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

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

environmental monitoring cleaning and disinfection

People

operator broth transfer tests hand cleaning and disinfection

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

Recovery Rates for TSA and SAB plates Cherwell plates (irradiated) Exposed for 3 hours in dispensary
60 50 40
cfu

30 20 10 0 0 1 2 3 4 5 6 7 Incubation Time - Days TSA 32oC SAB 25oC

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 less risk of false positives growth characteristics altered

Irradiated plates

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 0o, 45o and 90o

laminar air flow consideration

Surface Counts

methods

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

disruption of air flow - isokinetic probes quantitative comparison accessibility sensitivity

particle size sampling method

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.