1. Microbiological Validation
Mark Oldcorne
Wrexham Maelor Hospital

2. What is Validation?
The QA of any preparation activity is reliant on the satisfactory validation of the procedures
Validation should demonstrate that the overall process will reproducibly provide a product that complies with its specification

3. Action of proving (and documenting) that something ‘works’
What is validation?
Action of proving, in accordance with the principles of GMP, that any procedure, process, equipment, material, activity or system leads to the expected results
Action of proving (and documenting) that something ‘works’

4. 5 Pillars of GMP Premises People Processes Products Procedures
Profit!!!

5. Overview Microbiological validation of Premises People
environmental monitoring
cleaning and disinfection
People
operator broth transfer tests
hand cleaning and disinfection

6. Underpinned by Procedures
Overview
Processes
process broth tests
transfer spraying validation
gassing processes
Products
sterility tests
Underpinned by Procedures
(parametric release!)

7. Validation Master Plan
Part of
QA programme
Site Master File
Schedules
Methods
Responsibilities

8. Microbiological Validation Methods
Physical - AHU\filtration
Microbiological - Personnel activity

9. Microbiological Validation General Considerations 1
Biological systems
biological variability
imprecision of methods
low levels of contamination
Stressed / damaged organisms

10. Microbiological Validation General Considerations 2
Growth requirements of organisms
require universal growth media
TSA + Sabs Dex
Sequential temperature monitoring
vs use of multiple media

11. Microbiological Validation General Considerations 3
Incubation times
5-7 days
but must read after day 1

13. Microbiological Validation General Considerations 3
Incubation times
5-7 days
but must read after day 1
Incubation temperatures
Bacteria oC
Fungi 20-25oC

14. Microbiological Validation General Considerations 4
Sterile media
Aseptically prepared plates
Pre-incubation
Irradiated plates
less risk of false positives
growth characteristics altered
Fertility assessment

15. Microbiological Validation General Considerations 5
Viable but non-culturable organisms (VBNC)
Presence of non-recoverable organisms
Results are Retrospective
Prospective release
Retrospective release

16. Interpretation of Results
Facilities management
Investigational threshold (Alert limits)
Action limit
Counts
ID of Organisms
source of organism
consequences of contamination
presence of new organisms
failure of control measures

17. Trend analysis 1 Identify progressive and gross changes variables
Workspace
Room
Product
Operator (not just aseptic processing)
Operator – spraying in

18. Trend Analysis 2 Methods available
visualisation by charting and graphical methods
statistical analysis
population study software
exception reporting
follow up of problems
limitation of variables
number of exceptions (investigational or action limits) used as a measure of cleanliness

19. Microbiological Environmental Methods
Random Methods
Settle plates
Contact sampling
Finger dabs
Organised Methods
Active air sampling
*Assumptions*

20. Settle Plates method of collection sampling area sampling time
positioning
trend analysis
representative sample
laminar air flow consideration
0o, 45o and 90o

21. Surface Counts methods sampling area and location pickup efficiency
Rodac Plates
Swabs
Proprietary systems
sampling area and location
pickup efficiency
neutralisation
post sampling removal of media
application pressure
validation of disinfection processes

22. Finger Dabs sampling technique what does it assess? holes in gloves
poor transfer disinfection

23. Active Air Sampling methods sampling efficiency - recovery rates
impingement
filtration
sampling efficiency - recovery rates
particle size
sampling method
disruption of air flow - isokinetic probes
quantitative comparison
accessibility
sensitivity

24. Sterility Testing Ideal test? - product Test lacks sensitivity
Retest only available under certain conditions
Prospective vs retrospective
Appropriate levels of control
SAL of testing process

25. Broth Transfer Testing 1
Process transfer tests
worst case scenario
>batch size
x3 initially
6 monthly repeat
Operator transfer tests
Universal Broth Transfer Test
Adapted operator tests or process tests

26. Broth Transfer Tests 2 Ideal test? - QA of process
Statistical considerations to reach AQL
1 in 1000
or 1 in 10,000
Frequency?
Sensitivity - do we challenge the test?

27. Conclusions Microbiological validation indicates personnel involvement
All microbiological validation methods have limitations
Microbiological validation methods cannot be used in isolation and should be used holistically to gain a true picture of product assurance

28. The future Where does the validation cult go?
Next stage is validating the validation
Next stage to that is validating the validation of the primary validation and so on……….