Procedure for autoclave validation

STANDARD OPERATING PROCEDURE
Title:Procedure for autoclave validation
Department
:
Quality Control-Microbiology
  •     OBJECTIVE
  •          To lay down a procedure for autoclave validation.
  • 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 121o 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 55o C ±2o 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.
  •   Method-B
  •        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

 

  •    ABBREVIATION
  •   SOP      –  Standard operating procedure
  • IPA       –  Iso Propyl Alcohol
  •   SCDM     – Soybean Casein Digest Medium
  •     FORMAT
Sr. No.Title  Format No.
Log sheet for Autoclave validation Record
Log Sheet for Sterilization load no.
  •    DISTRIBUTION LIST
Sr. No.Copy No.Department
Quality Assurance
Quality Control-Microbiology
V.P.- Plant & Operations

                                              

                                                      Format -I

  • Equipment Detail:
Name of EquipmentVertical Autoclave
Equipment ID No

 

  •  Detail of Test:
Sr.NoDescriptionDetail
Temperature/pressure to be achieved121°C/15lbs
Biological indicator used Strip/Ampoule
Name of the test organismBacillus stearothermophilus ATCC-7953
Total population of strip/Ampoule
D value.
Media to be useSoybean Casein Digest broth
Incubation temperature55°C  to 60°C
Duration of Incubation

 

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

 

  • Details of Observation:
Sr.NoParametersObservationObserved 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

 

  •  Acceptance Criteria:

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

  •  Corrective action if observation does not meets the requirements:
               Corrective action taken:
               Observation after corrective action:

 

 

  •  Conclusion:
 

 

  •  Approval:

OBSERVATION SHEET

Sr. No   LocationDay 1Day 2Day 3Day 4Day 5Day 6Day 7Remark
Negative Control
Positive Control
Observed by

 

  •                                  Format -II

       Log Book of Sterilization(Autoclave) Load Number  

Instrument I.D.: ______________________ 

Sr.No.DateName of Sample     Load Number Done byChecked byRemark
    

 

   
    

 

   
    

 

   

 

 

 

About Reshma Verma

Check Also

Procedure of the replacement procedure for the Purified Water Pump

To implement and procedure of the replacement procedure for the Purified Water Pump. This SOP …