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Sterilisation

There are two main methods of sterilization and disinfection - physical and chemical. Physical methods include heat sterilization using dry heat like hot air ovens or moist heat like autoclaves. Chemical methods involve the use of chemicals like alcohols, aldehydes, halogens, phenols and gases which can kill microbes through various mechanisms like disrupting cell membranes or altering DNA. Proper sterilization and disinfection are important in healthcare, food production and other areas to prevent contamination and infection.

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Dr Md Suheeb Ahmer
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100 views

Sterilisation

There are two main methods of sterilization and disinfection - physical and chemical. Physical methods include heat sterilization using dry heat like hot air ovens or moist heat like autoclaves. Chemical methods involve the use of chemicals like alcohols, aldehydes, halogens, phenols and gases which can kill microbes through various mechanisms like disrupting cell membranes or altering DNA. Proper sterilization and disinfection are important in healthcare, food production and other areas to prevent contamination and infection.

Uploaded by

Dr Md Suheeb Ahmer
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Sterilisation & Disinfection

 Sterilisation : A process by which an article, surface or medium is free of all living


organisms either in vegetative state or spore state.
 Disinfection: Removal or destruction of all pathogenic organisms
 Asepsis: Prevention of infection by inhibiting the growth of bacteria.
 Decontamination: The process of rendering an article or an area free of contamination
including microbial, chemical, radioactive, etc.
 Bactericidal agents: Agents that kill bacteria.
 Bacteriostatic agents: Agents that prevent multiplication of bacteria.

Uses of sterilisation: To prevent contamination by extraneous organisms, to prevent sepsis in


surgery, for safety in food and drugs

CLASSIFICATION

Physical methods Chemical methods

Sunlight, drying. Alcohols - Ethyl alcohol, isopropyl alcohol


HEAT : 1) dry heat – red hot heating, Aldehydes - Formaldehyde, glutaraldehyde
flaming, incineration, hot air oven
2) Moist heat- below 100C Pasteurisation, Dyes
water bath/ vaccine bath, inspissations Metallic salts

Steam under normal pressure at 100C: Halogens


Arnolds steam sterilizer, tyndallisation.
Boiling
Steam under pressure (Above 100C): Phenols
autoclave

Filters, radiation Surface active agents

Ultrasonic and sonic vibration Gases: ETO, formaldehyde, betapropiolactone


• Physical methods

• Red hot : inoculating loops, wires, tip of forceps and needles are held in the flame of a
bunsen flame of a burner till they become red hot.

• Flaming: used to sterilize glass slides, scalpels & edges of test tubes by passing them
through Bunsen flame without allowing them to become red hot.

• Hot air oven:

• Most widely used method of sterilization by dry heat.

• The oven is electrically heated and fitted with a fan to ensure adequate and even
distribution of air. Also fitted with a thermostat to maintain the air at a chosen
temperature.

• Holding period – 160°C for 2 hour. Or 170°C for 1hr, or 180°C for 30 mins.

• Glassware like syringes, petridishes, flasks, pipettes and test tubes, surgical
instruments like scalpels scissors, forceps etc. liquid paraffin, dusting powder, fats,
grease, etc.

• Rubber, cotton avoided

• Incineration: Contaminated cloth, animal carcasses, pathological materials, infectious


hospital waste

MOIST HEAT

At Temp less than 100°C

 Pasteurisation- method of sterilization used to sterilise liquids such as vegetable/fruit


juices, milk etc.

2 methods – Holder method heating the liquid at 63°C for 30 min

– Flash method heating the liquid at 72°C for 20 sec


Followed by quickly cooling to 13°C

 Vaccine bath: used to sterilise vaccines- Heat inactivation by heating at 60°C for 1 hr

 Inspissation: LJ medium, Loeffler’s serum slope can be sterilized by heating at 80–85°C


for 30 min on 3 days

At 100°C

 Boiling

 Koch & Arnold steam steriliser:


media –90 min

 Tyndalisation: sugars, gelatin

 100°C – 20 min - 3 successive days

At temperature higher than 100°C

Autoclave
o Most widely used method of sterilization by moist heat.
o Uses steam under high pressure for sterilization

o Temperature used is 121°C for 15mins at 15 lbs pressure.


o Or 126°C with 20lbs pressure for 10mins
o Or 133°C at 30 lbs for 3 mins.
o Used to sterilize culture media, rubber materials, gowns, dressing, gloves etc.

Filtration

 Useful for sterilizing substances that get damaged by heating like sera, antibiotics,
enzymes, toxins.
 Also used for Separation of virus from bacteria

Types of filters

 Candle filters. ex: Berkfeld & Mandler: Made of kieselghur, graded porosity, sterilised
by steaming or autoclave
 Chamberland or Doulton: Made of unglazed ceramic

 Asbestos disc filters:

Seitz filter: - pad of asbestos - inserted into metal holder - discarded after use

 Sintered glass filters:

Used for air filtration, made of finely ground glass.


 Membrane filters:
o Cellulose nitrate
o Also called gradacol MEM
o To separate viruses, to know their size
o Cellulose acetate or millipore filters or oxoid MEM filters

Air filters: HEPA: High efficacy particulate air filters

Radiation

 Non-ionising radiation - Infrared and ultraviolet rays - low energy type

- Control of closed environment with UV lamp

- Bacteriological laboratory, inoculation hoods, laminar flow

 Ionising radiation - Gamma rays, X rays and cosmic rays lethal to penetrative
power

No appreciable increase in temperature – ‘cold sterilisation’

Use: Sterilising pharmaceuticals, medical, dental equipment, sterilising plastics,


syringes, swabs, catheters, oil grease

CHEMICAL METHODS

Alcohols

 Protein denaturation and lipid dissolution

 Bactericidal, fungicidal
 Not effective on spores and viruses

 Commonly used - Ethyl alcohol, isopropyl alcohol

 Concentration 60-90% in water

Aldehydes

Active against amino group in protein molecule

Formaldehyde

Used for sterilising instruments, heat-sensitive catheters, fumigation of wards, sick


rooms and laboratories

Glutaraldehyde

 Disinfection of medical equipment like cystoscopes and bronchoscope

 Effective against tubercle bacilli, fungi and viruses

Phenols

 Lister - antiseptic surgery

 Distillation of tar between 170°C and 270°C

 Damage to cell membrane, release cell contents and cell lysis

 Cresols, chlorhexidine, hexachloraphane

 Disinfection of floors, excreta, preoperative disinfection of skin

Halogens

Iodine – Skin disinfectant

 Bactericidal, sporicidal

 Iodophores more active


Chlorine – Used as hypochlorite

 Bactericidal

 Disinfect water supplies, swimming pools and used in diary industries

 Ethylene oxide

 - Kills microorganisms by altering their DNA by alkylation of amino carboxyl,


hydroxyl and sulphydryl groups in protein molecules

 - Used in sterilising packaged heat-sensitive devices

 - Respirators, heart–lung machines, dental equipment, sutures, books and


clothing

Formaldehyde

 Fumigation of theatres and rooms

 150 g of KMnO4 to 280 ml formalin for every 1000 cu ft.

 Doors sealed, opened after 48 hrs

Betapropiolactone (BPL)

 Condensation product of ketane and formaldehyde

 Hydrogen peroxide fogging

 Short cycle time, non-toxic

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