Laboratory Manual

For

Autoclave

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 Autoclave (Vertical)

Experiment:

To study the Autoclave and study its different parts

Theory:

This process involving heating in an autoclave with

saturated steam under pressure should be used whenever

possible for aqueous preparations and for surgical

materials. An aqueous product which is to be sterilized by

this method is distributed into suitable containers which

are then sealed so as to exclude micro-organisms. It is than

exposed to saturated steam for a time sufficient to ensure

that the entire contents of each container are maintained

for an effective combination of time and temperature to

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ensure sterility. The following combinations of temperature

and time are normally employed.

Holding temperature (ºC) Minimum holding time (minutes)

115 to 118 30

121 to 124 15

126 to 129 10

134 to 138 3

In this method micro-organisms are killed due to

coagulation or denaturation of proteins in living cell of

micro-organisms. It is more effective than dry heat because

saturated steam has more penetrating power than dry

heat. More over the thermal capacity of steam is much

higher than the thermal capacity of dry heat.

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Working:

Autoclave is a strong metallic chamber usually made up of

stainless-steel. The material to be sterilized is packed in a

clean fabric cloth and is placed in perforated chamber of an

autoclave. The water level is checked so that it should not

touch the bottom perforated metallic chamber. The lid is

closed with the help of nuts and bolts. Autoclave is

switched on and water is allowed to boil. Remove the

trapped air. Steam vent is then closed and pressure is

allowed to rise up to 15 lb/sq inch and at this pressure

saturated steam has a temperature of about 121oC. The

pressure is maintained for 30 min. the autoclave is switch

off allowed to cool, remove the steam and open it.

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Applications:

This method is used to sterilize bulk solutions, glassware,

surgical dressings, rubber gloves, culture media and

surgical instruments are efficiently sterilized by this

method. Solutions packed in sealed containers such as

SVP, LVP can be sterilize by this method.

Disadvantages:

1. It requires high temperature and moisture, unsuitable

for thermo labile substances.

2. This method cannot be used for oily injections, fats,

powders, ointments because steam cannot penetrate

them.

3. Moist medicaments, rubber and plastic articles are

destroyed by this method.

4. Preparations containing water, alcohol or other volatile

substances cannot be sterilized by this method.

5. It is unsuitable for surgical dressings

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