This SOP is applicable for the microbiology laboratory to achieve the accuracy in the microbiological results. Mainly Bacterial Endotoxin Test (BET), sterility / Bio Burden test are performed into the microbiology lab.
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1.0 PURPOSE:
The purpose of Good Laboratory Practices SOP is to provide the guideline for Good Laboratory Practices (GLP) for the microbiology laboratory.
2.0 SCOPE:
This procedure is applicable for Good Laboratory Practices at microbiology.
3.0 RESPONSIBILITY:
3.1 Microbiology
3.1.1 All employee working in the microbiology follow the Good Documentation Practices (GDP), ALCOA+, Good Laboratory Practices.
3.1.2 Prepare and review the specification, Standard test procedure (STP), test protocols, SOP, and standard formats.
3.1.3 Record and report the analytical results.
3.1.4 Maintain the laboratory as per the regulatory requirement.
3.2 Quality Assurance
3.2.1 Approve the specification, Standard test procedure (STP), test protocols, SOP, and standard formats.
3.2.2 Verify for the laboratory as per the regulatory requirement.
4.0 DEFINITIONS
Good Laboratory Practices (GLP)
Practices follow during analysis and clinical work for the reliable, accurate, consistent, reproducible analytical and research-oriented results to maintain the integrity of the product.
Refer SOP on GLP for Chemical (Wet) Analysis Laboratory
5.0 PROCEDURE
5.1 Bacterial Endotoxin Test (BET)
5.1.1 Always check the cleaning status of the LAF and working area of BET room before starting any analytical procedure.
5.1.2 The Laminar Air Flow (LAF) should be turned on 15 mins before starting the analysis.
5.1.3 Multi block heater should be turned on 1 hour before start the analysis.
5.1.4 Always use depyrogenated glassware for analysis of bacterial endotoxin test (BET).
5.1.5 Always check the logbook of Hot Air Oven for confirmation of glassware depyrogenation status.
5.1.6 After completion of depyrogenation cycle, drop down the oven temperature nearly about 60°C.
5.1.7 Hot glassware should be handled with care and always use heat resistant gloves for the handling hot glassware.
5.1.8 While performing the Bacterial Endotoxin Test (BET) used powdered free hand gloves to avoid the contamination.
5.1.9 BET reagents should be store in refrigerator at 2-8ºC.
5.1.10 Always use calibrated glassware’s for analysis.
5.1.11 Check the calibration status of instruments before start the procedure or analysis.
5.1.12 If required used Vortex shaker for mixing, solubility purpose.
5.1.13 Instruments should be used within its permitted calibration limit.
5.1.14 Keep the multi block heater on anti-vibrating table to avoid the disturbance in gel formation.
5.1.15 Check the temperature of WFI before reconstitution of lysate. It should be below 25ºC.
5.1.16 Reconstituted lysate should be used within 24 hrs.
5.1.17 Check the lysate reconstitution date and time before use of previously reconstituted lysate.
5.1.18 If reconstituted lysate is not used within 24 hrs then discard the unused reconstituted lysate.
5.1.19 If reconstituted lysate having foam or turbidity then do not use the lysate.
5.1.20 Handle depyrogenated glassware’s and items in such a way that their properties remain unaltered.
5.1.21 Use separate calibrated micropipette for handling of endotoxin.
5.1.22 Do not touch the tip of micropipette to any unwanted surface.
5.1.23 If pipette not used for analysis then tip of pipette always covered with aluminium foil and keep it into the pipette stand.
5.1.24 Observe and verify the results of BET in presence of second authorized person.
5.2 Sterility / Bio Burden (BB) testing:
5.2.1 During cleaning of LAF bench always keep the burner “OFF”.
5.2.2 Always wear hand gloves and mask during preparing as well as discarding media.
5.2.3 Use sterile utensils for analysis purpose.
5.2.4 All objects should be sterilized or disinfected before entering the clean room area.
5.2.5 Always use freshly prepared disinfectant solutions.
5.2.5 Check the cleaning status of area before entering into the area for analysis.
5.2.6 Follow entry and exit procedure while entering into the clean room area.
5.2.7 Check the pressure differential of testing area before going for analysis. If it is not within the specified limit then do not enter in to the clean room for analysis purpose.
5.2.8 During analysis or after touching the objects disinfect hands with 70% IPA.
5.2.9 Keep the burner off when Isopropyl Alcoho0l (IPA) is tested for the bioburden test.
5.2.10 Take the required culture suspension tubes from the refrigerator and keep a side till achieve to room temperature.
5.2.11 Culture media plates and tubes should be labeled with the help of marker pen prior entering into the area.
5.2.12 Follow aseptic techniques during whole analysis.
5.2.13 During the inoculating the filter paper into the media, keep near about 6 inches distance from the flame of burner to avoid wrong results.
5.2.14 Perform inoculation as early as possible. Don’t expose filters unnecessarily at the time of inoculation.
5.2.15 Disinfect the hand frequently as and when required during analysis.
5.2.16 Do not take any leaking samples for analysis in case of liquid, powder, semisolid sample.
5.2.17 Observe and verify the results of BET in presence of second authorized person.
6.0 ABBREVIATION
WFI – Water for Injection.
LAF – Laminar Air Flow
BET – Bacterial Endotoxin Test
IPA – Iso Propyl Alcohol
7.0 REFERENCES
7.1 21 CFR part 211 subpart I – Laboratory Control
7.2 Schedule L1
7.3 Handbook of Good Laboratory Practices.
8.0 HISTORY
Not Applicable
9.0 ANNEXURES
Not Applicable
FAQ:
1. Who is responsible for following good microbiological laboratory practice?
All employee working in microbiology department is responsible for good laboratory practices, good documentation practices.
2. What is microbiological laboratory practices?
Practices follow during microbiological analysis and testing for reliable, accurate, consistent, reproducible results to maintain the integrity of the product.
3. Why is it important to follow good laboratory practices in a microbiology laboratory?
1. To achieve reliable, accurate, reproducible results.
2. To maintain uniformity in work.
3. To avoid nonconformance, out of specification, out of trend, lab incidents etc.
4. To avoid cross-contamination, mix up etc. in the laboratory.