Basic Principles of GMP

1. Basic Principles of GMP WHO good practices for pharmaceutical microbiology laboratories Part 1 TRS 961, 2011. Annex 2

2. Microbiology Laboratories • Personnel • Environment • Premises • Environmental monitoring in the laboratory • Cleaning, disinfection and hygiene • Sterility test facilities • Validation of test methods • Equipment • Maintenance of equipment • Qualification • Calibration, performance verification and monitoring of use

3. Microbiology Laboratories • Reagents and culture media • Reagents • Media • Labelling • Organism resuscitation • Reference materials and reference cultures • International standards and pharmacopoeial reference substances • Reference cultures • Sampling • Sample handling and identification

4. Microbiology Laboratories • Disposal of contaminated waste • Quality assurance of results and quality control of performance • Internal quality control • Testing procedures • Test reports

5. Microbiology Laboratories Introduction and scope of document • Some of the activities in pharmaceutical microbiology laboratories include: • sterility testing; • detection, isolation, enumeration and identification of microorganisms (bacteria, yeast and moulds) and testing for bacterial endotoxins in different materials (e.g. starting materials, water), products, surfaces, garments and the environment; and • assay using microorganisms as part of the test system. •  The WHO guidelines cover recommendations for these activities • See also other guidelines and pharmacopoeia

6. Microbiology Laboratories Personnel

7. Microbiology Laboratories Personnel • Testing and supervision by experienced persons, qualified in microbiology or equivalent field • Training in microbiology and relevant practical experience • Current job descriptions for all personnel involved in tests and/or calibrations, validations and verifications should be maintained. • Records of all technical personnel, describing their qualifications, training and experience. • Opinions and interpretations of test results by authorized personnel with suitable experience and relevant knowledge 1.1 – 1.3

8. Microbiology Laboratories • All personnel to have adequate training e.g. • performance of tests • operation of equipment • basic techniques, e.g. plate pouring, counting of colonies, aseptic technique, media preparation, serial dilutions, and basic techniques in identification • Trained for containment of microorganisms within the laboratory • Trained in safe handling of microorganisms • Competence monitored - retraining where necessary 1.4 – 1.6

9. Microbiology Laboratories • There should be restricted access to the microbiological laboratory • Personnel should be made aware of: • the appropriate entry and exit procedures including gowning; • the intended use of a particular area; • the restrictions imposed on working within such areas; • the reasons for imposing such restrictions; and • the appropriate containment levels 2.1.5

10. Microbiology Laboratories Environment Premises

11. Microbiology Laboratories • Dedicated and separated: areas and support equipment (e.g. autoclaves and glassware) • Appropriate design for operations, sufficient space, avoid mix ups, contamination and cross-contamination • Suitable materials of construction to enable appropriate cleaning, disinfection and minimize the risks of contamination • Space for samples, reference organisms, media (if necessary, with cooling), testing and records. Separate storage locations for some media/samples 2.1.1. – 2.1.3.

12. Microbiology Laboratories • Separate air supply to laboratories and production areas with temperature and humidity control • Appropriate quality of air - not to be a source of contamination • Segregated, classified areas for sample preparation, media and equipment preparation and enumeration of microorganisms • Non-dedicated areas can be used based on risk management principles should be applied 2.1.4 – 2.1.6

13. Microbiology Laboratories • Classification should be based on criticality • Sterility testing should always be performed in a dedicated area • Equipment not moved between areas of different cleanliness class, and not be used outside the microbiology area, unless there are specific precautions in place to prevent cross-contamination 2.1.6 - 2.1.8

14. Microbiology Laboratories Environmental monitoring in the laboratory • Environmental monitoring programme where necessary e.g. in areas for sterility testing • Active air monitoring, air settling or contact plates, temperature and pressure differentials. • Alert and action limits; and trending of results Cleaning, disinfection and hygiene • Cleaning and disinfection programme with results • SOP for dealing with spillages. •  Hand-washing and hand-disinfection facilities 2.2.1 – 2.3.3

15. Microbiology Laboratories Sterility test facilities • Specific environmental requirements - clean-room requirements • Done under aseptic conditions • Class A (ISO 4.8) in Class B background (unidirectional airflow) - or within a barrier isolator/biosafety cabinet • Suitable design of the facility layout and room airflow patterns, to ensure that the unidirectional airflow patterns are not disrupted. • Premises, services and equipment subjected to the appropriate qualification process 2.4.1 – 2.4.3

16. Microbiology Laboratories

17. Microbiology Laboratories • Clean-room classification and air-handling equipment of the sterility test facilities should be requalified at least annually (or other frequency if no risk). Tests should include e.g.: • Non-viable and viable limits • HEPA filter integrity • Room airflows • SOPs to state mapping locations for sample points for routine monitoring, exposure duration, and frequency of all types of microbiological environmental monitoring • Garments appropriate, and operators trained and certified in gowning procedures 2.4.4, 2.4.9

18. Microbiology Laboratories • Grade A and B zones – supplied with terminal HEPA filters. •  Airflow alarms and pressure differentials and indication instruments • Room pressure readings taken and recorded • Pressure gauges should be labelled to indicate the area served and the acceptable specification. • Entry to the clean rooms via airlocks, suitable size • Final change room should be under “at rest” conditions of the same grade as the room it serves 2.4.5 – 2.4.8

19. Microbiology Laboratories • Environmental microbiological monitoring should reflect the facility used (room or isolator) and include a combination of air and surface sampling methods appropriate to the facility, such as: — active air sampling; — settle (exposure) plates; — surface contact — replicate organism detection and counting (RODAC) plates, swabs or flexible films; — operators’ glove prints. 2.4.11

20. Microbiology Laboratories

21. Microbiology Laboratories Microbial environmental monitoring of the sterility test zone: • during every work session • under operational (dynamic) conditions • written specifications • alert and action limits for microbial contamination 2.4.11

22. Microbiology Laboratories Validation of test methods • Compendial methods: Need to be shown to be suitable for use in recovering bacteria, yeast and mould in the presence of the specific product. Method verification needed and test method suitability including positive and negative controls • Non compendial or other recognized references: Validated before use and include e.g. accuracy, precision, specificity, limit of detection, limit of quantitation, linearity and robustness • Potentially inhibitory effects considered • Appropriate statistical methods used (see pharmacopoeias) 3.1 – 3.2

23. Microbiology Laboratories Equipment • Each item of equipment, instrument or other device used for testing, verification and calibration should be uniquely identified • Documented programme for the qualification, calibration, performance verification, maintenance • System for monitoring the use of its equipment. • SOP for maintenance of essential equipment at predetermined intervals • Detailed records kept 4.1

24. Microbiology Laboratories Qualification • For qualification of equipment see details in the training modules on Good practices for pharmaceutical quality control laboratories • DQ • IQ • OQ • PQ 4.2

25. Microbiology Laboratories Calibration, performance verification and monitoring of use • Instruments labelled (dates) • Frequency of calibration and performance verification determined • Intervals between calibration and verification should be shorter than the time the equipment has been found to take to drift outside acceptable limits • Equipment to perform to predefined acceptance criteria 4.3.1 – 4.3.3

26. Microbiology Laboratories Examples of maintenance of equipment • The next two slides provide information as examples • The frequency should be based on the need, type and previous performance of the equipment and on the recommendations in suppliers’ manuals

27. Microbiology Laboratories

28. Microbiology Laboratories

29. Microbiology Laboratories

30. Microbiology Laboratories Examples of equipment qualification and monitoring • This information is provided as an example and the frequency will be based on the need, type, previous performance and criticality of the equipment.

31. Microbiology Laboratories Temperature-controlledequipment (incubators,baths, fridges,freezers) Sterilizing ovens

32. Microbiology Laboratories  Type of equipment Requirement Suggested frequency Autoclaves

33. Microbiology Laboratories Isolators

34. Microbiology Laboratories Temperature measurement devices • Where temperature has a direct effect on the result of an analysis or is critical for the correct performance of equipment, temperature measuring devices should be of appropriate quality to achieve the accuracy required (e.g. liquid-in-glass thermometers, thermocouples and platinum resistance thermometers (PRTs) used in incubators and autoclaves). • Calibration of devices should be traceable to national or international standards for temperature 4.3.3

35. Microbiology Laboratories

36. Microbiology Laboratories Incubators, water-baths and ovens • The stability of temperature, uniformity of temperature distribution, time required to achieve equilibrium conditions in incubators, water-baths, ovens and temperature-controlled rooms should be established initially and documented, in particular with respect to typical uses (for example, position, space between, and height of, stacks of Petri dishes) • Recorded during initial validation, checked and recorded after each significant repair or modification • Operating temperature of this type of equipment should be monitored and records retained 4.3.4

37. Microbiology Laboratories Autoclaves, including media preparators • Autoclaves to meet specified time and temperature tolerances as well as pressure • Sensors used calibrated and timers verified • Validation to include temperature distribution studies for each operating cycle and each load configuration used; and heat penetration studies • In the case of media preparators – use two sensors • Validation and revalidation should consider heating, sterilisation and cooling 4.3.5

38. Microbiology Laboratories • Clear operating instructions (SOPs) • Acceptance/rejection criteria • Records of autoclave operations, including temperature and time, maintained for every cycle  • Monitoring may be achieved by one of the following: • using a thermocouple and recorder to produce a chart or printout; • direct observation and recording of maximum temperature achieved and time at that temperature. •  Use of chemical or biological indicators and autoclave tape • Separate autoclave for decontamination 4.3.5

39. Microbiology Laboratories Weights and balances • Regular calibration, traceable to standard weights and certified standard weights Volumetric equipment • Initial verification of volumetric equipment (automatic dispensers, dispenser/diluters, mechanical hand pipettes and disposable pipettes) - then regular checks thereafter • Checked for the accuracy of the delivered volume against the set volume (for several different settings in the case of variable volume instruments) and the precision - “single-use” also checked 4.3.6 – 4.3.7

40. Microbiology Laboratories Other equipment • Conductivity meters, oxygen meters, pH meters and other similar instruments verified regularly or before each use • Buffers used - storage and expiry dated • Where hygrometers are used – calibrated • Timers verified using a calibrated timer or national time signal • Centrifuges – verify rotations per minute (RPM). Where it is critical, the centrifuge should be calibrated 4.3.8