Autoclav

e
Dr. Navodit Goel

Autoclave
The autoclave is a piece of equipment used
for sterilizing various requirements in the lab
by wet sterilization method.
The name comes from Greek auto-,
ultimately meaning self, and Latin clavis
meaning keya self-locking device.
The word sterilizing means the destruction
of elimination of all forms of life (microbial,
including spores) present in inanimate
objects by means of physical, chemical or
gaseous procedures.

Operation Principle
When water is heated in a closed container,
saturated steam is produced under pressure.
According to Boyles Law, when volume of the steam,
is kept
constant, the temperature is directly
proportional to pressure.
If the pressure is reduced it boils at a lower
temperature. If the
pressure rises, it boils at a greater temperature.
At 15 lbs pressure 1210 C temperature can be
obtained.
Moist heat coagulates cell proteins of the
microorganisms and thus kills all the living entities
(including spores) in 15 to 20 minutes.

The process associated in achieving

sterile conditions of inanimate objects are
the following :

Cleaning
Decontamination
Inspection
Preparation and packing
Sterilization
Storage
Delivery of materials

Purpose of the Autoclave

In the laboratory, materials and
objects are
sterilized for the following
purposes:
1. To
prepare
materials
for
bacteriological cell
cultures (test
tubes, pipettes, Petridishes, etc.) in
order to avoid their contamination.
2. Prepare elements used for taking
samples.
(All must be in sterile
conditions:
needles,
tubes,

Two Type of Autoclave

1. Horizontal or downward
displacement
autoclave.

2. Vacuum assisted autoclave.

Horizontal or
downward
displacement
In a
displacement
autoclave
Downward cold air escapes
autoclave
through the bottom of the
chamber as steam displaces it
from above.
The valves should never be
obstructed and the chamber
must not be overfilled in order
for this system to function
efficiently.

Horizontal or downward
displacement autoclave contd..
This type of autoclave is electrically operated
and can be fully or semiautomatic.
The downward displacement autoclave
sterilizer is also
known as a gravity displacement unit.
It uses a heating element to heat up the
water and produce steam.
The steam, which is lighter than air, forces
the air inside the sterilization chamber to
move downward.
Eventually the air moves out through the
drain hole of
the sterilization chamber.
Once the temperature in the chamber is

Vaccum Assisted
Autoclave
A vacuum assisted
autoclave pulls
pressure from the
chamber.
Open
Steam

Steam

Vaccum Assisted
Autoclave
A vacuum autoclave is set up in a
similar fashion to a negative pressure
displacement unit in that there is a
vacuum pump to remove air from the
sterilization chamber and steam is
created in a second, separate
chamber or unit. The process begins
by the vacuum removing the air, then
a pulse of steam. This type of
autoclave is suitable for all types of

Heat
Exchan
ge

Steam
Generator

Vaccu
m
Pump

MAIN COMPONENTS
1.
2.

3.

4.
5.
6.
7.

8.
9.

Heating Elements
Temperature
Controller
Pressure Sensor
Chamber
Door gasket
Solenoid valve
Water level Sensor
Steam generator
Vaccum pump

Vacuum assisted or prevacuum

autoclave
Air is evacuated rapidly from the
sterilising chamber at the beginning of
the cycle creating a vacuum, so steam
penetration and sterilising occurs very
rapidly.
A second vacuum cycle withdraws
moisture after
sterilisation , facilitating drying of the
load.
Often, this autoclave is connected to a

Vertical Pressure
Cooker(Autoclave)
This cooker operates similarly to a
household pressure cooker. It is commonly
used in practice. The vertical pressure
cooker has an air vent at the top which is
manually closed after all the air has been
evacuated.
This is a possible disadvantage as some air
may remain trapped under the steam and
therefore cause a lower temperature to be
reached in this area.
As this is a manually operated steriliser,
human error can influence the effectiveness

Classification of
Autoclaves

Type "N" & Type "B Each

Type "N" units do not use a vacuum
to remove air from the sterilization
chamber. Type "N" autoclaves are
suitable for a specific type of load-for solid.
Type "B" units use a vacuum pump.
Type "B" autoclaves can be used on
wrapped and hollow instruments,
which means a piece of equipment

MAIN COMPONENTS
1. Heating Elements
2. Temperature
Controller
3. Pressure Sensor
4. Chamber
5. Door gasket
6. Solenoid valve
7. Water level Sensor

Preparation of Material
1. Syringes and needles
Syringes are wrapped in gauze and needles
are placed separately in small test tubes
that are plugged.
A pad of nonabsorbent cotton is placed at the
bottom of each tube to protect the tip of the
needle. Syringes and
needles are then
placed in a metal tray, which is placed
uncovered in the autoclave.
Glasswa such as specimen
tubes,
2. Glassware
specimen petri
collection
bottles are wrapped in
re
dishes,
brown paper
The
and Pasteur
are tied pipettes
with string.
are placed in large
tubes, which are then plugged.

What can be Autoclaved?

Surgical Instruments
Glassware
Plastic tubes and pipette tips
Culture and stocks of infectious
material
Discarded Live and attenuated
vaccine
Media and solutions
Waste

Preventive Maintenances
1.
2.
3.
4.
5.

Clean all the filters.(Water, Air )

Fill the water level of feed tank up to maximum limit
Check that the manometers function as expected.
Check the accuracy of temperature reading.
Activate the safety valves manually to verify that they
are operating well.
6. Conduct a general sterilization process write down
pressure, temperature, required times for completing
each phase of the cycle, put it on registration system.
7. Verify that it is functioning within tolerances defined by
manufacturer
8. Check any abnormal sound in solenoids.
9. Clean the chamber and remove limescale if any,
advice to use only distilled water.
10. Check the earth leakage current, protective earth.

Limesca
le

Limescale is the
hard, off- white,
chalky deposit
found in kettles, hotwater boilers
Add little amount of
any
acid heat the
chamber.
Deposited CaCO3
will be removed.

Autoclave: Performance
Indicators(Quality Control
How to know if autoclave is functioning
correctly:
Physic
al
Chemic
al
Biologic
al

-They check instruments control

parameters like temperature, time
and pressure.
-Heat

sensitive autoclave tape

- Not an indicator of successful
sterilization, useful
to keep track of autoclaved and
unautoclaved
- Tests ability ofitems
autoclave to sterilize effectively
- Geoacillus stearothermophilus spore strips often used
because they are
resistant to steam sterilization.

Precaution for
Autoclave Users
Eye Protection
Lab Coat,
Buttoned
Closed-toed
Shoes
Heat-resistant