Microbiological Validation Mark Oldcorne Wrexham Maelor Hospital

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

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’

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

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

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

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

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

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

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

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

Microbiological Validation General Considerations 3 Incubation times 5-7 days but must read after day 1 Incubation temperatures Bacteria - 30-32oC Fungi 20-25oC

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

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

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

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

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

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

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

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

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

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

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

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

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?

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

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……….