O MY LORD!

OPEN FOR ME MY
CHEST (GRANT ME SELF-
CONFIDENCE, CONTENTMENT, AND
BOLDNESS); EASE MY TASK FOR
ME; AND REMOVE THE
IMPEDIMENT FROM MY SPEECH,
SO THEY MAY UNDERSTAND
WHAT I SAY!
[SURAH TA-HA; 20:25-28]

In the name of Allah the most

gracious and the most merciful
1
 METHODS OF
STERILIZATION
(FILTRATION)
University College of Pharmacy,
University of the Punjab, Lahore

Presented by:
Ahmad Mehmood
Muhammad Muneeb
Presented to:
Dr. Amjad Hussain 2
 STERILIZATION
 Involves the destruction or removal of
all living microbes, spores and viruses on
an objective or in an area

 For this thing the object obtained after

sterilization is called sterile

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METHODS OF STERILIZATION
 Dry Heat Sterilization

 Moist Heat Sterilization

 Gaseous Sterilization

 Radiation Sterilization

 Filtration

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 FILTRATION
 Filtration is any of various mechanical,
physical or biological operations that
separate solids from fluids (liquids or gases)
by adding a medium through which only the
fluid can pass.

 The fluid that passes through is called

the filtrate.

 Physical Method, just removes the bacteria

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 Filtration occurs both in nature and
engineered system:
 Biological systems
 Geological systems
 Industrial forms

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 HISTORY
 Filters were used to trap inborne organisms
and sterile growth media.
 In microbiology – first use in 1890s
 Filter technology
 Charles Chamberland – Porcelain filter
 Julius Petri – Petri dish

(to cultivate growth media)

and developed sand filter
to separate bacterial cells
from the air

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 TYPES
1. Hot Filtration
Solids from a hot solution
2. Cold Filtration
Use of ice bath in order to rapidly cool down
the solution to be crystallized rather than
leaving it out to cool it down slowly in the
room temperature
3. Vacuum Filtration
Technique is most preferred for small batch
of solution in order to quickly dry out small
crystals
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FILTRATION STERILIZATION
 Used for sensitive pharmaceuticals and
protein solutions in biological research

with pore size 0.2 µm will effectively

 Filter
remove bacteria

 Ifviruses must also be removed, a much

smaller pore size around 20 nm is needed.

 Prions are not removed by filtration.

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 The filtration equipment and the filters
themselves may be purchased as presterilized
disposable units in sealed packaging.

 Or must be sterilized by the user, generally by

autoclaving at a temperature that does not
damage the fragile filter membranes.

 To ensure sterility, the filtration system

must be tested to ensure that the membranes
have not been punctured prior to or during use.

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 MECHANISM
 The major mechanisms of filtration are:
 Sieving
 Adsorption
 Trapping within the matrix of the filter material

 The potential hazard of microbial

multiplication within a depth filter and
subsequent contamination of the filtrate
(microbial grow through) should be recognized.

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 MATERIALS
 Useful for substances which get damaged by
heat
 To sterilize:
 Sera
 Sugar solution
 Antibiotic solution
 To obtain bacteria free filtrates of clinical
samples
 Purification of water

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TYPES OF FILTER PAPER
 Membrane filter

 Candle filters

 Asbestos disc filter

 Sintered glass filters

 Air filters

 Syringe filters
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MEMBRANE FILTER
 Most common
 Pore size 0.22 μm
 Known vol of sample
is used
 Consists of
 pad of nitrocellulose
acetate / polycarbonate
 Mounted in a holding
device

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Click icon to add picture

PROCESS OF
MEMBRANE FILTER

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MATERIAL USED:
 Cellulose
 Glass wood
 Faber glass mixture
 Polytetrafluroethyl-
ene (PTEE)

 Standards may vary

from lab to lab

(Fuel Filters)
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 Materials that can be Importance
filtered
 For softening of water
 Heat sensitive liquids filter beds are used
 Beverages  Bacterial cells trapped
 Some growth media
on filter can grow
 Toxicoids
 Blood solutions
 Can filter
 Gram-negative
 Pharmaceuticals
 micro-aerobic rod
 Injections
 IV drips  Microbiologist can
 Solutions count colonies to
 Parenteral determine the number
 Ophthalmic preparations of bacterial cells
originally present

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 Advantages Disadvantages
 Viruses can enter the
pores (200 nm)
 Cheap
 Particle entrapment may
take place as a result of
 Time saving adsorption
 Fluid must be relatively
 No complex method free of suspended
particulate material
 Require regular
 Easy to handle sterilization (MOIST) to
prevent microbial
colonization and grow of:
 Cladosporium sp.
 Stachyotrys sp.
 Pollutants
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NANO-WATER
FILTERS
(Bacteria from a marine water
sample are retained by this
membrane filter)

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HEPA FILTER
(HIGH-EFFICIENCY PARTICULATE AIR)
 Consists of mat of randomly arranged fibers
that trap:
 Particles
 Microorganisms
 Spores
 Part of Biological Safety Cabinet – filters
over 99% of all particles
 Range – 0.3 μm
 Filters air

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IMPORTANCE
 To ensure the best results,
pharmaceutical sterile filtration is
performed in a room with highly filtered
air (HEPA filtration) or in a laminar
flow cabinet or "flow box", a device which
produces a laminar stream of HEPA
filtered air.

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 HEPA filters are critical in the prevention
of the spread of airborne bacterial and
viral organisms and, therefore, infection.

 Typically, medical-use HEPA filtration

systems also incorporate high-energy
ultra-violet light units to kill off the live
bacteria and viruses trapped by the filter
media.

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Click icon to add picture

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APPLICATIONS
 Medical-use HEPA filtration systems also
incorporate:
 high-energy UV light units to kill off the live
bacteria
 and viruses trapped by the filter media

 To ensure air purity in:

 Respiratory
disease wards
 Pharmaceutical filling rooms

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 Advantages Disadvantages

 Recommended use in  Require constant

air dysfunction maintain and cleaning

 Can suck up second  Not helpful with odors

hand smoke and even
eliminate any smell in  complicated to free
the air
the air of odors and
gases
 99.9 percent dust
free environment  Quite expensive

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 Aseptic processing is  A process whereby a
the process by which product
a sterile (aseptic) is sterilized in its
product (typically final container or
food or packaging, which
pharmaceutical) is permits the
packaged in a sterile measurement and
container in a way evaluation of
that maintains quantifiable microbial
sterility. lethality.

Aseptic Sterilization Terminal Sterilization

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IMPORTANCE OF Filtration IN
PHARMACEUTICAL INDUSTRY
Epidural Injections:
 Epidural injections are frequently given
through a bacterial filter

Small Volume Injections:

 The drug is normally dissolved in the oil,
filtered under pressure and distributed into
ampoules. After sealing, the ampoules are
sterilized by dry heat.

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Eye-Ointments:
 Eye ointments are prepared in a semi-solid
base. The base is filtered when molten to
remove particles and sterilized at 160°C for
2 hours. The drug is incorporated prior to
sterilization.

Freeze-Dried Products:
 Freeze-drying consists of preparing the drug
solution, filtering through a bacteria-proof
filter, dispensing into containers, removing
water in a freeze-drier, then capping and
closing the containers. In the future it is
probable that many biotechnology.
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OTHER APPLICATIONS
 Sterilization of venting or displacement air in tissue
and microbiological culture (carbon filters and
hydrophobic membrane filters)

 Decontamination of air in mechanical ventilators

(glass fibre filters)

 Treatment of exhausted air from microbiological

safety cabinets (HEPA filters)

 The clarification and sterilization of medical gases

(glass wool depth filters and hydrophobic membrane
filters)
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