Sterilization and

Disinfection
 Introduction

Objective of sterilization
–Removal of microorganisms or destroy
them from materials or from areas since they
cause contamination, infection and decay.

In microbiology - to prevent contamination

Surgery - to maintain asepsis
Drug & food - for ensuring the safety
 Decontamination

 Any procedure that reduces pathogenic microbes to a level

where items are safe for handling & disposal.
 It can be achieved by cleaning, disinfection and
sterilization.

Cleaning

 A process that removes foreign material (dirt, organic

matter).
 Must precede disinfection and sterilization.
 Usually done with soap and water or detergent.
 Disinfection

 Destruction of most but not necessarily all pathogenic

microbes or their spores.

Sterilization

 The process by which an article, surface, or medium is

freed of all living microorganisms either in the vegetative or
spore state.
 Antiseptics

 A product that destroys or inhibits the growth of

microorganisms in or on living tissue. Chemical
compounds that could be applied topically on animate
surfaces.

Disinfectants

 Products used to kill microorganisms on inanimate

objects or surfaces. Disinfectants are not necessarily
sporicidal, but may be sporostatic, inhibiting germination or
outgrowth
 Definitions
• Antisepsis – used to indicate the prevention of
infection, usually by inhibiting the growth of
bacteria in wounds or tissues
SEPS ( A Greek word ) – PUTRID

• Bactericidal agents
• Bacteriostatic agents
• Cleaning
• Degerming
Spaulding System ( 1972 )

• Critical - penetrate/touch broken skin or

mucous membrane
- must be sterilized
• Semicritical -touch intact mucous membrane
- sterilize, high level disinfection
• Noncritical - surfaces do not touch mucous
membrane
- disinfection
 Selection of adequate level of contamination
methods Example Characteristics Item
Sterilization by Surgical Instruments that
autoclave, instruments, enter sterile
plasma gas needles, tissues, cavities or
sterilizer or catheters vascular system Critical
ethylene oxide
gas

High level Endoscopes, Objects that come

disinfection thermometers in contact with non
intact skin or Semi
mucous critical
membranes

Intermediate to Sphygmomano Objects that come

low level meter, bed in contact with Non
disinfection linen intact skin critical
12/22/2020
Methods Of Sterilization

Microbial
Control
Methods

Mechanical
Physical Chemical
Removal
Agents Agents
Methods
 Physical
Agents`
Heat Radiation

Non
Dry Moist Ionizing
Ionizing

Incineration Steam Under X-ray, Cathode, UV

Pressure Gamma
Sterilization
Hot Air Oven Sterilization Disinfection

Boiling Water/ Hot

Water Pasteurization

Disinfection
 Chemical
Agents

Gas Liquid

Sterilization Disinfection Animate Inanimate

Chemotherapy Sterilization

Antiseptics Disinfection
 Mechanical
Removal Methods

Filtration

Air Liquids

Disinfection Sterilization
 HEAT
Heat is the most common method of
sterilization
Factors determining the time & temperature for
sterilization

• Nature of heat – dry or moist

• Presence of organic matter
• Number of microorganisms present
• Characteristics of the organism
• Type of material from which the organisms
have to be eradicated
 PHYSICAL CONTROL WITH HEAT

• SUNLIGHT – Ultraviolet rays

Typhoid bacilli exposed to sun on pieces of
cloth were killed in 2 hours, where the controls
kept in dark were alive after 6 days

• DRYING - 4/5th of the bacterial cell is made-up

of water
 Dry Heat
DIRECT FLAME

Bunsen Burner
Simplest method is exposing the
item to be sterilized to the naked
flame e.g. Bunsen burner- for
sterilizing bacteriological loops,
knives, blades.
Incineration
 Methods of Sterilization
Dry Heat

Hot Air Oven

Hot air oven expose items to 160°C

for 2 hours. It has electric element in
the chamber as source of heat plus a
fan to circulate air for even distribution
of heat in chamber. Oven without fan
is dangerous.

Advantage - non corrosive method

 – ARTICLES THAT CAN BE STERILIZED
– Surgical instruments
– Glass ware
– Oils grease powders
– Certain pharmaceutical products

Test for efficiency

• Chemical method: Browns’ tube develops GREEN
spot at 160c after 1 hour
• Biological method:
• Clostridium tetani on a filter paper strip and place it
inside the hot air oven along with oven work for 160c
for 1hour.
• Then filter paper is innoculated and incubated…
 Moist Heat

• Temperatures below 1000C - pasteurization

• Temperatures at 1000C - boiling

• Steam at atmospheric pressure

 Moist Heat
PASTEURIZATION ( BELOW 1000C )
Purpose – To reduce the bacterial population of a
liquid such as milk
Spores are not affected by pasteurization

• Holding method
62.90C for 30 minutes (Mycobacterium
tuberculosis & Coxiella burnetti )
• Flash pasteurization – 71.60C for 15sec
• Ultra pasteurization – 820C for 3sec
 Moist Heat
BOILING WATER (1000C)

• Lower temperatures & less time of exposure are

required
• Denaturation of proteins
• Minimum exposure time – 30 minutes
• Less reliable
• Washing soda ( 2% conc.) may be added
Fractional sterilization ( steam at atmospheric
pressure )

• Tyndallization
( John Tyndall )

• Intermittent
sterilization
 Moist Heat
TYNDALLIZATION
Free flowing steam at 1000C for 30 minutes on
each of 3 successive days.
First day - steam kills all organisms except
spores, and it stimulates spores to germinate
vegetative cells

Second day – vegetative cells are killed

Third day – kills the remaining cells

 Moist Heat
AUTOCLAVING
• Standard sterilization method in hospitals.

• The Autoclave works under the same principle as

the pressure cooker where water boils at increased
atmospheric pressure i.e. because of increased
pressure the boiling point of water is >100°C.

• The autoclave is a tough double walled chamber in

which air is replaced by pure saturated steam
under pressure.
 Mechanism Of Action Of Autoclave

Moist heat In the form of pressurized steam

increase in the pressure of the gas

increase in the temperature

As the water molecules in steam becomes more
energized, their penetration increases
Pressure Temperatu Time (Min.)
re
15psi  121oC  15

20 psi  126oC  10

  32 psi  134oC  3
 AUTOCLAVING
 The air in the  evacuated  filled with
saturated steam.
 The chamber closed tightly  steam keeps
on filling  pressure gradually increases.

 The items to be sterilized gets surrounded

by saturated steam (moist heat) when on
contact with the surface of material to be
sterilized condenses to release its latent heat
of condensation eventually all the
microorganisms in what ever form – are
killed.

 The usual temperature achieved is 121 °C at

a pressure of 15 pps.i. at exposure time of
only 15-20 mins.
 Advantages of Autoclave
• Temperature is > 100°C therefore spores are
killed.

• The condensation also allows the steam to

penetrate rapidly into porous materials.

• Note: that autoclavable items must be steam

permeable. Can not be used for items that are
lacking water.
 Limitations Of Autoclave
• Plastic ware melts in high heat

• Sharp instruments become dull

• Oily substances cannot be treated

 ULTRASONIC WAVE STERILIZATION :
STERISONIC 1000C
– The equipment is designed to
perform functions like cleaning,
disinfecting and sterilizing.

– The solution is preheated to 1800F and

at this point the equipment is ready to
operate.
– The instruments are placed in the
tank for 5-15min.

– Function : produces high frequency,

high intensity cavitation in hot water
containing blood solvent.
Methods Of Sterilization
Radiation:
– U.V. light- Has limited sterilizing power  poor
penetration .
Generally used in irradiation of air in certain areas eg.
Operating Rooms and T.B. laboratories.

– Ionizing radiation- e.g. Gamma radiation: Source

Cobalt60  Used mainly in industrial facilities e.g.
sterilization of
disposable plastic syringes, gloves, specimens
containers and Petri Dishes.
 Sterilization By Chemical Methods
Ethylene Oxide Chamber:
may cause explosion if used
pure so mixed with an inert
gas e.g. Neon, Freon at a
ratio of 10:90

It requires high humidity and

is used at relative humidity
50-60% Temperature : 55-
60°C and exposure period 4-6
hours.
 Sterilization By Chemical Methods

Activated alkaline
Glutaraldehyde 2%:
Immerse item in solution for
about 20 minutes if organism
is TB.
 In case of spores, the
immersion period is extended
to 2-3 hours.
 DISINFECTANTS
 Factors influencing activity of Disinfectants
Directly proportional to concentration up to a
point – optimum concentration. After this level no
advantage in further increases in concentration.

Hypochlorites and Glutaraldehyde are

more active against hepatitis viruses than most
other disinfectants.
 Selection of antiseptics &
disinfectants
Prerequisites

• It should have a wide spectrum of activity

• Fast acting
• Active in the presence of organic matter
• Non-toxic to animals or humans ( antiseptic )
• Soluble in water
• It should not separate on standing
• Should have high penetrating power
• Surface compatibility
• Relatively inexpensive
Levels of disinfection

1. High - sterilizing agents. ex; ethylene

oxide gas
2. Intermediate - bactericidal agents
ex; formaldehyde, alcohols
3. Low - narrowest anti-microbial
activity. Ex: soaps, detergents
 Sterilizing pouch
 These sterilization pouches also have dual steam and EtO process
indicator arrows to visually show you that the instruments in
that pouch have been processed. For safe, easy instrument
sterilization and protection

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 DISINFECTANTS
Types of Disinfectants
Phenol and phenolics

• inactivate enzymes, or denature proteins.

• They are stable, persistent, and are not
sensitive to organic matter.
 DISINFECTANTS
Phenylphenol
• It is the main ingredient in most
formulations of Lysol.

Triclosan
• It is a widely used in many
household products.
• It has broad spectrum of activity,
especially against gram positive
bacteria.
• It is also effective against gram
negative bacteria and fungi.
 DISINFECTANTS
 Biguanides

• Chlorhexidine, a member of
the biguanide groupnot a
phenol but structure and
applications resemble
hexachlorophene.

• It is frequently used for

surgical skin preparation
and surgical hand scrubs.
 DISINFECTANTS
Halogens Chlorine  used as a gas or
Iodine is effective against  combination with other
bacteria, fungi, and some
viruses.
chemicals.
iodophore  combination of Chlorine  disinfecting
iodine and an organic municipal water supplies,
molecule. do not stain and swimming pools, and
are less irritating than iodine. sewage.
Ex:- Isodine and Betadine.
Sodium hypochlorite 
ordinary household bleach-
is good disinfectant.
 DISINFECTANTS
 Alcohols

• Concentrations of 60% to 95% are effective.

• They are bactericidal and fungicidal but are not effective

against endospores or non-enveloped viruses.
INSTRUMENT PROCESSING
The overall process consists of

• Holding ( presoaking )
• Precleaning
• Corrosion control, drying, lubrication
• Packaging
• Sterilization
• Sterilization monitoring
• Handling processed instruments
 HOLDING (PRESOAKING )
• Holding solution– detergent/water/enzyme
solution
• To prevent drying
• Perforated basket
• Extended soaking
 PRECLEANING
• Ultrasonic cleaning - reduces direct handling
- time saving
• Manual scrubbing - dangerous
- long handled brush
• Instrument washers
 Packaging
• Prevents contamination after sterilization,
during storage or when being distributed to chair
side

• Pouches, bags, cassettes ( stainless steel,

aluminum, and plastic/resin )

• Closed containers
 STERILIZATION

Universal sterilization
“if it can be sterilized, sterilize it”
Sterility assurance
- the correct performance of the proper
instrument processing steps and monitoring the
sterilization with biologic & chemical indicators
Small office sterilizer
Unsaturated chemical
vapor sterilization
( chemiclave )

Solution – 0.23% formaldehyde &

72.38% ethanol plus acetone,
ketone, water & other alcohols

Temperature - 2700 F( 1320 C )

Pressure - 25 psi ( 172 Kpa.)
Time - 20 min

Positive feature – corrosion is reduced or

completely eliminated

Negative feature – irritating fumes

 BARRIER TECHNIQUES
• Gowns & protective apparel:
• protective covering for street clothing
and exposed skin.

• Gowns, aprons, lab coats, clinic

jackets or similar outer garments,
either reusable or disposable, must be
worn as clothing so skin which is likely
to be exposed to body fluids or
infectious aerosols is covered.
•
• High necked, long sleeved, knee
length garments can provide
protection.

• Disposable head caps are used for

avoiding not coming into direct contact
with equipment or patient.
• Masks & face shield:
• It is used protection against
aerosolhave at least 95%
filtration efficiency for particles
3.0 to 5um in diameter & should
be changed for each patient.

• Face shield may be work in

addition to the mask for greater
protection against splatter. It
protects against contact with
larger splatter particles. It protect
from inhalation of aerosol
particles.

• Eye protection: Protective

eyewear with side shield is used.
 SURGICAL HAND SCRUB

• A 5 minute timed scrub will be used

surgical hand scrubs.
• Fingernails no polish , medium length.
• Clean fingernails with file.
• No jewelry on hand
• Wash hands and arms with solution to 2
inches above the elbow.
• Gloves:
• latex gloves better tactile
sensation more fragile 
frequent glove changes during
operation may necessary.

Gloves should always be worn

while touching:
• -Blood & body fluids
• -Mucous membrane.
• -Non-intact skin of all patients.
• -Items or surfaces soiled with
blood or body fluids to which
universal precautions apply.
GLOVING PROCEDURE
 SKIN PREPARATION
• 70% Isopropyl alcohol
• Acts by denaturing proteins
• Is bactericidal but short acting
• Effective against gram-positive and gram-
negative organisms
• Also fungicidal and virucidal
 SKIN PREPARATION (contd)
• 0.5% Chlorhexidine
• Acts by disrupting the bacterial cell wall
• Bactericidal but does not kill spore forming
organisms
• It is persistent and has a long duration of
action (up to 6 hours)
• More effective against gram-positive
organisms
 DRAPING

• Purpose is to isolate
the surgical
site/prepared area
from other parts of the
body that are not
prepared for the
surgery
 DEGLOVING
• If contamination occurs during any procedure,
both gown and gloves must be discarded and
new gown and gloves must be added.

• When removing gloves after a procedure is

finished, the gloves are removed after the gown
is removed inside out, using glove-to-glove, then
skin-to-skin technique.
 UNIVERSAL PRECAUTIONS
• Although the risk of contracting HIV/HbsAg in
the healthcare setting is low.

• Precautions to reduce the risk of exposure to

bloodborne pathogens.

• Each healthcare worker should assess their

possible risks and take precautions to reduce
these risks.
 UNIVERSAL PRECAUTIONS

• Gloves should be worn for touching blood and/or body

fluids, mucous membranes, non-intact skin, or
items/surfaces soiled with blood and/or body fluids.

• Gloves should be changed after contact with each

patient and hands washed after glove removal.

• Though gloves reduce the incidence of contamination,

they cannot prevent penetrating injuries from needles
and other sharp instruments.
 UNIVERSAL PRECAUTIONS
• Gowns or aprons should be worn during
procedures that are likely to generate splashes
of blood and/or body fluids onto clothing or
exposed skin.

• Masks and protective eyewear should be worn

during procedures that are likely to generate
droplets of blood and/or body fluids into the
mucous membranes of the mouth, nose, or
eyes.
 UNIVERSAL PRECAUTIONS
• Hands and skin surfaces should be washed after
contact with blood and/or body fluids, after
removing gloves, and between patient contact.

• Gloves should be worn to cleanup blood spills

the disinfectant should have a one minute
contact time and the area rinsed with tap water
 Use & Care of sharp instruments and
needles:
• Any sharps contaminated with patient blood & saliva
should be considered potentially infective & handled
with care to prevent injuries, double gloving, though
recommended & careful handling of instrument
required.

• Disposable syringes needles scalpel blades & other

sharp items should be discarded into puncture resistant
biohazard container that is easily accessible for
disposal.

• Exchange sharps always by dish and not hand to hand.

 FUMIGATION
•For 100 cubic feet of air space in a room, 50 ml 40% formalin is required. This gives
vapor of 2 mg gas per liter of air.
•Article and room exposed for a period of at least 4 hours.
•Diluted formalin may also be sprayed.
•Next the space is ventilated so that the poisonous gases are allowed to escape from
the space

Widely used fumigants include:

methyl isocyanate
hydrogen cyanide
formaldehyde
Iodoform
Methyl bromide

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 Glass Bead Sterilizer:
 TEMPERATURE-218oC-246oC
 It is used to sterilize:
-Absorbent points
-Broaches
-Files, reamers and other Root canal instruments

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 CONCLUSION
• The sterilization and asepsis is a one of the most
important step in the all minor as well as major surgical
procedures, so first most importantly; steps should be
taken to wash hands carefully and to wear gloves and
follow recommended aseptic practice.

• Whenever there is any contact between the hands

unsterilized tools/fluids care is taken not to contaminate
the surgical area.

• Surfaces, units & controls should be cleaned &

disinfected.