1.0 Objective:
To provide written procedure for
the operation and validation of Vertical Autoclave.
2.0 Scope:
This SOP cover operation and
validation of Vertical autoclave Model No. : 7451 FA in microbiology
department.
3.0 Responsibility:
Microbiologist:
Responsible for operation and validation of instrument as per procedure and
maintain the record.
QA Officer/QAM: Review the records and
governing the document.
4.0 Procedure:
4.1 Ensure that the instrument is clean
and free from dust and placed in a safe area.
4.2 Open the autoclave by unscrewing the
Wing Nuts.
4.3 Remove the baskets but leave the
Heater Cover X stand inside.
4.4 Ensure that the drain valve at the
bottom of the autoclave is closed.
4.5 Pour purified water direct into the
Autoclave, up to the heater cover X stand.
4.6 Load the Autoclave, close the lid, clamp
the Wing and tighten opposite fly nuts simultaneously with equal pressure.
4.7 Switch ‘ON’ the mains; the set temperature
and set time will be displayed on the Microprocessor controller in brackets. If
acceptable, press START. If change the temperature and time as per procedure
given in controller setting as below
4.8 Micro Processor Based (µP)
Controller Setting
4.8.1 When power is switched ON the display will
show status OK/EQUITRON for 3seconds and then the main screen will appear where
in the top line will display the process temperature, last set temperature (in
brackets) and bottom line will display time 00 and last set time (in brackets).
4.8.2 To change temperature press MENU, the
display will change to set temperature. Press Up or DOWN arrow keys. For the
temperature change permanent for all future cycles,
press SET, the display will
return to the main screen, for the temperature change only for
this particular cycle, press
MENU again, the display will change to set time.
4.8.3 To change time press MENU, if previous
time change, Press Up or DOWN arrow keys. For the time change permanent for all
future cycles, press SET, the display will return to the main screen, for the
time change only for this particular cycle, press MENU again, the display will
return to the main screen.
4.8.4 Press START to start the cycle. In case
of problems encountered, the buzzer will sound, the alarm LED will flash, and
the problem will be on the display.
4.8.5 After the start switch is pressed, to
change the set temperature and time incorporates the change, press START again and
the change will overwrite the earlier settings.
4.9 Allow the temperature reaches up to
100° C, then open the valve and release the vapour. Close the valve when
continuous steam comes out.
4.10 After this, the temperature and pressure
will gradually rise, which will be indicated by the µPC and pressure gauge.
4.11 Just before the set temperature the
Heater will be cut OFF and then gradually temperature is attain the set
temperature. The TIMER will start its countdown after the set temperature is
attained. During the countdown the temperature and hence the pressure will be
maintained by the µPC by switching ON/OFF the heater.
4.12 After the elapse of the set time. A
Buzzer will sound for about 20 seconds, the Heaters will be cut OFF and the
solenoid valve open to exhaust the pressure.
4.13 For faster evacuation of the pressure,
the manual steam release valve may be opened.
4.14 Unscrew the fly nuts only after the
pressure gauge shows ‘0’. Do not attempt to open the fly nuts at even 1 or 2
psi. Always wait up till the Gauge reads zero before opening the lid.
4.15 To avoid recontamination of material,
please remove the sterilized material at the earliest after total exhaust of
pressure. The Purge valve cum vacuum Breaker will allow air to
enter inside the chamber to
prevent a vacuum within.
4.16 Cleaning:
4.16.1 To clean the chamber, a drain valve is
provided at the bottom. Place a vessel of adequate size below it and open the
drain valve.
4.16.2 After the chamber is emptied, pour
additional water in the chamber so that residual matters / particles are
flushed out, taking care that the particles are not big enough to block the
valve.
4.16.3 Close drain valve and pour distilled water
for next cycle.
4.17 Switch ‘OFF’ the mains after 15 minutes
or as required.
4.18 Enter the details of usage in Autoclave
load register Annexure - 1
5.0 Validation:
5.1 Routine Validation:
5.1.1 Put a sterilization indicator strip in a
Petri plate along with each load of Autoclave.
5.1.2 If the colour white cross line of the
strip changes to brownish black it indicates the sterilization for the lot is
complete.
5.1.3 If the
colour of the cross line does not change to brownish black, then do not use
the items sterilized.
5.1.4 Paste the sterilization indicator strip
in the autoclave load register.
5.2 Quarterly Validation:
5.2.1 Sterilize one ampoule/ strip of Bacillus
stearothermophilus suspension (ATCC 7953 Bio indicator) at 121˚ C
for 15 minutes (15 lb/in2 pressure).
5.2.2 Remove the ampoule/ strip once
the sterilization is complete.
5.2.3 For Ampoule: Incubate at 43˚ to 45˚ C for 3 to 5 days
along with a positive control (Unautoclaved ampoule).
5.2.4
For Strips: After sterilization, aseptically transfer the
processed strip and one control strip into prepared sterilized Nutrient Broth.
Incubate at 55°C –60°C for 7 days along with a positive control ( Unautoclaved
strip / ampoule).
5.2.4 Observe the growth, purple colour should
turn to yellow in the positive control showing the presence of Bacillus
stearothermophilus. Sterilized ampoule should remain purple confirming the completeness
of sterilization.
5.2.5 Perform the observation 5.2.1 to 5.2.4
with different autoclave loads. (i) Empty
load.
(ii) Maximum load. (iii) Minimum load
5.3
Record the observations in the
autoclave validation register (Annexure - 2).
6.0 Routine maintenance:
6.1 Remove scaling from
the instrument and stand with 50 ml hydrochloric acid in 20 liter of water.
6.2 After cleaning with hydrochloric clean
instrument with distilled water.
7.0 Documentation:
7.1 Annexure – 1 - Autoclave usage log book. XXX/FRM/000
7.2 Annexure – 2 - Autoclave validation register. XXX/FRM/000
8.0 History of Revision:
Revision No.
|
Effective
Date
|
Revision
details
|
Reason for
revision
|
Annexure
- 1
|
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Date
|
Name of the Item
|
Autoclave Load No
|
Time
|
Indicator strip after
sterilization
|
Done by
|
Remarks
|
||
Start
|
Activate
|
Stop
|
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SOP No. : XXX/SOP/000-00 Format No. : XXX/FRM/000-00
|
Annexure
- 2
|
||||||||||||||||||||||||||||||
Validation of
Autoclave by Spore Strips
Instrument No :
SOP No. : XXX/SOP/000-00
Format No. :
|
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