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2. What is Aseptic Processing?
 The production of sterile drug products by bringing together
the product, container, and closure that have been subjected to
different sterilization methods separately, and assembled them
in an extremely high quality environment by skilled personnel
using the right tools.

3. Aseptic Processing: Essential Elements
Facility
Docume-
Equipment
ntation
Aseptic
Finish Pro Processing
duct Process
Testing
Control &
Personnel
Verification

4. Aseptic Processing: Essential Elements
 Facility
– Design
 Zoning, Differential Pressure
 Temperature
 Relative Humidity
 Personnel and Material Flow
– Air Filtration
 Equipment
– Material of Construction
– Sanitization
– Component Preparation/Sterilization

5. Aseptic Processing: Essential Elements
 Process
– Product Formulation
– Filtration
– Filling
– Lyophilization
– Capping
 Personnel
– Gowning Qualification
– Aseptic Technique
 Control and Verification
– Environmental and Personnel Monitoring
– Aseptic Filling Simulations (Media Fills)

6. Aseptic Processing: Essential Elements
 Finished Product Testing
– Sterility Testing
– Particulate Testing
– Container Closure Integrity Testing
– Other Final Product/Release Testing
– Stability Testing
 Documentation
– Media Fill Records
– Production Batch Records
– EM Trend Data
– Release Testing Batch Records
– Investigation
 Response to Excursions
 Corrective Actions

7. Environmental Monitoring Components
 Airborne nonviable particulate monitoring
 Airborne viable contaminant monitoring
 Viable contaminant monitoring of surfaces
 Viable contaminant monitoring of personnel
 Temperature and humidity monitoring
 Pressure differential monitoring

8. Environmental Monitoring Components
Water monitoring:
– Total organic carbon
– Conductivity
– Microbial Contaminants
– Endotoxin
Air monitoring:
– Microbial Contaminants.

9. Regulatory Basis for Environmental Monitoring
Program
 CFR GMP regulations
 FDA Guidance Documents
 USP Informational Chapter

10. Controlled Area
 Preparation or manufacturing area where nonsterile product,
in-process materials and product-contact equipment surfaces,
containers and closures are exposed to the environment
 Control nonviable and viable contaminants to reduce
product /process bioburden
 Class 100,000 or Class 10,000
 Capping areas are now considered controlled manufacturing
areas
– Should be supplied with HEPA filtered air
– Should meet class 100,000 conditions during static
conditions

11. Critical Area
 Aseptic processing area where sterile products, components or
in-process products are exposed to the environment and no
further processing will occur.
 Air quality must be Class 100 during processing
 Local Class 100 areas are often utilized during open
processing steps during drug substance manufacture.
• The area just preceding the sterile core should be one
classification higher than the core.

12. Microbial testing of air and water
Types of water used in pharmaceutical processes
1. Purified water
2. Water for Injections – PFW & WFI
3. Softened Water
4. Water for Final Rinse
5. Pure, or clean Steam
6. Water for cooling Autoclaves

13. Why purify raw water?
1. Although reasonably pure, it is always variable
2. Seasonal variations may occur in water
3. Some regions have very poor quality water
4. Must remove impurities to prevent product
contamination.
5. Control microbes to avoid contaminating products

14. Contaminants of water (1)
 There is no pure water in nature, as it can contain up to 90
possible unacceptable contaminants
 Contaminant groups:
1. Inorganic compounds
2. Organic compounds
3. Solids
4. Gases
5. Micro-organisms

15. Contaminants of water (2)
Micro-organisms :
1. Algae
2. Protozoa
– Cryptosporidium
– Giardia
1. Bacteria
– Pseudomonas
– Gram negative, non-fermenting bacteria
– Escherichia coli and coli forms

16. Contaminants of water (3)
Treatment depends on water’s chemistry and contaminants,
influenced by:
1. Rainfall
2. Erosion
3. Pollution
4. Dissolution
5. Evaporation
6. Sedimentation
7. Decomposition

17. Contaminants of water (4)
Problem minerals
1. Calcium and magnesium
2. Iron and manganese
3. Silicates
4. Carbon dioxide
5. Hydrogen sulfide
6. Phosphates

18. Contaminants of water (5)
Further problem minerals
1. Copper
2. Aluminum
3. Heavy metals
– Arsenic, lead, cadmium
1. Nitrates

19. Water For Injection
 Defined by USP
 Water purified by distillation or reverse osmosis
 Prepared from water complying with the U.S. EPA National
Primary Drinking Water Regulations
 Contains no added substance

20. Purified Water
 Defined in USP
 Obtained by a suitable process, usually one of the following:
– deionization
– reverse osmosis
– combination

21. Potable Water
 Meets National Drinking Water Regulations
 40 CFR Part 141
 Periodic monitoring in-house as well as periodic certificates
from municipality (if applicable)

22. Microbiological Testing
Objectives
 To review microbiological environmental and quality
control testing
– Microbiological Environmental Monitoring
– Container integrity testing
– Pre-sterilization testing.
– Media fill medium growth promotion testing
– Sterility Testing
– Other microbiological laboratory issues

23. Microbiological testing of water
Water
 Water should also be tested for presence of coli forms and/or
pseudomonad's if appropriate (may cause biofilm)
 Water used for parenterals should be tested for pyrogens
– limit is not more than 0.25 EU/ml
 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°C
 Sampling procedures should follow those used in production

24. Microbiological testing of Air
Sampling Locations
– Should be based on risk of microbiological contamination
– Should be clustered around areas where product or
components are exposed e.g.
 at filling heads on filling lines
 loading of product into lyophilizers
 stopper bowls
 where aseptic connections are made
 where there are high levels of operator activity (but
without impacting on production)

25. Methods
 Surface monitoring
– Product contact surfaces, floors, walls, and equipment
should be tested on a regular basis
– Touch plates - used for flat surfaces
 sample area of 25cm2
 medium protrudes above sides
 medium contains neutralisers
– Surface Swabs - used for irregular surfaces
 area approx 25cm2 is swabbed
 qualitative or quantitative

26. Active Air Monitoring:
 impaction, centrifugal and membrane (or gelatin)
samplers
 a certain volume of air is sampled (volume and
location should be meaningful)
 instruments should be calibrated

27. Passive Air Monitoring:
• Settle plates exposed for 30-60 minutes (longer may
result in agar drying out) and replaced for duration of
filling
• Media should be capable of growing a range of
bacteria and moulds. e.g. Soybean Casein Digest Agar
(SCDA)
• Should consider use of medium specific for moulds if
shown to be a problem in the environment
• Only give qualitative or semi-quantitative results
• Data generated considered in combination with active
air sampling results

28. Media
• Media used for media fills should be able to support the
growth of a wide range of microorganisms (bacteria
and moulds)
• Soybean Casein Digest Medium is usually used. An
anaerobic medium may also be substituted occasionally
if environmental monitoring indicates presence.
• Media used for microbiological testing should be
tested for its ability to support microbial growth

29. •After the media fill has been completed, it is important
to demonstrate that the media would have been able to
support the growth of organisms if they had been
present .
•containers with media should be inoculated with 10-
100 CFU of organisms such as Bacillus subtilis,
Staphylococcus aureus, Candida albicans, Aspergillus
niger. Environmental isolates should also be included

30.  Media types:
– Soybean Casein Digest medium (SCD), (also knows as
Trypticase Soy Broth(TSB)) and Fluid Thioglycollate
medium (FTM) is usually used (to detect aerobic and
anaerobic organisms)
– validation studies should demonstrate that the media are
capable of supporting growth of a range of low numbers of
organisms in the presence of product. May need to
incorporate inactivators
 growth should be evident after 3 days (bacteria), 5 days
(moulds)
– media may be purchased or made in-house using validated
sterilization procedures

31.  Incubation Period
– At least 14 days incubation
– 20-25°C for SCD/TSB, 30-35°C for FTM
– Test containers should be inspected at intervals
– temperatures should be monitored and temperature
monitoring devices should be calibrated
– if 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 further 7
days

32. Shukriyaa friends…