Sterilization: Sterility Test
Sterilization: Sterility Test
Sterilization: Sterility Test
➢ The ultimate goal in the manufacture of a sterile product is absolute absence of
microbial contamination.
➢ Inactivation of MOs by physical or chemical means.
➢ The D value is a single quantitative expression of the rate of killing of
microorganisms.
➢ This is the time required for a 90% reduction in the microbial population. Hence, the
time or dose it takes to reduce 1000 microbial cells to 100 cells is the D value.
➢ Sterilization Assurance limit (SAL): The product will be sterilized by a sterilization
process sufficient to produce a probability of non sterility of one out of 1 million
containers .
➢ SAL= 10-6(1 viable microorganism in 106 sterilized item of the final product).
➢ Whenever possible terminal sterilization is chosen, sterilized in its final container &
possible with heat stable particles.
❖ Sterility test
➢ Sterility test is applied to the Pharmaceutical preparations that are required
to sterile preparation like parenteral and ophthalmic preparation.
➢ To detect the presence of viable form of microorganism.
➢ Sterility test is carried out at aseptic condition(grade A laminar airflow
cabinet)
➢ The microorganism are placed in a culture medium which provides nutrition,
water, and favourable temperature/condition, microorganism will grow and
their presence detected in culture medium.
Method of sterilization:-
❖ Physical Method
➢ (a) Thermal (Heat) methods
➢ (b) Radiation method
➢ (c) Filtration method
❖ Chemical Method
➢ (a) liquid
➢ (b) Gases
1. Steam sterilization/moist heat sterilization/autoclave
● Used for aqueous & surgical materials.
● most widely used and reliable method of sterilization
● The efficiency to inactivate microorganisms is dependent upon the degree of
heat, the exposure time and the presence of water.
● Sterilization is carried out using saturated steam under pressure. Saturated
steam cause destruction of enzyme and other cellular component and protein.
● Autoclaving also kills virus
● not suitable for rubber, plastics, and equipment that would be damaged by
high temperatures
Holding temp. Holding time (minutes)
115-118 ℃ 30 minutes
134-138 ℃ 3 minutes
2. Dry heat sterilization
160 ℃ 2 hrs
170 ℃ 1 hrs
180 ℃ 30 minutes
3. Sterilization by filtration:
● Used for thermolabile/heat sensitive Drugs
● The size of filter medium pores to retain micro-organisms must be quite small. The
0.20- or 0.22 pore size filter media are considered to be capable of producing sterile
filtrates.
● Both clarification and sterilization of liquid preparation
● Filtration process does not destroy but removes the microorganisms by adsorption
and trapping within the matrix of the filter material.
● The bubble point test is a popular single-point physical integrity test for disc filter
membrane.A filter medium is wetted with a liquid, and test gas pressure is slowly
raised until a steady stream of bubbles appears from a tube or hose attached to the
downstream side of the filter and immersed in water (Fig. 9). The pressure at which
the bubbles first appear is recorded as the bubble point and is related to the largest
pores in the filter medium.
4. Ethylene oxide /(CH2)2O):
● Ethylene oxide is a colorless, odorless, and flammable gas.
● Cause alkylations of sulphydryl, amino, hydroxyl and carboxyl groups on proteins
and amino groups of nucleic acids/DNA.
● produce acute toxicity including irritation of the skin, conjunctiva and nasal mucosa.
Bio-logical indicators
● Biological indicators (BIs) are live spore forms of micro-organisms known to be the
most resistant living organisms to the lethal effects of the particular sterilization
process.
● Provide the effectiveness of a given sterilization process/validation of sterilization
process.
● Biological indicators are viable culture of known species of microorganism.
S. Methods of Material to be Biological indicator
No. sterilization sterilized
1 Moist Heat Saturated steam under pressure Aq. & Surgical Bacillus
(steam) items strearothermophilus