Sterilization

Sterilization: is a process by which an article, surface or medium is rid

of all living microorganisms including spores.
Classification
Physical agents
Sunlight - UV rays and heat has germicidal activity
Drying - Spores are unaffected
Heat - Most reliable, certain and rapid method of sterilization
Filtration - Used for biological fluids
Radiation - Ionizing and non-ionizing
Chemical agents
Phenols and cresols
Halogens
Metallic salts
Aldehydes
Alcohol
Dyes
Vapour-phase disinfectants
Surface- active disinfectants

Moist heat sterilisation

Moist Heat: Kills microorganisms by coagulating their proteins
Moist heat is more effective than dry heat
Temp below 100oC
At 100oC
Above 100oC
At temperature below 100oc

Heat labile fluids may be disinfected

Vaccine bath: Vaccines may be inactivated in a water bath at
60oc for one hour
Serum bath: Heat labile fluids such as serum may be
disinfected by heating at 56oc
Household utensils and patients clothing may be disinfected
by washing at 70oc-80oc
Pasteurization
Developed by Louis Pasteur to prevent the spoilage of beverages
Holder method: 63oc for 30 minutes
Flash method: 72oc for 20 seconds followed by rapid cooling at
13oC

 By this method nonsporing organisms like Mycobacteria, Brucella

and Salmonellae are destroyed
Coxiella burnetti (Q fever) not destroyed by Holder method
Ultra High TemperaturePasteurization
(UHT)
Milk is treated at 140oC for 3 seconds and then cooled very
quickly in a vacuum chamber
Advantage: Milk can be stored at room temperature for
several months
Inspissation
80-850 C for 30 mins , 3 consecutive days
Egg or serum containing culture media
Lowenstein Jensen medium & Loefflers serum slope
At 100oc
Boiling:
Boiling at 100oc for 10 to 30 mins
At this temp vegetative bacteria and some spores are killed
Metal, glass & rubber items are boiled, dried cooled and used
Free steam
Steam at normal atm pressure is at 100oc
It has latent heat which on condensing on the article
to be sterilised releases its latent heat

 One single exposure to steam for 90min ensures

complete sterilization
Tyndallization
An exposure of 100oc for 20 minutes on three
consecutive days
First exposure to steam kills all vegetative
bacteria and any spores present will germinate and
will be killed on subsequent occasions
Media containing sugar and gelatin
Arnold steam sterilizer
Above 100oc
It provides greater lethal action of moist heat
It is quicker in heating up the exposed articles
It can penetrate easily porous material such as
cotton wool stoppers, paper, cloth wrappers, bundles
of surgical linen and hollow apparatus
Dry Heat Sterilization
Heat: Kills microorganisms by denaturing their enzymes and
other proteins
Red Heat - inoculating loops and wires, points of forceps and
spatulas
Flaming- scalpel blades, needles, mouth of culture tubes, glass
slides and cover slips

 Incineration: Rapidly destroys contaminated material i.e. soiled

dressings, disposable items and biological waste
Hot air oven
Principle: Dry heat -killing effect is due to protein denaturation,
oxidative damage and toxic effect of elevated levels of electrolytes
Used for glass ware like glass syringes, test tubes, petridishes,
pipettes, and flasks, metal instruments such as forceps, scissors and
scalpels
Oils, jellies and powders and swab sticks packed in test tubes
Precautions
Should not be overloaded
Substances to be sterilized should be absolutely dry
Holding time at 160o C for 1hour
Oven should be allowed to cool for 2 hours before opening to
prevent cracking of glassware
Articles like rubber goods, fabrics, inflammable or volatile
substances should not be placed inside
Sterilization control
Autoclave
Principle
Water boils when its vapour pressure equals that of surrounding
atmosphere

 When pressure inside a closed vessel , the temp at which water

boils also
Saturated steam has greater penetrating power
When steam comes into contact with a cooler surface, it condenses
to water and gives up its latent heat to that surface.
Uses
Sterilisation by steam under pressure (autoclaving) is suitable for
culture media aqueous solutions, dressing materials, linen, gloves
etc
Satisfactory sterilization can be achieved at 15 pounds pressure per
square inch at 121oc for 20 min
Diagram

Sterilization controls
Biological control
Bacillus stearothermophilus
Chemical control
Brownes tube
Thermocouple