INDUSTRIAL STERILIZATION

ON LARGE SCALE

By
Mr. NILESH UTTAM UTPURE
Under the guidance of
Prof. BIDKAR J. S. sir
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.
 Its aim is to provide a product that is safe
and eliminates the possibility of
contamination.
WHY
To reduce amount of contaminants
present in environment, on surface of
containers, closures as well as
equipments and to achieve better sterile
condition.
SOURCES
Raw materials
Equipment & instruments
Manufacturing process
Container & closure system
Manufacturing environment
Workers
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 pcopoeia 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 121C for 15min
Can the product be
Use autoclaving at
sterilize by moist heat
121C for 15min
withF0 8 min

Can the formulation be

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

Can the product be

sterilized by dry heat with Use sterilization at 160C
an alternative combination for 120 min
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 e.g.
standard cycle achieving an
ionization
SAL of 10-6

Can the sterilized by Use a method different from

validated lower irradiation dry heat e.g. ionization
dose
 Can the formulation be Use the sterilized by
filtered trough a microbial validated lower irradiation
retentive filter 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 121C for 15 min.
An autoclave is commonly used for moist heat
sterilization.
Because it does not require as high a temp,
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
TempC 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 temp.
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 160 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
TempC 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 pceutical 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 10.8C
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 time(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
its routine work