INDUSTRIAL

STERILIZATION ON LARGE
SCALE

Deepankar ratha
Assistant professor,
CUTM,Rayagada
STERILIZATION
 Sterilization can be defined as the
process through which all forms of life
are destroyed, removed or
permanently inactivated.

 Sterilization is essential concept at

large scale as well as small scale for
the preparation of sterile
pharmaceutical products.
 It’s aim is to provide a product that is
safe and eliminates the possibility of
contamination.
WHY
 To reduce amount of contaminant’s
present in environment, on surface of
container’s, closure’s as well as
equipment’s and to achieve better
sterile condition.
SOURCE’S
 Raw materials
 Equipment & instrument’s
 Manufacturing process
 Container & closure system
 Manufacturing environment
 Worker’s
STERILIZATION
PROCESSES
 The sterilization is done by
1. Physical method
2. Chemical method
Physical Method
 Moist heat
1. saturated steam autoclave
2. superheated water autoclave
3. air over steam autoclave

 Dry heat
1. continuous tunnel sterilizer
2. hot air oven
3. Red heat
4. Flaming
 Radiation
1. Non-ionizing radiations
▪ Ultraviolet (UV) light
2. Ionizing radiations(cold sterilization)
▪ X-rays
▪ Gamma rays
▪ Cathode rays
 Filtration
1. Depth filter
▪ sintered glass filter
2. Screen filter
▪ particulate filter
▪ microbial filter
Chemical Method
 Chemical sterilization
◦ Gaseous sterilization
 Ethylene oxide
 Formaldehyde

◦ By using disinfectant or antimicrobial

agent
PROCESS SELECTION
 Process selection is most important
and sensitive point for preparation of
product.
 Those product intended to be sterile
should be terminally sterilized in their
final container as clearly stated in
EUROPIAN PARMACOPOEIA.
 Where it is not possible to carry out
terminal sterilization, choose the
alternative method.
 It is recognised that new terminal
sterilization process other than those
describe in p’copoeia may be
developed to provide sterility
assurance level equivalent to present
official method, & such process when
properly validated may offer
alternative approaches.
 The use of an inappropriate heat labile
packaging material can not in itself be
the sole reason for adoption of aseptic
processing, rather manufacturers
should choose the best sterilization
method achievable for a given
formulation & select the packaging
material accordingly.
 The choice of packaging material for
given product has to take into account
factors other than the method of
sterilization.

 In such cases these other factor need

to be clearly documented, explained &
scientifically justified.
PROCESS FOR AQUEOUS
PREPARATIONS
If the product is aqueous
Can product be sterilize by
moist heat at 121°C for 15min
Can the product be
Use autoclaving at
sterilize by moist heat
121°C for 15min
withF0 ≥8 min

Can the formulation

be filtered through a Use moist heat with
microbial retentive F0 ≥8 min
filter
 Use pre sterilized
Use combination of
individual component &
aseptic filtration &
aseptic compounding &
aseptic processing
filling
 If the product is non-aqueous, semi
solid or dry powder
Can the product be sterilized by
dry heat at 160°C for 120min

Can the product be

sterilized by dry heat with Use sterilization at
an alternative 160°C for 120 min
combination of time &
temp° so the standard
cycle achieving an SAL of
≤10-6
 Use dry heat with an
Can the product be
alternative combination of
sterilized by a method
time & temp° so the
different from dry heat
standard cycle achieving
e.g. ionization
an SAL of ≤10-6

Use a method different

Can the sterilized by
from dry heat e.g.
validated lower
ionization
irradiation dose
Can the formulation be Use the sterilized by
filtered trough a validated lower
microbial retentive filter irradiation dose

Use pre-sterilized
individual components & Use filtration & aseptic
aseptic compounding & technique
filling
SPECIFICATIONS
 MOIST HEAT STERILIZATION
◦ Micro organisms destroyed by cellular protein
coagulation
◦ The objects to be sterilized are exposed to
saturated steam under 1 atmosphere
pressure at a minimum temperature of 121°C
for 15 min.
◦ An autoclave is commonly used for moist
heat sterilization.
◦ Because it does not require as high a tempº,
moist heat sterilization cause less product
and equipment damage compared to dry
heat sterilization.
AUTOCLAVE
 Is a device to sterilize equipment and supplies
by subjecting them to high pressure saturated
steam at
Temp°C lb/sq.inch Time(min)
115-118 10 30
121-124 15 15
126-129 20 10
135-138 30 3

TYPES
 Portable autoclave (Bench autoclave)
 Stationary autoclave (Large sterilizer)
 Main Features
◦ Lid(door) fitted with clamps and asbestos
jacket , stationary autoclave may be double
doors at both ends one for loading and one
for unloading.
◦ Pressure gauge
◦ Thermocouple for measurement of tempº.
◦ Air vent to remove air before sterilization.
◦ Safety valve to permit escape of excess
steam to prevent explosion .
◦ Modern autoclaves are recording (record
pressure, temp during the whole process )
supplied with timer and are automatically
controlled .
 DRY HEAT STERILIZATION

◦ Is appropriate for materials that cannot

withstand moist –heat sterilization (e.g., oily
materials and powders)
◦ Objects are subjected to a temperature of at
least 160º for 120 minutes( if higher
temperatures can be used , less exposure
time is required)
HOT AIR OVEN
 Is a device to sterilize subject and supplies by
subjecting them to direct heat at
Temp°C Time(min)
170 60
160 120
150 150
140 180
 Is appropriate for the materials that can not
withstand steam sterilization (e.g. oily materials
& powders)
 If higher temperature can be used, less
exposure time is required.
 Main Features
◦ Door fitted with the clamps and asbestos
jacket has the single door
◦ Regulator for temperature control
◦ Fan attached inside for air circulation
◦ Perforated shelf for keeping subject inside
 RADIATIONS

◦ Energy transmitted through space in variety

of forms is generally called radiation
◦ This method is also called as cold
sterilization because it produce relatively little
heat
◦ Thus, it is possible to sterilize heat sensitive
materials such techniques are being in food
industries mainly
 Non ionizing radiations (UV)
◦ UV in region of 2537 A° has been shown to
posses the greatest activity in destroying MO
◦ Commonly employed in reduction of air-
borne contamination in the maintenance of
aseptic areas & rooms
◦ Source of artificial UV radiation s is UV lamps
(generally called sterilizing lamp or germicidal
lamp)
◦ UV light is absorbed by the nucleic acid of
the cell where it does the greatest damage
 Ionizing radiations (cold sterilization)

◦ X-rays, gamma rays & cathode rays are

lethal to DNA & other vital cell constituents
◦ They have high penetration power &
considerable energy
◦ The factors that effect the lethal activity of
ionizing radiations are oxygen effect,
protective compounds, sensitizing agents, pH
of cultures, freezing, moisture & recovery
conditions
 Filtration

◦ This is a non-thermal method of sterilization

used widely in the p’ceutical industry where
heat labile solutions are to be sterilized
◦ This is useful for large volume solutions, eye
drops, antibiotic solutions, sera &
carbohydrate solutions
◦ This also useful for separation of
bacteriophages & bacterial toxins from
bacteria for the isolation of MO which are
scanty in fluids
◦ 3 main stages involved in filtration

1. Passage of the solution through a

previously sterilized bacteria-proof filter
unite
2. Aseptic transference of filtrate to sterile
containers then sealed aseptically
3. Testing of sample for sterility
 Gaseous sterilization

◦ The destruction of all living MO with chemical

in gaseous or vapours state
◦ When material is affected by the dry or moist
heat then the gaseous sterilization is used
◦ All these gases are toxic to human being
above certain conc. and may exhibit other
undesirable side effect
◦ Ethylene oxide is most widely used gaseous
sterilization agent in pharmaceutical industry
◦ In addition to these, various glycols, methyl
bromide and alcohol have been used for
room sterilization
 Ethylene oxide
◦ It is colorless liq. with BP 10.8°C
◦ Highly inflammable and may be explosive
when mixed with air in conc. Greater than
3%. Its mixture with CO2 in certain
proportion makes it inflammable
◦ Conc. & time relationship commonly for
sterilization is as below
conc.(mg/lit) exposure
ti e(hrs.)
44 24
88 10
442 4
884 2
 Disinfectant or antimicrobial agents

◦ Chemical agents most commonly used as

disinfectant ant antiseptic e.g. phenols,
alcohols, halogens, dyes, aldehyde etc.
DEVELOPMENT & VALIDATION
OF PROCESS & EQUIPMENT

Process validation:
• It is analysis of data gathered
throughout the design &
manufacturing of product in order to
confirm that the process can reliably
output products of determined
standard.
• Regulatory authorities like EMA & FDA
have published guidelines relating to
process validation
 The purpose of validation is to ensure
varied inputs lead to consistent & high
quality outputs
 Process validation is an ongoing
process that must be frequently
adapted as manufacturing feedback is
gathered
 End to end validation & production is
essential in determining product
quality because quality can not always
be determined by finished product
inspection
 Process validation can be broken in to 3
steps
1. Process design
2. Process qualification
3. Continued process verification

 Process design:
In this data from the development
phase are gathered & analyzed to define
the commercial manufacturing process.
The data used to establish benchmark for
quality & production control
 Process qualification:
In this stage the process design
assessed to conclude if the process is able
to meet determine manufacturing target.
All the process & manufacturing
equipment is proofed to confirm quality &
output capabilities

Continued process validation:

It is the ongoing monitoring of all
aspects of production cycle. It aims to
ensure that all levels production are
controlled & regulated
EQUIPMENT VALIDATION/
QUALIFICATION
 Equipment validation is divided into

1. Design qualification
2. Installation qualification
3. Operational qualification
4. Performance qualification
 DQ:
It define the functional &
operational specification of the
instrument & details for the continues
design in selection of supplier

 IQ:
Demonstrates that the process or
equipment meets all specifications, is
installed correctly, and all required
components and documentation
needed for continued operation are
installed and in place.
 OQ:
Demonstrates that all facets of the
process or equipment are operating
correctly.
 PQ:
It is the process of demonstrating
that an instrument consistently
performed according to specification
appropriate for it’s routine work