Sec.

6
STERILIZATION and
DISINFECTION
RADIATION
B. Ionizing rays
A. Non-ionizing rays 1) X-rays
1) Ultraviolet (UV) light 2) Gamma rays
3) Beam of electrons
Ultraviolet (UV) light

UV light is an electromagnetic

radiation that has a wavelength
longer than X-Rays but shorter
than visible light
Germicidal UV lamps are used
Bacterial spores are resistant and
require a dose up to 10 times
greater than vegetative bacteria.
Uses
1. Sterilize spaces since the covid-19
pandemic especially in hospitals.
2. Sterilizing operation rooms.
3. In laboratories for sterilization
purposes.
4. Water disinfection
Ionizing radiations:

Penetrate deep into the objects.

It is called cold sterilization because it does not produce
heat.
It sterilizes by damaging bacterial DNA and proteins.
It will destroy bacterial endospores and vegetative cells;
however ionizing radiation is not always effective against
viruses.
Gamma rays
Irradiation of product using
photons from radioisotope (Cobalt
60)
Uses:
1-Pharmaceuticals (e.g. drugs and
hormones)
2-Plastic disposables such as syringes
3-Human tissue grafts
X-rays
• Higher energy and penetrating
power.
• Kill mainly by the production of free
• Radicals, these highly reactive
radicals can break covalent bonds
in DNA.
• Used for medical devices and
pharmaceuticals.
 UV Gamma X-ray
Mode of Interferes with Damages DNA of Damages DNA of
action bacterial replication microorganism microorganism
Penetration Lower High penetration High penetration
power materials can power so materials can
be sterilized inside be sterilized inside
container container

Time Longer Rapid Rapid

Safety Exposure is harmful Exposure is harmful Exposure is harmful

Beam of electrons
• They can sterilize materials at room temperature with brief
exposure.
• Limited pentation power
• Used for surgical supplies, drugs
FILTRATION
Preferred method of sterilizing solutions with heat-sensitive
components.
In the past, intravenous solutions were autoclaved, but heat-
resistant endotoxin (LPS) in the cell walls of the dead gram
negative bacteria caused fever in recipients of these solutions.
Therefore, solutions are now filtered to make them pyrogen-free
before to autoclaving.
 LPS Filter membrane

Intrav Pyrog
FILT en-
enous
RATI free
soluti solutio
ON
ons n
Heat-resistant Nitrocellulose pore
endotoxin size of 0.22 um
The most commonly used filter is
composed of nitrocellulose and has a
pore size of 0.22 um.

Filters work by physically trapping

particles larger than the pore size and
by retaining somewhat smaller
particles via electrostatic attraction of
the particles to the filters.
Chemical agents
Membrane disrupting agents

1. Ethanol
• Widely used as a skin antiseptic before immunization or
venipuncture.
• It acts by disrupting the cell membrane, and denatures proteins.
• Ethanol requires the presence of water for maximal activity
(i.e., it is more effective at 70% than at 100%).
• But, ineffective against spores
2. Detergents
Detergents disrupt the cell membrane of the microorganism.

Example:
• Quaternary ammonium compounds (e.g.,
benzalkonium chloride) are used for skin antisepsis.
• Benzalkonium chloride is the active ingredient in Lysol,
a commonly used disinfectant for floors and other
surfaces.
3. Phenols
Phenol was the first disinfectant used in the operating room (by
Lister in the 1860s), but it is rarely used today because it is
irritating.
Phenols disrupts cell membranes, and denature proteins.
Examples
• Chlorhexidine is a chlorinated phenol that is used as a hand
disinfectant before surgery and in the cleansing of wounds.

• Hexachlorophene, used in germicidal soaps, but it has limited

use due to toxicity.
MODIFICATION OF PROTEINS

1. Chlorine
• It is the active component of hypochlorite
(bleach, Clorox), which is used as a
disinfectant in the home and in hospitals.
• Chlorine is used to purify water and to treat
swimming pools.
2. Iodophor compound: Betadine ®

Iodine is the most effective skin

antiseptic
 A water soluble compound
A complex of iodine and a solubilizing
carrier.
Uses
1) Hand disinfection
2) Skin preparation before invasive
procedures:
 (a) Surgical procedures
 (b) Non surgical procedures: insertion of
intravascular catheters and venepuncture
3) Antiseptic irrigation

Iodine is effective against Staphylococcus

epidermidis which is skin flora.
3. Heavy Metals
Mercury and silver have the greatest antibacterial activity of
the heavy metals and are the most widely used in medicine.
Examples
• Merbromin (Mercurochrome), contains mercury, used as skin
antiseptic.
• Silver nitrate drops
• Silver sulfadiazine is used to prevent infection of burn wounds.
4. Hydrogen Peroxide
Used as an antiseptic to clean
wounds and to disinfect contact
lenses.
5. Formaldehyde & Glutaraldehyde
Formaldehyde (formalin 37% formaldehyde solution),
denatures proteins and nucleic acids.

Glutaraldehyde
• 10 times more effective than formaldehyde.
• Less toxic.
Uses:
Sterilize respiratory therapy equipment, endoscopes, and
hemodialysis equipment
Ethylene oxide gas
Surgical instruments and plastics
that can be damaged by moist heat
are usually sterilized by exposure to
ethylene oxide gas.

It is classified as a mutagen and a

carcinogen.
Acids & Alkalis
• Weak acids, such as benzoic,
propionic, and citric acids
• Used as food preservatives
because they are
bacteriostatic.
Activities of commonly used disinfectants

Disinfectant Bacteria Spores Fungi Enveloped

viruses
Phenol Good Nil Good Good
Hypochlorite Good Fair Good Good
Alcohols Good Nil Good Good
Glutaraldehyde Good Good Good Good
Iodophor Good Nil Good Good
MODIFICATION OF NUCLEIC ACIDS
A variety of dyes not only stain microorganisms, but also inhibit
their growth.
Examples
Crystal violet used as a skin antiseptic.
Malachite green:
• A component of Lowenstein-Jensen’s medium, which is used
to grow M. tuberculosis.
• The dye inhibits the growth of unwanted organisms in the
sputum during the 6-week incubation period.
Testing efficiency of sterilization

Biological indicators
Mechanical indicators
Chemical indicators
Biological indicators

The most effective method of monitoring

sterilization because they assess the
sterilization process directly by killing known
highly resistant microorganisms (e.g. Spores of
Geobacillus stearothermophilus.

2 vials are used, one “test vial” and the other

as a “control vial”.
 Spore test
Test vial Spores are Killed Spores are not Killed
Control vial Spores are alive Spores are alive
Efficiency test PASS FAIL

Spore test Negative Positive

Result No color change color change
Interpretation Spores are Killed Spores are alive
Chemical indicators
Uses sensitive chemicals that change color when exposed to
high temperatures or combinations of time and temperature.
Examples; tapes, strips and special markings on packaging
materials.
Results are obtained immediately following the sterilization
cycle and therefore they are faster than a spore test.
Mechanical indicators
It involves checking the sterilizer gauges, computer displays,
or printouts, and documenting in your sterilization records
that pressure, temperature, and exposure time
Unlike mechanical or chemical indicators, spore
testing determines that spores – the most resistant
microorganisms – have been killed during sterilization.
SELF-ASSESSMENT QUESTIONS

Place T for true statements and F for false ones.

1) Seventy percent alcohol is a better antiseptic than iodine, so
70% alcohol should be used to disinfect the skin prior to
drawing a blood culture rather than iodine. (F)

2) Disinfectants kill both bacterial cells and bacterial spores.

(F)
3) During sterilization by autoclaving, the temperature must
be raised above boiling in order to kill bacterial spores. (T)

4) Transmission of milk-borne diseases can be prevented by

pasteurization, which kills both bacterial cells and spores. (F)

5) Ultraviolet light used in the operating room to disinfect the

room kills bacteria primarily by causing oxidation of lipids in
the cell membrane. (F)
Which one of the following chemicals is used to sterilize
heat-sensitive materials, such as surgical instruments, in
the hospital?
(A) Benzalkonium chloride
(B) Lysol
(C) Ethylene oxide
(D) Povidone-iodine
Complete the following statements:
1) Biological controls used for autoclave is spores of
Geobacillus stearothermophilus
2) Sterilization cycle operated by autoclave utilizes temperature …
121⁰C and pressure …15 psi…
3) Autoclave is used for sterilization of … Surgical instruments...
and ……Culture media…
4) Main difference between sterilization and disinfection is… all
micro-organisms are destroyed by sterilization while disinfection
destroys micro-organisms that may cause disease only …
5) Moist heat kills microorganism by … denaturation of
proteins...while dry heat kills by…… Oxidative damage….
Give reason
1) Intravenous solutions are now filtered to make by membrane
filter before autoclaving.
2) Shorter time is needed in moist heat sterilization than dry heat.
3) Gamma rays can be used in medical devices factories to
sterilize closed containers.
4) Heat-sensitive surgical instruments and plastics are sterilized by
ethylene oxide gas
5) An autoclave is ineffective if the spores from the test vial are
not killed during monitoring sterilization
Answers
1) Membrane filters removes heat-resistant endotoxin (LPS)
which may cause fever in recipients of these solutions
2) Moist heat has higher pentation power and higher efficiency
3) Gamma rays has high pentation power
4) Ethylene oxide gas can sterilize them at room temperature
unlike other methods that uses heat it may destroy Heat-
sensitive surgical instruments and plastics
5) Because spores of Geobacillus stearothermophilus are highly
resistant to high temperatures if not killed then sterilization
fails.