Procedure for autoclave validation

STANDARD OPERATING PROCEDURE
Title	:	Procedure for autoclave validation
Department	:	Quality Control-Microbiology

SCOPE
- This SOP is applicable for autoclave validation in microbiology laboratory .
RESPONSIBILITY
- Officer / Executive-QC – responsible for Implementation of SOP.
ACCOUNTABILITY
- Manager – Quality Control
PROCEDURE
- Method-A:

Clean and load the autoclave for sterilization with accessories and glassware’s.
Prepare the required number of (10ml) tubes of Soya bean casein digest medium.
Take require nos. of strips of spores of Geobacillus stearothermophilus ATCC 7953.
Put in petriplates and label it, arrange the petiplates at different location no.
Keep one strip unexposed for positive control.
Run the cycle at 15 lbs. pressure and 121^o C temperature for 15 min.
After completion of cycle remove all tubes and Petri dishes.
Mark location no.,date on all the SCDM tube .
Transfer strips aseptically to respective labelled tubes.
Transfer one unexposed strip to one SCDM tube and label it as positive control.
Keep one SCDM tube uninoculated and label it as negative control.
Incubate all the tubes at 55^o C ±2^o C for 7days
At the end of 7 days all the the inoculated SCDM tubes should show no growth.
Negative control tube should show no growth.
Positive control tube should show growth with in 72 hrs.
The above observations will demonstrate satisfactory validation of the autoclave.
Record the observation in Format-I.

Validation is done by the bioindicator system consists of ampoule containing spores of Geobacillus stearothermophillus ATCC 7953.
These ampoules are stored under refrigerator when not in use. On the day of validation
Take three ampoules and allow them to come at room temperature.
Load the autoclave and place the ampoule in different locations in autoclave within glass beakers to avoid accidental breakage of ampoule during sterilization.
Run the sterilization cycle as per procedure. (validated sterilization condition 121°C for 15 min at 15 PSI.)
Unload the chamber and remove the ampoule, mark them and incubate at 55°C ±2°C for 48 hour.
The third non sterile unexposed ampoule is incubated along with the sterilized ampoule as a Positive control.
At the end of 48 hour days.
Positive control – Original purple colour gradually changes to deep yellow with turbidity.
Exposed ampoule – The Original purple colour remains as it is at the end of incubation period.
The above observations will demonstrate satisfactory validation of the autoclave.
Record the result in Format-I

Sr. No.	Title	Format No.
	Log sheet for Autoclave validation Record
	Log Sheet for Sterilization load no.

Sr. No.	Copy No.	Department
		Quality Assurance
		Quality Control-Microbiology
		V.P.- Plant & Operations

Format -I

Name of Equipment	Vertical Autoclave
Equipment ID No

Sr.No	Description	Detail
	Temperature/pressure to be achieved	121°C/15lbs
	Biological indicator used	Strip/Ampoule
	Name of the test organism	Bacillus stearothermophilus ATCC-7953
	Total population of strip/Ampoule
	D value.
	Media to be use	Soybean Casein Digest broth
	Incubation temperature	55°C to 60°C
	Duration of Incubation

Details of last Validation:
Validation On
Validation Due on
Report No.
Detail of current Validation:
Date of Validation
Validation Due on
Report No.

Sr.No	Parameters	Observation	Observed by
	Temperature/Pressure achieved
	Name of the test organism
	Media used
	Incubation temperature
	Duration of Incubation
	Lot number of Strip/Ampoule
	Manufacturer of strip/Ampoule

There should not be any growth in form of turbidity in any of the tubes except Positive control.

Corrective action taken:

Observation after corrective action:

OBSERVATION SHEET

Log Book of Sterilization(Autoclave) Load Number

Instrument I.D.: ______________________