sop for calibration and validation of micro autoclave

1.0 OBJECTIVE
1.1 To describe a procedure for calibration and validation of laboratory autoclave.
2.0 SCOPE
2.1 This procedure is applicable for calibration and validation of laboratory autoclave in Microbiology Laboratory
3.0 RESPONSIBILITY
3.1 Officer Microbiologist
3.2 Executive Microbiologist
4.0 ACCOUNTABILITY

4.1 Head – QC
5.0 PROCEDURE

5.1 Calibration

5.1.1 Put biological indicator spore strip (Bacillus stearothermophilus) inside the autoclave
with sample load at various position.
5.1.2 Operate the autoclave for 15 minute at 15 psi and 1210C temperature.
5.1.3 After sterilization cycle is over Switch ‘OFF’ the main supply, after the specified period and open the steam release valve & wait for cool the autoclave, transfer aseptically the spore strips one by one to 100-100 ml Sterile Soyabean Casein Digest Medium containing tubes and incubate at 50ºC-55ºC for 7 days.

5.1.4 After incubation period, if no growth is observed, autoclave shall be considered calibrated.

5.1. 5 Calibration Frequency: Once in 6 month or after major maintenance.

5.2 Validation
5.2.1 Temperature calibration (Heat distribution in empty chamber autoclave)

5.2.1.1 Place thermocouples at different positions in empty chamber of autoclave.
5.2.1.2 Close the autoclave and operate the cycle. After achieving the 121°C temperature record the temperature of

the probes at five minutes intervals for 15 minutes.

5.2.1.3 Carryout the study of uniform heat distribution in the empty autoclave chamber for three consecutive runs.

5.2.2 Temperature calibration (Slow to Heat point distribution)

5.2.2.1 Place thermocouples at different positions in half load of autoclave.
5.2.2.2 Close the autoclave and operate the cycle. After achieving the 121°C temperature, record the temperature of the probes at five minutes intervals for 15 minutes.

5.2.2.3 Carryout the study of slow to heat point determination in the half load of autoclave for three consecutive runs.

5.3 Final Validation
5.3.1 Select five thermocouples showing minimum temperature out of ten thermocouples used in study of slow to heat point determination.

5.3.2 Place five thermocouples at the different cold points in full load and sixth thermocouple at the drain point.

5.3.3 Place biological spore’s strips (Bacillus stearothermophilus) at all five cold points and one at the drain point.

5.3.4 Close the autoclave and operate the cycle. After achieving the 121°C temperature, record the temperature of the probes at five minutes intervals for 15 minutes.

5.3.5 Carryout the study of slow to heat point determination in the full load of autoclave for three consecutive runs.

5.3.6 After autoclaving is over take out the spore strips.

5.3.7 Incubate the autoclaved spore strips at 50°C-55°C and one un-autoclaved strips as positive control for 7 days.

5.3.8 Above said calibration shall be performed for by authorized out source calibration party once a year and whenever a major maintenance is done.

5.3.9 Frequency: Once in a year.

6.0 REFERENCE
In House
7.0 FORMATS
Annexure : Autoclave Calibration & Validation Report
8.0 ABBREVIATIONS
QCM : Quality Control Microbiology

SOP : Standard Operating Procedure
QA : Quality Assurance
QC : Quality Control
0C : Degree centigrade
9.0 REVISION HISTORY