SOP FOR OPERATION AND VALIDATION OF PORTABLE AUTOCLAVE

1.0       Objective:

               To provide written procedure for the operation and validation of Portable Autoclave.

2.0          Scope:

               This SOP covers operation and validation of Portable autoclave  in microbiology.

3.0          Responsibility:

               Microbiologist: Responsible for operation and validation of instrument as per procedure and maintain the record.

              QA Officer/QAM: Review the records and governing the document.

4.0          Procedure:

4.1          Ensure that the instrument is clean and free from dust and placed in a safe area.

4.2          Open the lid and remove the dressing drum –basket and ensure that the chamber is clean.

4.3          Pore the purified water in the vessel up to the level to get heaters immersed in the water.

4.4        Place the carriers in it’s original position after loading the samples or may be empty as may require to test initially.

4.5      Close the lid carefully and securely tight the fly nuts of opposite direction one after another to ensure the proper sealing of vessel.

4.6          Switch ON the mains, allows the unit to heat up and from the steam in the vessel.

4.7        Observe the pressure gauge. Soon it starts indicating the increment of pressure gauge turn back to Zero reading.

4.8         Allow exhausting the air pocket in the vessel for a minute to ensure the proper purging the dry air.

4.9          Now again close the steam release valve.

4.10        Allow the pressure to reach to it’s maximum capacity.

4.11     The safety valve is pre-settled to activate at 15 psi pressure so as to maintain the operating pressure at 15 psi constant.

4.12        Run the test for the desired duration to complete the sterilization cycle.

4.13     On completion of test cycle, switch off the supply.

4.14   Open the steam release valve to purge out the steam pressure of the vessel.

4.15    Observed the zero reading on the gauge and then open the lid carefully using the asbestos or safety gloves to remove the samples to observe the sterilization result with the help of indicator. 

4.16                Enter the details of usage in Autoclave load register Annexure – 1

5.0          Validation:

5.1          Routine Validation:

5.1.1       Put a sterilization indicator strip in a Petri plate along with each load of Autoclave.

5.1.2       If the colour white cross line of the strip changes to brownish black it indicates the sterilization for the lot is complete.

5.1.3       If the colour of the cross line does not change to brownish black, then do not use the  items sterilized.

5.1.4       Paste the sterilization indicator strip in the autoclave load register.

5.2          Quarterly Validation:          

5.2.1 Sterilize one ampoule/Strip of Geobacillus stearothermophilus (ATCC 7953 Bio indicator) at 121˚ C for 15 minutes (15 lb/in2 pressure).

5.2.2   Remove the ampoule/Strip once the sterilization is complete.

5.2.2    For Ampoule:  Incubate at 43˚ to 45˚ C for 3 to 5 days along with a positive control (Unautoclaved ampoule).

5.2.3  For Strips:  After sterilization, aseptically transfer the processed strip and one control strip into prepared sterilized Nutrient Broth. Incubate at 55°C –60°C for 7 days along with a positive control ( Unautoclaved strip / ampoule).

5.2.4       Observe the growth, purple colour should turn to yellow in the positive control showing the presence of Bacillus stearothermophilus. Sterilized ampoule should remain            purple confirming the completeness of sterilization.

5.2.5       Perform the observation 5.2.1 to 5.2.4 with different autoclave loads. (i)  Empty load.

              (ii)      Maximum load.           (iii)       Minimum load

5.3        Record the observations in the autoclave validation register (Annexure - 2).

6.0          Routine maintenance:

6.1                 Remove scaling from the instrument and stand with 50 ml hydrochloric acid in 20 liter of water.

6.2      After cleaning with hydrochloric clean instrument with distilled water.

7.0          Documentation:

7.1          Annexure – 1 -  Autoclave usage log book.                    XXX/FRM/000-00

7.2          Annexure – 2 -  Autoclave validation Format.               XXX/FRM/000-00

8.0          History of Revision:

Revision No.	Effective Date	Revision details	Reason for revision

Annexure - 1
Date	Name of the Item	Autoclave Load No	Time			Indicator strip after sterilization	Done by	Remarks
			Start	Activate	Stop
SOP No. : XXX/SOP/000-00                                                          Format No. : XXX/FRM/000-00

Annexure - 2

Validation of Autoclave by Spore Strips Instrument No : Date of validation:                                                          Date of observation: Autoclave Load No.: Indicator used Biological:  Geobacillus stereothermophillus ATCC – 7953 Batch no: Mfg date: Exp. Date Media used Soyabean casein digest broth 20 ml Sterilization condition Temperature: 121°c Pressure       : 15lb Time            : 30 min. Position / point Top, middle & bottom Incubation At  55 to 60°c for 7 days Observation No growth/growth observed after 7 days of incubation Conclusion Autoclaved is fit/not fit for use and said to be validated / not validated Next validation due Date: Validated by : Dept. head: SOP No. : XXX/SOP/000-00                  Format No. : XXX/FRM/000-00