Operation and Calibration of Vertical Autoclave
This Document describes (Standard Operating procedure) SOP for Operation, Cleaning and Calibration of Vertical Autoclave
SOP for Operation, Cleaning and Calibration of Vertical Autoclave
I. Purpose & Scope:
- The purpose of this SOP is to lay down the procedure for Operation, Cleaning and Calibration of Vertical Autoclave
- This Standard operating procedure shall be applicable for Operation, Cleaning and Calibration of Vertical Autoclave
II. Responsibilities:
All Quality Control and Quality Assurance personnel shall be responsible to follow and implement this SOP.
III : Introduction and Procedural Part:
Precautions while handling of vertical Autoclave:
- Check electrical power socket. Avoid loose connection.
- Ensure electrical earthing is present.
- Fill the purified water in autoclave above heater level. Check level before every cycle.
- Ensure lid fitting bolts are evenly tightened diagonally.
- Check and ensure temperature controller is set at required sterilization temperature and time.
- Do not make physical contact with equipment when in use during sterilization cycle.
- Avoid excess load of media into Autoclave. For load configuration and diagrammatic representation refer Annexure II, Annexure III and Annexure IV.
- Before opening the lid fitting bolts, ensure that pressure is ‘0’ (zero) in pressure gauge. If necessary, open the steam release valve and evacuate all steam pressure from the chamber. Then and then only open the lid.
- Avoid the sharing of chemicals/reagent around equipment.
General Instructions while handling of vertical Autoclave:
- Place equipment at designated location on leveled floor. Maintain sufficient distance around the equipment for operator movement.
- Use purified water to pour into the chamber and place the stand and basket inside.
- For list of articles and diagrammatic standard load configuration of autoclave, refer Annexure-II, Annexure-III and Annexure-IV.
- All accessories and Media bottles/ tubes should be kept at designated place in the Autoclave. However the locations of accessories and media bottles/ tubes may vary depending on the number of articles in the particular load.
- Connect the autoclave unit to electric supply; switch ‘ON’ the mains.
- Set the temperature and hold time required for sterilization as per validation of autoclave.
- For sterilization cycle at set temperature 121.6°C, lower limit is 121.1°C and upper limit is 124°C during Sterilization hold period.
- Set sterilization hold time for sterilization Autoclave is 20 minutes and for Destruction Autoclave is 30 minutes.
- Record the cycle details of sterilization Autoclave and Destruction Autoclave in Annexure-II and Annexure-III respectively.
- Observe the temperature on display and pressure on the pressure gauge. The pressure should be above 15 lbs corresponding to the temperature of 121.6 °C during sterilization.
- Observe that, the set conditions are achieved correctly.
- Record the cleaning, calibration and maintenance/repair activity details of Autoclave of Vertical Autoclave for Media Sterilization and Vertical Autoclave for Media Destruction.
Operation Procedure for Vertical Autoclave:
When switch on the mains, display shows the following screens for 5 seconds by serially as shown below. Follow the procedure given below.
| Fully Automatic
Autoclave logger |
| Fully Automatic
Autoclave Controller |
| PV : 33.2 o C 20 Min
Ready / Print 15 Records |
- Then press PROG. Button and display will show the sterilization temperature, adjust the required temperature by using arrow buttons. Then press enter; now display will show sterilization hold time (minutes). Adjust the required time by using arrow buttons. Then press enter. After setting the time and temperature (Example 121.6°C for 20 Minutes), press enter after every setting till the following type of screen displayed on controller display.
| PV : 33.2°C 20 Min
Ready / Print 15 Records |
- Then press PRN/ACK button, now display will show the following type of screen.
| PV : 33.2°C 20 Min
Wait Printing On |
- After some time display will show the following type of screen, it indicates the controller is ‘Ready’ and cycle can be started.
| PV:33.2°C 20 Min
SP:121.6°C 20 Min/Ready |
- Then press start/stop button, heating and cycle will start. Display will show the following type of screen. Operation of autoclave will proceed as flow given below.
| PV:33.2°C 20 Min
SP:121.6°C 20 Min /Heating |
- After achieving the sterilization temperature (121.6°C), display will show the following type of screen.
| PV : 121.6° C 20 Min
SP : 121.6 °C 20 Min /Sterilization |
- After completion of hold time (20 min), exhaust will start and display will show the following type of screen.
| PV : 120.9°C 20 Min
EXHAUST |
- Then pressure will release and temperature will fall down and cycle will end. Display will show the following type of screen.
| PV : 96.8°C 20 Min
End Of Cycle. |
- Vertical Autoclave:
- Switch on the mains. Then display will show the following screens serially.
| AUTOCLAVE VERSION 4.03 |
| PV: 26.3°C
PRESS START |
- Then press ‘Menu’ button, display will show the sterilization temperature, adjust the required temperature by using arrow buttons. Then press enter; now display will show sterilization hold time (minutes). Adjust the required time by using arrow buttons. Then press enter. After setting the time and temperature (Example 121.6°C for 30 Minutes), press enter after every setting till the following type of screen displayed on controller display.
| PV: 26.3°C
PRESS START |
- Then press start/stop button till the heater and cycle starts. Once the cycle started, the display will show the following type of screen. Operation of autoclave will proceed as flow given below.
| PV: 26.3°C ET: 30 Min
SP : 121.6°C ST: 30 Min |
- After achieving the sterilization temperature (121.6°C), display will show the following type of screen.
| PV: 121.6°C ET: 30 Min
SP : 121.6°C ST: 30 Min |
- After completion of hold time (30 min), exhaust will start and display will show the following type of screen.
| PV : 120.9°C 30 Min
EXHAUST |
- Then pressure will release and temperature will fall down and cycle will end.
| PV : 98.3°C 30 Min
End Of Cycle |
Verification/Calibration Procedure for vertical Autoclave: ( Using Biological Indicator) :
- Media required: Sterile Soybean Casein Digest Medium ( SCDM)
- Take three clean, dry petri dishes and Lable them individually as Top, Middle and Bottom.
- Place Geobacillus stearothermophilus (ATCC7953) spore strips/ampoules (having a spore population of atleast 106 cfu per unit) in each of three petri dishes.
- Ensure that the water level in the vertical autoclave is at sufficient level.
- Place ‘petri dish 3’ at the bottom part of the SS basket in the autoclave.
- Load the material to be sterilized inside the basket up to the central part of the autoclave. Place ‘petri dish 2’ on the load at approximate central part of the autoclave.
- Load rest of the material up to the top and place ‘petri dish 1’ over it.
- After loading the material and biological indicators, switch ‘ON’ the vertical autoclave, after completion of the sterilization cycle remove all the three petri dishes from the autoclave and transfer to LAF for further testing.
- Aseptically transfer the Geobacillus stearothermophilus strips to the separate tube of presterilized 10 ml SCDM (Soybean casein digest medium).Mark the tubes appropriately.
- Aseptically transfer non-sterilized Geobacillus stearothermophilus strip into a tube of presterilized 10 ml SCDM to serve as positive control.
- Mark one tube of presterilized 10 ml SCDM of same lot as negative control.
- Incubate all SCDM tubes at 55-60°C for 7 days and observe them every day for any visual evidence of growth (turbidity).
- If Geobacillus stearothermophilus ampoules are used then after sterilization directly keep the ampoules at 55-60°C for 7 days. Keep negative control ampoule and one ampoule as non-sterilized to serve as positive control along with test.
- Record the observation in the report format as per Annexure-I.
- If the autoclave cycle is effective then all the spores will kill and no turbidity will be observed in test. The positive control will show turbidity due to growth of spores.
- If there is no evidence of growth in any of the tubes except positive control, then it can be concluded that instrument is valid for further use.
- If evidence of growth is not found in positive control tube or ampoule then test is invalid. Repeat the verification procedure after permission of Department. Head. Check the GPT of used media.
- If growth is observed in any of the three tubes / ampoules, repeat the verification procedure immediately.
- If evidence of growth is detected once again, report to Quality Assurance Manager and notify the defect to Engineering Department. Affix ‘UNDER MAINTENANCE LABEL’ on the instrument and do not use it.
- Frequency of verification by using biological indicators is ‘Quarterly’ and after any major maintenance job performed.
- Ensure that pressure gauge, safety valve, solenoid valve & temperature controllers are to be calibrated by outside party once in a year and after major maintenance.
- Frequency of Outside Calibration: Once in a year and after major maintenance.
Cleaning Procedure for vertical Autoclave:
- Clean the autoclave with liquid soap & purified water
- For this, remove water from the autoclave. Add 2% soap solution and scrub the bottom and walls of autoclave with the help of smooth scrubber. Rinse thoroughly with water to remove the soap traces. Finally rinse with 2.5 % Dettol or 2.5 % savlon solution.
- After cleaning, add the purified water in autoclave upto the sufficient level.
IV: Annexure :
| Annexure | Title of Annexure |
| Annexure-I | Verification of Vertical Autoclave with Biological Indicator |
| Annexure-II | List of Standard load configuration of Vertical Autoclave |
| Annexure-III | Diagrammatic Standard Load Configuration of Vertical Autoclave |
| Annexure-IV | Diagrammatic Standard Load Configuration of Vertical Autoclave |
Annexure I
Verification of Vertical Autoclave with Biological Indicator
Page No.: 1 of 1
| Date of Verification | Instrument ID No. | ||
| Autoclave started at: | Cycle Temp & time | ||
| Sterilization hold period | Cycle end time: | ||
| Analysis Started On: | Analysis Completed On: |
Details of Process:
Details of Biological indicator Used:
| Lot No. | Mfg. date | ||
| Mfd. By | Exp. Date |
Details of Media:
| Name of Media | Media
Lot No. |
Incubation Period | Instrument
ID No. |
|
| Temp | Time | |||
| Soybean Casein Digest medium | ||||
Observation:
| Strip/Ampoule No. | Location | Observation | |||||||
| 1st day | 2nd day | 3rd day | 4th day | 5th day | 6th day | 7th day | 8th day* | ||
| 1 | |||||||||
| 2 | |||||||||
| 3 | |||||||||
| 4 (Positive control) | |||||||||
| 5 (Negative control) | |||||||||
Note: +ve: Growth observed, -ve: No Growth observed, H: Holiday/Weekly off
* : Column to be used for recording observation if 7th day is holiday/Weekly off
Conclusion: The sterilization process by Autoclave is effective / non-effective as observed from above results.
Observed By: Checked By:
Date Date
Annexure II
List of Standard load configuration of Vertical Autoclave
Page No.: 1 of 2
- A) Equipment & Accessories Cycle Load Pattern I:
[Sterilization Cycle at 121.6 °C for 20 minutes]
| Name of item | Total number of item |
| 1) Membrane filtration cup | 08 no. |
| 2) Manifold | 01 no. |
| 3) Dressing drum (Plate/Garment) | 01 no. |
| 4) Air sampling Sieve | 01 no. |
| 5) Scissors, forceps, spatula | 05 no.(each) |
| 6) Sampling bottles | 12 no. |
| 7) Spreader, Swab sticks | 10 no.(each) |
| 8) Pipette tips box | 01 no. |
| 9) 0.45µ Size Membrane filters in Glass Petri Plate | 01 Glass Petri Plate |
- B) Agar/Broth Medium Cycle Load Pattern II-A: [Sterilization Cycle at 121.6°C for 20 minutes]
| Name of item
|
Total number of item |
| 1) Test tubes with Media in stand | 24 X 100ml |
| 2) 250ml Flask with Media | 04 X 150ml |
| 3) 500ml Flask with Media | 06 X 300ml |
| 4) 1000ml Flask with Media | 01 X 600ml |
| 5) Beakers with test tubes containing media. | 20 X 10 ml & 20 X 20ml |
| Total | Approximately 6 Liters Media. |
Page No.: 2 of 2
C) Agar/Broth Medium Cycle Load Pattern II-B: [Sterilization Cycle at 121.6°C for 20 minutes]
| Name of item
|
Total number of item |
| 1) 500ml Flask with Media | 01 X 200ml |
| 2) Beaker with test tubes containing media. | 1 X 20 ml |
| Total | Approximately 220 ml Media. |
- D) RVSEB (Rappaport vassiliadis salmonella enrichment broth) Broth Cycle Load Pattern III:
[ Sterilization Cycle at 115°C for 15 minutes with lower limit 115°C and Upper limit 116°C during sterilization hold period.]
| Name of item
|
Total number of item |
| 1) Beakers with test tubes containing broth Media | 58X10ml , 1X20 ml |
| Total | Approximately 600 ml Media. |
