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SOP for Sterilization by Autoclave for Microbiological Analysis

Standard operating procedure to sterilize the culture media, glassware, dresses, Sterility utensils, filtration assembly by steam sterilization in autoclave.


       To lay down a Procedure is to provide guidelines for Sterilization by Autoclave for Microbiological testing.

2.0  SCOPE:

       This procedure is applicable for the sterilization of culture media, glassware, dresses, Sterility utensils, filtration assembly etc. in Microbiology Laboratory of Quality Control department.




       HEAD QA & QC


5.1  Prepare culture media as per SOP for preparation of Culture Media and dispense the medium as desired.
5.2  Properly plug the dispensed media with non-absorbent cotton. Filtration assembly outlet must be plugged with non-absorbent cotton and then wrap with butter paper to cover the whole assembly. Sampling bottles must be wrapped with aluminium foils till neck of bottles.
5.3  Label the media containers, filtration assembly and glassware
5.4  Place the wrapped containers in SS tray and transfer to autoclave room.
5.5  Wear safety heat resistant gloves and then load the autoclave by media along with a chemical Indicator strip and same procedure will follow for the hard accessories and garment cycle.
5.6  Operate the autoclave as per SOP for Operation of Autoclave
5.7  Observe the autoclave cycle and make entry in Autoclave logbook the time when temperature is reached at 121°C (Sterilize the media at 121°C and 15 psi for 20 minutes and glassware, garments and filtration assembly at 121°C and 15 psi for 30 minutes), Hold the cycle for 20 or 30 minutes as per the type of load and note down the holding time.
5.8  Open the door (Sterile side) when pressure becomes 0 psi and load material must be unloaded from sterile side (Cooling zone).
5.9  Check the chemical indicator strip for color change from white to black. Simultaneously check the printout of temperature probes, if a color change of chemical indicator is not proper or satisfactory and temperature in printout is out of range, immediately inform to your superior for further action and discard the media as per SOP for destruction of Microbial Waste.
5.10  If the color changes of chemical indicator is satisfactory and temperature in printout is within the prescribed limit, the autoclaved media is suitable for microbiological testing.
5.11  Label the chemical Indicator slip and printout of temperature with Autoclave cycle number and attach to record.

5.12  Precautions

5.12.1  During loading and unloading of an Autoclave always wear a heat resistant hand gloves.
5.12.2  Reconstitute only the required amount of medium. Avoid preparing excess medium and remelting multiple times.
5.12.3  Sterilization should be strictly done as per stipulated instructions.


SOP : Standard Operating Procedure
QA : Quality Assurance
QC : Quality Control
NA : Not Applicable
SS : Stainless steel

Also see: Low Temperature Sterilization Process (115°C)

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