Quick Reference USD 2453 Rev A 1.

00

Membrane Cassette Care and Use Procedures

Centramate and Centrasette Cassettes —
T-Series Omega Membrane
 Table of Contents

Table of Contents
Safety Notice ...........................................................................................................2
Learn About Safety .................................................................................................3
TFF Process Flowchart ............................................................................................4
Introduction and Key to Diagrams ..........................................................................5
Step 1: Installation ................................................................................................6
Step 1a: Measuring Critical Volumes ......................................................................................... 8
Step 2: Flushing...................................................................................................10
Step 3: Sanitization..............................................................................................12
Step 4: Normalized Water Permeability (NWP)..................................................14
Step 5: Integrity Testing......................................................................................16
Step 6: Buffer Conditioning ................................................................................18
Step 7: Optimization ...........................................................................................20
Step 8: Concentration..........................................................................................22
Step 9: Diafiltration.............................................................................................24
Step 10: Product Recovery.....................................................................................26
Step 11: Buffer Flush.............................................................................................28
Step 12: Clean in Place (CIP) ................................................................................30
Step 13: Storage.....................................................................................................32
Torque Recommendations.....................................................................................34
Operating Specifications ........................................................................................35
Temperature Correction Factors ............................................................................36
Diafiltration — Continuous vs. Discontinuous......................................................36
Conversion Tables .................................................................................................37

Table of Contents Page 1

Safety Notice

Page 2 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Learn About Safety

Learn About Safety

Please read and follow the safety instructions in this User Guide.
Important — Read First
Storage solutions recommended for sanitizing, cleaning, flushing, and storage may be hazardous or corrosive.
Follow proper safety procedures when preparing, mixing, and handling these reagents. Refer to Material Safety Data
Sheets (MSDS) — available from your supplier — to learn about the specific characteristics, necessary precautions, and
suitable remedies for each reagent used.
1. Always wear protective clothing including safety glasses and gloves when working with membrane cassettes,
equipment, samples, and reagents.
2. Provide sufficient space for assembling all system components and operating the system.
3. Disconnecting a system component or dismantling an installed cassette holder without first isolating and
depressurizing it can result in personal injury and equipment damage. Depressurize a TFF System and cassette
holder before dismantling any component.
4. Some system components may be very heavy. Take proper precautions when moving or lifting equipment to
prevent personal injury. In some cases, hoists or other lifting equipment may be required.
5. You should always use pressure gauges or pressure sensing devices in your system so you can monitor the system
pressure and differential pressure across the membrane cassette.
6. Take suitable precautions before sterilizing the holder with stream or using the holder with hot fluids.
7. Do not use your cassette holder as a coded pressure vessel.
8. Wipe up spills promptly to prevent injury from contact or slipping.
9. Complete the following safety procedures:
a. Read about the operating limits of the membrane cassette and the proper methods for use detailed in this
T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide.
b. Ensure that your process and cleaning conditions do not exceed the operating limits of the cassette holder,
membrane cassette, and sealing materials.
c. Check that your process equipment and the cassette holder meet local safety codes.
d. Inspect the holder and seals regularly to detect damaged components.

Safety Conventions in this Manual

Safety information is identified in this instruction manual by the following convention:

Information

Identifies important information about the current topic.

Safety Warning
Warning: Safety concerns are identified in a box with a red border. The symbol identifies a dangerous or
potentially dangerous situation that may cause irreversible damage to equipment and poses a safety risk
that can cause serious personal injury.

Learn About Safety Page 3

TFF Process Flowchart

TFF Process Flowchart

Pre-use Post-use
Processing
Conditioning Conditioning

1. Installation 7. Optimization 11. Buffer Flush

Page 6 (*Once only) Page 28
Page 20

2. Flushing 12. Clean In

Page 10 8 and 9. Place (CIP)
Concentration Page 30
and/or
3. Sanitization Diafiltration
Page 12 Pages 22 and 24 2. Flushing
Page 10

2. Flushing 10. Product

Page 10 Recovery 4. Normalized Water
Page 26 Permeability
Page 14
4. Normalized Water
Permeability (NWP)
Page 14 13. Storage
Note: The specific steps and
Page 32
order of steps for processing
are application specific and
5. Integrity Test
must be established by the
Page 16 user. Basic guidelines are
shown in this abridged
manual. Pall can assist with
optimizing your process.
6. Buffer
Conditioning
Page 18

Page 4 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Introduction and Key to Diagrams

Introduction and Key to Diagrams

This manual supplements the T-Series Membrane Care and Use Procedures (USD 2433 Rev A), which can be downloaded
from the Pall website at www.pall.com. Refer to page 35 for recommended flow rates and operating specifications for the
Pall range of membrane cassettes.This manual contains operating guidelines. All process steps should be validated by a
qualified individual.

Gauge —
Valve open Zero pressure

Gauge —
Valve partially Low pressure
closed/open e.g. 0.5 bar/7 psi

Gauge - Low pressure

e.g. 1 bar/15 psi
Valve closed

Gauge —
Medium pressure
PUMP e.g. 1.5 bar/22 psi
Pump running
4
Gauge —
Medium pressure
PUMP
Pump stopped e.g. 2 bar/30 psi
´
Pipe full (Water)
Pipe empty
for buffer

Pipe closed Pipe full (Product)

Key to Diagrams

Introduction and Key to Diagrams Page 5

Installation

Step 1: Installation

1. Installation

Safety Information

Wear gloves to avoid 1.1

contact with storage Remove cassettes and Please refer to
solutions. gaskets from packaging. T-Series Membrane
Cassette Care and
Use Procedures
(USD 2433).
1.2
Remove protective
sheet from gaskets,
rinse with WFI.

1.3
Inspect holder, gaskets
and cassettes
and remove any foreign
material which may affect
sealing.

Install multiple
Ensure that a gasket 1.4 cassettes with the
is positioned Install cassettes in printed cassette
holder taking care to align information facing all
between each on the same side and
interface as shown the holes on the holder,
direction, so that it
in Figure 1. gaskets and cassettes. can be read.

1.5
Apply the correct torque
(Table 6, Page 34) to the holder in
a criss-cross pattern, ¼ turn at a
time as shown in Figure 2.

Measuring Volumes
Proceed to Step 1a
(page 8) to measure
system volumes

Page 6 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Installation

Hardware End-plate
Gasket
Pall Omega 30 kDa
Centrasette Cassette
Gasket
Pall Omega 30 kDa
Centrasette Cassette
Gasket
Hardware Manifold

Figure 1: Example of Multiple Cassette Installation

¼ turn ¼ turn ¼ turn

Step 1 Step 2 Step 3

¼ turn
Step 4 Step 5 etc
Figure 2: Example of ¼-turn Torque Procedure

Installation Page 7
Installation

Step 1a: Measuring Critical Volumes

1a. Measuring Volumes

Safety Information

1a.1 Hold-Up Volume

To measure
Add a measured volume (greater than total hold-up volume)
volumes you will
to a suitable graduated cylinder. Note down volume need appropriate
(Use Table 1 for noting all values) (Volume A). sized graduated
cylinders.
1a.11 Non-
Recoverable Volume 1a.2 Recirculate
Pump/drain all water Place feed and retentate lines into
from system into the graduated cylinder. This procedure is
vessel and measure a guideline only
Close permeate valve, run pump and will not cover
volume (Volume E). slowly to fill all pipework on all set-ups.
Minimum working upstream side.
volume - Volume E = Depending on
system set-up,
Non-recoverable this procedure
volume. 1a.3 may need to be
Pull tubing up to top of fluid. adjusted (e.g., if
Adjust tubing does not affect the the feed vessel is
1a.10 Determine fluid level. Measure and note the connected directly
Minimum Working new volume in graduated cylinder to the pump).
Volume (Volume B).
Upstream Hold-up
Volume + Volume D =
Minimum working 1a.4 Upstream Hold-Up
volume. Note value. Volume A - Volume B = Upstream Hold-up
volume. Note value.
1a.9 Minimum Working Volume
Add measured volumes to the
1a.5 Downstream Hold-Up
vessel such that the volume is just
Open Permeate line to fill with fluid.
enough to stop air being pulled
Measure and note the new volume in the
into the pipework/system at
graduated cylinder (Volume C).
recommended process crossflow.
Note volume added.
(Volume D).
1a.6 Downstream Hold-Up
Volume B - Volume C = Permeate
1a.8
(downstream) hold-up volume. Note value.
Move/connect the feed,
retentate and permeate lines
(still filled with fluid) to the actual 1a.7 Total Hold-Up
feed vessel to be used during Volume A - Volume C =
processing. Total Hold-up volume.

Page 8 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Installation

RETENTATE RETENTATE

A PERMEATE PERMEATE

B
GRAD. GRAD.
CYLINDER CYLINDER

PUMP PUMP
´ FEED 4 FEED

Figure 3: Setup for Hold-up Volume — Figure 4: Volume B (1a.3)

Volume A (1a.1)

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED
C GRAD.
VESSEL
CYLINDER

PUMP PUMP

4 FEED
4 FEED

Figure 5: Volume C (1a.5) Figure 6: Minimum Working Volume at Process CFF

(1a.8)

Table 1: Measured Volumes

Volume Tag Description Measured / Noted Value
Volume A Initial Volume
Volume B Vessel volume after 1a.3
Upstream Hold-up Volume Volume A - Volume B
Volume C Vessel volume after 1a.5
Permeate Hold-up Volume Volume B - Volume C
Total Hold-up Volume Volume A - Volume C
Volume D Volume added in 1a.9
Minimum Working Volume Upstream Hold-up Volume + Volume D
Volume E Volume recovered in 1a.11
Non-recoverable Volume Minimum Working Volume - Volume E

Installation Page 9
Flushing

Step 2: Flushing

2. Flushing

Safety Information
Re-use or
Take appropriate Initial Flush or
Post Before the
safety measures to Pre-sanitization
Sanitization Flushing
avoid injury or procedure it is
contact with fluids. recommended
that system hold-
2.1 Setup up and minimum
Set up the system with permeate working volumes
and retentate streams to waste. be established.
See Procedure 1a
for example.

2.2 Flush to Waste

Fill feed vessel with flush water and
start pump to generate flow through Recirculation
times and
retentate line to waste/drain. volumes are
application
specific and must
Stop Pump be validated.
and proceed 2.3 Flush to Waste Flush system with
to Step 3. Apply retentate pressure (partially pharmaceutical-
Sanitization. close retentate valve) to generate grade water.
permeate flow to waste/drain. Use of a lower
quality water may
introduce
inorganic
2.6 impurities that
Repeat could affect
recirculation 2.4 Recirculation membrane
and drain steps To reduce water usage, recirculate performance and
to achieve product recovery.
solution back to feed vessel
desired end for 5 – 30 minutes.
point.
During flushing it
is also
recommended to
2.5 Flush to Waste calibrate the
Drain system by directing pump (determine
the speed settings
retentate and permeate lines to to achieve
waste/drain (as per Figure 8). required process
and CIP crossflow
rates).

Page 10 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Flushing

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE WASTE

FEED FEED
VESSEL VESSEL

WASTE WASTE
PUMP PUMP
´ FEED 4 FEED

Figure 7: Set-up for Retentate and Permeate to Figure 8: Retentate and Permeate to Waste (2.2 and 2.5)
Waste (2.1)

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE

FEED
FEED VESSEL
VESSEL

WASTE
PUMP PUMP
4 FEED 4 FEED

Figure 9: Flush to Waste with Pressure (2.3) Figure 10: Recirculation with Pressure (2.4)

Flushing Page 11
Sanitization

Step 3: Sanitization

3. Sanitization

Safety Information

Take appropriate 3.1 Drain System

Ensure you use a
safety measures to Drain system and reservoir by large enough
avoid injury or pumping to waste. volume to enable
contact with fluids. an initial purge to
waste followed by
recirculation.
3.2 Add Sanitization Fluid
Fill reservoir with suitable volume
Recirculation
of sanitization fluid (see Table 2). times and
volumes are
application
specific. Must be
3.3 Short Flush to Waste validated.
Run pump to flush approximately
one or two system hold-up Please refer to
volumes to waste. T-Series
Membrane Care
and Use
procedures
3.4 Recirculate (Open) (USD 2433).
Check the retentate and permeate
valves are open. Adjust pump to Sanitization
give recommended crossflow retentate flow rate
(See Table 7, page 35). should be 1.5 x
process retentate
flow rate.

3.5 Recirculate (Pressure)

Apply retentate pressure (partially
close retentate valve). Check
pump is still at recommended
crossflow (See Table 7, page 35).

3.7 Flush
3.6 Recirculate solution
Repeat flushing
for 30 – 60 minutes.
procedure (Step 2).

Page 12 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Sanitization

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE

FEED
FEED VESSEL
VESSEL

PUMP WASTE PUMP

4 FEED
4 FEED

Figure 11: Retentate and Permeate to Waste Figure 12: Recirculation with Permeate and
(3.1 – 3.3) Retentate Open (3.4)

RETENTATE
PERMEATE

FEED
VESSEL

PUMP

4 FEED

Figure 13: Recirculation with Pressure (3.5)

Table 2: Recommended Sanitization Solutions and Conditions

Membrane Type Choose an Appropriate Solution from the List Below

Omega™ 0.1 – 0.5 N NaOH + 10 – 50 ppm NaOCl

0.1 – 0.5 N NaOH
200 – 400 ppm NaOCl pH 6 – 8

0.1 N Acetic Acid (HAc) 0.1 N Phosphoric Acid (H3PO4)

Temperature 25 – 45 °C

Sanitization Page 13
Normalized Water Permeability (NWP)

Step 4: Normalized Water Permeability (NWP)

4. Normalized Water
Permeability
Safety Information

4.1 Water quality

Take appropriate
With system in recirculation, fill should be Water
safety measures to For Injection
feed vessel with water and
avoid injury or (WFI) or at a
increase pump speed to achieve
contact with fluids. minimum 0.2 µm
recommended crossflow rate filtered DI.
(Page 35).

The initial
4.2 Remove Air NWP20°C of the
Partially close the retentate valve to increase feed pressure cassette is
essential to
then open immediately. Repeat several times.
calculate because
Set pump speed to zero. it is used as the
basis to determine
membrane
recovery.
4.3 Set Initial TMP Value
Fully close the retentate valve, then increase pump speed
slowly to obtain desired TMP.
Be sure to use
calibrated
equipment that is
accurate in the
4.4 Measure Permeate Flow measurement
Open, then close the retentate valve to release trapped air. range.
Measure the permeate flow rate (typically mL/min) and
convert to LMH. Measure water temperature.
Typical TMP
Value

4.5 Plot Permeate Flux vs. TMP (Figure 18). bar (psi)
UF: 0.5 to 1.5
(7 – 22)
MF: 0.1 to 0.3
4.6 Increase TMP Value (2 – 5)
(See Typical TMP for typical values)
Increase pump speed slowly to increase
TMP. Repeat steps 4.4 – 4.5. 4.7 Repeat Step 4.6 at next TMP.

4.8 Determine NWP at a single point easily measured on the plot.

This can be used as a quick reference for future (See Figure 18).

Page 14 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Normalized Water Permeability (NWP)

RETENTATE RETENTATE

PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PUMP PUMP

4 FEED
4 FEED

Figure 14: Recirculation (4.1) Figure 15: Pressure Increase to Remove Air (4.2)

Equations:

TMP = ((PF + PR)/2) - PP

RETENTATE Water Permeate Flux Rate (LMH)

=
PERMEATE perm TMP (bar or psi)
LMH
FEED NWP20°C = X TCF*
VESSEL TMP

Membrane NWP after cleaning

= X 100%
Recovery NWP initial
PUMP

4 FEED
Conversions:
1 bar = 14.5 psi

Figure 16: Using Pump to Obtain Desired TMP (4.3) TCF = Temperature Correction Factor
See Table 9: Temperature Correction Factor

Determining Membrane NWP Water Permeability = 110 LMH @ 10 psi

= 11 LMH/psi
Permeate Flux (LMH)

NWP20°C = 110/10 X TCF

Temp = 16°C
150 X Temperature Correction Factor = 1.109
(Correcting the water temperature
100 X from 16 °C to 20 °C)
50 X Normalized Water Permeability (NWP20°C)
0 = 11 X 1.109
5 10 15 = 12.2 LMH/psi
Transmembrane Pressure (psi)
Figure 17: Determining NWP for a Membrane Cassette

Normalized Water Permeability (NWP) Page 15

Integrity Testing

Step 5: Integrity Testing

5. Integrity Test

Safety Information

Take appropriate 5.1

Ensure membranes are wet and Before performing
safety measures to air flow integrity
avoid injury or system is drained. tests (IT), the
contact with fluids. membrane
cassettes must be
completely wetted
5.2 out.
Membrane Integrity Test Typically IT
Close the retentate valve, open permeate valve and run follows flushing
pump slowly to generate air pressure (see Table 3) on the and NWP, so the
feed/retentate side. Stop pump cassettes should
be fully wetted.

5.2a These procedures

Monitor feed/retentate pressure to assess rate of assume a positive
pressure drop. displacement
pump (e.g.,
peristaltic or
diaphragm) is
Membrane Pressure drop used to generate
and System No > Limit (see page 17) air pressure.
Integral Refer to T-Series
Membrane
Yes Cassette Care
and Use
5.2b Procedures
(USD 2433) for
Run pump slowly to maintain pressure. Measure air more detailed
flow on permeate using an inverted beaker or alternative protocol.
device (at correct test pressure)

Membrane Air flow >

and System No
Specification
Integral
Yes

5.3 Check system for leaks

(cassette holder, clamps and seals). Repeat from 5.2b.
If air flow is greater than specifications, cassette fails IT.

Page 16 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Integrity Testing

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PUMP
PUMP
4 FEED
´ FEED

Figure 18: Membrane Integrity Test — Figure 19: Membrane Integrity Test — Monitoring Air
Generating Air Pressure (5.2) Pressure (5.2a)

Limit (guideline) for pressure drop:

200 mbar/min or 3 psi/min
RETENTATE
PERMEATE
(Note: this is a guideline only. Direct
measurement of diffusive flow should
FEED
VESSEL be used where necessary)

PUMP
´ FEED

Figure 20: Measuring Air Flow

with Inverted Beaker (5.2b)

Table 3: Recommended Air Flow Test Pressure and Specifications

Membrane MWCO Test Pressure Allowable Air Forward Flow Rate per Unit of Membrane

≤ 500 kDa 2.0 bar (30 psi) < 1600 sccm/m2 (< 150 sccm/ft2)

Integrity Testing Page 17

Buffer Conditioning

Step 6: Buffer Conditioning

6. Buffer Conditioning

Safety Information

Take appropriate 6.1

Ensure you use a
safety measures to Drain system large enough
avoid injury or Direct retentate and permeate to volume to enable
contact with fluids. waste and run pump. an initial purge to
waste followed by
recirculation.

6.2 Buffer Purge

Fill feed vessel with buffer and start pump to generate flow Buffer
through retentate and permeate lines to waste/drain conditioning
removes trapped
air from the filter
and system and
establishes
6.3 Buffer Purge conditions for pH,
Purge lines for a few seconds (may need some retentate ionic strength and
pressure to force flow to permeate). Stop pump. temperature to
match the process
sample.
6.4 Buffer Conditioning
Place system in recirculation. Ramp up pump speed to It is important to
know the system
recommended crossflow (Page 35). hold-up volume
when calculating
concentration
factors, because
this volume of
6.5 Remove Air buffer will dilute
Partially close the retentate valve to increase feed pressure the product
then open immediately. Repeat several times. slightly when
product is added
to the feed vessel.
See 1a,
Pages 8 and 9.
6.6 Buffer Recirculation
Recirculate for 10 – 15 minutes for pH and ionic stability In the diagrams
and where necessary until temperature has stabilized. opposite, buffer is
shown as water.
In a real process
this will be the
6.7 Remove Excess buffer from feed vessel buffer in which the
Direct permeate to waste. Stop pump when fluid in vessel product is in at the
reaches bottom, keeping feed/retentate flowpath flooded start of the
with buffer. process.

Page 18 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Buffer Conditioning

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE WASTE

FEED FEED
VESSEL VESSEL

WASTE PUMP WASTE

PUMP

4 FEED
4 FEED

Figure 21: Drain to Waste via Pump (6.1) Figure 22: Retentate and Permeate to waste (6.2)

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PUMP WASTE
PUMP
4 FEED
´ FEED

Figure 23: Recirculate with Pressure (6.4, 6.5) Figure 24: End of Buffer Conditioning (6.7)

Buffer Conditioning Page 19

Optimization

Step 7: Optimization

7. Optimization

Safety Information

Take appropriate 7.1 Add Sample Hold-up volume,

Following buffer conditioning, minimum working
safety measures to volume and pump
avoid injury or check torque, add sample to
feed vessel. speed settings for
contact with fluids. process and CIP
should already
have been
determined during
7.2 Recirculate Sample the earlier flushing
Start pump and ramp up to required retentate flow rate (or steps. See
crossflow rate). Recirculate for a few minutes. Procedures
1a and 2,
pages 8 – 10.

7.3 Record Parameters

Measure permeate flow (using stopwatch and Optimum TMP:
graduated cylinder). Return permeate volume
The optimum
collected to feed vessel. Record flow rate and TMP for a given
all other parameters on logging sheet or in a process is
suitable spreadsheet. determined from
the plot of
permeate flux vs.
transmembrane
7.4 Plot Data pressure.
Either using graph paper or a spreadsheet, plot each
data point to create a graph of Flux Rate vs. TMP.
See Figure 29. For UF it is
typically the
pressure at which
the curve begins
to plateau or level
7.5 Increase TMP out (see example
Increase TMP by partially closing retentate valve. in Figure 29).
Pump speed may need to be increased slightly to
maintain crossflow. Leave recirculating for a few
minutes to stabilize conditions. In some cases
transmission of a
solute may also
be a requirement,
and in these
7.6 Identify Optimum TMP cases a high TMP
Repeat Steps 7.3 – 7.5 until the optimum TMP is reached. is generally not
See Figure 29. Leave system running at the optimum TMP the optimum
ready for the concentration procedure. condition

Page 20 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Optimization

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PERMEATE PUMP PERMEATE

PUMP
´ FEED
VESSEL
4 FEED
VESSEL

Figure 25: Add Product Sample to Feed Vessel Figure 26: Recirculate Sample to Stabilize Conditions (7.2)
(7.1)

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PUMP PERMEATE PUMP PERMEATE

4 FEED
VESSEL
4 FEED
VESSEL

Figure 27: Measuring Permeate Flowrate (7.3) Figure 28: Increasing TMP by Partially Closing
Retentate Valve (7.5)

Plotting Flux Vs TMP Equations:

Permeate Flux (LMH)

Optimum TMP
60 TMP = ((PF + PR)/2) - PP
X X To convert mL/min/ft² to
40 X
LMH:
X
20 mL/min/ft² x 0.6456 = LMH
X
Conversions:
0
0.5 1.0 1.5 2.0 2.5
1 bar = 14.5 psi
Transmembrane Pressure (bar)
Figure 29: Example of Optimization

Optimization Page 21
Concentration

Step 8: Concentration

8. Concentration

Safety Information

8.1 Begin Concentration It is

Take appropriate recommended
safety measures to With system running at optimum that target
avoid injury or TMP, direct permeate to collection concentration
contact with fluids. vessel, start stopwatch to time factor is exceeded
concentration step. to allow for
pooling of product
recovery samples
to achieve
8.2 Record Parameters at Time = 0 maximum
Measure permeate flow (using stopwatch and graduated recovery and
cylinder). Record flow rate and all other parameters on target
logging sheet or in a suitable spreadsheet. concentration
factors.

During
8.3 Record Parameters Ongoing concentration the
At a suitable interval (collected volume or elapsed time) system can
measure permeate flow (using stopwatch and graduated always be placed
in recirculation
cylinder). Record flow rate and all other parameters on mode as a safe
logging sheet or in a suitable spreadsheet. condition if
necessary. If this
is done, the
stopwatch should
8.4 Plot Data be paused until
Either using graph paper or a spreadsheet, plot each data concentration is
point to create a graph of Flux rate Vs log Volume re-started.
Concentration Factor (VCF). See Page 23.

During
concentration it is
8.5 Continue until End-Point normal for the
Continue concentrating until desired end-point. This may process flux rate to
be a target concentration factor, a maximum pressure or a gradually decrease
with increasing
target point (max pressure or VCF) to begin diafiltration. concentration
factor. If crossflow
is maintained
8.6 Maintain Recirculation throughout,
Proceed to Diafiltration or Product Recovery, as necessary, pressures are also
likely to increase.
or the system can be placed in recirculation before The plot of Flux
moving to diafiltration or product recovery. Do not stop vs. log VCF should
the pump until you are ready to recover the product be a straight line.
from the system.

Page 22 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Concentration

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PUMP PERMEATE PUMP PERMEATE

4 FEED
VESSEL
4 FEED
VESSEL

Figure 30: Concentration Configuration (8.1) Figure 31: Recirculate at target end-point
(8.5 – 8.6)

Plotting Flux Vs VCF

Permeate Flux (LMH)

60 X CG =
X Gel Layer Concentration
X
X CD =
40 X Diafiltration Concentration
X CG (Optimum)
X X
X CG
20 X CD =
e1.0
0 CD = 0.368 x CG
1 2 3 4 5 6 7 8 910 20 30 40 50 60 70 100

Volume Concentration Factor (VCF) (log scale)

Figure 32: Example of Concentration Plot

Equations:
Sample Volume
Initial Sample Concentration =
Sample Volume + Hold-up Volume

Volume Concentration Factor Total Initial Volume (sample + hold-up)

=
(VCF) Total Initial Volume - Permeate Volume collected

Actual (theoretical) Product = Initial Sample Concentration x VCF

Concentration

Figure 33: Equations for Concentration Factor and Product Concentration

Concentration Page 23
Diafiltration

Step 9: Diafiltration

9. Diafiltration

Safety Information

9.1 Diafiltration
Before starting diafiltration decide what type of diafiltration Volume (DV)
(continuous or discontinuous) that you are going to perform One diafiltration
and how many diafiltration volumes you will use. volume (DV) is
equal to the total
volume on the
upstream side
9.2a(i) Continuous 9.2b(i) Discontinuous prior to
Begin continuous diafiltration Begin discontinuous diafiltration.
using a vacuum on the feed diafiltration by adding one
vessel or a pump to diafiltration volume of
introduce buffer to the feed buffer to the feed vessel.
vessel. Direct permeate to Direct permeate to Continuous
collection vessel. collection vessel. Diafiltration
Diafiltration buffer
is drawn into the
feed vessel by
9.2a(ii) Continuous 9.2b(ii) Discontinuous vacuum created
Measure permeate volume Concentrate sample back as permeate is
until required end-point to original volume. Add removed from the
(number of DVs or other further DVs and repeat as system or added
using a transfer
e.g., target conductivity). required. pump matched to
the permeate
9.3 Record Parameters Ongoing flow rate.
At a suitable interval (collected volume or elapsed time)
measure permeate flow (using stopwatch and graduated
cylinder). Record flow rate and all other parameters on
Discontinuous
logging sheet or in a suitable spreadsheet. Diafiltration
One diafiltration
volume of buffer is
9.4 Plot Data added to the feed
Either using graph paper or a spreadsheet, plot each data vessel (sample is
point to create a graph of Flux rate vs. Number of diluted) and the
total volume
Diafiltration Volumes (DVs). See Page 25. is then
re-concentrated
back to the initial
9.5 Maintain Recirculation volume. This is
repeated several
At the end of diafiltration the system should be placed in times, until the
recirculation mode. Do not stop the pump until you are desired end point
ready to recover the product from the system. is reached.

Page 24 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Diafiltration

BUFFER BUFFER
VESSEL VESSEL
Qbuffer = Qpermeate
Qbuffer

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL
1 DV

Qpermeate

PUMP PERMEATE PUMP PERMEATE

4 FEED
VESSEL
4 FEED
VESSEL

Figure 34: Continuous Diafiltration (9.2a) Figure 35: Discontinuous Diafiltration,

Adding 1 DV (9.2b)

Note: Some form of mixing in the feed

vessel will typically be required during
RETENTATE
diafiltration to ensure the added buffer
PERMEATE
mixes thoroughly with the product.
FEED
VESSEL

PUMP PERMEATE

4 FEED
VESSEL

Figure 36: Recirculate at Target End-point (9.5)

Plotting Flux vs DVs

Permeate Flux (LMH)

60

40

20 X X X X X X

0
1 2 3 4 5
Number of Diafiltration Volumes (DVs)
Figure 37: Example of Diafiltration

Diafiltration Page 25
Product Recovery

Step 10: Product Recovery

10. Product Recovery

Safety Information

The procedure
10.1 Maximize Recovery shown is an
With system in recirculation from previous step (either example only.
concentration or diafiltration), open the retentate valve fully.
Product recovery
should be
optimized for each
process

10.2 Close Permeate

Close the permeate valve fully.
To maximize
product yield at
the desired final
10.3 Adjust Crossflow concentration, it is
Adjust pump speed to achieve desired process crossflow. recommended that
Recirculate for 10 minutes. target
concentration
factor is exceeded
to allow for pooling
10.4 Purge Vessel of product recovery
samples to
Stop pump. Direct retentate to collection vessel. Run pump achieve maximum
slowly to purge volume from feed vessel into the collection recovery and target
vessel (pipework/tubing remains full). Stop pump. concentration
factors

10.5 Purge Pipework/Tubing

Add one upstream hold-up volume of buffer to feed vessel. Run
pump slowly until vessel is just empty (hold-up volume purge).
Stop pump.

10.6 Additional Recovery

Introduce buffer into feed vessel such that total system volume
= minimum working volume. Repeat steps 10.3 to 10.5.
Use pump to air purge lines. Collect recovered sample in a
separate container.

Page 26 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Product Recovery

RETENTATE RETENTATE

PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PUMP PUMP

4 FEED
4 FEED

Figure 38: Recirculation with Screen Channel Figure 39: Purge Vessel (10.4)
Cassettes (10.3)

RETENTATE RETENTATE
PERMEATE
PERMEATE

FEED
VESSEL FEED
VESSEL

PUMP PUMP
WASTE
4 FEED 4 FEED

Figure 40: Purge Pipework/Tubing (10.5) Figure 41: Additional Recovery,

Blow out Feed/Retentate Line (10.6)

Product Recovery Page 27

Buffer Flush

Step 11: Buffer Flush

11. Buffer Flush

Safety Information

11.1 Buffer flush is

Take appropriate
Drain the vessel by running the recommended to
safety measures to remove foulants
pump and directing retentate and
avoid injury or that can be easily
permeate to waste.
contact with fluids. washed out prior
Stop the pump. to introducing
chemicals.
Alternatively,
11.2 users can go
Introduce a suitable volume of straight to Step
buffer or WFI into feed vessel to 12. CIP.
allow flushing to waste through the
retentate and permeate lines.

Recirculation
times and
11.3 volumes are
With permeate closed, pump application
buffer through retentate to specific. Must be
waste. Stop pump after validated.
approximately 2 total hold-up
volumes have passed to waste.

11.4
Open permeate valve. Start pump
and apply retentate pressure to
purge the permeate line to waste.

11.5
Drain System (Figure 42) ready for
Step 12. CIP.

Page 28 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Buffer Flush

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE WASTE

FEED FEED
VESSEL VESSEL

WASTE WASTE
PUMP PUMP
4 FEED ´ FEED

Figure 42: Drain via Pump, Retentate and Figure 43: Retentate and Permeate to Waste (11.2)
Permeate to waste (11.1)

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE WASTE

FEED FEED
VESSEL VESSEL

WASTE WASTE
PUMP PUMP
4 FEED 4 FEED

Figure 44: Retentate to Waste Figure 45: Permeate Flush to Waste under Pressure (11.4)
Permeate Closed (11.3)

Buffer Flush Page 29

Clean in Place (CIP)

Step 12: Clean in Place (CIP)

12. Clean In Place (CIP)

Safety Information

Take appropriate 12.1 Drain System

Ensure you use a
safety measures to Drain system and reservoir by large enough
avoid injury or pumping to waste. volume to enable
contact with fluids. an initial purge to
waste followed by
recirculation.

12.2 Add CIP Fluid

Fill reservoir with suitable volume
Recirculation
of CIP fluid (see Table 4). times, volumes,
sanitization agent,
and temperature
are application
specific. Must be
12.3 Flush to Waste validated.
Run pump to flush approximately
one or two total hold-up volumes CIP retentate
to waste. flowrate should be
1.5 x process
retentate
flow rate.
12.4 Recirculation If CIP solution
Before feed vessel runs empty, looks dirty after
place system in recirculation only a few
mode. minutes, stop
cleaning, and
begin again at
12.1.

12.5
Close permeate valve during Permeate valve may be closed or
CIP. open during cleaning. If more than
one cleaning cycle — close during
first cycle and then leave open.

12.6 12.7 Repeat NWP

Recirculate CIP solution for Repeat NWP (Step 4)
45 – 60 minutes. Drain and flush to determine %
with WFI/equivalent. membrane recovery.

Page 30 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Clean in Place (CIP)

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE

FEED
FEED VESSEL
VESSEL

WASTE
PUMP PUMP

4 FEED
4 FEED

Figure 46: Retentate and Permeate Figure 47: Recirculation with Permeate Open (12.4)
to Waste (12.1 and 12.3)

RETENTATE RETENTATE
PERMEATE PERMEATE

FEED FEED
VESSEL VESSEL

PUMP PUMP

4 FEED
4 FEED

Figure 48: Recirculation with Permeate Closed (12.5) Figure 49: Recirculation with Permeate Open, Retentate
Partially Closed (12.5)

Table 4: Recommended CIP Solutions and Conditions

Membrane Type Choose Suitable CIP Solution Temperature

0.1 – 0.5 N NaOH

Omega 25 – 45 °C
0.1 – 0.5 N NaOH + 200 – 400 ppm NaOCl

Clean in Place (CIP) Page 31

Storage

Step 13: Storage

13. Storage

Safety Information

Take appropriate 13.1 Drain

safety measures to Drain system and add storage
avoid injury or agent to feed reservoir. Ensure you use a
contact with fluids. large enough
volume to enable
an initial purge to
waste followed by
13.2 Flush to Waste recirculation.
Set up the system with retentate
and permeate directed to waste.

13.3 Flush to Waste Recirculation

Pump storage solution through times and
both retentate and permeate to volumes are
waste (see Table 5). application
specific. Must be
validated.

13.4 Recirculation During

Before feed vessel runs empty, recirculation,
place system in retentate flow rate
recirculation mode. should be 1.5 x
process retentate
flow rate.

13.5 Recirculation
Apply some retentate pressure
and recirculate storage solution for
3 – 5 minutes.

13.6a
Storage in Holder
Close all feed, retentate and permeate
valves. If auto-torque, reduce AT
pressure by 25%. Check for leaks.
13.6b
Storage out of Holder
Place cassettes in sealed plastic
bags. Place bags into container with
lid. Store out of light at 4 – 15 °C.

Page 32 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Storage

RETENTATE RETENTATE
PERMEATE PERMEATE
WASTE
FEED
VESSEL
FEED
VESSEL

WASTE
PUMP PUMP

4 FEED
4 FEED

Figure 50: Retentate and Permeate Figure 51: Recirculation with Pressure (13.4 and 13.5)
to Waste (13.2 and 13.3)

Table 5: Recommended Storage Solutions and Conditions

Membrane Type Omega

< 6 months 0.05 – 0.1 N NaOH

> 6 months 15% Glycerin + 0.05% Sodium Azide

0.05 - 0.1N NaOH*

Temperature 4 – 15 °C Optimal
25 °C Maximum

* If stored in NaOH for more than 6 months, it is recommended that the pH of solution in the cassettes is checked at 6 month intervals.
Cassettes can be flushed with fresh storage solution and then left for another 6 months.

Storage Page 33
Storage

Torque Recommendations
Periodically check the torque on each bolt for MT holders. The cassette will compress initially
upon installation. Increasing the temperature of the feed solution will cause expansion of the
cassettes, causing the clamping force to increase. Reduce torque accordingly if the system is to
contain heated solution.

Table 6: Recommended Membrane Cassette Torque Values

Recommended Torque Range for Manual Torque Cassette
Holders
Holder Type No. of Bolts on Holder Omega TFF Membrane Cassettes

inch-lb Nm

Centramate LV 4 70 – 90 8 – 10

Centramate 4 70 – 90 8 – 10

Centrasette 5 4 350 – 550 40 – 60

Centrasette LV 4 350 – 550 40 – 60

Centrasette 10 2 700 – 1100 80 – 120

Centrasette P 2 700 – 1100 80 – 120

Centrasette 10 4 350 – 550 40 – 60

Recommended Pressure Range for Auto-torque Cassette

Holders
No. of Hydraulic Pistons on
Holder Omega TFF Membrane Cassettes

psig barg

Centrasette LV AT 4 750 – 1100 52 – 76

Centrasette 5 AT 4 750 – 1100 52 – 76

Centrasette 10 AT 2 1500 – 2250 103 – 155

Centrastak 10 AT 2/level 1500 – 2250 103 – 155

Page 34 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Storage

Operating Specifications

Recommended Cross Flow Flux (CFF) rates for Pall Membrane Cassettes
Table 7 below lists the recommended CFF rates for operating Pall Membrane Cassettes.
CFF = retentate flow rate (L/min) / membrane area (m² or ft²)

Table 7: Recommended Crossflow Rates

Recommended CFF
Holder Type Units Minimum CFF Process Recommended CFF Process Cleaning (CIP)

Screen Channel Cassettes Screen Channel Cassettes Screen Channel Cassettes

L/min/m2 3.0 5.0 – 8.0 7 – 12

Centramate or
Centrasette
L/min/ft2 0.3 0.5 – 0.8 0.7 – 1.2

Table 8 gives the operating limits for pressure, temperature and pH. Exceeding any of these limits
may cause permanent damage to the cassette and pose a safety hazard for the operator.

Table 8: Cassette Operating Limits for Pressure, Temperature, and pH

Maximum Recommended Operating Pressure pH Range

Membrane Type Temp bar psi

4 – 23 °C 6.0 90
Omega 2 – 14
24 – 55 °C 4.0 60

Storage Page 35
Storage

Temperature Correction Factors

Table 9: Temperature Correction Factor (TCF)
Temp Temp TCF Temp Temp TCF Temp Temp TCF

°C °F °C °F °C °F

60 140 0.47 40 104 0.65 20 68 1.00

59 138 0.47 39 102 0.66 19 66 1.03

58 136 0.48 38 100 0.68 18 64 1.05

57 135 0.49 37 99 0.69 17 63 1.08

56 133 0.50 36 97 0.70 16 61 1.11

55 131 0.50 35 95 0.72 15 59 1.14

54 129 0.51 34 93 0.73 14 57 1.17

53 127 0.52 33 91 0.75 13 55 1.20

52 126 0.53 32 90 0.76 12 54 1.23

51 124 0.54 31 88 0.78 11 52 1.27

50 122 0.55 30 86 0.80 10 50 1.30

49 120 0.56 29 84 0.81 9 48 1.34

48 118 0.56 28 82 0.83 8 46 1.38

47 117 0.57 27 81 0.85 7 45 1.43

46 115 0.58 26 79 0.87 6 43 1.47

45 113 0.59 25 77 0.89 5 41 1.52

44 111 0.61 24 75 0.91 4 39 1.57

43 109 0.62 23 73 0.93 3 37 1.62

42 108 0.63 22 72 0.95 2 36 1.67

41 106 0.64 21 70 0.98 1 34 1.73

Page 36 T-Series Membrane Cassette Care and Use Procedures Quick Reference Guide
 Storage

Diafiltration — Continuous vs. Discontinuous

Table 10: Continuous vs Discontinuous Diafiltration Table
Percent Transmission Continuous Diafiltration Discontinuous Diafiltration
(passage) of a Salt
Solute Removal Number of DVs Solute Removal Number of DVs

63.21 1 50.00 1

86.27 2 75.00 2

100% Passage 95.02 3 87.50 3

98.17 4 93.75 4

99.33 5 96.88 5

55.07 1 42.57 1

79.81 2 67.01 2

80% Passage 90.93 3 81.05 3

95.92 4 89.12 4

98.17 5 93.75 5

Conversion Tables
Table 11: Conversion Tables
Area Pressure Flow Volume

ft2 to m2 psi to bar m3/hr L/min Gal to Liter

0.1 0.0092 1 0.07 1 to 16.7 1 3.78

0.2 0.0186 10 0.7 m3 Liter

1 0.0929 30 2 1 1000

2 0.186 bar to psi

5 0.46 1 14.5

m2 to ft2 bar to kPa

1 10.76 1 100

Storage Page 37
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All data, specifications and information contained in this publication are based on information we
believe reliable and represent values in effect at the time of printing. Pall Corporation reserves the right
to make changes without prior notice.

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