Qualification Test Methods

1, Bacterial Endotoxins Test: A solution of filter extract is mixed with

Limulus Amebocyte Lysate. A reaction at different dilutions indicates the
maximum level of endotoxin present. The test results determine the level of
endotoxins contained within the filter. The filters should contain less than 0.25
endotoxin units per milliliter (EU/mL) per USP Bacterial Endotoxins Test

2, Hydraulic Stress Test at 25°C, 80°C and 135°C: This

hydraulic stress test is designed to determine mechanical strength in actual use
at ambient and elevated temperatures Filters were sterilized by steaming at 135
°C for 30 minutes prior to testing.
The filters were then plugged with a colloidal plugging agent to increase the
water pressure differential across the devices. Dynamic hydraulic stress was
then applied in the forward and reverse directions. An air/water diffusion test and
an air/water bubble point test was performed after the various hydraulic stresses
were applied to monitor any change in filter integrity

3, Oxidizable Substances:
USP Oxidizable Substances Tests were performed to determine the presence or
absence of oxidizable substances contained in the filters. The filters were
autoclaved for one cycle of 60 minutes at 126 °C. The filters were then flushed at
a flow rate of 200 mL/min with purified water. The filter effluent was collected in
200 mL aliquots which were then tested for oxidizable substances per current
USP methodology.

4, Flush Test for Total Organic Carbon (TOC) and

Conductivity:
Flushing tests were performed to monitor the reduction of total organic
and conductivity with flush volume. The filters membrane were autoclaved for
one cycle of 60 minutes, 126 ºC prior to running the flush test. The filters were
flushed with purified water (WFI quality) at a flow rate of 500 milliliters per
minute. The filter effluent was sampled at 500 ml intervals and tested for
conductivity and total organic carbon
5, Non-Fiber Releasing:
Fiber releasing tests were performed to show that the filters membrane do not
shed fibers. Filters were autoclaved for one cycle of 60 minutes at 126 °C. Filters
were then flushed with three 5 L aliquots of 0.1 μm filtered water at a pressure of
2.07 bar (30 psi) for a total of 15 liters. The filtrate was passed through
0.8 μm black gridded disc filter to collect any fibers removed from the cartridge
filter.
Each disc was examined under a microscope for the presence or absence of any
fibers. A system blank/background (test system fibers counted with no sample
filter) was run prior to running the filters. Backgrounds are subtracted from filter
results.

6, Particle Releasing:
This test was performed to show that the filters membrane do not release
particles into the filtrate during use Filters were autoclaved for one cycle of 60
minutes at 126 °C then flushed with a total of 10 liters of 0.1 μm filtered water at
a flow rate of 1 L per minute. A portion of the filtrate was monitored for size and
quantity of particles.

7, Safety Test :
The USP <88> Safety Test was conducted to confirm that filters membrane are
nontoxic. The test articles were extracted in 0.9% USP Sodium Chloride for
Injection at no less than 85 °C for one hour