COLLEGE OF HEALTH AND ALLIED PROFESSION

PHARMACY DEPARTMENT

METHODS OF
STERILIZATION
DAVID S. HINOLAN, RPH
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

 SIGNIFICANCE OF WHY STERILIZATION IS

PERFORMED
 METHODS OF STERILIZATION
 MACHINES AND CHEMICAL AGENTS
CATEGORIZED UNDER A SPECIFIC
METHOD
 ACTUAL APPLICATION OF THESE
CONCEPTS TO THE HEALTHCARE
PRACTICE
COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

SIGNIFICANCE OF
STERILIZATION
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

1. Protection of our patients

2. Prevention of the spread of infection
3. Prevention of a sudden change in a product’s
microbiological properties
COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

METHODS OF
STERILIZATION
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

A. Moist Heat Sterilization/Heat Sterilization

B. Dry Heat Sterilization
C. Radiation
D. Physical Separation
E. Sterilization through chemical agents
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

A bacteria’s structure is
highly composed of
proteins, glycoproteins and
fatty acids.

The outer structure (cell

wall and biofilms) of the
bacteria is the one
responsible of protecting it.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Moist Heat Sterilization

Mechanism: Protein Denaturation
1. AUTOCLAVE
• 121 Degrees Celsius
• 15-16 psi
• 15 minutes

This method is only applicable to materials that are not water

labile. A widely used sterilization method because of its ability to
kill not only bacteria, fungi and virus, but also, spores from spore
forming bacteria such as Clostridium and Bacillus family.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Dry Heat Sterilization

Mechanism: Oxidation
1. Oven
• 170 Degrees Celsius
• 2 Hours

This method is highly applicable to your raw materials (dried

extracts, powdered active ingredients and granules) that are not
water labile and those which cannot be sterilized using moist heat
method.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

DRY HEAT
STERILIZATION
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

DRY HEAT
STERILIZATION
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Radiation
Mechanism: Genetic Mutation
1. Ionizing Gamma (y) Irridiation Cobalt-60
2. Ultraviolet Radiation
3. Electron Beam Cathode

The type of sterilization method is used to plastic and other

materials that are heat- and water-labile. The ionizing light kills the
microorganism through the formation of free radicals, and by
exciting DNA/RNA’s covalent bonding, thus leading to a drastic
chemical reaction, impeding the replication process inside the cells.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Radiation
Disadvantages
1. Shadowing Effect
2. Varied dosing, in kGy, for different materials
3. May potentially affect stability of active ingredient and
excipients, thus lowering the absorption, potency and efficacy.

What is kGY?
• It stands for Kilogray or 1000 gray. Gray is, on the other hand,
defined as 1 Joule/Kg heat absorbance on a specific object.
• Quantified via Dosimeter.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Cobalt-60

A radioisotope of
Cobalt-60, created by
bombarding Cobalt-59
with neutron.

SAL: 25 – 32 kGy
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

UV Radiation
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Electron Beam Cathode

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Physical Separation
Mechanism: Filtration
1. Adsorption
2. Sieve Retention

Three Types of Filter

3. Porcelain Filter Cartridges
4. Abestos Cellulose Layer
5. Microporous Membranes (Zgismondy, first brand of
commercially available and successful filter)
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Physical Separation
Membrane Retention Challenge Test
- Ability to remove 1 x 107 per cm2 of Brevundimonas diminuta

This type of sterilization method is useful for substances that are

heat-labile and are easily degraded by radiation. This is commonly
seen in the sterilization procedure of vaccines, parenteral and
suited biological products.
COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Porcelain Filter
Cartridges
COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Abestos Cellulose
Filter
COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

0.1 um

Microporous or
Membrane
0.45 um
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Sterilization through chemical agents

Mechanisms
• Alkylation
• Protein Denaturation
• Halogenation
• Oxidation
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Alkylation
Agent used: Ethylene Oxide

Ethylene oxide causes alkylation of a

microorganism’s nucleic acid, thus ceasing
the proper linking of protein bonds, thus
leading to an eventual cellular death.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Protein Denaturation
Agent used: Ethanol

Ethanol alters the proteins found in the

cell wall and membrane of a
microorganisms, lowering the cell
membrane integrity, thus causing the
leakage of cytoplasmic fluid. It also
denatures the lipid membranes.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Halogenation
Agent used: Povidone Iodine

Interacts with the nucleotides, fatty acids

and proteins both in the cytoplasm and
cytoplasmic membrane. It denatures the
structures, thus leading, to the inability to
perform specific function, impeding
respiration and leading to eventual cell
death.
 COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

Oxidation
Agent used: Diluted Potassium Permanganate

Acts by oxidizing the cells, attacking the genetic

integrity thus leading to cell death.
COLLEGE OF HEALTH AND ALLIED PROFESSION • PHARMACY

THE
END