Pharmaceutical Sterilization

Alaaldin M. Alkilany, PhD

Professor of Pharmaceutics and Nanotechnology

College of Pharmacy
Qatar University 1
Sterilization Methods
• A sterilization process should represent a compromise between
achieving target SAL and maintaining product stability.
• Validation based on the intended application and continuous
monitoring of a sterilization process is always required.
• Sterilization is not an alternative to GMP; it is only the final stage in
a program of microbiological control.
• 5 methods of sterilization are recognized in Eur Ph:
1. Steam sterilization (heating in an autoclave)
2. Dry heat
3. Gas sterilization
4. Ionizing radiation
5. Filtration
Heat Sterilization
• Heat is the most reliable and widely used means of sterilization
• Heat sterilization is limited to thermostable products.
• For moisture-resistant products: moist heat (121-134 °C) sterilization
is used
• For moisture-sensitive products: dry heat (160-180 °C) sterilization is
used.

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Heat Sterilization Processes
• Heat sterilization cycle contains a heating-up stage, a holding stage and a cooling
stage.
• Holding stage is the main stage for killing. However, during both the heating-up and
cooling-down stages, the product is held at an elevated temperature and these
stages contribute to the overall biocidal potential of the process.

The most commonly employed standard temperature/time cycles for bottled fluids and
porous loads (e.g. surgical dressings) are 121 °C for 15 minutes and 134°C for 3 minutes,
respectively.
Moist Heat Sterilization
Applications:
• culture media
• containers and closures
• aqueous injections
• ophthalmic preparations
• irrigation fluids
• surgical and diagnostic equipment
• dressings, sheets
• processing (decontamination) of soiled
and contaminated items
 Dry Heat Sterilization
• The instrument used is a hot air oven with perforated
shelves to allow for heat & air flow
• Higher temperatures in the range 160-180 °C are used
at normal air pressure
• Its application is generally restricted to glassware and
metal surgical instruments, non-aqueous
thermostable liquids and thermostable powders
• The major industrial application is in the sterilization
of glass bottles which are to be filled aseptically, and
here there is another advantage which is destroying
bacterial endotoxins (pyrogens) which are difficult to
be eliminated by other means.
• For Depyrogenation of glass, temperatures of
approximately 250 °C are used.
 Filtration Sterilization
• The process of filtration is unique amongst
sterilization techniques in that
- it removes, rather than destroys, microorganisms
- It eliminates both viable and non-viable particles
thus it can be used for both the clarification and
sterilization of liquids and gases
• Applications :
- Sterilization of heat-sensitive injections and
ophthalmic solutions and biological products
- Treatment of air and other gases for supply to
aseptic areas
- They may act as part of venting systems on
industrial machines such as fermenters,
centrifuges, autoclaves and freeze-dryers
Note: Membrane filters are used in sterility testing
where they can be employed to trap and concentrate
contaminating organisms from solutions under test.
These filters are then placed on the surface of a solid
nutrient medium and incubated to encourage colony
development
Filtration Sterilization
• The major mechanisms of
filtration are sieving,
adsorption and trapping
Cellulose acetate filters
within the matrix of the filter
material.
• Synthetic membrane filters
(derived from cellulose esters
or other polymeric materials),
Fibrous pads, sintered glass Sintered glass filters
and sintered ceramic products
can be used.

Ceramic filters
Filtration Sterilization - Liquids
• Membrane filters of (0.2-0.22 µm)
nominal pore diameter are most
commonly used.
• Two filters of 0.2 mm pore diameter
from different manufacturers will not
behave similarly, because, in addition to
the sieving effect, trapping, adsorption
and charge effects all contribute
significantly towards the removal of
particles.
• Consequently, the depth of the
membrane, its charge and the
tortuosity of the channels are all factors
which can make the performance of
one filter superior to that of another.
Filtration Sterilization - Liquids
• Membrane filters, in the form of discs, can be
assembled into
- Pressure-operated filter holders for syringe mounting and
in-line use
or
- Vacuum filtration tower devices
Filtration Sterilization - Gases
• The principal application for filtration sterilization of gases
is in the provision of sterile air to
ØAseptic manufacturing sites
ØHospital isolation units
ØSterilization of venting air in tissue and microbiological culture
ØDecontamination of air in mechanical ventilators
ØTreatment of exhausted air from microbiological safety cabinets
ØThe clarification and sterilization of medical gases

HEPA
filter can remove 99.97% min of particles
greater than 0.3 mm
Sterilization Control & Sterility Assurance
• Currently, awareness of the limitation of sterility
testing in terms of their ability to detect low
microbial count resulted in relying on satisfactory
quality standards/practices during the whole
manufacturing process.
• i.e. the quality is assured by process monitoring &
performance criteria, which are considered under
four subjects:
1. Bioburden determination (API, excipients and
packaging)
2. Environmental monitoring (manufacturing area and
regular check!)
3. Validation & in process monitoring of sterilization
procedures (chemical and biological indicators)
4. Sterility test
1. Bioburden Determination
• It is important to have a low pre-sterilization
bioburden by:
• having high quality of raw materials,
• the environment does not encourage microbial growth
of the m.o already present in raw material,
• low microbial contamination during manufacture.
• Therefore, manufacturing process may utilize
adverse temp, extreme pH, organic solvent in order
to prevent increase in the microbial load.
2. Environmental Monitoring
• The level of microbial contamination in manufacturing
areas is monitored regularly so as not to exceed certain
levels.
• M.O. in atmosphere are monitored by settle plates &
air sampler.
• Contamination on surfaces or manufacturing
equipment is measured by swabs or contact plates
(Rodac-replicate organism detection & counting-plates)
which are petri dishes overfilled with agar media
• Operators in manufacturing areas are monitored.
samples from clothes (face mask, gloves & fingerprint)
are taken

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3. Validation & In-Process Monitoring of
Sterilization Procedures

• Validation: demonstrating that a process will

consistently produce the results that it is intended to.
• Validation of steam sterilizer include various elements
such as:
• Calibration of all physical instruments used to monitor the
process (thermocouples, pressure gauge, timers..etc)
• Use of process efficiency indicators (alone or with bioburden
m.o.)
• Show repeatability of the process (at least repeat 3 times)
• Documentation for all the previous steps
• Usually records of temp, pressure, time, humidity are kept for
each batch of sterilized product
 3. Validation & In-Process Monitoring of
Sterilization Procedures
• Biological indicators:
• Used for thermal, chemical or radiation sterilization
• they consist of standardized bacterial spore preparations
which are either suspension in water or culture medium
or spores dried on paper, Al or plastic carrier.
• After sterilization, the spores are aseptically transferred
to nutrient medium, incubated & periodically examined
for signs of growth.
• Selection of BI:
Ø Must be non-pathogenic
Ø Should possess above-average resistance
to the particular sterilization
4. Sterility Test
• A test which assesses whether a sterile
pharmaceutical or medical product is free from
contaminating m.o. or not.
• To be sure that no organism is present; a universal
growth medium that supports the growth of all
types of m.o. should be used, which is not available.
Practically, media capable of supporting non
fastidious bacteria, yeast & mould are used
• Pharmacopeial tests do not look for viruses, which
can pass thru sterilization filters.
4. Sterility Test
• Disadvantages of Sterility Test:
• Destructive test
• Questionable suitability for testing large
expensive or delicate products
• It is a statistical process where part of a batch is
randomly sampled & batch is released based on
this sample
• The procedure intends to demonstrate a
negative! i.e. failure to detect m.o. could be a
consequence of the use of unsuitable media or
inappropriate cultural conditions.
Sterile Manufacturing: Essential
requirements to minimize contamination
• Separated from other units
• Special construction
requirement (material and
design to prevent
contamination and allow
ease of cleaning and
disinfection and
maintenance).
• Entry through airlocks (for
both people/ goods)
• Clean rooms (A, B, C and D)
(HEPA filters)
• cGMP and adherence to
regulatory guidelines
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Regulatory guidelines
 Sterile Manufacturing unit: Essential
requirements to minimize contamination
• HEPA filters on the ceiling
• Exhaust vents on the floor
• Laminar air flow
 Sterile Manufacturing unit: Essential
requirements to minimize contamination
• Seamless and rounded floor to wall junctions
• Readily accessible corners Floors, walls and ceilings constructed of
smooth hard surfaces that can be easily cleaned
• Limited equipment, personnel
• Layout of equipment to optimize movement of operators
• Airlock and interlocking doors to control air balance and avoid
contamination
Airlock systems
• Goal: Preventing the classified area from the
contamination that may occur during the entry and
exit of personnel and material
• Minimize contamination of dust and particulates
• The airlock consists of a relatively small chamber
with two airtight doors, which do not open
simultaneously (interlock system with alarm if both
are open)
• Doors open to higher pressure side
• High air change (min 20 changes per hour)
 Types of airlock

15 Pa 25 Pa 30 Pa 15 Pa 30 Pa 15 Pa
+ ++ +++ + +++ +

Cascade airlock bubble airlock

30 Pa 15 Pa 30 Pa
+++ + +++

sink airlock
More essentials…
Air showers

Gowning rooms
Types of sterile manufacturing
• Sterile manufacturing with terminal sterilization
• Aseptic manufacturing
vProduction in a controlled environment in order to
prevent any microbiological contamination
vMaterials used in the process are previously
sterilized and introduced in the manufacturing area
as sterile
vThis approach is designed to avoid any
contamination during the production and filling
processes for products which cannot be produced
by terminal sterilization (sterilized at the end of
production process using available sterilization
techniques due to instability)
Types of sterile manufacturing