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

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

Rapid microbiological methods

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.

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?)

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.

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.

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

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?

Current guidance European Pharmacopeia “Alternative Methods for Control of Microbial Quality” (Ph. Eur. 5.1.6). 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, 610.12 (21 CFR 610.12) (2012 update)

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?

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?

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?

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

Risk assessment

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.

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)?

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.

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?

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.

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

Contamination within a cleanroom Practical example Contamination within a cleanroom

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.

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.

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.

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

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

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.

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

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

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.