STERILIZING GRADE FILTER

&
FILTER VALIDATION

Ajeet Kumar Singh

 STERILIZING GRADE FILTER
“ A filter which, when challenged with the micro-
organism Pseudomonas diminuta at a minimum
concentration of 107/cm2of filter surface, will
produce a sterile effluent” ………… (1987)

“ A filter that, when appropriately VALIDATED, will

remove all micro-organisms from a fluid stream,
producing a sterile effluent” ………..FDA(2004)
 STERILIZING GRADE FILTER REQUIREMENTS
BP2007 &
PARAMETERS USP 28 FDA
EC-GMP
Retentive membrane Retentive membrane with a Bacteria retentive
with a nominal pore size nominal pore size 0.2µm or membrane with a
Nominal pore size
0.2µm or 0.22µm less nominal pore size of
0.22 mm or less
Pre-filtration is not Yes. Pre-filtration
Pre-filtration No recommended but setting of bioburden limits – Not
Bioburden pre-filtration limit for More Than 10 cfu/100
bioburden is to be established ml
Integrity test
Before use Yes Yes Yes
After use Yes Yes Yes

Cited Integrity test

Yes Yes Yes
Bubble point
Yes No Yes
Pressure hold

A challenge of not less A challenge concentration of A challenge of at least

Microbial challenge than 107 P.dimunata at least 107 organisms per cm2 107 cfu of P. diminuta
test suspension per sq. com of effective filtration area of per cm2 of active filter
of filter surface area B. diminuta surface
 PORE SIZE RATING

o 0.22 µm sterilizing filter membranes : Capable

of retaining 100% of a culture of 107 P.diminuta
/cm2 of membrane surface under a pressure of
NLT 30 psi (2.0 bar).………….USP-28
 Use of Multiple Sterilizing Grade
Filters in Aseptic Processes ……..
US PERSPECTIVE

o Use of redundant sterilizing filters should be

considered in many cases. This additional filter must
be satisfactorily tested before use, but does not
require post-use integrity testing unless the primary
filter fails……………PDA Technical report
 Use of Multiple Sterilizing Grade
Filters in Aseptic Processes ……..
EU PERSPECTIVE

o Potential additional risks as compared with other

sterilization processes → can do second filtration
(sterilized microorganism retaining filter) immediately
prior to filling.

o The final sterile filtration should be as close as

possible to the filling point.………(EU GMP)
 Multiple filters US Vs EU
US EU
Filter #1 passes integrity testing Filter #1 (Bioburden reduction
post-use, batch can be released filter)
Filter #2 is discarded without Must pass the post integrity
testing test

If filter #1 fails integrity testing

post-use, than filter #2 is tested Filter #2 (Final Sterilizing

If filter #2 passes, batch can be filter)

released Must pass the post integrity

test
 VALIDATION FO ASEPTIC PROCESSING
AND STERILLIZATION
FDA GUIDANCE NOTE

21 CFR 211.113(b) states that "Appropriate written

procedures, designed to prevent microbiological
contamination of drug products relate to be sterile,
shall be established and followed. Such procedures
shall include validation of any sterilization
process."
 Requirements/Responsibility for Filter
Validation
Filter Data Provided by
Parameter Filter User
Manufacturer Millipore
Recommend Operate within the -
Sterilization sterilizing procedure prescribed limits.
Procedure with temperature Validate the process
and cycle time limits application
Provide filter usage Validate for their Filter usage limits with
Integrity limits process and solution water provided in the
Testing guide. Product specific
limit to be set by filter user
Meet non-fibre Document the Manufacturer certifies that
Fibres release claim [21 appearance of the the filter/cartridge is non-
CFR 210.(3b)] filtered product fiber shedding
Provide information Validate with their --
Sterilizing on release criteria process solution
grade filter
 Requirements/Responsibility for Filter
Validation (contd….)
Parameter Filter Manufacturer
Provide extractables data –
Extractables type and amount, Pass USP
test
Provide compatibility
Compatibility
tables
Provide test data showing
Toxicity product meets USP plastic
test requirements
Adsorption Provide known information
Data Provided by
Filter User
Millipore
Data provided with
Evaluate with their
water. Done on model
system and solution
stream systems
Evaluate with their Data provided for
solution different solvents
Document -
Evaluate with their
solution (both for active -
as well as preservatives)
 FILTER VALIDATION

o DEMONSTRATION OF BACTERIAL RETENTION

o COMPATIBILITY STUDIES

o INTEGRITY TESTING
 BACTERIAL RETENTION STUDY FOR
STERILIZING GRADE FILTER

VIABILITY STUDY
Most appropriate method of delivering the challenge of 107/cm2
Product viability study with B. Diminuta--------Direct Inoculation
↓
Viability decrease greater than or equal to 1 log
Or
Oil based product

PRODUCT IS CONSIDERED ANTAGONIST

Surrogate fluid e.g. Saline lactose Broth ….for bacterial retention
 BACTERIAL RETENTION STUDY

PRODUCT CONDITIONING STAGE

Product is continuously filtered at desired flow rate for the
max. processing time after that filter is rinsed to remove
the product from the filter—Dead ended challenge filtration

CHALLENGED STAGE
B. Diminuta is suspended in product/media and Product is
continuously filtered at desired flow rate for the max.
processing time

Analysis of filtrate for the the presence of B.diminuta

 BACTERIAL RETENTION STUDY (contd…)

ACCEPTANCE CRITERIA

o 0.22µm filter must demonstrate complete retention.

o 0.45µm filter (Positive control) must demonstrate

complete recovery of test organism.

o Sterility of the test system shall be demonstrated

o 0.22 & 0.45µm filter must not fail in the post integrity
test.
 COMPATIBILITY ASSESSMENT

Chemical and Physical resistance

Filter is subjected to:
o Microscopic inspection for the absence of damage
which might influence the filter performance
o Dimensional measurements
o Extractable and Leachable analysis
 STRUCTURAL COMPATIBILITY

SWELLING OF FILTER PLASTICIZING EFFECT

Pore space tighter Release the polymer
Flow rate decrease stress
Bubble point ( ) Pore space open
Flow rate increase
Bubble point ( )

o To ensure the structural compatibility recording of

Bubble point is necessary after contact with the
product
 ADSORPTION

o Formulations containing low concentration of

actives
o Product containing preservatives
o Specific nature of actives ingredients
 INTEGRITY TESTING
Objective: To ascertain whether changes in the filter may
have occurred during filtration
Conform the correctness of the filter manufacturer’s pore-
size rating
If fails in the final integrity test ------filtrate should be re-
filter or discarded
Laplace law 4γ Cosθ
P=
D

P: Pressure necessary to expel the water

D: Diameter γ: Surface tension
θ: Angle of wetting
 Bubble Point

o Liquid is held in the pores of the filter by surface

tension and capillary forces.
o The minimum pressure required to force liquid out
of the the pores………..Bubble point
o Bubble point is a function of wetting liquid
surface tension, pore size → Specific to
membrane polymer
Procedure for Bubble point determination
Filter → wetted with Purified water
(approx. 200 ml (Temp.---, Time 15-20 mins)

Record the bubble point

Repeat the test with rewetting the membrane until

stable bubble point in three consecutive tests

Repeat the test with test fluid

Calculate the Bubble point ratio

(BPR)
Manually determination of Bubble point
 Bubble Point

If Bubble point value lower than the specification:

o Non-integral membrane
o High temperature
o Integral filter,but wrong pore size
o Incomplete wetted membrane
 Diffusive flow/Forward flow test

o Pressure at which gas molecules that migrate from

water-filled pores of a wetted membrane.

o Directly proportional to the differential pressure

and the total surface area of the filter.

o Filter → Wetted with liquid → Upstream side of the

filter pressurized → diffusive flow is measured
 Pressure Hold/ Decay Test

o Indirect method → diffusive low/forward flow testing

o Filter housing pressurized → Gas flow across the
membrane quantitatively measured as decay in the
pressure over a specified period of time
o Conform the integrity of the entire filter assembly
o Pressure decay below the max. allowable value →
Passes the physical integrity test
Filter validation sample requirements

Test Sample Size

200 ml from three different
Bubble Point
lot

Viability study 50 ml from one lot (Sterile)

Bacterial retention
study & 2000 mL from one single lot
Compatibility
 MAPPING OF ACTIVITY
DRF MFG. SITE

Establish lead formula based on

Pre-formulation Studies
Batch Size,
Operating
Integrity Compatibility
Conditions
Testing
(200 ml
non-sterile
sample of 3
lots) Adsorption

50 ml sterile sample for

viability testing

Extractables – Model Solvents

•FILTER MFR.
2000 ml Bacterial Retention
INFORMATION NECESSARY FOR ACCESS QUESTIONNAIRE

o Aq/Non aq.
o Biological/Non Biological
o pH
o Specificity Gravity
o Viscosity
Product Description o Detail unit formula along with CAS No.
o MSDS
o Neutralizing agent
o Sensitivity if any
o pH modification possible
o Product description (Preservative)
INFORMATION NECESSARY FOR ACCESS QUESTIONNAIRE
Contd...

o Processing Temp. for Filtration

o Minimum Batch Size (Min, Actual and
Critical Process max.)
Description o Filtration (Continuous/Intermittent)
o Flow Rate (mL/min)
Test gas o Compressed Air / Nitrogen/Other
Type o Membrane/Cartridge
Process type o Continous / intermittent
 CONCLUSION
Ensure that the following information is
available on the filter

o Compatibility

o Bacterial Challenge Testing

o Extractable Testing

o Product Specific Integrity Testing Values