Autoclave Operation SOP Template

1.Purpose

To sterilize laboratory materials and waste reliably using the autoclave, while protecting operators from steam and pressure hazards and confirming each cycle achieves sterilization.

2.Scope

Applies to steam autoclaves used to sterilize glassware, media, instruments and biohazard waste. Chemical disinfection of surfaces is covered by the lab cleaning SOP.

Definitions

Gravity cycle

A standard sterilization cycle relying on gravity displacement of air by steam, used for most solid goods.

Liquid cycle

A slower-exhaust cycle used for liquids to prevent boil-over from rapid pressure release.

Biological indicator

A test vial containing heat-resistant spores used to confirm a cycle actually achieved sterilization.

3.Responsibilities

Lab Technician

Loads, runs and unloads the autoclave, and records cycle parameters and indicator results.

Laboratory Manager

Reviews weekly biological indicator results and approves the autoclave's return to service after a failure.

Maintenance Technician

Services the autoclave, checks water levels and gaskets, and repairs faults.

RACI matrix

R = Responsible, A = Accountable, C = Consulted, I = Informed

4.Materials and PPE

Materials, tools and systems

• →Autoclave-safe bags and containers
• →Chemical indicator tape or strips
• →Biological indicator vials and incubator
• →Autoclave cycle log
• →Heat-resistant gloves and tray for unloading

Personal protective equipment

• →Heat-resistant gloves
• →Face shield or safety glasses
• →Closed-toe shoes

5.Procedure

1. 5.1

   Check the water reservoir

   Lab Technician

   Check the autoclave's water reservoir level and top it up with the manufacturer-specified water type before starting a cycle.

2. 5.2

   Inspect the door seal

   Lab Technician

   Inspect the door gasket for cracks, debris or misalignment, since a damaged seal will prevent the chamber from reaching pressure.

3. 5.3

   Load items correctly

   Lab Technician

   Place items in autoclave-safe containers or bags, venting biohazard bags loosely and avoiding overpacking so steam can circulate around every item.

   Warning: Never seal a biohazard bag tightly closed; steam must be able to penetrate the load.

4. 5.4

   Apply a chemical indicator

   Lab Technician

   Place a chemical indicator strip or tape on or inside each load so a color change confirms exposure to autoclave conditions.

5. 5.5

   Select the correct cycle

   Lab Technician

   Choose the gravity, liquid or waste cycle matching the load type, since liquids and waste need slower exhaust to avoid boil-over.

6. 5.6

   Close and lock the door

   Lab Technician

   Close the chamber door and confirm it is fully locked before starting the cycle, following the unit's specific locking procedure.

7. 5.7

   Start the cycle and record parameters

   Lab Technician

   Start the selected cycle and record the temperature, pressure, run time and operator on the autoclave cycle log.

8. 5.8

   Monitor for completion

   Lab Technician

   Check the display or printout at the end of the cycle to confirm the sterilization phase reached and held the required temperature and time.

   Checkpoint: The cycle printout or display confirms the sterilization set point was reached and held for the full cycle time.

9. 5.9

   Allow pressure to release before opening

   Lab Technician

   Wait until the chamber pressure gauge reads zero and any manufacturer-specified cooling time has passed before unlocking the door.

   Warning: Never force open the door while the chamber shows any residual pressure.

10. 5.10

    Unload with heat-resistant gloves

    Lab Technician

    Open the door slowly to let residual steam escape away from your face, then remove items using heat-resistant gloves and a tray.

11. 5.11

    Verify sterilization results

    Lab Technician

    Check the chemical indicator color change on each item, and run and read a weekly biological indicator test against a positive control.

    Checkpoint: The weekly biological indicator shows no growth compared with the positive control before the autoclave continues routine use.

12. 5.12

    Log the cycle and report failures

    Lab Technician

    Complete the cycle log entry with indicator results, and report a failed cycle or failed biological indicator to the laboratory manager immediately, holding the load as non-sterile.

6.Quality checks

• →Every load carries a chemical indicator that shows the expected color change.
• →Cycle logs show the sterilization temperature and time were reached and held.
• →A biological indicator test is run and read at least weekly.
• →Failed loads are held as non-sterile and reprocessed, not released.

7.Records

• →Autoclave cycle log
• →Biological indicator test results
• →Chemical indicator records
• →Maintenance and repair history

8.KPIs

• →Percentage of cycles passing biological indicator verification
• →Number of failed or aborted cycles per month
• →Average autoclave downtime for repair
• →Percentage of weekly biological indicator tests completed on schedule

9.Common mistakes

• →Sealing a biohazard bag tightly so steam cannot reach the contents.
• →Opening the door before the pressure gauge reads zero.
• →Overpacking the chamber so steam cannot circulate around every item.
• →Skipping the weekly biological indicator test when the schedule is busy.

10.Revision history