SOP FOR THE OPERATION AND VALIDATION OF VERTICAL AUTOCLAVE

1.0 Objective:

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

2.0          Scope:

               This SOP cover operation and validation of Vertical autoclave Model No. : 7451 FA in microbiology department.

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 autoclave by unscrewing the Wing Nuts.

4.3          Remove the baskets but leave the Heater Cover X stand inside.

4.4          Ensure that the drain valve at the bottom of the autoclave is closed.

4.5          Pour purified water direct into the Autoclave, up to the heater cover X stand.

4.6     Load the Autoclave, close the lid, clamp the Wing and tighten opposite fly nuts simultaneously with equal pressure.

4.7     Switch ‘ON’ the mains; the set temperature and set time will be displayed on the Microprocessor controller in brackets. If acceptable, press START. If change the temperature and time as per procedure given in controller setting as below

4.8          Micro Processor Based (µP) Controller Setting

4.8.1      When power is switched ON the display will show status OK/EQUITRON for 3seconds and then the main screen will appear where in the top line will display the process temperature, last set temperature (in brackets) and bottom line will display time 00 and last set time (in brackets).

4.8.2      To change temperature press MENU, the display will change to set temperature. Press Up or DOWN arrow keys. For the temperature change permanent for all future cycles,

               press SET, the display will return to the main screen, for the temperature change only for        

               this particular cycle, press MENU again, the display will change to set time.

4.8.3      To change time press MENU, if previous time change, Press Up or DOWN arrow keys. For the time change permanent for all future cycles, press SET, the display will return to the main screen, for the time change only for this particular cycle, press MENU again, the display will return to the main screen.

4.8.4       Press START to start the cycle. In case of problems encountered, the buzzer will sound, the alarm LED will flash, and the problem will be on the display.

4.8.5      After the start switch is pressed, to change the set temperature and time incorporates the change, press START again and the change will overwrite the earlier settings. 

4.9          Allow the temperature reaches up to 100° C, then open the valve and release the vapour. Close the valve when continuous steam comes out.

4.10       After this, the temperature and pressure will gradually rise, which will be indicated by the µPC and pressure gauge.

4.11       Just before the set temperature the Heater will be cut OFF and then gradually temperature is attain the set temperature. The TIMER will start its countdown after the set temperature is attained. During the countdown the temperature and hence the pressure will be maintained by the µPC by switching ON/OFF the heater.  

4.12      After the elapse of the set time. A Buzzer will sound for about 20 seconds, the Heaters will be cut OFF and the solenoid valve open to exhaust the pressure.

4.13        For faster evacuation of the pressure, the manual steam release valve may be opened.

4.14        Unscrew the fly nuts only after the pressure gauge shows ‘0’. Do not attempt to open the fly nuts at even 1 or 2 psi. Always wait up till the Gauge reads zero before opening the lid.

4.15       To avoid recontamination of material, please remove the sterilized material at the earliest after total exhaust of pressure. The Purge valve cum vacuum Breaker will allow air to

               enter inside the chamber to prevent a vacuum within.

4.16        Cleaning:

4.16.1     To clean the chamber, a drain valve is provided at the bottom. Place a vessel of adequate size below it and open the drain valve.

4.16.2   After the chamber is emptied, pour additional water in the chamber so that residual matters / particles are flushed out, taking care that the particles are not big enough to block the valve.

4.16.3     Close drain valve and pour distilled water for next cycle.

4.17        Switch ‘OFF’ the mains after 15 minutes or as required.

4.18        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 Bacillus stearothermophilus suspension (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.

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

5.2.4              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

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

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