SOP for Good Laboratory Practices (GLP) in Microbiology, Good Laboratory Practices (GLP) for the Bacterial Endotoxin Test (BET) and Sterility / Bio Burden test

SOP for Good Laboratory Practices (GLP) in Microbiology, Good Laboratory Practices (GLP) for the Bacterial Endotoxin Test (BET) and Sterility / Bio Burden test

This SOP is applicable for the microbiology laboratory to achieve accuracy in the microbiological results. Mainly Bacterial Endotoxin Test (BET) and sterility / Bio Burden tests are performed into the microbiology lab.

SOP for Good Laboratory Practices (GLP) in Microbiology, Good Laboratory Practices (GLP) for the Bacterial Endotoxin Test (BET) and Sterility / Bio Burden test

1.0 PURPOSE:

Refer SOP on GLP for Chemical (Wet) Analysis Laboratory

2.0 SCOPE:

Refer SOP on GLP for Chemical (Wet) Analysis Laboratory

3.0 RESPONSIBILITY:

Refer SOP on GLP for Chemical (Wet) Analysis Laboratory

4.0 DEFINITIONS

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 to nearly about 60°C.

5.1.7 Hot glassware should be handled with care and always use heat-resistant gloves for handling hot glassware.

5.1.8 While performing the Bacterial Endotoxin Test (BET) used powdered free hand-gloves to avoid 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 starting 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 the 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 doesn't 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 the handling 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 the 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 a 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 a 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 cleanroom area.

5.2.5 Always use freshly prepared disinfectant solutions.

5.2.6 Check the cleaning status of area before entering into the area for analysis.

5.2.7 Follow entry and exit procedure while entering into the clean room area.

5.2.8 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.

5.2.9 During analysis or after touching the objects disinfect hands with 70% IPA.

5.2.10 Keep the burner off when Isopropyl Alcoho0l (IPA) is tested for the bioburden test.

5.2.11 Take the required culture suspension tubes from the refrigerator and keep a side till achieve to room temperature.

5.2.12 Culture media plates and tubes should be labeled with the help of marker pen prior entering into the area.

5.2.13 Follow aseptic techniques during whole analysis.

5.2.14 During the inoculating the filter paper into the media, keep near about 6 inches distance from the flame of burner to avoid wrong results.

5.2.15 Perform inoculation as early as possible. Don’t expose filters unnecessarily at the time of inoculation.

5.2.16 Disinfect the hand frequently as and when required during analysis.

5.2.17 Do not take any leaking samples for analysis in case of liquid, powder, semisolid sample.

5.2.18 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

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