This document discusses microbiology activities related to sterility assurance for pharmaceutical products. It outlines the microbiological testing done on raw materials, processes, water systems, environments and finished products. This includes bioburden testing, pathogen testing, endotoxin testing, and sterility testing. It also discusses environmental monitoring programs for sterile facilities and aseptic filling areas. Key requirements outlined include pre-filtration bioburden limits, media fill qualifications, aseptic process simulations, and staff performance monitoring. Recent issues with increased out of limit environmental monitoring results are also summarized.

1. Microbiology and Sterility
Assurance
Dr. Tim Sandle
www.pharmamicroresources.com

2. Microbiology Activities
• Laboratory:
– In-coming raw materials
• Bioburden
• Specific pathogens
– Processing of plasma
• Hygiene swabs
• Bioburden
– Intermediate product
• Bioburden
• Endotoxin
– Finished product
• Sterility
• Endotoxin
• Pyrogens
• Toxicity

3. Microbiology Activities
• Testing of environment
• Viable samples
• Particle counting
• Testing of water
– WFI
– Purified
– Mains
• Microbial identification
– What?, Why?, Where?

4. Impact on Sterile Supplies
• Sterile Supplies is an EU GMP Grade C facility
(ISO 14644 class 8)
– Certified cleanroom: annual
– Environmental monitoring, based on risk:
• Types of activities
• Whether there are water sources
• Room history
– Water samples are taken from outlets

5. Impact on Aseptic Filling
• Routine monitoring programme of support rooms
– Monitored according to risk e.g. corridors monitored
more often than infrequently used rooms
– Monitoring is:
• Twice weekly
• Weekly
• Fortnightly
• Monthly
– Monitoring is for:
• Viable counts
• Particles
• Fungi

6. Impact on Aseptic Filling
• Batch monitoring
– Filtration of the product
– Unloading stoppers
– Machine set-up
– Pre-fill period (target 4 hours)
– Filling
– Post-fill monitoring
– Machine clean down

7. Environmental Monitoring
• Samples
– Settle plates (deposition rate / 4 hours)
– Active air (cubic metre)
– Contact plates
– Swabs
– Finger plates (staff)
– Gloveport plates
– Gown plates

8. Environmental Monitoring
• Routine, support samples taken by
Microbiology
• Batch samples taken by Sterile Operations
• Samples contain tryptone soya agar
– Some contain neutraliser
– Samples incubated at a regime designed to
encourage the growth of bacteria and fungi.

9. Important Points #1
• Pre-final filtration TVAC
– Taken in Sterile Operations
for certain samples
– Bioburden assessment of
product prior to
sterilisation through 0.22
µm filter
a) Sample must be placed
at 2-8oC within 2 hours
b) Samples must be tested
within 24 hours
c) Bioburden must not
exceed 10 CFU / 100mL
• Any of the above will lead
to batch rejection.

10. Important Points #2
• Media fills
– Qualify the filling line and the people
– Different vial sizes and numbers in rotation
• Maximum broth fill size sets maximum batch size
– Require a set number of aseptic interventions
– Contain the maximum stoppage
– Run every six-months per line
– Each person must participate once per year and
carry out an intervention

11. Important Points #3
• AFS qualification
– Gown test
• Visual check of change procedure against SOP
• Samples relating to environment and individual
– Aseptic qualification test
• Test of aseptic technique
– Conducted
• Three times for new staff or after a prolonged absence
• Six-monthly for qualified staff
• Zero tolerance for staff who miss due dates

12. Important Points #4
• Data is reviewed every month and summaries
are sent out to Sterile Ops Management
• Includes staff performance
– Each staff member is rated green, amber or red
– Includes:
• Gown test results
• Finger plate trends
• Exit suit plate trends
• Behavioural issues

13. Current issues
• There is currently an upward trend in the AFS
relating to environmental monitoring data.
• Trend began after the winter 2013 shutdown.
• Primarily relates to people, behaviours and
aseptic practices.
• Some issues linked to cleaning.

14. Current issues
0
2
4
6
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12
14
Jan-12 Feb-12 Mar-12 Apr-12 May-12 Jun-12 Jul-12 Aug-12 Sep-12 Oct-12 Nov-12 Dec-12 Jan-13 Feb-13 Mar-13 Apr-13 May-13 Jun-13
Total number of samples OOL: batch and routine 2012 - 2013
No. OOL 2012 average 3 per. Mov. Avg. (No. OOL)

15. Current issues
• Importance of:
– Good gowning practices.
– Not being the AFS for more than four hours (four hour
gown change policy).
– Slow, deliberate movements.
– Spraying gloves before each activity.
– Using the 1 minute hand rubbing procedure.
– Spraying gloves every 10 minutes.
– Reporting broken hand dispensers and fabric issues to
a supervisor.
– Cleaning and disinfecting surfaces to standard.

16. Thank you
Any questions?