1. Tim Sandle Pharmaceutical Microbiology: www.pharmamicroresources.com
Application of rapid methods for the risk assessment of pharmaceutical processes
Tim Sandle
Pharmaceutical Microbiology:

2. Introduction Introduction to rapid methods Advantages of rapid methods
Putting rapid methods in place
Risk assessment of pharmaceutical processes
Application of rapid methods for risk assessment:
Example of an aerosol based Instantaneous Microbial Detection System

3. Rapid microbiological methods

4. What is a ‘rapid method’?
Rapid microbiological method technologies aim to provide more sensitive, accurate, precise, and reproducible test results when compared with conventional, growth-based methods.
They normally involve some form of automation.
They normal capture data electronically.

5. What is a rapid method?
Microbiological testing generally falls into one of four groups:
Qualitative tests for Presence/Absence (e.g. is E. coli in the water?)
Quantitative tests for Enumeration (e.g. how many bacteria are in a product sample?)
Quantitative tests for Potency or Toxicity (e.g. what level of endotoxin is in the sample?)
Identification tests (e.g. which species of bacteria was found in the sample?)

6. What is a rapid method? Several types:
Growth based methods: Measurement of biochemical or physiological parameters that reflect the growth of microorganisms. Require growth, so that a detectable signal is usually achieved by a period of subculture.
Viability based methods: use viability stains and laser excitation for the detection and quantification of microorganisms without the need for cellular growth (e.g. flow cytometry)
Cellular component based: the detection and analysis of specific portions of the microbial cell, including ATP, endotoxin, proteins and surface macromolecules.

7. What is a rapid method? And:
Optical spectroscopy methods: utilize light scattering and other optical techniques to detect, enumerate and (possibly) identify microorganisms (e.g. ‘real time’ airborne particle counters)
Nucleic acid amplification technologies: such as PCR-DNA amplification, RNA-based reverse-transcriptase amplification, 16S rRNA typing, gene sequencing and other novel techniques.
Micro-Electrical-Mechanical Systems (MEMS): utilize microarrays, biosensors, and nanotechnology, to provide miniaturized technology platforms.

8. What is wanted from a rapid method?
Time
To prepare the test
Conduct the test
Sample throughout
Time to result
Reduction in the time taken to conduct complimentary tests
Less time for data analysis
Simpler results reporting

9. What is wanted from a rapid method?
Accuracy
Reduction in human error
Reduction in subjectivity
To detect more accurately in comparison to a conventional method e.g. a cultural method?
To detect what a cultural method cannot?
Other areas:
Electronic capture of data?
Automation?
Connecting apparatus?

10. Current guidance
European Pharmacopeia “Alternative Methods for Control of Microbial Quality” (Ph. Eur ).
USP<1223>“Validation of Alternative Microbiological Methods”
These chapters outline the validation requirements.
FDA PAT initiative
FDA's cGMPs for the 21st Century: A Risk-Based Approach
ICH
ICH Q8; Pharmaceutical Development, and ICH Q9; Quality Risk Management encourage PAT
EU ‘real time release’
Generally encouraged but no specific guidance
CFR on sterility testing
Paved the way for a rapid sterility test.
Title 21 Code of Federal Regulations, (21 CFR ) (2012 update)

11. What is involved in validation / justifying a rapid method?
First, consider:
What do I want to achieve?
How much budget do I have?
Which technologies are available?
Which technologies are ‘mature’? (who else is using them?)
How ‘rapid’ is the rapid method?
What papers have been published? (are these ‘independent’?)
What have regulators said?

12. What is involved in validation / justifying a rapid method?
Then select the technology...and pose more questions:
How much will the validation cost?
How long will the validation take?
How many personnel will the validation require?
How many tests will I need to run for the validation?
Does the validation require a comparison with another (existing) method?
How will the data be analysed and reported?

13. What is involved in validation / justifying a rapid method?
Considerations for gaining approval:
Internal change control process.
Regulatory process:
Which agencies do I need to approach? (multiple markets?)
What data is needed for a submission?
How long will it take to get approval?
Europe: normally a Type 2 change
FDA: normally the Prior Approval Supplement process.
What will be needed for a regulatory inspection?

14. General validation steps
Risk Assessment
Validation Master Plan (VMP)
User Requirements Specifications (URS)
Design Qualification (DQ)
Supplier Assessment/Audit
Functional Design Specifications (FDS)
Requirements Traceability Matrix (RTM)
Training and Standard Operating Procedures (SOPs)
The Test Plan
Factory Acceptance and Site Acceptance Testing (FAT and SAT)
Installation Qualification (IQ)
Operational Qualification (OQ)
Performance Qualification (PQ)
Validation Summary Report

15. Risk assessment

16. Importance of risk assessment
The use of risk assessment in the pharmaceutical industry is an expectation of regulatory authorities.
The risk assessment involves either a quantitative or qualitative determination of one or more risks.
Risks relate to a situation where a recognized hazard may result in harm.
A hazard, in this context, is any circumstance in the production, control and distribution of a pharmaceutical product, which can cause an adverse health effect.

17. What is ‘risk’?
Risk is defined as the combination of the probability of occurrence of harm and the severity of that harm i.e.
What might go wrong?
What is the likelihood (probability) it will go wrong?
What are the consequences (severity)?

18. Microbiological risks
Microbiological contamination in the product, which might cause patient harm.
From:
Equipment
Air e.g. Cleanrooms:
Air filtration
Air direction
Air movement (pressures)
People
Water
Central issue is contamination transfer.

19. What does a ‘risk’ mean?
Is the risk acceptable and what controls are available to mitigate the risk?
Is the risk above an acceptable level?
What can be done to reduce or eliminate risks?
What is the appropriate balance among benefits, risks and resources?
Are new risks introduced as a result of the identified risks being controlled?

20. Use of risk assessment Risk assessment can be:
Proactive
Reactive
Focused on process improvements
There is no such thing as ‘zero risk’
A decision is required as to what is ‘acceptable risk’.
Risk Assessment is not an exact science.
Different people will have a different perspective on the same hazard.

21. Basics of risk assessment
Formal risk approaches normally share four basic concepts, which are listed below:
Risk assessment,
Risk control,
Risk review,
Risk communication.

22. Contamination within a cleanroom
Practical example
Contamination within a cleanroom

23. Cleanroom contamination
Points of contamination:
adjacent areas;
supply air;
cleanroom air;
surfaces;
people;
machines;
ancillary equipment;
materials;
containers;
packaging;
liquids
Focus on:
The reinstatement of a cleanroom following a period of production downtime.

24. Method of assessment
Azbil BioVigilant's Instantaneous Microbial Detection System (IMDA).
Uses advances in light scattering, optics and special software.
Provides real-time data about particles and biological activity in air.
Suited for cleanrooms and controlled environments.

25. IMDA In real-time: Air is passed through a laser.
The instrument counts the numbers of particles in a sample of air (inert and biological) through two detectors.
Biological particles are detected using a fluorescence detector, looking for three biological markers: NADH, riboflavin, and DPA.
These metabolites cause fluorescence and the presence of a biological microorganism.

26. Cleanroom use Cleanrooms: As built At rest In-operation
Downtime for maintenance
E.g. HEPA filter replacement, new equipment, drilling
Reinstatement
Cleaning and environmental monitoring

27. Cleanroom reinstatement
Problem:
Waiting for results through environmental monitoring
3 – 10 days, depending upon media and incubation parameters.
Solution
IMDA can provide an assessment in less than one hour

28. Study Need to understand normal operating conditions
Setting a baseline
Observing events
Understand state of the area in relation to viable monitoring, but a direct comparison is not possible.

29. Study Assessment during normal operations
Assessment during shutdown period
Highest Bio reading = 180
Highest Bio reading = 2,500

30. Study Assessment post-shutdown
Assessment of cleaning and recovery period
Assessment post-shutdown
Highest bio reading = 370
Highest bio reading = 220

31. Assessment IMDA provides the means to:
Understand what is ‘normal’ in terms of biological activity & set alert and action profiles.
Understand the impact of people in the area, equipment movements etc.
Understand what happens when an area is declassified.
Understand the impact of cleaning.
Compare the cleanroom following reinstatement with the previous status.