SterileProduct

Manufacturing

Introduction
Togiveanoverviewoftheprinciplesinvolvedinthe
To give an overview of the principles involved in the
manufactureofsterileproducts
Theoverallobjectiveistoproduceproductthathasahigh
assuranceofsterility(andwhichmeetsallotherquality
parameters)
Thispresentation:
Summarisesthegeneralapproach
Givesaframeworkforotherdetailedguidesonspecificaspectsof
sterilisation&sterilemanufacturing
Illustratestheunderlyingprinciples
y gp
p
Providesadviceandgivesrecommendations.

GeneralPrinciplesofSterile
p
Manufacturing

M i t H t St ili ti
MoistHeatSterilization
DryHeatSterilization
AsepticProcessing
Aseptic Processing
EnvironmentalMonitoring
EthyleneOxideSterilization
Ethylene Oxide Sterilization
SterileFiltration
Water systems validation
Watersystemsvalidation
Sterilitytesting
Radiation Sterilization
RadiationSterilization
VisualInspection

Fundementals

S ili is
Sterility
i the
h absence
b
off living
li i organisms
i
This is an absolute definition

The p
probabilityy of achievingg sterilityy depends
p
on the overall p
process

It is generally accepted that a terminally sterilized product should have a probability of

nonsterility of less than 106 (i
non
(i..e., a lower probability than one in a million of having a non
non
sterile
t il unit)
it)

This is often expressed as an SAL Sterility Assurance Level of 106

This is a worst
worstcase figure (with a challenge more resistant than product bioburden)
bioburden). Real
confidence levels are generally very much higher

A figure
fi
that
th t has
h sometimes
ti
b
been
quoted
t d for
f aseptically
ti ll filled
fill d product
d t is
i probability
b bilit off non
non

3
sterility of less than 10 . However, this is harder to analyse as contamination does not
follow a clear statistical distribution.
distribution. Potential contamination sources are not randomly
distributed..
distributed

Why Validate and Control?

WhyValidateandControl?
The
Th test for
f sterility
ili cannot confirm
fi
that
h the
h whole
h l
batch is sterile
It is performed on a sample from a batch and has
statistical limitations
It can miss contamination if only a proportion of units are
nonsterile
non

It is thus necessary to recognize and understand every

aspect that could lead to loss of sterility assurance
Such conditions should be prevented by the
application of carefully designed barriers and/or
control measures.
measures.

Development Validationand
Control
It is
i important
i
that
h the
h product
d
and
d process are
designed to maximise sterility assurance
Wherever possible, the product should be
developed to withstand sterilization in the final
container
i
Once the product design is defined, a suitable
production process must be developed
This is installed and validated
The process must then be tightly controlled to
assure reliabilityy and consistency.
consistencyy.

Product Design Considerations

ProductDesignConsiderations
For New
Ne Products:
Prod cts:
Products

Define product and processing requirements

Consider stability of product to the sterilization conditions
Base the process on achieving the required sterility assurance level
Where possible choose terminal sterilization in final container
Define process flow and the important microbiological aspects
Ensure changes are subject to strict change control

For reviewing existing (marketed) products:

products:

Establish the process description and assess in detail

Preferably, sterilization should be by compendial procedures
Where other procedures are registered, assess SAL
Where necessary (if existing SAL is too low) may need to improve
process and maybe re
reregister
Require justification & validation.
validation.

Facility Design
FacilityDesign
M
Mustbeincompliancewithcompanypoliciesandprocedures,for
st be in compliance ith compan policies and proced res for
example:
Mustminimisetheriskofcontaminationatallcriticalstages
R
RequiredGradesofCleanRooms:needtobeappropriateforthe
i dG d
f Cl
R
d b
i
f h
process e.g.forTerminalSterilizationorAsepticFill
process
PersonnelAccessandMaterialFlow
Restrictedaccess,correctgowning
R
i d
i
Materialsflow,airlocks,decontamination,segregation
HVAC
HVACSystem
Segregation/DedicatedHVACofcorrectstandard
RequirescontrolofFiltration/P/AirFlow/Temp./Pressure/Humidity
Airflowpatternsdemonstrated
f
p
NosinksanddrainsinZoneA/Bareas,airbreakstodrainsinothers
Surfacesandeaseofcleaning:smoothunbrokenimpervioussurfaces

Cleaninganddisinfectionofthe
Facility
Cleaning
Cleaninganddisinfectionisimportantinenvironmentalcontrol
and disinfection is important in environmental control
Efficacyneedstobevalidated
Validatedprocedures,conductedconsistently

InclassA&Bareas,thecleaninganddisinfectantmaterialsmustbe
I l
A&B
h l
i
d di i f
i l
b
sterilized
Andneedtominimisecontaminationriskinotherareas

Operatingproceduresmustinclude,atminimum:
i
d
i l d
i i

Preparationofcleaningmaterials(andsterilizationifapplicable)
Exactprocedureofcleaning&disinfection.
Responsibility&scheduling.
Typeandconcentrationofdetergentsanddisinfectants.
Typeofcleaningtools.

Trainingisrequiredforcleaninganddisinfectionof
Trainingisrequiredforcleaninganddisinfectionofcleanrooms
cleanrooms
Routinedecontaminationusingformaldehydegasshouldbeavoided.

Water
All
Allwatersystemsrequiregooddesignandvalidation
t
t
i
dd i
d lid ti
yp
y,
pharmacopoeial
p
p
ggrades,validationincludes
,
Typically,forpharmacopoeial
Typically,for
Twostudiesoveratotalof4weekstoassessagainsttheacceptancecriteria,
Additional11monthstoverifythatthesystemremainsundercontrol

Mustdemonstrateconsistentproductionofwaterofthe
Must demonstrate consistent production of water of the
requiredquality
Physico
Physicochemical,
Microbiological,
Biological(
Biological(endotoxin
endotoxin,whereapplicable)
,whereapplicable)

Watersystemsmustberegularlymonitoredfollowinga
Water systems must be regularly monitored following a
definedwrittenmonitoringplanbasedonresultsofthe
validationstudies.

Categories of Water
CategoriesofWater
Water
W t for
f Injections
I j ti
(WFI)
For injectables formulation
Final rinse water ffor p
productcontact items (f
product
(for injectables
injectables)
j
)
Freshly prepared or from a validated hot (e
(e..g., >75
75C) storage
/distribution system or otherwise protected from microbial
contamination
o a
a o

Highly Purified Water (HPW)

To European Pharmacopoeia

Purified Water (PW)

For initial washing of product
productcontact items
Prepared,
Prepared suitably stored and distributed to maintain quality
and prevent microbiological proliferation, following the
relevant company procedures
procedures..

Gasses and Vacuum

GassesandVacuum
Gases
G
Specificationequivalenttotheroomairqualitywhereitistobeused
Inasepticapplications,gasesaretobefiltersterilized
p
pp
,g
f
Considersterilefilteringnon
Considersterilefilteringnonproductcontactgasesforaseptic
applications.(But,notesafetyconsiderations,e.g.avoidanceof
leakage)
Allgasfilterstobeintegritytestedoninstallationandatdefined
intervals

VacuumSystems
y

Sometimesusedforcleaninganddustcontrol
Maybemobileunits,fittedwithexhaustHEPAfilters
Or
Ormayhavecentraldustcollection
may have central dust collection
Onthese,usededicatedvacuumpumpsprotectedagainstback
Onthese,usededicatedvacuumpumpsprotectedagainstbackflow
Designtopreventunprotectedrouteintotheasepticsuite.

Equipment (1)
Equipment(1)
EquipmentQualification
E i
t Q lifi ti
Toincludethecriticalaspectsforsterileproductprocessing
Qualificationofcriticalaspectsofmoistheatsterilization,aseptic
f
f
p
f
,
p
processing,dryheatsterilizationetc.

CleaningandSanitizationofEquipment
Equipmentdesignedforeasycleaningandsanitization
Equipment designed for easy cleaning and sanitization
ForTerminalSterilizationapplications,lowmicrobialchallenge.
Wherepossible,criticalsurfacesshouldbesterilized
Forasepticwork,thecritical(productcontact)surfacesmustbe
For aseptic work the critical (product contact) surfaces must be
sterilizedbeforeuse.Inexceptionalcaseswherethisisnotpossible
(e.g.,somestopperbowls),theyshouldbesanitizedbyavalidated
method
Cleaningvalidationmustshoweffectivenessandabsenceofresidues.

Equipment (2)
Equipment(2)
Equipment
i
S ili i and
Sterilization
d handling
h dli
Sterilization must follow a validated procedure
Aseptic processes designed to minimise aseptic
assembly and intervention
Unavoidable
U
id bl aseptic
i assembly
bl needs
d clear
l
& precise
i
procedures
Aseptic assembly must be simulated (worst
(worstcase) in
media fill simulation trials

Sterilization In Place is a good method where

possible must be validated.
validated.

Personnel
Training
T i i
personnell appropriately
i t l trained
t i d for
f
sterile
t il
processing, including assessment and documentation:
documentation:

Basic GMP
Fundamentals
d
l off microbiology
b l
Personal hygiene, health and cleanliness
Behaviour and aseptic working techniques
G
Gowning
i and
d entry
t procedures
d
Cleaning and disinfection
Sterilization procedures, validation and routine operation
Emergency procedures to protect product quality (e
(e..g. loss of HVAC System,
System
loss of power, equipment interventions etc
etc..)

Personnel participating in aseptic processing must have

practical training in aseptic techniques before doing aseptic
manipulations
They must have participated in a successful media fill run.
run.

Gowning and Aseptic Technique

GowningandAsepticTechnique

Gowning
Personnelmustcorrectlywearappropriatecleanroomgarments
Detailed,easilyunderstood,gowningprocedure(preferablyillustrated)

AsepticTechniques
Personnelintheasepticmanufacturingarea,mustunderstandtheprinciples
of aseptic procedures
ofasepticprocedures
Theymustonlybeconsideredqualifiedafterappropriatetraining,working
undersupervisionanddemonstrationofcompetence
Thesupervisorshouldobservetechnique&correctasnecessary
Allpersonneldirectlyinvolvedinasepticprocessingmustparticipateina
mediafillatleastonceperyear

Gl
Glovedisinfection
di i f ti
Steriledisinfectantsmustbeavailable(e.g.,alcoholbased)
Glovedisinfectionmustbereasonablyfrequent,definedinSOP.

Environmental Monitoring (1)

EnvironmentalMonitoring(1)
Thescopeofenvironmentalmonitoring
includes:
Nonviableparticulates,
Viable (microbial) counts
Viable(microbial)counts
Differentialpressures
Temperatures
Humidities
Airflows

Environmental Monitoring (2)

EnvironmentalMonitoring(2)
MonitoringDuringRoomQualification
Monitoring D ring Room Q alification

OperationalQualification(OQ)atrestconditionstoverifyoperation
PerformanceQualification(PQ)inworstcaseoperationalconditions
ActionlevelsshouldmeetUSPorEuroGMPasapplicable
Alertlevelstightenoughtodetectdeterioration,butnotsotightthat
theybecomemeaninglessduetofrequenttransgression
PQmustcoverasufficientperiodtoestablishconsistency

RoutineMonitoring
Ensuresarearemainssatisfactory.Resultsshouldbewithinalertlevel
f
y
Resultsabovealertlevelsneedreviewandperhapscorrectiveactions
Aboveactionlevels,musttriggerappropriateactions(describedin
g
guide),
),
Resultsmustbeassessedfortrendssothatprogressiveorsudden
changesintheresultsmaybeobserved.Thisshouldbereviewed
regularly.

Environmental Monitoring (3)

EnvironmentalMonitoring(3)

Deviation Reports and Failure Investigations

DeviationReportsandFailureInvestigations

RecommendedMethodsforRoutineMonitoring

Thedatamustbeanalysed
Wherenecessaryfurtherinvestigationsinitiated
Possible contamination sources to be assessed and eliminated
Possiblecontaminationsourcestobeassessedand,eliminated
Outcomeanddetailmustbereported
Physicalmeasurementsoftheairsupply
Physical
measurements of the air supply
Physicalandmicrobiologicalmonitoringoftheenvironment
Particles(viableandnonviable)intheair
Microorganisms settling out of the air
Microorganismssettlingoutoftheair
Microorganismscontaminatingsurfaces
Presenceofmicroorganismsonthehandsandgarments

Monitoring Plan
MonitoringPlan
Definedmonitoringplans:tests,locations,alert/actionlevels&frequencies
Maycontaindetailsofwater,compressedgascleansteamtesting
Areviewofenvironmentaldataisarequirementforbatchrelease.
A review of environmental data is a requirement for batch release.

Bioburden andComponents
and Components

Active Ingredients Excipients,Additives

ActiveIngredients,Excipients
ActiveIngredients,
Excipients,Additives
Additives
Allingredientsshouldhaveappropriatebiologicalspecifications
Anylimitationstosterilizationmustbedefined
Descriptionoforigin(e.g.virological
i i
f i i (
i l i l /prion
/ i risk)
i k)

MaterialsUsedintheProcess
Whereappropriate,determinebioburden (e.g.,ionexchangematerials)

PrimaryPackagingComponents
Containerandtheclosureandcleaning/sterilizationtobeclearlyspecified
Stepssuchassiliconization
p
mayneedmonitoring
y
g
Ifcleaning/sterilizationisbysupplier,sameexigenciesapply

Containerclosureintegrity
Container
Theintegritymustbevalidated
The integrity must be validated
Simulate,whereappropriate:stressfromprocessing
Methodappropriatetocontainer/closuresystem

Weighing,Compunding and
Sterilization

Weighing
Weighingandcompoundingmustbeinsuitablyclassifiedrooms
and compounding must be in suitably classified rooms

Vesselsmustbecleaned,andsterilizedorsanitisedasappropriateandstoreddry
inawaytopreventmicrobialcontamination

Storageofpresterilizationintermediatestobecontrolled&timelimited
Storageofpre

Followingaspectstobeconsidered:
Prefiltrationbioburden
Pre filtration bioburden (filtersterilizedmaterial)
(filter sterilized material)
Presterilizationbioburden
Appropriateinprocesscontrols

Sterilizationofproductandproductcontactmaterials
SelectionofasuitablesterilizationprotocolmustbebasedonSAL
Methodmustalsoconsiderthestabilityoftheproduct
Method must also consider the stability of the product
Validationalwaysrequired
Changecontrolisvital;evenapparentlyminorchangemustbeassessed

Terminal Sterilization
TerminalSterilization

Steam Sterilization
SteamSterilization

Byfarthemostcommonmethodforaqueousbasedpharmaceuticals
PreferredcycleisthePharm Eur referencecycleis15minutesat121C
The sterilization cycle chosen must be compatible with product stability
Thesterilizationcyclechosenmustbecompatiblewithproductstability
Sterilizationparametersclearlydefined
Inconjunctionwithothercontrols,therequiredSALmustbedemonstrated
Validationtoconfirmsterilizationconditionsconsistentlythroughouttheload

SterilizationbyIonizingRadiation
Commonformedicaldevices,butnotforpharmaceuticals.
Pharm.Eur.referencecondition,25KiloGray (kGy),hasbeenwidelyaccepted.Other
conditions may be used if validated and accepted by the regulator
conditionsmaybeusedifvalidatedandacceptedbytheregulator
Importanttoconsidersusceptibilityoftheproducttoradiationdamage

DryHeatSterilization
Lower
Lowerantimicrobialefficacythanmoistheat,thushighertemperaturesand/orlonger
antimicrobial efficacy than moist heat thus higher temperatures and/or longer
exposures.Pharm Eur referencecycleis2hours@160C
Rarelyusedforterminalsterilizationofpharmaceuticals;inrarecasesheatresistantnon
aqueousproductsmaybeterminallysterilized.

SterilizationofItemsforAsepticFill
(1)

Steam Sterilization
SteamSterilization
Widelyused,butcarefulvalidationneeded particularlycomplexitems
Broadlysimilartoterminalsteamsterilization,buttwoaspectsarecritical
Qualityofsaturatedsteam
Quality of saturated steam
Removalofairandsubsequentsteampenetration

SterilizationbyIonizingRadiation
Maybeusedfortemperaturesensitiveprimarypackagingorcomponents
Usedfordisposablesforsterileareasandsterilitytestingareas
Validationincludesdosimetry, correct,even,irradiationoftheitems

DryHeatSterilization/
DryHeatSterilization/Depyrogenation
Depyrogenation
Sterilization/depyrogenation ofheatresistantprimarypackagingmaterials
Pharm Eur notesthattemperaturesinexcessof220oChavebeenfrequently
used,theUSPsuggests250 15oC
Validationmustincludeendotoxin
V lid ti
ti l d
d t i challengestudies
h ll
t di
Dryheatmaybeusedtosterilizenonaqueouspreparations(e.g.Ointment
bases)atlowertemperature/timerelationships,withoutdepyrogenation.

SterilizationofItemsforAsepticFill
(2)
EthyleneOxideSterilization
Eth lene O ide Sterili ation
Quitewidelyusedtosterilizeheatlabilecomponents
EuropeanPharmacopoeiaandtheEuropeanGMPguideindicatethat
thi
thismethodshouldonlybeusedwherethereisnosuitable
th d h ld l b
d h
th
i
it bl
alternative
Hazardous toxic,potentiallycarcinogenic,flammable,potentially
explosive
Generallyconductedbyspecializedcontractors
Therearestrictregulatorylimitsonmaximumpermissibleproduct
residues
Bulkpacksforsterilizationmustbegaspermeable,butsealed
againstmicrobialingress
Sterilizationmustconsiderpackaging,loadpattern,gaspenetration
Sterilization must consider packaging load pattern gas penetration
(ethyleneoxide&watervapour),bulkpackintegrity
ValidationandroutinemonitoringmustincludeBiologicalindicators.

Sterilization by Filtration (Liquids)

SterilizationbyFiltration(Liquids)

Principle:
Contaminatingorganismsarenotkilled,butareretainedonthefilters.Anyfaultsinthe
filterstructure,maycompromisethis

Validationincludes:
Retentionofbacterialchallenge:B.diminuta at107percm2
Thisiscorrelatedwithanintegritytestvalue

Validationshouldaddress:

Filtersuitability toxicity,extractables,sheddingofparticles
Adsorptionofproduct
Compatibilitywithproductsolvents
Therequiredfiltersizeandsuitabilityofthefiltrationequipment
h
i d fil
i
d i bili
f h fil
i
i
RetentionofB.diminuta intheactualproductunderprocessconditions
Parametersforthephysicalintegritytest

RoutineFiltration
Routine Filtration
Conductedinlinewiththevalidatedparameters
Checkintegritytesting,processtime,differentialpressure,flowrates,sterilizationandreuse
offilters.

PerformanceQualificationofAseptic
Manufacturing
Based on simulating the risk of contamination in all
aseptic operations
For a new process, a minimum of three consecutive
satisfactory media filling trials
For aqueous liquid products, simulation trials use a
liquid microbiological medium
For solid dosage forms, a powder placebo is used,
followed by aseptic reconstitution into a liquid
microbiological
b l
l medium
d
The following slide gives a general overview
overview....
....

AsepticProcessSimulation
(MediaFillTrial)
MediaFillTrials(MFTs)
Media Fill Trials (MFTs)

Allprocessstagessimulatedascloselyaspossible
Particularlyinterventionsandmanualmanipulations
Mustfollowroutineproceduresandincludeallinterventions
Regularinterventionssimulatedwiththesamefrequencyasactualprocess
Ineachcase,theworstcaseeventualitymustbecovered
Processmustbesuccessfullyvalidatedbeforeproductfillingispermitted
Revalidationbymediafillmustbeconductedeveryhalfyear(eachline)

ManufacturingEnvironment
g
Microbiologicalmonitoringmustbeperformedduringthetrial

FillingConditionsandEquipment
Allaccordingtoroutineoperatingconditionsandatnormaltimesofday
All according to routine operating conditions and at normal times of day
Containersmustbepassedthroughallstages.

Thank You
Any Questions