SNS COLLEGE OF ALLIED HEALTH SCIENCES

SNS Kalvi Nagar, Coimbatore - 35

Affiliated to Dr MGR Medical University, Chennai

TOPIC : ASEPTIC FILTRATION OF LIQUIDS AND AIR LIQUIDS

 FILTRATION

• Preferred method of sterilizing heat-sensitive liquid and gases without exposure to

denaturing heat.
• Rather than destroying contaminating microorganisms, it simply removes them.
• Sterilize antibiotic solutions, toxic chemicals, radioisotopes, vaccines, and
carbohydrates, which are all heat-sensitive.
• Capable of preventing the passage of both viable and nonviable particles and can
thus be used for both the clarification and sterilization of liquids and gases.
• The principal application of sterilizing grade filters is the treatment of heat-sensitive
injections and ophthalmic solutions, biological products and air and other gases for su
pply to aseptic areas.

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 FILTERS

• The liquid or gas is passed through a filter, a device with pores too small for the
passage of microorganisms, but large enough to allow the passage of the liquid or gas.
Materials Name of the filter
Asbestos pad Seitz filter
Diatomaceous earth Berkefeld filter
Procelain Chamberland-Pasteur filter
Sintered glass disks Sintered glass filter
Cellulose Membrane filter
Borosilicate glass fiber HEPA filter
Clay, mud Candle filter
 FILTERS

• Filters may also be required in industrial applications where they become part of venting
systems on
1. Fermenters,
2. Centrifuges,
3. Autoclaves
4. Freeze-driers.
• Certain types of filter (membrane filters) also have an important
role in sterility testing, where they can be employed to trap and concentrate
contaminating organisms from solutions under test.
• These filters are then placed on a solid nutrient medium or in a liquid medium and
incubated to encourage colony growth or turbidity.
 FILTERS SELECTION

• The selection of filters for sterilization must account for the size range of the
contaminants to be excluded.
• Most commonly used filter is composed of nitrocellulose and has a pore size of 0.22μm .
• The size of the bacteria ranges from 0.5 to 5.0 μm whereas the size of the viruses ranges
from 100 nm to 0.36 μm.
• Thus a filter of 0.22μm retains all bacteria and spores but not all viruses.
 WORKING MECHANISM OF FILTRATION

• The major mechanisms of filtration are

1. Sieving,
2. Adsorption and
3. Trapping within the matrix of the filter material.
• Of these, only sieving can be regarded as absolute as it ensures the exclusion of all
particles above a defined size.
• It is generally accepted that synthetic membrane filters, derived from cellulose
esters or other polymeric materials, approximate most closely to sieve filters.
• Filters work by physically trapping particles larger than the pore size and by
retaining somewhat smaller particles via electrostatic attraction of the particles to
the filters.
 MEMBRANE FILTERS

• It is 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.
• Most common type of filters used for liquid sterilization in the microbiology laboratory.
• Composed of high tensile strength polymers such as cellulose acetate, cellulose
nitrate, or polysulfone.
• Prepared as circular membranes of about 150μm thickness
• Contain millions of microscopic pores of uniform diameters; the size of which is adjusted
based on requirements, during the polymerization process.
• Porosities of membrane filters range from 0.1μm to 10μm
• Most commonly used membrane filter has the pore size of 0.22μm and 0.45μm.
• Filtration under pressure most suitable, as filling at high flow rates directly into the final
containers is possible without problems of foaming, solvent evaporation or air leaks.
• The membranes are held in special holders and often preceded by depth filters made of
glass fibers to remove larger particles that might clog the membrane filter.
• The solution is pulled or forced through the filter and is collected in previously sterilized
containers.
• Uses of membrane filter
• Sterilization of fluid materials (pharmaceuticals, ophthalmic solutions, antibiotics, and
other heat-sensitive solutions in laboratories and industries
• Identification and enumeration of microorganisms
 DEPTH FILTERS

• A depth filter is a fibrous sheet or mat made from a random array of overlapping
paper or borosilicate (glass) fibers.
• The depth filter traps particles in the network of fibers in the structure.
• Fibrous pads, sintered glass and sintered ceramic products can be
regarded as depth filters relying principally on mechanisms of adsorption and
entrapment.
Uses
• Filter sterilization of air in industrial processes
• Forced air heating and cooling systems used in houses contains simple depth filter to
trap dust, spores, and allergens
• Use in Biological safety cabinets contain a filter known as high-efficiency particulate
air (HEPA) filter, which is a type of depth filter.
 Applications of Depth Filters
• Sterilization of air filters in industrial processes
• Forced air cooling and heating systems that are used in homes have a
simple filter that traps dust, spores and allergens
• Utilization in biosafety-related applications, like biosafety cabinets.
• The cabinets for biological safety contain the HEPA, or high-efficiency
particle air (HEPA) filter it is a form of depth filter.
DEPTH VS MEMBRANE
 AIR FILTRATION METHOD

• Air can also be cleaned through filtering.

• The air is cleared of disease by passing it through a the High Efficiency Particle Arrester
(HEPA) filter.
• Laminar flow biological safety cabinets are among the most crucial air filters.
• HEPA filters typically remove 0.3 μm test particles with an efficiency of atleast
99.97% including most microorganisms, from the airstream.
• Certain operating theaters and rooms that are occupied by patients with burns are
filtered to reduce the number of microbes that are a part of the air.
• HEPA filters can eliminate almost all microorganisms that are greater than 0.3mm within
diameter.
 HEPA FILTER

• A typical HEPA filter is a single sheet of borosilicate glass fiber that has been
treated with a water-repellent binder.
• The filter, pleated to increase the overall surface area, is mounted inside a rigid,
supportive frame.
• HEPA filters come in various shapes and sizes, from several square centimeters
for vacuum cleaners to several square meters for biological containment hoods
and room air systems.
HEPA Filter
Control of airborne particulate materials with HEPA filters allows the construction
of “clean rooms” and isolation rooms for quarantine, as well as specialized
diagnostic/research laboratories.
 FILTRATION STERILIZATION OF GASES

• The principal application for filtration sterilization of gases is in the provision of

sterile air to aseptic manufacturing suites, hospital isolation units and some operating
theatres.
• Filters employed generally consist of pleated sheets of glass microfibres separated and
supported by aluminium framework; these are employed in ducts, wall or ceiling panels
or laminar airflow cabinets.
• In practice, their microorganism removal efficiency is rather better as the majority
of bacteria are found associated with dust particles and only the larger fungal spores
are found in the free state.
• Air is forced through HEPA filters by blower fans, and prefilters are used to remove
larger particles to extend the lifetime of the HEPA filter.
Advantages of Filtration Sterilization
• Less capital intensive
• Suitable for heat-sensitive liquids (infusions, vaccines, hormones, etc).
• Large volume of liquids can be filtered reasonably fast

Limitations of Filtration Sterilization

• Only liquids and gases can be sterilized by this process
• Filters are expensive to replace, especially nano-filters
• Clogging may occur
 FILTER INTEGRITY TESTING

• Tests carried out to verify and assure the quality and readiness of the filter
membrane regarding the regulatory requirements are called filter integrity tests.
• If failed, shows that the filter is no longer re-usable, and also the previously filtered
batch needs inspection - filters require testing both before and after use
• Filter integrity testing is performed to assure a contamination-free product.
• Integrity of the filter membrane, disk filters, capsules/cartridges are checked.
• The machine that performs this task is called filter integrity machine
• The first filter has a 0.45 µm size followed by 0.22 µm. Later, they get
sterilized with steam for another batch process.
• The diameter or size of the pore is uniformly maintained on the filter membrane.
This is effective in removing bacteria and not viruses.
The two cases to perform filter integrity testing are:
• New Filters
• Filters that require re-use
• In the first case, filter manufacturers themselves perform the testing
along with certificates and documentation.
• If so, it is not mandatory for drug makers to test new filters.
• Meanwhile, the second case often requires testing to check the
intactness of the pore size of the membrane.
FILTER INTEGRITY TESTING

• Generally, common components installed

on the testing unit are housing, HMI,
printer, tubings, alarms, TC Clamp
connections, etc.
• Proper connections at the inlet and outlet
are made prior to starting the test
sequence must be checked before
initiating.
 Classification of Filter Integrity Testing

The testing is classified into two categories:

• Destructive Testing
• Non-Destructive Testing

• Destructive Testing
• Destructive Testing is by and large carried out at the filter manufacturer’s location from
each lot as per their pre-defined sampling plan.
• Destruction of filters takes place to ensure that no bacteria can pass through the
membrane. Simply put, filters retain bacteria.
 Non-Destructive Testing
• Filters used in the previous batch may be damaged.
• Reusing a filter adopts non-destructive testing where it is checked for its
intactness and quality for the next batch.
• If filter integrity testing fails, the previous batch may need re-testing/processing
as per suitable immediate corrective action.
• The following are the different methods for non-destructive filter integrity
testing:
• Bubble Point Test
• Diffusion Test
• Pressure Drop Test
• Water Intrusion Test
• These tests were developed to physically make sure the integrity of a filter
cartridge.
 Bubble Point Test

• This test works on the principle of capillary action and surface tension.
• Liquid when passed through the filter pores fills inside the pores with the help of
surface tension and capillary action.
• Pressure is applied to measure the diameter of a pore.
• The minimum amount of pressure required to remove the liquid from the pores
helps in measuring the pore size.
• The minimum amount of pressure at which the first bubble of liquid comes out
from a pore of the membrane, known as the bubble point for that filter.
 Procedure

• Filter is wetted with suitable solvent (‘water for hydrophilic filters’ and ‘water+alcohol
for hydrophobic filters’) uniformly.
• Pressure is applied to the upstream of the filter.
• The amount of minimum pressure expected from the recommendations provided by
filter manufacturers.
• Pressure increases slowly until bubbling occurs downstream.
• If bubbling appears at a lower pressure than the set pressure, the test is considered a fail.
• The pressure at which bubbling occurs downstream of the filter is its Bubble Point.
 Diffusion Test

• This test works on the principle of Fick’s Law of Diffusion,

• Where the rate of diffusion is directly proportional to the membrane’s surface area.
• The effectiveness of the diffusion test depends on the downstream gas flow rate.
• Typically, the pressure used to perform this test is 75-80% of the bubble point of
that membrane.
• The manufacturer provides requirements/specifications for this test.
 Procedure

• Filter is wetted uniformly with a suitable solvent.

• Pressure is applied upstream of the membrane, gradually increased up to the
recommended value and allowed for stabilization.
• Downstream gas flow rate is measured with the help of a suitable flow meter for 60
seconds.
• If the value of the gas flow rate comes higher than the recommendation, the test failed.
 Pressure Drop Test

• While dealing with different housing arrangements, bubble point or diffusion tests
becomes ineffective.
• Then, pressure drop tests are performed on the automated filter integrity testing units.
• Because this test is performed upstream of the assembly, the downstream manual
operations get eliminated.
• As manufacturers do not specify the exact test requirements, this test relies on system
volume data.
• They measure changes in upstream pressure because of diffusion via the application of
an appropriate and accurate pressure gauge.
 Water intrusion test

• This test is typically performed to detect the quality of defects in hydrophobic filters.
• These filters are generally installed as air vent filters.
• Water with pressure penetrated inside the filter upstream to compare the outcomes with
the manufacturer’s specification limits
THANK YOU

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