KULLIYYAH OF ENGINEERING

DEPARTMENT OF BIOTECHNOLOGY
BTE 3212 | BIOTECHNOLOGY ENGINEERING LAB IV

Semester I 14/15

Cross Flow Filtration Process

By
Group members (Section 1):

1127380

Fatin Nabilah binti Murad (introduction & objectives)

1121448

Nurfarhana binti Abdul Hanid (procedures)

1123746

Suhadahafiza binti Shafiee (results & calculation)

1124650

Nurhafatin Emira binti Shahrul Nazron (discussion)

1126494

Nurul Alia binti Fazil (conclusion)

Introduction
Filtration is a method that use of a medium to separate solids from liquid. There are many
methods to separate solids from liquid. However, in this case, it uses the function of pressure
driven by using membrane filtration to separate solids from liquid. Filtration process can be
divided into two different operational modes that are Normal Flow Filtration and Tangential
Flow Filtration (TFF). Tangential Flow Filtration is also called as Cross Flow Filtration (CFF).
Normal Flow Filtration operates to increase process efficiency from early phase product
development through to production by protecting downstream operations or by polishing final
products. In NFF, the feed stream moves perpendicular to the membrane and purified liquid
passes through the membrane. Membrane filters primarily retain matter due to size differences
between the molecules and the pores in the membrane. Particulates and aggregates are retained
in the pores or form a filter cake that reduces flow and increases pressure over time. Generally,
normal flow filters are used when clarification or bioburden reduction is desired in relatively low
solid streams, such as during final polishing of a product.
Meanwhile, Cross Flow Filtration operates when a liquid comprising circulating the
liquid tangential to a charge modified organic polymer microporous filter membrane. The
membrane consist of organic polymer microporous filter membrane that has micro-structure
throughout the membrane and an amount of a charge modifying agent bound to the substantially
all of the membrane micro-structure without substantial pore size reduction or pore blockage
(Marinaccio et. Al, 19 Dec 1989). In CFF, an incoming feed stream passes across the surface of a
cross flow membrane and generated two exiting streams. The permeate stream is the portion of
the fluid that passes through the membrane. This filtered fluid will contain some percentage of
soluble or insoluble components from the initial feed stream that are smaller than the membrane
removal rating. The remainder of the feed stream, which does not pass through the cross flow
membrane, is known as the retentate stream.

The rate of flow of fluid refers to the resistance of the filter to flow of fluid and resistance
that trapped with formula below:
Permeability (Flux) = V/ (t x Ax P)
Whereas the filtrate flux can be calculated by using the Hagen-Poiseulle Equation that show the
liquid flow through cylindrical pores as below:
J = ( . r2 . P ) / ( 8. . X )
The Hagen-Poiseulle Equation explains about the cell concentration and temperature that control
trans-membrane pressure which is inversely proportional to liquid viscosity. The increase in
pressure or temperature will increase the flux. Moreover, when the filter cake is high, a gel layer
is formed and filtrate flux will decrease because the density of the filter cake and concentration
of retained-cell will increased.
Objectives
a) To study the cross flow filtration mechanism either in normal flow filtration or tangential
flow filtration (cross flow filtration)
b) To know the factors affecting during filtration process

Procedure
A) Microfiltration
1. The clean water was used and 0.1 to 0.5N NaOH solution was made-up in order to
enhance the result.
2. Solution 50oC was recirculated, 5 to 10 psig inlet and 1 to 5 psig outlet for 30 to 60
minutes. The NaOH solution was been sure it contacts all areas within the cartridge.
3. The cartridge was thoroughly drained.
4. Purified water was used to flush to drain, in order to make sure the permeate and
retentate were neutral.

B) Ultrafiltration
1. Clean water was used (WFI or 10000 NMWC UF permeate).
2. The transmembrane pressure of cartridge was adjusted to 15 psig for 1000 NMWC
and 3000 NMWC where 10 psig for 5000 NMWC through 30000 NMWC pre sizes
and 5 psig for larger pore sizes.
3. The retentate flow rate was be curtained 1/10th of the permeate flow.
4. The retentate and permeate was discharged to drain.
5. Hot water with 50oC was used to rinse the whole system. To enhance the glycerol
removal 100 ppm of NaOCl was added.
6. The rising was continued for 90 minutes in order to make sure NaOCl has been rinsed
out before introducing process solution.
C) Experiment run
1. The membrane was inserted at its position. The equipment was checked to make sure
it can be run well and smoothly without leaking.
2. The membrane was wetted by flushing 1 L deionized water into system. The filtration
was ran and the time was measured for every 50ml water from permeate tubing. The
pH was checked in order to make sure it was neutral.
3. The system was the filtration system was run by using a sample recommended
condition.
4. By putting a volumetric cylinder at the end of permeate pipe the permeate rate was
measured and the time was taken for every 50ml sample liquid.
5. According to the table the filtration rate was measured and graph was drawn.
6. After finished, the cross flow was cleaned by 0.5 NaOH for 30 to 60 minutes.
7. The permeate rate and water pH was checked after it was flushed with many times of
deionized water. Cleaning was confirmed when the permeate rate was achieved 80%.

Result
Run 1

Vol(ml)

Run 2
Time(sec
)

flow rate
(mL/s)

50
100

315 0.158730
601 0.166389

150

783 0.191571

200

1054 0.189753

250

1310 0.190840

300

1584 0.189394

350

1837 0.190528

400

2123 0.188413

450

2400 0.187500

500
550

2678 0.186706
2936 0.187330

600

3200 0.187500

650

3482 0.186674

700

3765 0.185923

750

4036 0.185828

800

4313 0.185486

850

4605 0.184582

900

4863 0.185071

950

5174 0.183610

1000

5462 0.183083

Flux
(L/m2.hr
)
5.19480
5
5.44547
6.26959
2
6.21010
9
6.24566
3
6.19834
7
6.23546
3
6.16623
1
6.13636
4
6.11039
4
6.13079
6.13636
4
6.10934
2
6.08475
2
6.08162
9
6.07044
2
6.04086
5
6.05686
7
6.00906
6
5.99181
1

Vol(ml
)

Time(sec
)

50 172
100 324
150 475
200 633
250 789
300 947
350 1106
400 1121
450 1438
500 1609
550 1782
600 1964
650 2147
700 2334
750 2528
800 2725
850 2982
900 3117
950 3480

flow
rate
(mL/s)
0.29069
8
0.30864
2
0.31578
9
0.31595
6
0.31685
7
0.31679
0
0.31645
6
0.35682
4
0.31293
5
0.31075
2
0.30864
2
0.30549
9
0.30274
8
0.29991
4
0.29667
7
0.29357
8
0.28504
4
0.28873
9
0.27298
9

Flux
(L/m2.hr
)
47.5687
1
50.5050
5
51.6746
4
51.7018
5
51.8492
9
51.8383
4
51.7836
6
58.3894
3
51.2074
9
50.8503
3
50.5050
5
49.9907
4
49.5405
9
49.0768
9
48.5471
8
48.0400
3
46.6435
47.2482
3
44.6708
5

Run 3
flow rate
Flux
Vol(ml) Time(sec) (mL/s) (L/m2.hr)
50
27 1.851852 31.74603
100
39 2.564103 43.95604
150
103 1.456311 24.96533
200
146 1.369863 23.48337
250
206 1.213592 20.80444
300
230 1.304348 22.36025
350
252 1.388889 23.80952
400
319 1.253918 21.49575
450
338 1.331361 22.82333
500
400 1.250000 21.42857
550
425 1.294118 22.18487
600
448 1.339286 22.95918
650
511 1.272016 21.80598
700
533 1.313321 22.51407
750
558 1.344086 23.04147
800
620 1.290323 22.11982
850
645 1.317829 22.59136
900
707 1.272984 21.82259
950
731 1.299590 22.27868
1000
800 1.250000 21.42857
Run4

Vol(ml
)

Time(sec
)

50

100

10

150

21

200

34

250

47

300

58

350

71

400

84

450

95

500

108

550

121

600

135

650

147

700

158

750

171

800

183

850

196

900

209

950

221

1000
Run 1

233

flow
Flux
rate
(L/m2.hr
(mL/s)
)
7.14285 306.122
7
4
10.0000 428.571
0
4
7.14285 306.122
7
4
5.88235 252.100
3
8
5.31914 227.963
9
5
5.17241 221.674
4
9
4.92957 211.267
7
6
4.76190 204.081
5
6
4.73684 203.007
2
5
4.62963 198.412
0
7
4.54545 194.805
5
2
4.44444 190.476
4
2
4.42176 189.504
9
4
4.43038 189.873
0
4
4.38596 187.969
5
9
4.37158 187.353
5
6
4.33673 185.860
5
1
4.30622 184.552
0
3
4.29864 184.227
3
5
4.29184 183.936
5
2

Graph of flux (L/m2.hr) vs. filtration time(min.)

Flux (L/m.h)

7
6
5
4
3
2
1
0

filtration time (min.)

Run 2

Graph of filtration rate(ml/s) vs. filtration time(min.)

filtration rate (ml/s)

70
60
50
40
30
20
10
0

filtration time (min.)

Run 3

Graph of filtration rate(ml/s) vs. filtration time(min.)

3
2.5
2
1.5
1
filtration rate (ml/s)

0.5
0

filtration time (min.)

Run 4

Graph of filtration rate(ml/s) vs. filtration time(min.)

12
10
8
6
4
2
0

filtration rate (ml/s)

filtration time (min.)

Total Protein (mg/l)

run1
run2
run3
run4

blank
0.602
0.601
0.56
0.572

TOTAL
OD
PROTEIN
Before After
Before After
filtratio filtratio filtratio filtratio
n
n
n
n
1.157
0.629
2.915
0.275
1.188
0.652
3.075
0.395
2.053
0.98
7.605
2.240
1.716
1.307
5.860
3.815

DISCUSSION
This experiment is about the mechanism of cross flow filtration and factors affecting
during the process of filtration. Filtration is a pressure driven separation process that uses
membranes to separate components in a liquid solution or suspension based on their size and
charge differences. Cross flow filtration is also known as tangential flow filtration. This type of
filtration can be divided into two different categories based on the different sizes of components
being separated which are microfiltration and ultrafiltration. Microfiltration can reduce bigger

size components between 0.1 to 2.5 whereas ultrafiltration is used to reduce for much fine
products.
The experiment will begin with microfiltration then follow with ultrafiltration as because
larger components need to be removed first before the smaller components been removed. But in
this experiment, we only use microfiltration. Before we put the sample into the filtrate, first we
need to rinse the filtrate by using NaOH solution to sterilize the filtrate and avoid it from
contamination. Based on the results between run 1 and run 2, the total time to filtrate 1000 ml of
sample for run 1 is 5462 seconds while for run 2 the total time to filtrate 950 ml is 3480 seconds.
There is much differents in time because for run 1 the pressure that was used is 5psi whereas for
run 2 the pressure that was used is 10 psi. This can be said that the higher the pressure, the least
time was used to filtrate the sample. For run 2, the total volume that has been filtrate was 950 ml.
Supposed, we have to filtrate 1000 ml of sample. This happened because there was a leakage at
the beginning. Therefore, because of the leakage, it has reduced the total volume of sample of
run 2.
For run 3, the total time to filtrate 1000 ml of sample was 800 seconds while run 4
completed the process with much more faster which in 233 seconds. For Thursday session,
which is run 3 and run 4, the total time that they need to filtrate 1000 ml of sample was faster
than Tuesday sesssion which is run 1 and run 2 because they used different size of the tube. For
run 1 and run 2, we used smaller tube as compared to run 3 and run 4. Therefore, we can said
that different size of the tube also can affect the time to filtrate the sample. The bigger the size of
the tube, the least time that we need to use to filtrate the sample.
For run 1 and run 2, the results of the flow rate and flux have been calculated. It shows
that run 1 has smaller flow rate and flux compared to run 2. The results for run 1 at 950 ml are
0.18361 mL/s and 6.009066 L/m2.hr both for flow rate and flux respectively while the results for
run 2 at 950 ml are 0.272989 mL/s and 44.67085 L/m2.hr respectively. This is because for both
of these run 1 and run 2, we have used different value of pressure. Run 2 used higher flow
pressure which is 10psi compared to run 2 which is 5psi. This will affect both of the flow rate
and flux results since from the theory, we already known as the pressure increases, the value of
the flow rate and flux will also increases.
While based on the results of run 3 and run 4, we can also see that run 3 has smaller flow
rate and flux compared to run 4. The results of flow rate and flux for run 3 at 1000 ml are 1.25

mL/s and 53.57143 L/m2.hr respectively while the results for run 4 at 1000 ml are 4.291845mL/s
and 183.9362 L/m2.hr respectively. From what we see, we can concluded based on the total time
taken itself. Run 3 has longer time taken compared to run 4. So, we can concluded as the more
longer time taken to filtrate the sample, the smaller its flow rate and flux. This also can be seen
for run 1 and run 2 as run 1 has longer time taken compared to run 2 and their flow rate and flux
are smaller than run 2.
The results for protein concentration were also calculated. As we can see, the result of
protein concentration for all the runs before filtration is much more higher than after the
filtration. This is because before the filtration the protein concentration is still high compared to
after the filtration. This occurs because the protein was degraded into less concentration after it
has been filtrate since the protein has been going through the flow pressure over time. The
pressures used were the same throughout the process. However, the flow rates were increasing
over the time. Hence, this will make the concentration of the protein lesser after filtration occurs
because of more heat were produced.
Based on the previous experiment on cell rupture, the result of protein concentration for
run 2 was compared between sonication, homogenization and filtration process. For sonication
process which is before the protein was homogenized, the protein concentration was 5.385 g/L.
After it has been homogenized, the protein concentration was 4.355 g/L. Meanwhile, after
filtration process, the protein concentration was 0. 395 g/L. Based on the result, the protein
concentration during sonication process was more higher compared to after homogenization.
This is because during sonication there was no heat produced. After it has been homogenized, the
tprotein concentration reduce since more heat produced at higher pressure. By comparing the
protein concentration during homogenization and filtration process, we can see that filtration
process reduce more protein concentration compared to homogenization process. This is
probably because filtration took longer time than homogenization and it results in redcuing the
concentration of protein since the higher the time taken during the filtration process, the more
higher the flow rate will be and more heat will produced.
There are some errors occurred during the experiment. For run 2, there was a leakage at
the beginning. It has reduced our volume of the sample from 1000ml to 950ml. This is happened
because the clamp was not tight enough. To avoid this from happening, we should be more extra
careful and clamped it tightly. Next, there may be error during taking the OD. We may

accidentally put less or more volume in the centrifuge tube while using micropipette than what
we supposed to put and it may effect the reading of the optical density (OD).
Some precautions should always been taking care of are we have to always check
whether there is leakage or not before we start. Always check the leakage at the place that we
need to clamp since it has higher possibility to leak there. We should always rinse the filtrate
before and after the process to avoid it from contaminate and keep it sterilize. We also need to
use micropipette correctly to get the right volume so that we can get the correct reading of the
OD.

Conclusion
The higher the flow pressure, the higher the flow rate and flux. This can be seen in Run 1
and Run 2 where the same area of tube used with different flow pressure, which Run 2 gives
much higher values of flow rate and flux than Run 1 which have flow pressure of 10 psi and 5
psi respectively.
Filtration tube with larger surface area will give off larger value of flow rate and flux.
This can be seen when comparing the values between Run 3 and Run 4 to Run 1 and Run 2.
The values concentration of total protein for all Runs is much smaller after filtration
because only small sized proteins can pass through the filter. Comparing the total protein
concentration using sonicator, homogenizer and filtration from last experiment, we can conclude
that the concentration is highest when using homogenizer, provided that the heat generation is
not large enough to denature the protein. Next is using sonicator, where it is suitable for small
scale experiment and lastly the concentration of protein using filtration, where only small sized
protein can pas through the filter.
References
Daniel Christodos, Ph.D., P.E., Principle Munipal Engineer, Tangential Flow Filtration to enable
high solids concentration, Improved process throughput, Capacity and Cost, 23 rd Annual
2013.

Retrieved

November

2014,

from

http://www.slideshare.net/njcnews777/tangential-flow-filtration-tff-membrane-designfor-industrial-and-municipal-applications
GE Healthcare Life Sciences, Bioprocess Filtration, Normal Flow Filtration. Retrieved 2
November

2014,

from

http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/en/GELifeSciences/pro
ducts/AlternativeProductStructure_16139/
Marinaccio et. al, 19 Dec 1989, Cross-flow filtration, US 4888115 A, Dec 19, 1989
Pall Corporation, How cross flow filtration works. Retrieved 2 November 2014, from
http://www.pall.com/main/graphic-arts/how-crossflow-filtration-works.page