Seminar on Aseptic Processing operation - PowerPoint Presentation

1. Seminar onAseptic Processing operationbyRanjith Kumar kankala.M.Pharm (I sem)Department of PharmaceuticsBLUE BIRDS COLLEGE OF PHARMACYAffiliated to Kakatiya universityWarangal2009

2. Schedule (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.Conclusion References

3. Aseptic 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. Progression of SymptomsFeverDecreased Blood PressureRapid Breathing and Heart RateSkin LesionsSpontaneous Blood Clotting Organ FailureDeath“Sepsis is a serious medical condition characterized by a whole-body inflammatory state caused by infection.”

5. Causes of sepsisSterile 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.”

6. Asepsis 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.

7. Producing drug products byTerminal 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. Aseptic processing 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.

8. Terminal SterilizationSterile Drug Product !

9. Aseptic Processing Drug ProductSterilization ProcessContainerClosureExcipientSterilization ProcessSterilization ProcessSterilization ProcessSterile ClosureSterile ExcipientAseptic ProcessingSterile DrugProductSterileContainerSterile Final ProductCan use multiple sterilization processes each optimized for the individual component

10. Bacteria, 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. Contaminating agents

11. Humans 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.

12. Sources of Contamination: Personnel born contaminantsPoor or improper Sanitization: Procedures deficient, or poorly executed Air 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.

13. Control1st step – eliminating the source of contamination ! 2nd Step - Reduce the Risk of contamination through: Sterile barriersSurface monitoring Aseptic technique

14. Gowning (sterile barrier)If people are a major source of contamination we avoid contaminating the product while we process it.

15. Surface MonitoringTouch or Contact plates - RODAC Plates (Replicate Organism Detectionand Counting)Swabs

16. Aseptic 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 gloves Keep the entire body out of the path of unidirectional airflow Approach a necessary manipulation in a manner that does not compromise sterility of the product

17. What’s wrong with this picture?

18. CORRECT

19. Horizontal airflow Vertical airflowwww.ors.od.nih.gov/ds/pubs/bsc/graphics/fig3.gif Unidirectional airflowThe operator should never come between the air source and the product. pressure differential b/n critical area from external environment (17.5-50 Pa)

20. DisinfectantsISOPROPYL 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.,

21. Sporicidal agentsGlutaraldehydeFormaldehydesodium hypochloriteIodine and iodophorsPeroxygensEthylene oxideP- Propiolactone

22. IsolatorsAdvantage:No direct contact between operator & product.

23. Microbial identification should extend to the species level. Routine traditional techniques phenotypic and biochemical.Genotypic techniques are suggested for failure investigations.Microbial Environmental Monitoring: Identification

24. Phenotypic technique Gram StainIdentifying Microbes

25. Staphylococcus xylosus Reduction of Tetrazolium Violet Biochemical Assays

26. Genotypic MethodsUse DNA sequence (often ribosomal RNA genes rDNA) to identify organismFaster, and more accurate then traditional biochemical and phenotypic techniques

27. QC Micro: Identifying Microbes Genotype Based Assay:PCR: Polymerase Chain Reaction

28. Extremely heat stable – recommended conditions for inactivation are 180 0 C for 3 hours. Endotoxin: a pyrogenic (fever inducing) substance (e.g. lipopolysaccharide) present in the bacterial cell wall. Endotoxin reactions range from fever to death. Endotoxin TestingLAL Assay (Limulus amoebocyte lysate)ENDOTOXIN LIMIT FOR WFI IS 0.25 EU/ml

29. Microbiological testing of waterUniversal 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

30. Microbiological testing of airCompressed 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.)

31. Characterization of aseptic process The four pillars of a robust * aseptic process Personnel training & monitoringEnvironmental monitoringFacilities design Media fills

32. Personnel Training & Monitoring Avoiding contamination means knowing the potential sources of contaminationPersonnelEquipmentAir/liquidsDrug productContainers/closures Outside environmentAnything Brought in contact with, or in the vicinity of, the product is a potential source of contamination!

33. Environmental 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.

34. 1.2.3. 4.5.6.7.8.9. 10.11.13. 12.Environmental Monitoring Critical (processing) areas Sampling Sampling of adjacent classified areas (aseptic corridors, gowning rooms, etc) will provide trenddata and may help identify sources of contamination.

35. Facilities: General Clean room DesignHEPA/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 operators

36. Facilities: Clean room Classification

37. Class 10,000 clean roomhttp://www.americancleanrooms.com/am/photogallery_08.htmlClass 100 clean roomFacilities: Clean room Classification

38. Facilities: HEPA Filtershttp://people.deas.harvard.edu/~jones/lab_arch/nano_facilities/hepa.gifHigh Efficiency Particulate Air filtersMinimum particle collection efficiency: 99.97% for 0.3µm diameter particles.Disposable Filter made of pleated borosilicate glass

39. Media Fill testUsed to validate the aseptic processUse 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.

40. Media and Incubation conditionsSoybean 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 monitored product 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

41. Theoretical EvaluationWhyte mathematical model contamination is due to air borne microbesd = equivalent particle diameterA= area of container opening (cm2)t = time (sec) Cont rate (c) = 0.0032.d2.A.t

42. PostScript (conclusion)The challenge in aseptic processing is always personnel:􀂄 As a source of microbial andParticle contamination.􀂄As a brake on the implementation ofImproved technology.

43. REFERENCESEncyclopedia of pharm.technologyRUSSELL A. D.. Bacterial Spores and Chemical Sporicidal Agents. clinical microbiology reviews. 3(2): 99-119 (1999) .http://www.fda.gov/cber/gdlns/steraseptic.pdf http://www.emedicinehealth.com/images/4453 http://pathmicro.med.sc.edu/fox/lps.jpg http://micro.med.harvard.edu/faculty/rudner.html

44. ThanQ