STERILIZATION

By: Rahul Malik

January, 2016

Definition
Sterilization is the freeing of an article from
all living organisms, including bacteria and
their spores.
Sterilization of culture media, containers and
instruments is essential in microbiological
work for isolation and maintenance
of microbes.

In surgery and medicine, the sterilization

of instruments, drugs.

Sterilization can be effected in a variety of

ways, which can be conveniently
categorized as follows:

I. PHYSICAL METHODS
II. CHEMICAL METHODS

I. PHYSICAL METHODS
A. Heat :
1. Dry heat
2 Moist heat
B. Radiations:
1. Ultraviolet radiations
2. Ionizing radiations
C. Filtration

STERILIZATION BY HEAT
Heat can be applied in two forms.
1. The dry heat
2. Moist heat.

Mechanism of killing by dry heat

Dry heat kills the organisms by destructive
oxidation of essential
cell constituents
Killing of the most resistant spores by dry
heat requires a temperature
of about 160 C for 60 minutes

 Dry heat is employed for glassware;

syringes, metal instruments
and paper wrapped goods, which are not
spoiled by high
temperatures.
It is also used for anhydrous fats, oils and
powders that are
impermeable to moisture

Mechanism of killing by moist

heat
Moist heat kills the organisms by
coagulating and denaturing
their enzymes and structural protein.
Sterilization by moist heat of the most
resistant spores generally
requires 121 C for 15-30 minutes.

Moist heat is used for the sterilization

of culture media, and all other materials
through which steam can penetrate
Moist heat is more effective than dry
heat
Sterilization can be done at lower
temperatures in a given time
at a shorter duration at the same
temperature

METHODS OF STERILIZATION
BY DRY HEAT
1. RED HEAT
Inoculating wires, points of forceps and
searing spatulas are sterilized by
holding them in the flame of Bunsen
burner until they are seen to be red-hot.

2. FLAMING
This method is used for sterilizing
scalpel, mouth of culture tubes, glass
slides etc.
It involves passing of an article through
Bunsen flame without allowing it to
become red-hot

3. HOT AIR OVEN

This is the main means of sterilization by
dry heat. Exposure at a temperature of 160
C for 1 hour is generally employed.
4. INFRARED RADIATIONS
Source employed is an electrically heated
element, the infra red rays are directed on
to the object to be sterilized and
temperature of 180 C can be obtained

METHODS OF STERILIZATION BY MOIST

HEAT
Moist heat can be employed at
1. Temperature below 100 C
2. Temperature of 100 C
3. Temperature above 100 C

MOIST HEAT BELOW 100 C

EXAMPLES
1. Pasteurization of milk
In Pasteurization of milk the temperature
employed is either63 C for 30 minutes
or 72 C for 20 seconds. All nonsporeforming pathogens
in milk like
Salmonellae, M.tuberculosis are killed.

MOIST HEAT ABOVE 100C

1. Sterilization in an autoclave
Autoclaving is the most reliable method
It is the method most widely used for
sterilization of culture media and surgical
supplies

When water is boiled within a closed

vessel at an increased pressure, the
temperature at which it boils and the
steam it forms will rise above 100 C
This principle is used in the autoclave
Normally autoclaving is done at 15 lbs.
(pounds per sq. Inch pressure) and 115
C for 15 minutes

STERILIZATION BY FILTRATION
When fluids are passed through
bacteria stopping filters, they are made
free from bacteria.
It is useful for making preparations of
soluble products of bacterial
growth such as toxins

Liquids that would be damaged by heat

such as serum and
antibiotic solutions can be sterilized by
filtration
Efficient filters should be able to retain
Serratia marcescens

TYPES OF FILTERS
There are different kinds of filters
1. Earthenware candles - called
Berkfield & Chamberland filters
2. Asbestos and asbestos-paper
discs filters - called Seitz filters
3. Sintered glass filters
4. Cellulose membrane filters
5. Fibre glass filters.

MERITS AND DEMERITS OF

HEAT STERILIZATION
Advantages of heat sterilization
1. Sterilization is very effective
2. Instruments are standardized
deliver the required effective heat

to

3. Heat deliver system can be monitored

effectively with various controls like
pressure gauge, temperature meters etc
4. Established quality control methods
available

Disadvantages
1. Steam impermeable materials like
fats, oils and powders can not be
sterilized by autoclaving.
2. Heat sensitive materials can not be
sterilized by heat

Examples:
1. Serum can not be sterilized
2. Antibiotics
3. Plastic materials
4. Vaccines
5. Rubbers

3. Presence of organic matters interfere

with effective sterilization
4. Dangers of explosion when high
pressure is used.

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