The document provides operating instructions and maintenance procedures for an autoclave. It describes how to prepare materials for sterilization, operate the autoclave including setting temperature and timers, and perform safety, quality control and maintenance checks. Key steps include using steam or chemical indicators to confirm sterilization, testing spore ampoules weekly for sterility, and documenting maintenance and test results in a log sheet. The autoclave sterilizes equipment and supplies using high pressure saturated steam to decontaminate items.

1. AUTOCLAVE
OPERATING
INSTRUCTION AND
MAINTENANCE
Prepared and Presented by: Mr. Say Sinith
Date: 15 June 2016
Bacteriolog
y Section

2. Content
 Introduction
 Materials and Equipment
 Procedure
 Materials Preparation
 Operation of Machine
 Caution
 Quality Control and Maintenance
 Daily Verify
 Weekly Verify
 Spore Ampoules Test Result
 Autoclave Maintenance Log Sheet
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3. Introduction
3
 Autoclave is a pressure chamber used elevated
temperature and pressure to perform sterilization. (121-
134°C)
 It used to sterilize surgical equipment, laboratory
instruments, pharmaceutical items, and other materials.
It can sterilize solids, liquids, and instruments of
various shapes and sizes.
 Many autoclaves are used to sterilize equipment and
supplies by subjecting them to high pressure saturated
steam at 121°C for 15–20mn depending on the size of
the load and contents.
 The autoclave was invented by Charles Chamberland in
1879, from Steam digester created by Denis Papin in

4. Materials and Equipment
 Autoclave Machine
 Autoclave Indicator (Steam Indicator)
 Bacillus QC or Spore Ampoules
 Heat Block Incubator 60oC
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5.  Only autoclavable materials (check with the manufacturer
instruction manual, MSDS files, or safety officer)
 Liquids in an autoclavable bottle ≥ 2 times the volume to be
autoclaved (E.g. 1L of media in 2L flask)
 Caps must be slightly loose
 Waste must put in biohazard bags
 Sharps containers
 Placed in an autoclave tray or chamber
 Stick steam indicator and put Spore ampoules
 Warning: Use of non-autoclavable trays or chamber and
containers may result in injury to the user and damage to the
equipment.
Procedure: Materials
Preparation5

6. Procedure: Operation of
Machine
 Switch power breaker “ON”, Power switch to “ON”
 Swing door close clockwise as tight you can open it
when complete.
 Loading appropriate amount of content in the tray or
chamber.
 Set Sterilizing temperature with appropriate timer.
 Press the mode switch which you like to sterile
(Unwrapped, Wrapped, and Solutions)
 Set the dry timer for
 After you press the mode switch the sterilizer will
automatically progress through cycle if the pressure of
steam generator has reached the setting pressure.
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7. Procedure: Operation of
Machine7
 When cycle is complete, completion pilot lamp displays
and buzzer sounds, the power be to “OFF” position
automatically.
 Wait the pressure of jacket and chamber to become zero
then press mode switch again to “OFF” position.
 Swing door open anticlockwise and allow it to cool
 Observe Steam indicator for proper changing to dark
color.
 Remove all autoclave contents and transfer into “After
autoclaved” bin with black bag.
 Send the Spore Ampoules back to Bacteriology section.
 Incubate the Spore Ampoules at 55-60oC in heat block

8. Procedure: Operation of
Machine8
Load content waste
into autoclave
chamber and close
the door tightly
Switch on electric power
supply
Observed “Water” “Heat”
and “Door” to close
properly
“ Power” button turn
Green and “Complete”
button
Set “Time” as required, “Ster.
Timer” and “Dry Timer”
Select for normal waste, for
mental, and for glassware and
liquid
After complete observed interior
pressure and exterior chamber till it
point “0”
Stand behind the
door way and
open gently for
safety
Urgent
Stop
Process

9. Caution
 Do not overload, Avoid contact of load components with the wall of the
chamber.
 Closed the door properly
 Check digital display corresponds to the time set properly.
 Once the cycle started ensure autoclave is functioning properly (alarm).
 Only open autoclave door until the chamber pressure gauge is point at (0)
pressure.
 When open the door:
 Turn the handle slowly and standing behind the door.
 Never rush open the door (liquids boil over or bottles break).
 Remove all trays from the autoclave properly with heat resistant gloves.
 If there is any spills or leaks or experience any problems with the autoclave
 Report to responsible person immediately to take proper actions.
 Clean up any mess you have caused.
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10. Quality Control and
Maintenance
 Daily Verify:
 Assess Steam indicator or Chemical indicator to
assess steam sterilization. Ensure that it changes to
a proper color indicated.
 Inspect if there is any problem encountered and
complete the maintenance log sheet.
 Maintenance log sheet
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11. Autoclave Maintenance Log
Sheet11
 Daily Verify:

12. Quality Control and
Maintenance12
 Weekly Verify:
 Assess efficacy of steam sterilization by take
isolated colonies of Bacillus species/ Spore
Ampoule from Bacteriology section every Tuesday
to integrate with autoclave process.
 Inspect and verify the cleaning before and after duty
week
 Assess the softener and salt solution tank
 Clean and Flush steam generator

13. Autoclave Maintenance Log
Sheet13
 Weekly Verify:

14. Spore Ampoules Test Results
 Weekly Verify:
 Record negative (no growth) results after 48h
incubation.
 No color change in the purple media indicates
proper sterilization.
 Any positive (turbidity and/or color change from
purple to yellow) result should be reported
immediately to a supervisor.
 Corrective action is considered
 Returned the autoclaved back to hazard bin
14
Passe
d
Failed

15. Quality Control and
Maintenance15
Drainage
valve

16. References
 Medsource TC series Floor Type Steam
Sterilizer User’s manual
 N2-SOPBAC-007-00: Autoclave
 N2-FC-BAC-090-00: Autoclave waste
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