SOP calibration of lab autoclave.
OBJECTIVE:
Objective of this SOP is to define the validation procedure of lab autoclave by using biological indicator carrier.
SCOPE:
This procedure is applicable for validation of lab autoclave using biological indicator carrier in Microbiology laboratory.
REFERENCE:
In house
RESPONSIBILITY:
Microbiologist: to follow the process given as per SOP.
Q.C Head: to ensure the proper execution of the SOP.
Q.A: to ensure that the SOP is being properly implemented
ABBREVIATIONS:
Q.C: Quality control
Q.A: Quality assurance
psi: pounds per square inch
SCDM: Soyabean casein digest medium
INSTRUMENT DETAILS:
A) Instrument name: Lab autoclave
Make: Metalab
Sr. no: 11367
Instrument code no.: STSP/QC/AUT/01
Location: Sterilization room, Microbiology laboratory.
B) Instrument name: Lab autoclave
Make: Visco enterprises
Instrument code no.: STSP/QC/AUT/02
Location: Sterilization room, Microbiology laboratory.
PROCEDURE:
A. By using Biological indicator:
1. Open the lid of the autoclave with the help of peddles lifting arrangement.
2. Fill the autoclave with water until the heating coils are completely immersed in it.
3. Arrange the media and glassware’s in the autoclave.
4. Place the indicators in the areas of load least accessible to steam. Places such as geometrical centers and the upper & lower regions i.e. top and bottom of the load to be sterilized are considered suitable areas for placement of these indicators.
5. Close the lid of the autoclave and tighten the knobs of the opposite side simultaneously. Plug in the mains and put on the main switch.
6. Run the autoclave on full load at pressure 15 psi and temperature 121°C for 20 min.
7. On completion of sterilization cycle, remove the indicators from the test loads. These indicators should be tested further by following procedure.
8. Firstly using sterile scissors, cut open one end of envelope. Thereafter remove the indicator with sterile tweezers and aseptically transfer it to a test tube containing sterile 10ml SCDM. Secondly, incubate these for 7days at 55-60° C.
9. Incubate one indicator strip which has not been autoclaved as positive control and one un-inoculated SCDM as negative control.
10. If turbidity is observed in any of the test tubes when compared to the positive control, failure of the sterilization process is indicated and hence the autoclave should be re-validated.
11. If no growth is observed even after 7 days of incubation, it indicates that there is homogenous dispersion of temperature across the autoclave during full load and hence, we can conclude that the autoclave is validated.
12. Each envelope consists of Bacillus stearothermophilus (Geobacillus stearothermophilus) spores impregnated on chromatography paper strips. No. of spores present per strip is 106.
B. By using Chemical indicator strips:
1. Open the lid of the autoclave with the help of peddles lifting arrangement.
2. Fill the autoclave with water until the heating coils are completely immersed in it.
3. Arrange the media and glassware’s in the autoclave.
4. Arrange three indicator strips inside the autoclave in such a manner that one strip is kept at the bottom, one in middle and one at the top.
5. Close the lid of the autoclave and tighten the knobs of the opposite side simultaneously.
6. Plug in the mains and put on the main switch.
7. Run the autoclave on full load at pressure 15 psi and temperature 121°C for 20 min.
8. After autoclaving, remove the strips from the autoclave and observe the change in the color of indicator strips from yellow to dark brown or black.
9. This change in color of the indicator strips to dark brown or black indicates that there is homogenous dispersion of temperature across the autoclave during full load and hence, we can conclude that the autoclave is calibrated.
Vertical Autoclave
FREQUENCY:
Monthly
ANNEXURE:
I. Autoclave calibration-Log book
II. Autoclave calibration record by using chemical indicator
III. Autoclave calibration record by using biological indicator
I. Autoclave calibration-Log book
Sr. no Date Time Strip no. Autoclave load no. Done by
From to Bottom middle top
Annexure II: Autoclave validation record by using chemical indicator
Equipment: Lab autoclave
Equipment no.:
Make:
Frequency: Monthly
Date of calibration:
Due date:
Sterilization time from:
Sterilization time to:
Sterilization at (⁰ C/ psi):
Autoclave load no.:
Observation:
Sr. no Autoclave strips Observation
1 Strip 1 (bottom)
2 Strip 2 (middle)
3 Strip 3 (top)
Acceptance criteria: Color of the strip changes from yellow to dark brown or black.
Conclusion: The autoclave is showing/ not showing homogenous temperature dispersion across it during full load. Hence, it can be concluded that the autoclave is calibrated/ not calibrated.
Analyzed by Checked by Approved by
Date: Date: Date:
Annexure III: Autoclave validation record by using biological indicator
Equipment: Lab autoclave
Equipment no.:
Make:
Frequency: Monthly
Date of calibration:
Due date:
Sterilization time from:
Sterilization time to:
Sterilization at (⁰ C/ psi):
Autoclave load no.:
Biological indicators manufactured by: Batch no:
Media used and incubation condition: Soyabean casein digest medium incubated at 55-60°C for 7 days.
Each envelop consists of 106 viable spores of Bacillus stearothermophilus.
Observation:
Sr. no Autoclave strips Observation
1 Strip 1 (bottom)
2 Strip 2 (middle)
3 Strip 3 (top)
4 Strip 4 (side left)
5 Strip 5 (side right)
6 Positive control
7 Negative control
Acceptance criteria: No turbidity should be observed even after 7days of incubation as compared to the positive control.
Conclusion: No turbidity/ turbidity are observed in any of the tube as compared to the positive control. Hence, it can be concluded that the autoclave is validated/ not validated.
Analyzed by Checked by Approved by
Date: Date: Date:
Issue No :03
Supersedes: 02
Issue Date: 6/3/2013 Effective Date: 7/3/2013
Review Date: 5/3/2015
Equipment: Lab autoclave
Equipment no.:
Make:
Frequency: Monthly
Date of calibration:
Due date:
Sterilization time from:
Sterilization time to:
Sterilization at (⁰ C/ psi):
Autoclave load no.:
Observation:
Sr. no Autoclave strips Observation
1 Strip 1 (bottom)
2 Strip 2 (middle)
3 Strip 3 (top)
Acceptance criteria: Color of the strip changes from yellow to dark brown or black.
Conclusion: The autoclave is showing/ not showing homogenous temperature dispersion across it during full load. Hence, it can be concluded that the autoclave is calibrated/ not calibrated.
Analyzed by Checked by Approved by
Date: Date: Date: