Presentation on theme: "Seminar on Aseptic Processing operation"— Presentation transcript:
1Seminar on Aseptic Processing operation by Ranjith Kumar kankala.M.Pharm (I sem)Department of PharmaceuticsBLUE BIRDS COLLEGE OF PHARMACYAffiliated to Kakatiya universityWarangal2009
2Schedule (contents) Introduction to aseptic processing, Aseptic Processing vs. Terminal Sterilizationcontamination: Sources and control,Microbial environmental monitoringMicrobiological testing of air and waterCharacterization of aseptic process,Media and incubation conditions.ConclusionReferences
3Aseptic ProcessingAseptic Processing is the processing of drug components ( drug product, containers, excipients, etc.) in a manner that makes impossible of microbiological contamination of the final sealed product.
4“Sepsis is a serious medical condition characterized by a whole-body inflammatory state caused by infection.”Progression of SymptomsFeverDecreased Blood PressureRapid Breathing and Heart RateSkin LesionsSpontaneous Blood ClottingOrgan FailureDeath
5Causes of sepsis Sterile drug manufacturers should have a keen awareness of the public health implications of distributing a non-sterile product. Poor cGMP conditions at a manufacturing facility can ultimately pose a life-threatening health risk to a patient.”
6Asepsis is the practice to reduce or eliminate contaminants (such as bacteria, viruses, fungi, and parasites) from entering the field to prevent infection. Ideally, a field is "sterile" — free of contaminants — a situation that is difficult to attain. However, the goal is elimination of infection.
7Producing drug products by Aseptic processingTerminal sterilizationProduct containers are filled and sealed under high-quality environmental conditions designed to minimize contamination, but not to guarantee sterility.Product in its final container is subject to a sterilization process such as heat or irradiation.Drug product, container, and closure are subject to sterilization separately, and then brought together.Because there is no process to sterilize the product in its final container, it is critical that containers be filled and sealed in an extremely high –quality environment.
8Terminal Sterilization Drug ProductContainer / ClosureExcipiantsSterilization ProcessSterile Drug Product !
9Aseptic Processing Drug Product Sterile Drug Product Sterile Sterile Sterilization ProcessSterileDrugProductSterileFinalProductContainerSterilization ProcessSterileContainerAsepticProcessingClosureSterilization ProcessSterile ClosureExcipientSterilization ProcessSterileExcipientCan use multiple sterilization processes each optimized for the individual component
10Contaminating agentsBacteria, virus, fungi and other viable microbes cause a serious contamination. Bacterial spores and endotoxins Non viable Particles like dust, fibers, or other material are suspended in the air and may contaminate product.
11Humans and bacteriaOver 200 different species of bacteria are found associated with humans.Bacteria are found in the intestines, eyes, nares, mouth, hair and skin.Dry skin can have 1000’s of microbes / mm2 !Staphylococcus epidermidis Scanning EM. CDC.
12Sources of Contamination: Personnel born contaminantsPoor or improper Sanitization: Procedures deficient, or poorly executedAir born contaminants.Inadequate HEPA seal (over 90% vials contaminated)Velocity through HEPA Filters: Variable velocities between filters. Inadequate laminar flow resulted. Low or undetectable velocity at work surface.Mechanical failure of filling tank; main pump failure; cooling system leaks at joints.30
13Control 1st step – eliminating the source of contamination ! 2nd Step - Reduce the Risk ofcontamination through:Sterile barriersSurface monitoringAseptic technique
14Gowning (sterile barrier) If people are a major source of contamination we avoid contaminating the product while we process it.
16Aseptic Technique (skill) Contact sterile materials only with sterile instruments:Operators should not contact sterile products, containers, closures, or critical surfaces with any part of their gown or glovesKeep the entire body out of the path of unidirectional airflowApproach a necessary manipulation in a manner that does not compromise sterility of the product
19Unidirectional airflow The operator should never come between the air source and the product.pressure differential b/ncritical area from external environment ( Pa)Vertical airflowHorizontal airflow
20Disinfectants ISOPROPYL ALCOHOL (70%) Powerful disinfectant Effectively kills bacteria and fungiMode of action: denatures proteins, dissolves lipids and can lead to cell membrane disintegration.But does not inactivate spores!e.g., phenols, Alcohols, Aldehydes etc.,
22IsolatorsAdvantage:No direct contact between operator & product.
23Microbial Environmental Monitoring: Identification Microbial identification should extend to the species level.Routine traditional techniques phenotypic andbiochemical.Genotypic techniques are suggested for failure investigations.
25Reduction of Tetrazolium Violet Biochemical AssaysReduction of Tetrazolium VioletStaphylococcus xylosus25
26Genotypic MethodsUse DNA sequence (often ribosomal RNA genes rDNA) to identify organismFaster, and more accurate then traditional biochemical and phenotypic techniques
27QC Micro: Identifying Microbes Genotype Based Assay:PCR: Polymerase Chain Reaction27
28Endotoxin Testing LAL Assay (Limulus amoebocyte lysate) Endotoxin: a pyrogenic (fever inducing) substance (e.g. lipopolysaccharide) present in the bacterial cell wall. Endotoxin reactions range from fever to death.LAL Assay (Limulus amoebocyte lysate)ENDOTOXIN LIMIT FOR WFI IS0.25 EU/mlExtremely heat stable – recommended conditions for inactivation are C for 3 hours.28
29Microbiological testing of water Universal solvent ,Used as Vehicle and used to rince and cleaning of apparatusWater should also be tested for presence of coliforms and/or pseudomonads if appropriate (may cause biofilm)Water should be tested using R2A agar (low nutrient for the recovery of water borne organisms) incubated for at least 5 days at 30-35°CSampling procedures should follow those used in production
30Microbiological testing of air Compressed Air/Nitrogen/CO2Air sampling should be done and tested for the presence of non-viables and viables by exposure to the environment.Pressure control orifices should be used to provide a steady stream of air.Fall out plateSlit sampler(slit-to-agar sampler) Slit Sampler (New Brunswick Scientifics Model STA-230 Slit-to-Agar Air Sampler.)
31Characterization of aseptic process The four pillars of a robust Characterization of aseptic process The four pillars of a robust * aseptic processPersonnel training & monitoringEnvironmental monitoringFacilities designMedia fills
32Personnel Training & Monitoring Avoiding contamination means knowing the potential sources of contaminationPersonnelEquipmentAir/liquidsDrug productContainers/closuresOutside environmentAnything Brought in contact with, or in the vicinity of, the product is a potential source of contamination!
33Environmental Monitoring The goal of the environmental monitoring program is to provide meaningful information on the quality of the aseptic processing environment during production as well as environmental trends.
34Environmental Monitoring Sampling18.104.22.168.22.214.171.124.126.96.36.199.12.Critical (processing) areasSampling of adjacent classifiedareas (aseptic corridors, gowningrooms, etc) will provide trenddata and may help identifysources of contamination.
35Facilities: General Clean room Design HEPA/ULPA filters on ceilingExhaust vents on floorAirlocks and interlocking doors to control air balanceSeamless and rounded floor to wall junctionsReadily accessible cornersFloors, walls, and ceilings constructed of smooth hard surfaces that can be easily cleanedLimited equipment, fixtures and personnelLayout of equipment to optimize comfort and movement of operators35
38Facilities: HEPA Filters High Efficiency Particulate Air filtersMinimum particle collection efficiency:99.97% for 0.3µm diameter particles.DisposableFilter made of pleated borosilicate glass38
39Media Fill test Used to validate the aseptic process Use microbial growth media instead of drug product-any contamination will result in microbial growth.It doesn’t provide a direct relation for sterility but gives an adequate evaluation for operational processing steps.
40Media and Incubation conditions Soybean casein digest medium (SCD)Fluid thioglycollate medium (FTM) for anerobesInoculated with < 100 cfu challengeAt least 14 days incubation30-35°C for SCD, 20-25°C for FTMtemperatures should be monitoredproduct produces suspension, flocculation or deposit in media, suitable portions (2-5%) should be transferred to fresh media, after 14 days, and incubated for a futher 7 days
41Theoretical Evaluation Whyte mathematical modelcontamination is due to air borne microbesd = equivalent particle diameterA= area of container opening (cm2)t = time (sec)Cont rate (c) = d2.A.t
42PostScript (conclusion) The challenge in aseptic processing is always personnel: As a source of microbial andParticle contamination.As a brake on the implementation ofImproved technology.
43REFERENCES Encyclopedia of pharm.technology RUSSELL A. D.. Bacterial Spores and Chemical Sporicidal Agents. clinical microbiology reviews. 3(2): (1999) .