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Objective: To set up a general guideline for the operation and cleaning of Double Door Autoclave.
Scope: This SOP is applicable for operation and cleaning of Double Door Autoclave which is used for sterilization of media, petri-plate, glassware (e.g. glass rod, conical flasks etc.) filtration funnel, micro-pipette tips, glass pipette etc.
Responsibility
=>> Operating personnel is responsible for carrying out the procedure.
=>> Overall responsibility for training, implementation & follow-up with the QC Manager or his nominee.
Precautions:
=>> Check that the redial locking is properly tightened.
=>> Avoid contact with hot part.
=>> Cheek the load configuration displayed with the autoclave for prepared loading.
The maximum and minimum load is defined and is given bellow.
Minimum load.
1. 6 nos. filtration funnels
2. 2 nos. of 500 ml conical flask with agar media (1 litre).
3. 10 nos. of 500 ml empty conical flask.
4. 2 nos. of micropipette tips box.
Maximum load.
1. 6 nos. filtration funnels
2. 19 nos. of empty 500 ml conical flask.
3. 8 nos. of 500 ml conical flask with agar media (4 litres).
4. 10 nos. of 100 ml culture bottle with broth media (2 litres).
5. 20 nos. of screw cap test tubes.
6. 2 nos. of micropipette tips box.
7. 4 nos. of 500 ml conical flask with purified water (2 litres).
Procedure:
=>> Operation
=>> Connect the instrument to the mains plug. Connection is indicated by the red light that glows when connected (on the control box).
=>> Un-tight the redial lock and open the door.
=>> Turn Multi control valve to the ‘VAC DRY’ position.
=>> Open water fill valve and fill the steam generator / Boiler with purified water to ¾ the of it’s capacity indicated by the water level indicator.
=>> After filling the water turn Multi control valve to the “OFF SLOW EXH” position.
=>> Put toggle switch to ON position. The yellow indicating lamp on the control box will glow indicating heater on.
=>> When the jacket pressure reaches 15 psi, indicated on the jacket pressure gauge, load the material in the chamber and close the door well by tightening the redial lock.
=>> Turn Multi-Control Valve to the’ STER’ position. Steam will now be admitted into the chamber and will displace the air inside through the discharge line. When the dial thermometer indicates a temperature of 121ºC that is the start of the sterilizing period, wait 20 minutes to complete sterilization.
=>> After completion of the sterilizing period, switch off the heater by putting the toggle switch to OFF position and turn the Multi – control valve to the ‘FAST EXH’ position, for removing steam.
=>> When the compound gauge will indicate zero pressure, turn Multi- control valve to the ‘VAC DRY’ position and wait until the temp comes to 80 °C.
=>> For removing the sterilized material, open the second door from post sterilization site (Buffer room), and remove the materials by wearing heat proof gloves.
=>> After removing the material close the second door and switch off the machine.
=>> Keep the record of operation and cleaning in specific Log Book (Annexure-I).
=>> Cleaning
=>> Clean the chamber with purified water if any visible dirt is present, before that, ensure the chamber is cool.
=>> Clean outer surface and all parts of instrument carefully after every seven days.
