,fixtures,andpartitionexposedcolumns,wallstuds,bracing,
pipes,andsoonareunacceptable.Theneedforclean-abilityalso
eliminatestheopenfloorsystemcommonlyusedinthemicroelectronics
industryforlaminarairflowrooms.Thegoalofthedesigner,when
creatingthedetailsforthearchitecturalfinishesandjoiningmethods,is
toeliminatealledgesorsurfaceswithintheroomwheredirtmay
accumulate.
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19. 3.LIGHTNINGFIXTURES:
Lightingfixturesshouldbereducedflushwiththeceiling.Sincemost
lightingfixturesarenottightlysealed,thediffusershouldbesealed
integrallywiththeceiling,andthelampschangedfromoutsidethe
room.Eitherrecessedorsurfacemountedfixturescanbeused.Special
“wash-down”fixturesarewellsealed,butprotrudeobtrusivelyintothe
roomandhaveclipsandsealinglipswhicharedifficulttosanitize.
AreashavingafullHEPAceilingobviouslycannotaccommodate
recessedlightingfixtures.Intheseareas,fixturesareofaspecial
“teardrop”shapewhichminimizesdisruptiontothelaminarairflow
pattern.
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20. 4.CHANGEROOMS:
Personnelaccesstoallcontrolledareasshouldbethroughchange
rooms.Changeroomsconceptsandlayoutsvaryfromsingleclosetsize
roomstoexpensivemulti-roomcomplexes.
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21. 5.PERSONNEL FLOW:-Themovementofpersonnelshouldbe
plannedduringthedesignofindividualplantareas.Eachindividual
productionareamayhaveasmoothandefficientpersonnelflowpattern,
adiscontinuousorcrowdedpatternmaydevelopwhenseveral
individualproductionareaplantsarecombined.Theseparationof
peopleandproductsisgreatlyfacilitatedbytheuseofthethird
dimension.Securityconcernsaboutpersonnelflowmayinclude
minimizingaccesstocontrolledsubstancesandminimizingthe
personneltrafficinornearworkareaswherecontrolledsubstancesare
handled.
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4 3
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4 2
3
NO YES
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22. STERILITY TESTING OF PARENTRALS
The Basic quality control tests which are performed on sterile parenteral
products include:-
1) Sterility Tests.
2) Pyrogen Tests.
3) Leaker Tests.
4) Particulate matter testing.
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23. 1)Sterilitytests:-SterilityisthemostimportantandAbsolutelyEssentialcharacteristicsofParenteralproducts.
SterilitymeanscompleteabsenceofallviableMicro-organism.Itisanabsoluteterm.Themethodswhichareusedto
performsterilitytestsare
•a)Directtransfermethod.
•B)membranefiltrationmethod.
•A)DirectTransfermethod:–itisantraditionalsterilitytestmethodwhichinvolvesadirectinoculationofrequired
volumeofasampleintwoteststubecontainingaculturemediumthatisFTM,SCDM.
•Thismethodissimpleintheorybutdifficultinpracticewhenthedemandforrepetitioninopeningcontainer,sampling
Transferring,andmixingincreasescausespotentialerrorinoperatortechnique
•B)MembraneFiltrationmethod:–ItisofficialinU.S.P.1970.ThismethodbasicallyinvolvesfiltrationofSample
throughmembranefiltersofporosity0.22micronandDiameter47mm.ThefiltrationisassistedunderVacuum,after
filtrationcompletionthemembraneiscutinto2halvesandonehalveisplacedintwotesttubescontainingFTM,SCDM
medium.
•*Interpretation:–Ifnovisibleevidenceofmicrobialgrowthinculturemediumintesttubethenitisinterpretedthatthe
samplerepresentinglotiswithoutintrinsiccontamination
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24. 2)PyrogenTest:–PyrogensareproductsofmetabolisminmicroorganismsGm-ve
bacteriaproducesmostpotentpyrogens.Thesearelipopolysacchrideschemicallyand
heatstableandarecapableofpassingthroughbacteriaretentivefilter.Whenthese
pyrogensareintroducedintoabodytheyproduceamarkresponseoffeverwithbody
acheandvasoconstrictionwithinanonsetof1hour.Basicallytherearetestperformedto
detectthepresenceofpyrogensinsterileparenteralproductstheyareC)RabbitTestD)
LALTest.
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25. •C)Rabbittest:–ThistestbasicallyinvolvestheinjectionSamplesolutionwhichistobetested
intoaRabbitsWhichareuseastestanimalsthroughearvein.TheTemperaturesensingprobe
(ClinicalThermometer,Thermistororsimilarprobe)intoarectumcavityofRabbitatthedepthof
7.5cm,thetestsolutionmustbewarmedat37degreespriortoinjection.ThenRectaltemperature
isrecordedat1,2,3hrsubsequenttoinjection.Thistestisperformedinseparateareadesigned
solelyforthispurposeunderenvironmentalconditionssimilartoanimalhouseshouldbefreefrom
disturbancesthatlikelytoexcitethem.Initiallythistestisperformedon3Rabbitsbutifrequired
resultsarenotobtainedthistestisrepeatedon5additionalRabbitswithsamesamplesolution
administertoinitial3rabbits.Priorto1hrofinjectingsamplesolutionsthecontroltemperaturesof
rabbitsaredetermined.Useonlythoserabbitswhosecontroltemperatureisnovarybymorethan1
degreeCelsius.
•*Interpretation:-Thesolutionisjudgedtobenonpyrogenicifnosinglerabbitshowrisein
temperatureof0.5degreeCelsiusbutifthisconditionisnotmetthenthetestifrepeatedon5
additionalrabbitswithsamepreparationadminister
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26. D)LALtest:–Itisanrecentlydevelopedinvitrotestmethodforpyrogenutilizing
gellingpropertyoflysatesofamebocytesoflimulusPolyphemuswhichisfound
onlyatspecificlocationsalongtheeastcoastofNorthAmericaandalong
southeastAsia.Itisderivedfromhorseshoecrab;thebasicprocedureisthe
combinationof0.1mloftestsamplewithLALReagentafterincubationfor1hrat37
degreeCelsiusthemixtureisanalyzedforthepresenceofGelclot.TheLALTestis
positiveindicatingthatthepresenceofendotoxin.Itsapplicationsaremainlyto
Pharmaceutics,Biological,devices,diseasestates,food,andvalidationofheatcycles.
ThismethodhasseveraladvantagesofRabbittesttheyareGreatersensitivityand
reliabilityspecificity,lessvariation,widerapplication,lessexpensiveandsimplicity.
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27. 3) LeakerTest:–Theleakertestisintendedtodetectincompletelysealed
ampoules,sothattheymaybediscarded.Tipsealedampoulesaremoreproneto
leakthanpullsealed.Inadditiontothatcrackmypresentaroundsealoratthebase
ofampouleasaresultofimproperhandlingleakersareusuallydetectedby
producingnegativepressurewithintheincompletelysealedampouleusuallyintoa
vacuumchamberwhilethoseampoulesaresubmergedintoacoloreddyesolution
of0.5to1%methyleneblue.Vialsandbottlesarenotsubjectedtosuchleakertest
becauserubberclosureisnotrigidhoweverbottlesareoftensealedwhilevacuumis
pulledsothatbottleremainsevacuatedduringitsshelflife.
Thepresenceofvacuumisdetectedbystrikingatthebaseofbottlesharplywiththe
heelofhandtoproducetypicalwaterhammersound.Anothertestistoapply
asparktesterprobeoutsidetothebottlemovingformliquidlayerintoairspacea
bluesparkdischargeoccurisairspaceisevacuated.
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28. 4)Particulatemattertesting:–
Particulatematterisprimaryconcernintheparenteralproducts
givenbyI.V.Route,allparenteralproductsshouldbefreefrom
insolubleparticle.FurtherU.S.P.statesthatGMPRequiresthatall
containersbevisuallyinspectedandthatwithvisibleparticlebe
discarded.Thevisualinspectionisdonebyholdingtheampulebyits
neckagainsthighlyilluminatedscreens.Whitescreensforthedetection
ofblackparticleandblackscreensforthedetectionofwhiteparticlesto
detectheavyparticlesitmaybenecessarytoinvertcontainerbutcare
mustbeexercisedtoavoidairbubble.Theinstrumentalmethodsare
basedonprinciplesoflightscattering,lightabsorption,electrical
resistanceasincoultercounter.Amethodwhichutilizesavideoimage
projectioncoulddetectamovingparticlewithoutdestructionofproduct
unit.
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29. THANK YOU
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