RESEARCH IIi
TFF SYSTEM
®
KrosFlo Research IIi TFF System
Product Information and Operating Instructions
advancing the science of separation™
TRADEMARKS AND WARRANTY
Spectrum’s KrosFlo® Research IIi TFF System meets strict quality control standards and is warranted against
defects in material and workmanship for a period of two (2) years from date of shipment.
The information contained herein is believed to be accurate and is offered in good faith for the convenience of
the user. PRODUCTS ARE FURNISHED UPON THE CONDITION THAT THE USER ASSUMES ALL RISKS
AND LIABILITIES AND THAT NEITHER THE SELLER NOR MANUFACTURER SHALL BE LIABLE FOR ANY
LOSS OR DAMAGE, DIRECT OR CONSEQUENTIAL, ARISING FROM THE USE OF THESE PRODUCTS.
Spectrum®, KrosFlo®, MicroKros®, MidiKros® and MiniKros® are registered trademarks of Spectrum
Laboratories, Inc. Microsoft®, Windows® and Excel® are registered trademarks of Microsoft Corporation. CFlex® and Pharmapure® are registered trademarks of Saint-Gobain. MasterFlex® is a registered trademark of
Cole-Parmer. Trademarks bearing the ® symbol in this publication are registered in the U.S. and in other countries.
This document copyright © 2015 by Spectrum Laboratories, Inc. All rights reserved. Reproduction prohibited
except by permission of the copyright owner.
ii
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
TABLE OF CONTENTS
Table of Contents
System Overview and Operation
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2. Safety Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
4. System Configuration and Major Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
4.1 KrosFlo® Research IIi Pump Drive, Pump Head, & Integrated Pressure Monitor . . 5
4.2 KrosFlo® Research IIi Mounting Hardware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4.3 KrosFlo® Research IIi Octopus Cable . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4.4 Flowpath Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4.5 Starter Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
5. System Options and Parts List . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
6. Materials of Construction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
7. KrosFlo Research IIi Pump/Pressure Monitor Set Up and Operation. . . . . . . . . . . . . . 8
7.1 Installation and Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
7.2 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
7.2.1 Turning on the System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
7.2.2 Control Panel. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
7.2.3 Home Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
7.2.4 Setup Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
A. Pressure Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
B. Alarm Settings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
C. Tubing Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
D. Hardware Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
E. Diagnostics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
7.2.5 Troubleshooting and Error Messages. . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
7.3 Updating KRIIi Firmware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
8. KrosFlo Research IIi Pump Head Set Up and Operation . . . . . . . . . . . . . . . . . . . . . . 17
8.1 Explanation of Symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
8.2 Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
8.3 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
8.4 Installation and Removal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
8.5 Tubing Loading and Unloading . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
8.6 Multi-Channel Mounting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
8.7 Maintenance and Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
9. KF Comm Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
9.1 KF Comm License . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
9.2 Installation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
9.2.1 KF COMM Software Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
1
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
TABLE OF CONTENTS (continued)
9.2.2 Drivers for USB Connection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
9.3 Instructions for Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 5
9.3.1 Opening the Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
9.3.2 Data Collection Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
9.3.3 Navigating the Worksheets and Charts . . . . . . . . . . . . . . . . . . . . . . . . . 27
9.3.4 Compliancy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
10. Basic Concepts of Tangential Flow Filtration (TFF) . . . . . . . . . . . . . . . . . . . . . . . . . . 31
10.1 Dead-end Filtration Tangential Flow Filtration (TFF) . . . . . . . . . . . . . . . . . . . . . . 31
10.2 Assembling the Flowpath . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
10.2.1 Constant Volume Mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
10.2.2 Batch Mode. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
10.3 Integrity Testing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
10.3.1 Leak Test. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
10.4 KRIIi as a System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
10.4.1 Batch Concentration (Product in the Retentate). . . . . . . . . . . . . . . . . . . 42
10.4.2 Batch Clarification (Product in the Permeate). . . . . . . . . . . . . . . . . . . . . 42
10.4.3 Constant Volume . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
10.4.4 Diafiltration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
10.5 Operating the System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
10.5.1 Instructions for Batch Concentration/Clarification . . . . . . . . . . . . . . . . . 45
10.5.2 Instructions for Constant Volume & Diafiltration . . . . . . . . . . . . . . . . . . . 46
10.6 Pre-Straining . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
10.7 Process Optimization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
10.8 In-Process Module Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
10.8.1 Pump Off Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
10.8.2 Forward Flushing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
10.8.3 Reverse Flushing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
10.9 Module Selection and Scale-up . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
11. Optional Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
11.1 Permeate Scale Compatible with the KrosFlo Reserch IIi System . . . . . . . . . . . 52
11.1.2 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
11.1.3 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
11.2 Automatic Backpressure Valve for KrosFlo Reserch IIi System . . . . . . . . . . . . . 56
11.2.1 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
11.2.2 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
12. Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
13. Contact Information - Worldwide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
2
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
INTRODUCTION
1. Introduction
The KrosFlo® Research IIi Tangential Flow Filtration (TFF) System is the ideal system for small-scale
process or R&D studies for microfiltration and ultrafiltration applications. The system consists of the
KrosFlo® Research IIi Pump and Pump Head, with an integrated KrosFlo® Digital Pressure Monitor,
and a disposable flow path, all of which have features that ensure efficient and reproducible TFF
processes. The Pump Drive used in conjunction with the KrosFlo® Research IIi Pump Head provides
flow rates from 0.01 to 2300 ml/min. The system has both audible and visual low and high pressure
alarms for the inlet and permeate, and a high pressure pump shut-off that help maintain membrane
integrity and achieve high product recovery. The KrosFlo Research IIi TFF system comes standard
with KF Comm, a software program that automatically downloads and graphs the run data — including inlet, retentate and transmembrane pressures and flow rates — into an MS Excel® spreadsheet.
The disposable flowpath — including the filter, pressure transducers, tubing and fittings — eliminates
the possibility of cross contamination and allows samples to be concentrated down to as low as 1
ml. Other standard features include: a digital readout of the recirculation flow rate, the easy-to-use
KrosFlo® Research IIi Pump Head, and adjustable holders attached to the pump drive that secure the
process vessels, SpectrumLabs.com’s HF modules and flowpath components.
The membrane surface areas range from 5 cm2 to 1050 cm2 and are available in 0.5, 0.75 or 1.0 mm
ID Hollow Fibers made of Polysulfone (PS), Modified Polyethersulfone (mPES), Polyethersulfone
(PES), or Mixed Cellulose Ester (ME). Pore sizes range from 1 kD up to 0.65 µm for both Ultrafiltration
and Microfiltration applications.
The KrosFlo® Research IIi TFF System offers many conveniences over traditional cross-flow membrane systems. The hollow fiber (HF) membrane modules and cross-flow filtration system provides
faster and gentler separation that helps avoid membrane fouling and maximizes product recovery.
The disposability of the modules eliminates not only the potential for cross-contamination and costs
associated with cleaning and rinsing, but also the difficulties associated with validating reusable membranes. Used in conjunction with HF membrane modules, the KrosFlo® Research IIi TFF System
offers the following advantages:
•
Faster process times
•
Superior filtration dynamics
•
Module disposability
•
Lower costs
•
Direct and easy scale-up for production volumes
2. Safety Precautions
3
DANGER:
High voltages exist and are accessible in the KrosFlo® Research IIi System.
Use extreme caution when servicing internal components.
WARNINGS:
Tubing breakage may result in fluid being sprayed from pump. Use
appropriate measures to protect operator and equipment.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
SAFETY PRECAUTIONS
Turn drive off before removing or installing tubing. Fingers or loose clothing may
get caught in drive mechanism.
CAUTION:
Power must be turned off before connecting the external Octopus cable
to prevent damage to the drive.
CAUTION:
To avoid electrical shock, the power cord protective grounding conductor
must be connected to ground. Not for operation in wet locations as defined
by EN 61010-1.
CAUTION:
Do not contaminate the lubricant in the container, on the shaft or on the seal
with foreign material. Failure to observe this precaution may result in damage
to the seal and premature failure of the seal.
CAUTION:
Risk of crushing. Keep fingers away from rotor while pump is in operation.
Stop pump before loading or unloading tubing.
Explanation of Symbols
CAUTION:
Risk of Danger. Consult Operating Instructions for nature of hazard and
corrective actions.
CAUTION:
Risk of electric shock. Consult Operating Instructions for nature of hazard and
corrective actions.
WARNINGS:
PRODUCT USE LIMITATION
This product is not designed for, nor intended for use in patient connected applications; including, but not limited to, medical and dental use, and accordingly has not
been submitted for FDA approval.
3. Specifications
Output
Speed:
0.1 to 600 rpm
Torque output, Maximum:
180 oz-in (13 kg•cm)
540 oz-in Starting
4
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
SPECIFICATIONS
Speed regulation:
Line ±0.1% F.S.
Load ±0.1% F.S.
Drift ±0.1% F.S.
Display:
128 x 64 LCD w/ LED Backlight
Pressure Sensor Limits:
-9.99 to 75 psi
Supply voltage limits:
90 to 260 Vrms @ 50/60 Hz
Input
(Universal Input)
Current, Maximum:
2.2A @ 115 Vrms, or 1.1A @ 230 Vrms
Construction
Dimensions (L × W × H):
10.5 in × 8 in × 8 in
(267 × 203 × 203 mm)
Weight:
13 lb (5.9 kg)
Enclosure Rating:
IP 33 per IEC 60529
Environment
Temperature, Operating:
0° to 40°C (32° to 104°F)
Temperature, Storage:
–25° to 65°C (–13° to 149°F)
Humidity (non-condensing):
10% to 90%
Altitude:
Less than 2000 m
Pollution Degree
Pollution Degree 2
(Indoor use - lab, office)
Chemical Resistance:
Exposed material is aluminum,
ABS plastic and vinyl
Pressure Sensors either
Polycarbonate or Polysulfone
Compliance:
(For ETL Mark);
UL 61010-1, CAN/CSA C22.2 No. 61010-1
(For CE Mark):
EN61010-1 (EU Low Voltage Directive) and
EN61326 (EU EMC Directive)
RoHS
4. System Configurations and Major Components
4.1 KrosFlo® Research IIi Pump Drive, Pump Head, & Integrated Pressure Monitor
The KrosFlo® Research IIi TFF System comes with the Digital Pressure Monitor integrated into the Pump
Drive as one unit. The Programmable Logic Controller (PLC) runs both the Pump and Pressure monitor
functions. The Precision Pump Head (ACR2-H3S-01N) can accommodate tubing sizes from #13 (1/16”
HB) to #18 (3/8” HB) and can handle rates from 0.006 ml/min up to 2300 ml/min. The High Performance
5
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
SYSTEM CONFIGURATIONS AND MAJOR COMPONENTS
Pump Head (ACR2-H4S-01N) can accommodate tubing sizes #15 (3/16” HB) to #36 (3/8” HB) and can
handle rates from 1.7 ml/min up to 2900 ml/min.
4.2 KrosFlo® Research IIi Mounting Hardware
The KrosFlo® Research IIi TFF System comes with an attached mounting plate for the KrosFlo®
Research IIi Pump Head and a stainless steel mounting bracket to connect the stainless steel hoop
post. The stainless steel hoop supports the Module Trilobites and Reservoir Trilobites that come with
the Starter Kit. The module Trilobites can accommodate SpectrumLabs.com’s MicroKros®,
MidiKros®, MiniKros® Sampler, and some of the MiniKros® hollow fiber filtration modules. The reservoir Trilobites can secure reservoirs from 15 ml up to 2 L. A standard lab clamp is also included to
support larger reservoirs or filters as necessary.
4.3 KrosFlo® Research IIi Octopus Cable
The system comes with an ‘Octopus cable’ that has the following connections: 3 pressure transducers (feed, retentate, permeate), a USB port for capturing data on a computer, an RS232 port for connecting a permeate Scale (optional) and a secondary RS232 port to connect the Automatic
Backpressure Valve (optional).
4.4 Flowpath Kit
The Universal Flowpath Kit includes many different sized fittings for #13 tubing up to #17 tubing to
accommodate the three sizes of filtration modules used in the system: MicroKros® (1 ml – 100 ml),
MidiKros® (100 ml – 3 L), and MiniKros® Sampler (3L – 15 L). One each of 15 ml, 50 ml, and 250 ml,
conical bottom reservoirs are included. Packages of size #14 and #16 tubing are also included.
4.5 Starter Kit
The starter kit includes other items needed to construct and support the flowpath, including reusable
items such as: Trilobites, flow restrictors, tie wraps and a tie wrap gun.
5. System Options and Parts List
The available universal system with corresponding catalog number is listed below:
KrosFlo® Research IIi TFF System Selection Guide
Part Number
Pump Capacity
Tubing
SYR2-U20-01N
2.3 LPM, 110/220V
Pharmapure®, #14, #16
OPTIONAL EQUIPMENT:
ACPC-F16-01N
ACPC-F17-01N
ACR2-SC5-01N
ACR2-S35-01N
ACR2-SC4-01N
ACR2-SC2-01N
ACR2-SC8-01N
ACR2-SC6-01N
ACR2-SC9-01N
ACR2-SC7-01N
ACR2-H4S-01N
6
Automatic Backpressure Valve for use with #13,14 or 16 tubing
Automatic Backpressure Valve for use with #17 tubing
Ohaus® Scout Pro Scale 400g (0.01g), 110V
Ohaus® Scout Pro Scale 400g (0.01g), 220V
Ohaus® Scout Pro Scale 4000g (0.1g), 110V
Ohaus® Scout Pro Scale 4000g (0.1g), 220V
Ohaus® Navigator Scale 10000g (1.0g), Universal
Ohaus® Navigator Scale 21000g (0.10g), Universal
Ohaus® Navigator Scale 20000g (1.0g), Universal
Ohaus® Navigator Scale 51000g (0.5g), Universal
KRII Pump Head for use with High Performance Tubing #15, 24, 35 or 36 tubing
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
SYSTEM OPTIONS AND PARTS LIST
System Parts List
Part Description
Quantity per System
KrosFlo® Research IIi Pump Drive w/ Integrated Pressure Monitor
Stainless Steel Hoop Posts w/ Mounting Screws
Attached Mounting Bracket for Pump Head
Octopus Cable
USB Extension Cable
Power supply Cable
Digital Pressure Transducers
KF COMM Data Collection Software Disc
KrosFlo® Research II Precision Pump Head
Flowpath Kit
Plastic Fittings Box w/ Accessories
#14 Extended Life Pharmapure® Pump Tubing
#16 Extended Life Pharmapure® Pump Tubing
Starter Kit
Tie Wrap Gun
Tie Wraps
Module Trilobite
Reservoir Trilobite
Pressure Control Valve
5/16” to 3/8” OD Tube
5/32” to 1/4” OD Tube
Reservoirs
15 ml Conical Bottom with C-Flex® Molded Seal
50 ml Conical Bottom with C-Flex® Molded Seal
250 ml Conical Bottom with C-Flex® Molded Seal
1
1
1
1
1
1
3
1
1
1
25 ft
25 ft
1
100
2
2
2
2
1
1
1
6. Materials of Construction
Part Description
Material
KrosFlo® Research IIi TFF System (Product Contact Surfaces)
C-Flex® / Pharmapure®
Tubing / Reservoir Closures
Reservoirs
Polypropylene
Disposable Pressure Transducers
Polysulfone
Plastic Fittings
Polypropylene / Polycarbonate
Hollow Fiber Module (Module Sold Separately)
Membrane
Housing
Potting
End-caps
7
ME, mPES, PES or PS
Pigmented and Non-pigmented Polysulfone
Polyurethane / Epoxy
Pigmented Polysulfone
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PUMP/PRESSURE MONITOR SET UP AND OPERATION
7. KrosFlo Research IIi Pump/Pressure Monitor Set Up and Operation
7.1. Installation and Setup
Before Starting Drive
•
The drive should be mounted on a flat horizontal surface, with no more than two (2)
Pump Heads mounted.
•
The ambient air temperature should not exceed 104° F (40° C) and adequate air flow
should be provided for.
•
The Octopus Cable should be attached to the back connector on the drive and the
pressure sensors should be connected.
•
Tubing should be clean and routed so that bend radii are at a minimum four (4) times
the tube diameter and as short as possible.
•
Use a tube size of appropriate diameter for the flow rate and viscosity required.
•
To maintain the best accuracy of flow rates, re-calibrate tubing regularly.
•
For tubing selection and compatibility, see Tubing Selection Guide provided with the
pump head.
•
When cleaning or performing maintenance, please remove power from the drive.
7.2. Operation
7.2.1 Turning On the System
1.
Plug the power cord into the IEC Connector, located on the rear of the drive. Plug the
opposite end of the power cord into an electrical outlet.
2.
Flip the power switch located on the rear of the drive.
3.
After the flash screen appears, the home screen will appear on the LCD screen.
7.2.2. The Control Panel
•
8
To navigate all menus on the drive, use the directional pad directly to the right of the
LCD screen.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
THE CONTROL PANEL
•
The
(ENTER) key, located in the middle of the directional pad, is used to
enter a highlighted field or option. This key is often referred to as the ENTER key in
this manual.
•
The
(START/STOP) key, located at the top right of the control panel, is used to
start and pause the drive. This key is functional at any time, even when the display is
in a setup screen. This key is often referred to as the START/STOP key in this manual.
•
The ALARM OFF key, located at the bottom right of the control panel, is used to
silence the audible alarm.
7.2.3. Home Screen
9
A.
Feed Pressure – Measurement from pressure sensor on the inlet or feed of the TFF filter.
Units can either be psi, mbar, or bar.
B.
Permeate Pressure – Measurement from pressure sensor on the permeate of the TFF
filter. Units can either be psi, mbar, or bar.
C.
Retentate Pressure – Measurement from pressure sensor on the outlet or retentate of the
TFF filter. Units can either be psi, mbar, or bar.
D.
TMP (Transmembrane Pressure) – Calculated measurement of the average of the Feed
and Retentate pressures minus the Permeate pressure.
E.
Feed Flow Rate – Recirculation flow rate based on calculation of pump rpm and selected
tubing size. Press the ENTER key when this is highlighted to change pump speed. When
the Feed Flow Rate screen is shown, the four pressures will still display on the right side
of the screen. Units are ml/min.
F.
Setup – Pressing the ENTER key on this icon goes to the Setup screen. The Setup
screen has calibration functions, alarm settings, hardware setup and diagnostics. Please
refer to the Setup section below.
G.
Pump Direction – Pressing the ENTER key on this icon toggles between clockwise and
counterclockwise flow direction.
H.
Tubing Size – Pressing the ENTER key on this icon will select the tubing size (#13, #14,
#16, #25, HP15, #17, HP24, #18, HP35, #36, HP36).
I.
Cursor Idle Place Holder – Place the cursor on this icon when cursor is idle to avoid any
unintended changes.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
THE CONTROL PANEL
7.2.4. Setup Screen
A. Pressure Calibrate
1.
Ensure that positive air source is calibrated
2.
Connect pressure transducers to KRIIi Octopus Cable’s pressure transducer ports
3.
After readings stabilize, tare KRIIi’s pressure readings
4.
Turn on positive air source and adjust to dispense 5 psi of pressure, then turn positive
air source back off
5.
Connect Feed pressure transducer to be in-line with positive air source — cap the
remaining open end of the pressure transducer
6.
Turn on positive air source
7.
Wait for Feed pressure reading to stabilize
8.
If Feed pressure reading differs from calibrated source measurement, navigate to
Settings menu
9.
Select Pressure Calibrate
10. Select source of pressure (in this case, “Feed”)
11. Adjust “Set:” pressure reading to reflect that of calibrated source
12. Save changes by selecting “Cal”
13. Repeat steps 4-11 with the Inlet and then Outlet ports
Pf/Pp/Pr: The current reading for Feed/Permeate/Retentate Pressure
10
Set:
Press the ENTER button to change the current reading to the new calibrated
value, as shown on the calibrated pressure gauge.
Clear:
Press the ENTER button to clear all previous calibration values for the Feed sensor. A confirmation screen will appear.
Cal:
Press the ENTER button to confirm and save the new values.
Quit:
Return to Setup Screen
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
THE CONTROL PANEL
B. Alarm Settings
To set the pressure alarms, press the ENTER button on the highlighted alarm. The pressure setting
for the alarm can then be changed using the arrows. Zero sets the alarm to off.
Pf Hi Stop:
Secondary alarm that stops the pump, sounds a continuous beep and flashes the screen when the Feed pressure reaches the setpoint
Pf Hi Alarm:
Primary alarm that sounds a discontinuous beep and flashes the screen
when the Feed pressure reaches the setpoint
Pf Lo Alarm:
Alarm that sounds a beep and flashes the screen when the Feed pressure
goes below setpoint
Pp Lo Alarm:
Primary alarm that sounds a beep and flashes the screen when the
Permeate pressure goes below setpoint
Pp Lo Stop:
Secondary Alarm that stops the pump, sounds a beep and flashes the
screen when the Permeate pressure goes below setpoint
Silent Alarm:
Option to silence all audible alarms
C. Tubing Calibration
To Calibrate individual tubing sizes, follow these directions:
11
1.
Mount Pump Head to drive
2.
Insert appropriate tubing into Pump Head
3.
Insert tube inlet into supply fluid
4.
Insert tube outlet into measuring container; container should be a graduated container or
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
THE CONTROL PANEL
a container placed on a scale for increased accuracy
If using a scale, an acceptable weight-to-volume conversion for water is 1 gram = 1 ml.
5.
Turn on drive using power switch located on the rear of the drive.
6.
Go to the Setup Menu by selecting the SETUP icon and pressing the ENTER key. Use
the UP and DOWN keys to highlight TUBING CALIBRATE in the Setup Menu and press
the ENTER key.
7.
Set the drive for the desired flow direction, tube size, and flow rate. Note that these settings are retained and transferred to other mode screens when entering or leaving the
TUBING CALIBRATION screen.
•
Note that the calibration volume is fixed and cannot be changed.
8.
Divert tubing outlet into drain, then press and hold the prime key on the drive console to
fill tubing lines. Priming will stop when key is released. Direct tubing outlet back into measuring container.
9.
Highlight the START field and press the ENTER key. The drive will run based on the
default volume at the estimated flow rate selected.
10. Upon completion of the calibration run period, the CAL VOLUME field will be highlighted.
Press the ENTER key and adjust the CAL VOLUME to the measured quantity. Press
ENTER to SAVE setting and EXIT the field.
A lower case “c” will display when the calibrated tubing size is selected. The volume units
are dependent on the flow rate units. For example, defining the flow rate unit as ml/min
will change the volume units to ml; defining the flow rate unit as oz/min will change the
volume units to oz.
11. To clear the calibration, select the CLEAR option on the tubing calibration screen when
the tubing to be cleared is selected.
Tubing Calibration Notes
•
If the drive is stopped during calibration, empty the container and restart the
procedure.
•
Calibration time at maximum allowable flow rate (default max flow rate) is 5-10
seconds and at minimum allowable flow rate (approximately 4% of the maximum flow rate) is 4 minutes.
•
Minimum and maximum flow rates will change after a tubing calibration due to
a re-calculation of the vol/rev.
•
Optimum results are best obtained after tubing has been broken in by running
in pump for at least 10 minutes. Steps 8-10 can be repeated as necessary to
optimize the accuracy of the tubing cal.
Calculate Run Time Formula
•
12
60 / (flow rate [ml/min] / cal volume [ml]) = cal run time (seconds)
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
THE CONTROL PANEL
Invalid Calculated Run Time Example
•
#13 tubing flow rate range is 0.006 ml/min – 36.0 ml/min
•
At flow rate of 1 ml/min, cal run time calculation is as follows:
60 / (1 ml/min / 6 ml) = 360 seconds
360 seconds exceed the max run time of 4 minutes (240 seconds)
D. Hardware Setup
Auto Tare:
Turn on or off the auto tare function during system start up
Pressure Units:
Select between psi, mbar, or bar
Time:
Change system time
Date:
Change system date
Key beep:
Toggle key beep on/off
Display contrast:
Raise or lower the screen contrast
Psi Decimals:
Number of decimals for psi units; decimal digits for mbar and bar are fixed
Key Timeout:
Select “Yes” or “No” to toggle on/off key timeout; when “Yes,” the key
timeout will restore system to Home screen after 10 minutes of inactivity
Running Change: When on, the tubing size and pump direction can be changed while the
pump is running. If off, the pump needs to be stopped before the tubing
size or the direction can be changed.
Open Detect:
13
The open detect function prevents the drive from turning when pump head
is in the open position. Pump head must be oriented vertically. In order to
use the pump head with the inlet/outlet to the left or right, the open detect
must be turned off.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
TROBLESHOOTING AND ERROR MESSAGES
E. Diagnostics
7.2.5. Troubleshooting and Error Messages
No Power
Description:
Motor does not rotate; Display does not light
Error Condition(s): No Power
Actions:
Check fuse; replace, if necessary; check that the
unit is plugged into a live line; check connection of
power cord; check the cord for continuity; replace if
defective; return for servicing
Motor Error #1: Motor OverCurrent
Description:
Motor is drawing too much current for a short
duration of time
Error Condition(s): The motor current is above 4.0 A peak
Actions:
Drive will stop immediately; determine if pump
head is not binding and the load is above recommended maximum load; if error persists, consult
factory
Motor Error #2: Mains Overvoltage
Description:
The measured AC voltage reported by the drive is
too high
Error Condition(s): The drive voltage is above 260V AC
Actions:
Pump will stop immediately; check supply line
voltage, if error persists, consult factory
Motor Error #3: Mains Undervoltage
Description:
The measured AC voltage reported by the
drive is too low
Error Condition(s): The drive voltage is below 90V AC
14
Actions:
Pump will stop immediately; check supply
line voltage
NOTE:
This error when is normal if displayed during
power down; if error persists, consult factory
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
TROUBLESHOOTING AND ERROR MESSAGES
Motor Error #4: Motor OverTemp
Description:
The motor is overheating
Error Condition(s): The temperature value from motor is above
given threshold value
Actions:
Pump will stop immediately; determine if
ambient air temperature is less than 104° F
(40° C); pump turns freely, and airflow is
unrestricted; if error persists, consult factory
Motor Error #5: Communications Lost
Description:
Communication to the motor is incorrect,
has disappeared, or some other communication error
Error Condition(s): Motor does not receive data from serial port
Actions:
cycle
Drive will attempt to stop pump; power
drive; if error persists, consult factory
Checksum Error: ROM Checksum Wrong
Description:
Run-time checksum (checked at power-on)
contains a bad checksum value.
Error Condition(s): Checksum at power-on displays invalid
value
Actions:
Power cycle the drive; if error persists, consult factory
HW Error #1: Invalid Interrupt or Address
Description:
Software jumps to an invalid address, invalid
interrupt, or other abort/exception (i.e., Data
Fetch Abort, Software Prefetch Abort,
Undefined Instruction, or FIQ Event)
Error Condition(s): May occur due to invalid pointer references,
RAM memory corruption, or other computerrelated issues; the Abort Exception/Interrupt
within the CPU is failing to branch out to its
respective exception handler functions
Actions:
15
Power cycle the drive; if error persists, consult factory
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
TROUBLESHOOTING AND ERROR MESSAGES
HW Error #2: Watchdog Timeout
Description:
Program has stopped running (i.e., Software
Locked Up) because Watchdog has not
been
updated
Error Condition(s): Interrupt triggered when the Watchdog has
not been updated.
Actions:
Power cycle the drive; if error persists, consult factory
7.3 Updating KRIIi Firmware
1.
Connect the pump to the computer using the USB cable on the Octopus Cable and
power on pump
2.
Ensure both the ProgramKRIIi.exe and the KR2iSource.cod file are in the same directory
3.
Start the ProgramKRIIi.exe by double-clicking on it
4.
Click on the Start button to begin the process
5.
Either click the Find Comm button, or select the Comm port used by the pump from the
dropdown list; it will be the same as the Comm port utilized by KFComm in Excel.
6.
Click on the Go button and the pump display will go dark and the pump will begin to
update. The update will take approximately 10 minutes. After update is completed, the
update utility screen on the computer will display a confirmation.
7.
To verify a successful update, restart the pump. The boot-up sequence should now display the most current version.
If the pump is interrupted while updating, the display will remain dark. Restart both the
pump and ProgramKRIIi.exe and proceed as before. If the display remains dark for an
extended period of time while programming, restart the process. The pump will not operate until the program updates successfully.
16
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PUMP HEAD SET UP AND OPERATION
8. KrosFlo® Research IIi Pump Head Set Up and Operation
8.1 Explanation of Symbols
WARNING
or DANGER:
This word and symbol are needed in safety messages for hazards that are not immediately
accessible, but that present a probability of serious personal injury if the hazards are not
avoided. These safety messages describe precautions that must be taken to avoid hazards.
WARNING
Warning or Danger with this symbol indicates probability of serious injury exists if crushing
hazard is not avoided.
WARNING:
To reduce risk of electrical shock, connect only to a properly grounded receptacle.
Turn drive off before removing, adjusting or installing tubing. Fingers or loose clothing may
get caught in the rollers.
(The rotor is partially exposed when the Actuator lever is in the open position.)
Do not remove back cover. The strong spring inside may cause injury.
Tubing breakage may result in fluid being sprayed from pump. Use appropriate measures
to protect operator and equipment.
Risk of crushing. Keep fingers away from rotor while pump is in operation. Stop pump before
loading or unloading tubing.
Fingers may get trapped between rollers on the pump rotor.
17
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PUMP HEAD SPECIFICATIONS
8.2 Specifications
Part Number
Roller and Bearing Material
Number of Rollers
MasterFlex® L/S® Tubing
ACR2-H3I-01N
Stainless Steel
3
L/S® 13, 14, 16, 25, 17, 18
Typical Flow, Pressure and Vacuum Data—3 roller pumps
Flow rate*
@ 600 rpm
mL/min
Discharge Pressure*
Continuous
Intermittent
psig (bar)
psig (bar)
Vacuum*
@ 600 rpm
in (mm) Hg
Suction Lift*
@ 600 rpm
ft (m) H20
®
MasterFlex
LS®
L/S®
Tubing
Tubing
@ 1 rpm
mL/Rev
L/S® 13
L/S® 14
L/S® 16
0.06
0.22
0.80
36
130
480
25 (1.7)
25 (1.7)
25 (1.7)
40 (2.7)
40 (2.7)
40 (2.7)
26 (660)
26 (660)
26 (660)
29 (8.8)
29 (8.8)
29 (8.8)
L/S® 25
L/S® 17
L/S® 18
1.7
2.8
3.8
1000
1700
2300
20 (1.4)
15 (1.0)
10 (0.7)
35 (2.4)
20 (1.4)
15 (1.0)
26 (660)
20 (510)
20 (510)
29 (8.8)
22 (6.7)
22 (6.7)
Part Number
Roller and Bearing Material
Number of Rollers
MasterFlex® L/S® Tubing
ACR2-H4S-01N
Stainless Steel
3
L/S® 15, 24, 35, 26
Typical Flow, Pressure and Vacuum Data—3 roller pumps
®
MasterFlex
LS®
L/S®
Tubing
Tubing
L/S®
L/S®
L/S®
L/S®
18
15
24
35
36
Flow rate*
@ 1 rpm
@ 600 rpm
mL/Rev
mL/min
1.8
3
4.3
5.8
1100
1800
2600
3400
Max.
Discharge Pressure*
psig (bar)
40
40
35
20
(2.7)
(2.7)
(2.4)
(1.4)
Vacuum*
@ 600 rpm
in (mm) Hg
26
26
26
24
(660)
(660)
(660)
(610)
Suction Lift*
@ 600 rpm
ft (m) H20
29
29
29
27
(8.8)
(8.8)
(8.8)
(8.3)
Number of rollers:
3
Maximum pump speed (rpm):
600
Maximum torque load—Starting:
400 oz-in (29 kg-cm)
Maximum torque load—Running:
Up to 180 oz-in (13 kg-cm)
Housing materials:
Glass-filled polypropylene (PP), polyphenylene
sulfide (PPS), nylon (PA)
Roller materials:
Stainless steel (SS)
Bearing materials:
Sealed stainless steel
Rotor materials:
Stainless steel
Chemical resistance:
Most substances, except strong acids or alkalis,
organic solvents or hydrocarbons.
Temperature, Operating ‡:
32°F to 104°F (0°C to 40°C)
Temperature, Storage:
–49°F to 149°F (–45°C to 65°C)
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PUMP HEAD SPECIFICATIONS
Humidity:
10% to 90% (non-condensing)
Altitude:
2000 m or less
Dimensions (W x H x D):
(Excluding shaft and cosmetic cover)
Operating
3.45" x 4.75" x 3.08" (8.8 cm x 12.1 cm x 7.8 cm)
Open
3.45" x 5.65" x 3.08" (8.8 cm x 14.4 cm x 7.8 cm)
Weight:
1.1 lb (0.5 kg)
*For NORPRENE , PHARMED , and TYGON
®
®
®
tubing. Values will be less with silicone, C-FLEX®, and Viton-®. Flow rate and discharge
pressure will vary based on tubing size, formulation, and operating temperature. The chart above is only a guide.
‡ Use in this temperature range for continuous duty operation with no decrease in performance or product life. Pump Heads will work
outside this range with some possible reductions in performance or product life.
19
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PUMP HEAD INTRODUCTION
8.3 Introduction
The KrosFlo® Research II Pump Head provides a simple, easy-touse peristaltic pump system when combined with a KrosFlo®
Research IIi Pump Drive. The pump head accepts several different
tubing sizes for a wide range of flow rates. The unique lever actuator design and automatic tubing retention allow for quick tubing
changes. The Pump Drive is designed to transfer or dispense liquids of various viscosities, as well as solids suspended in liquids. It
is NOT intended to pump flammable or explosive substances.
(4) Screws
A mounting kit adapts this pump head to the Pump Drive. The kit
Mounting Plate
includes a mounting plate and four screws, which are used to attach Bayonet Tabs
the mounting plate to the drive. The pump head is quickly and easily Figure 1 Attaching mounting plate to
drive
attached to the mounting plate or another pump head by means of
the bayonet feature on the back of the pump head. The pump head
Pump Shaft with Tang
may be oriented with tubing entering and exiting to the left, right, up
or down. In order to mount the pump head in the left, right, or up oriBayonet
entations, the Open Detect function must be turned off. See the
Features
Hardware Setup section under the Setup menu. Mounting orientation options may be limited with some drives. Pump heads are easily
removed using the one-finger release lever (Bayonet Lock Lever) on
Bayonet Lock
the upper left side of the pump head (see Figure 2). The tang on the
Lever, locked
pump head shaft couples the pump head to standard pump drives
from rear
position
to transmit power.
Each KrosFlo® Research IIi Pump Drive is supplied with one mounting kit pre-assembled.
Figure 2 Back of Research II Pump
Head
8.4 Installation and Removal
WARNING: Stop the pump drive before installing or
removing the pump head from the drive.
Approximately 30°
1.
If mounting plate is not attached to the pump drive, attach it using
the provided four Phillips head screws (see Figure 1).
2.
Orient the pump head with its back facing the drive and insert the
tang on the pump head shaft into the shaft’s slot on the drive. Align
the bayonet features on the back of the pump head with the bayonet tabs on the front of the mounting plate (see Figures 1, 2 and 3).
The pump head should be tilted about 30° counterclockwise from
the intended installed orientation. Press pump head firmly against
the drive and rotate clockwise until no more rotation is possible (see
Figure 4). The bayonet lock lever will automatically snap toward the
back of the pump, locking it to the mounting plate.
3.
Remove the pump head from the drive by holding the bayonet lock
lever forward while rotating the pump head as far as possible in the
counterclockwise direction, then pull the pump head away from the
drive to detach it. The actuator lever should be in the far right position when removing the pump head (see Figure 5).
Figure 3 Position for engaging bayonet
feature for horizontal mounting
Figure 4 Bayonet feature locked in
horizontal pump orientation
20
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PUMP HEAD TUBING LOADING AND UNLOADING
8.5 Tubing Loading and Unloading
Bayonet Lock
Lever-Pull
Forward
to Release
WARNING: Stop the pump drive before installing or
removing tubing from the pump head.
1.
To load tubing, open the pump head by moving the actuator
lever counterclockwise (see Figures 5 and 6). Insert a loop of
tubing into one open tubing retainer, between the occlusion
bed and the rollers and into the other tubing retainer (see
Figure 7). Position the tubing so that it is firmly centered
against the rollers. While holding the tubing ends, move the
actuator lever back to the far clockwise (right) position, as
shown in Figure 5. The pump head will automatically grip the
tubing. Approximately 5 pounds of force must be applied to
the actuator lever to fully close the pump head and place the
lever in its locked position (far right position) or to fully open
the pump head (far left position).
Actuator
Lever-Far
Right Position
Figure 5 Pump head in fully closed
position
Actuator LeverFar Left Position
Occlusion Bed
NOTE: It is unnecessary to have an end of the tubing free to load or
unload tubing from the pump head. A length of tubing may be
loaded into the pump without disconnecting it from adjacent
devices.
Roller
Figure 6 Pump head in fully open position
2.
Before unloading tubing
from the pump head, first
turn off the drive. Open
the pump head by moving the actuator lever
counterclockwise (left),
as described above. This
will automatically open
the tubing retainers, as
well as lift the occlusion
bed away from the tubing. Pull the tubing away
from the pump head.
Occlusion
Bed
NOTE: When pump is not being
used, store with actuator
lever half way between far
left and far right positions
(see Figures 5 and 6).
Tubing
Retainer
(one per side)
Figure 7 Tubing path through pump head - during loading
21
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PUMP HEAD MULTI-CHANNEL MOUNTING
8.6 Multi-Channel Mounting
1.
The KrosFlo® Research II Pump Heads can be mounted in
tandem (see Figures 8, 9 and 10). Once the mounting plate
is attached to the pump drive, no other mounting hardware
is required.
2.
Install the first pump head according to the mounting
instructions in Step 8.4
3.
To install a second pump head, the cosmetic cover must be
removed from the first pump head. Grasp the cover by the
notches and pull it off (see Figure 8). Align the second pump
head to the first, as if the first pump head were the drive, and
continue to follow pump head mounting instructions in step
8.4 (see Figures 9 and 10).
Bayonet Tabs
on Pump Front
Cosmetic
Cover
Notch (here and
on opposite edge)
Figure 8 Preparation to mount a second
pump head
NOTE: The tubing on the
inner pump head(s)
can be changed
without removing
the outer pump
head(s) from the
drive.
Figure 9 Engaging bayonet of second
pump head to bayonet tabs
on first pump head
Figure 10 Drive with both pump heads
locked in position
CAUTION: Be sure that bayonet features on back of each pump head are fully engaged with
bayonet tabs on the mounting plate or adjacent pump head before operating
pump drive. Bayonet lock lever (see Figure 5.) will snap back when bayonet
features engage completely.
8.7 Maintenance and Cleaning
No lubrication is required for the KrosFlo® Research II Pump Head. Only use a mild detergent solution or
70% isopropyl alcohol to clean the pump head. Do not immerse nor use excessive fluid. Pump head
requires no maintenance beyond cleaning. There are no user serviceable or replaceable parts inside.
22
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM SOFTWARE
9. KF Comm Software
9.1 KF Comm License
Copyright © 2015 Spectrum Laboratories, Inc., Rancho Dominguez, USA. All rights reserved.
Redistribution and use in source and binary forms, with or without modification, are permitted provided that
the following conditions are met:
• Redistribution of source code must retain the above copyright notice, this list of conditions
and the following disclaimer.
• Redistribution in binary form must reproduce the above copyright notice, this list of conditions
and the following disclaimer in the documentation and/or other materials provided with the
distribution.
• Neither the name of Spectrum Laboratories, Inc. nor the names of its contributors may be
used to endorse or promote products derived from this software without specific prior written
permission.
THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS “AS IS” AND ANY
EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF
MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL
THE COPYRIGHT OWNER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT
OF SUBSTITUTE GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION)
HOWEVER CAUSED AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR
TORT (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE POSSIBILITY OF SUCH DAMAGE.
24
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM SOFTWARE - INSTALLATION
9.2. Installation
9.2.1 KF COMM Software Installation
The KrosFlo® Research IIi TFF System is accompanied with a disc of software that can be used on Windows®
PCs. The software requires Microsoft® Windows 95 or newer and Microsoft® Excel® 32-bit 2003 or newer.
KF COMM will work properly on a 64-bit computer if Excel® 32-bit is used to run the software.
The supplied software includes an ActiveX control which allows other applications to communicate with the
Pressure Monitor, a spreadsheet template for automatically recording your process separation conditions, and
a spreadsheet for assigning COM ports. The Pressure Monitor and KrosFlo® Research IIi TFF System accessories are connected to the computer with a USB connection. The USB connection creates a virtual serial
(COM) port that the software connects to.
While there are ways to share serial ports across computers, SpectrumLabs.com has not tested doing so. The
spreadsheet templates installed with KF COMM may be copied and moved to other locations on the
Windows® PC, and will continue to retain its KF COMM capabilities. However, if the spreadsheets are moved
to a Windows® PC without KF COMM installed, the spreadsheets will not have KF COMM capabilities. The
spreadsheet can be viewed and manipulated like a typical Excel® sheet on computers without KF COMM.
NOTE: If the user does not have administrator privileges on the computer, the installation
will fail without giving notive that it has not completed sucessfully.
1.
To start the software installation, put the software disc into the CD drive and setup will
load automatically. If it does not start, double-click on the icon for the CD drive and then
double click on the Setup.exe file.
2. After the installation begins, accept the terms of the software license to continue.
3.
You will be prompted to select the installation location (typically C:\Program
Files\SpectrumLaboratories). One copy of each spreadsheet will be installed in this location. The KF Comm control will be installed in your windows system directory (typically
C:\Windows\System).
9.2.2 Drivers for USB connection
To use the USB connector, attach the USB extension cable from the Octopus cable wire labeled
“Computer” to a PC. Windows® will automatically search for the proper driver if the system is turned on. If
not found, a prompt will be displayed to find the driver location. The driver is located in the CD that comes
with the system (folder: FTDI USB Drivers). The FTDI driver can also be downloaded and installed from
http://www.ftdichip.com/Drivers/VCP.htm (the device is FT2232). Windows® will then perform a similar
search for the driver for the permeate scale even if the scale is not attached or not going to be used.
9.3. Instructions for Use
9.3.1 Opening the Template
The workbook template used to record the process parameters is named TFF Trial Template. This
Excel® workbook contains programming that automatically collects and graphs real-time run data.
Other files located in the folder include: the KrosFlo Research IIi System Manual, and an Excel® file
called Override. The Override file manually assigns the COM port for the USB data connection from
the KrosFlo Research IIi System to the PC. This is done if the COM ports cannot be detected from
the TFF Trial Template spreadsheet.
25
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM SOFTWARE - INSTRUCTIONS FOR USE
The preset macros record pressure and flow readings during the experiment. Depending on the
security settings and the version of Excel®, a prompt may ask for permission to run the macros
upon opening the template. If the Excel® security level is High, check the “Always trust macros from
this source” box before clicking the “Enable Macros” button. It may be necessary to have an
administrator accept the certificate used to allow loading of the macros.
If the macros are not enabled, the data will not be collected. If opening a workbook that already
contains data, the data remains accessible like an Excel workbook whether or not macros are
enabled.
9.3.2 Data Collection Tools
The TFF Trial Template contains both data collection worksheets and automatic graph worksheets. When
one of the data collection worksheets is open and the KrosFlo Research IIi System is connected to the
computer, the first row of the spreadsheet will display the current readings (the yellow background cells
next to the “Now”). If the system is connected and on, yet the first line remains blank, use the configure
control to find the COM port.
The first four icons on the standard toolbar can be used to control the data collection. The same items will
also appear on the Data menu at the top of the window. For newer versions of Excel, the icons appear in
the Add-Ins menu.
Configure Collection: Used to set how the data is collected. Clicking on this will bring up a
dialog.Use this dialog to set the number of seconds per line collected (default is 60 seconds per line). A 3 hour run at 1 line per second would produce about 10,000 data points.
To collect fewer points, a larger number would need to be set for the seconds per line
parameter. An Excel sheet can only contain about 65,000 rows; this limits collection to
about 18 hours at 1 second per line.
This control also can be used to find the COM port for the USB connection from the
Octopus Cable. If the COM port is unknown, click the Find Comm button. The control will
cycle through the free COM ports on the computer to identify the KrosFlo Research IIi
System. If the system is turned on and connected to the computer through the USB or
Serial connection (from the Valve serial connector), the COM port should be found within 1
cycle of the available ports. The control will continuously seek the Pressure Monitor until
one is found or until the control is cancelled. If another program is being used to communicate with the system, you'll need to close that program before selecting the Configure tool
so that the serial port becomes available to the workbook.
26
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM SOFTWARE - INSTRUCTIONS FOR USE
Start Collection: Used to start collecting data. Once the first row in the worksheet is properly
displaying the real-time pressure information, click on this icon to begin saving pressure data.
After clicking on the Start Collection button, the Configure Collection will be disabled until Stop
Collection is pressed.
Stop Collection: After clicking on Start Collection to start saving data, the Stop Collection function is used to suspend data collection. If you select Start, Stop, and then Start again, collection will resume where it left off; it does not begin anew at the top of the sheet.
End Collection: Used to end data collection. Certain functions in Excel® need to be disabled
for the data in the TFF Trial Template to be accessible. Clicking this will stop the spreadsheet
from updating the first line, stop the data collection, and disable the other controls. The data
can then be accessed for analysis.
If the End Collection button is clicked once and then the workbook is saved, data collection will
still be available the next time the workbook is opened. If the End Collection button is clicked
twice and then the workbook is saved, data collection will no longer be available the next time
the workbook is opened.
9.3.3 Navigating the Worksheets and Charts
1. Worksheets Overview
In the TFF Trial Template workbook, there are three different worksheets for data collection (orange tabs at bottom of Excel® workbook). These three worksheets are used for
collecting data for different stages of a filtration process:
!
Integrity Test – For performing test to ensure membrane integrity
Module Characteristics – For analyzing initial water flux
Trial Data – For collecting run data
NOTE: Data is only collected in the worksheet that is open.
The six remaining worksheets with the green tabs are automatic graphs. Here is a brief overview:
Graph
Description
NWP (Normalized Water Permeability)
Graphs data from ‘Module Characteristics’ work
sheet for analyzing initial water flux
CF and Pressures vs. VT
Flux Rate vs. Time
Flux vs. CF
Feed Flow Rate vs. Time
Pressures vs. Time
27
Graphs data from ‘Trial Data’ worksheet to analyze
process conditions
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM SOFTWARE - INSTRUCTIONS FOR USE
2. Header Information and Options
NOTE:
For the data collection worksheets, there are three different types of columns:
collected data (yellow), calculated data (light green) and user input data (tan)
!
The data collection worksheets have a header (see above) with various inputs for filter selection, run
options, and run data — including:
a.
Date
b.
Filter Module part number
•
c.
Serial number
d.
Tubing size through the pump head and Tubing Calibration
•
Correct tubing size must be selected for proper feed flow rates to be captured
in the spreadsheet.
•
If the pump is calibrated (see 7.2.4 section C), then the correction factor will
need to be adjusted to determine the correct feed rate. This is done while the pump
is running. Click on the icon next to the Pump Calibration value and enter the value
in the displayed flow rate on the pump display.
e.
The starting amount of material for Concentration Factor calculation
f.
The method for permeate collection – by weight or by volume
•
When using the optional permeate scale for automatic data collection, the
“By Weight” button must be selected.
g.
Filtration application, objective and notes
h.
PSI vs. mbar units for pressure
i.
Shear rate guidelines
•
Feed rate @ 4000s-1 is the recommended maximum feed flow for shear sensitive
applications. Feed Rate @ 12000s-1 is the recommended maximum feed flow for
fouling applications.
j.
Average Permeate Flux and Permeate Flow over the entire filtration run
k.
Module Table Date
•
28
When the filter part number is entered, the filter information — including the surface
area, approximate fiber count, fiber length, shear rate guidelines and membrane type —
will be automatically entered.
Date reference for when the filter module database created. Database will be
updated as new filters and filter membranes are released.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM SOFTWARE - INSTRUCTIONS FOR USE
3. Module Characteristics Worksheet
The ‘Module Characteristics’ worksheet collects data to measure the NWP of the filter.
The header for the data collection in the worksheet looks like this:
!
Breakdown of the abbreviation descriptions:
Column
Abbreviation
Description
Time
Time (H:MM:SS)
Pinlet
Inlet Pressure
Pretentate
Retentate Pressure
Ppermeate
Permeate Pressure
TMP
Calculated Transmembrane Pressure
DP
Pressure Drop through filter
Qinlet
Feed Flow (calculated from Pump RPM and tubing size)
Qpermeate
Permeate Flow (measured and entered by user at specific times)
Temp
Temperature (measured and entered by user at specific times)
TCF
Temperature Correction Factor (calculated from temperature)
Water Flux
Liters of permeate per m2 of surface area per hour
(calculated from permeate flow/SA)
Water Flux @ 20°C
Corrected Water Flux
WP
Water Permeability
NWP
Normalized Water Permeability
After a filter has been properly rinsed and/or wetted according to the instructions on the
insert titled, “Hollow Fiber Module Preparation & and Instruction Guide,” the initial water
flux data can be measured. After making sure that the first row is reading the current
pressure data, click on the Start Collection button. At the set time interval, make a permeate flux measurement by using a graduated cylinder and input the data in the Qpermeate
column. Continue collecting data points at different transmembrane pressures (TMP’s).
Input the temperature. The ‘NWP’ worksheet will automatically graph the NWP vs. TMP.
29
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM SOFTWARE - INSTRUCTIONS FOR USE
4. Integrity Test Worksheet
The ‘Integrity Test’ worksheet measures the inlet pressure at set time intervals and calculates the ∆P over time. If the psi/min change is not > 0.5 psi/min then the filter is fit for use.
A sharp decrease in pressure over time indicates either a broken filter or incompletely wetted filter. Follow the instructions in the “Hollow Fiber Modlue Preparation & Instruction
Guide” insert and/or Section 10.3 for proper wetting and integrity testing procedures.
5. Trial Data Worksheet
The Trial Data worksheet is used to log and calculate the filtration run data (see sample
below).
!
The columns are the same as the ‘Module Characteristics’ worksheet with the following additions:
Column
Abbreviation
Description
Qretentate
Retentate Flow
Mpermeate
Total mass of permeate (user inputted). This can be changed to total
volume of permeate by clicking on the ‘By Volume’ button on the header.
LMH
Filtrate rate in liters per m2 of membrane per hour
VT
Volumetric Throughput
(a measure of the total permeate per cm2 of SA)
Pump
RPM of the pump
Temp
Temperature (user entered)
Conc. Factor
Concentration Factor
Shear
Shear rate of the fluid through the fibers (s-1)
NOTE:
The permeate volume/mass are to be measured and entered by the user at the corresponding times. The spreadsheet will then calculate the permeate flow rate, the LMH and
the VT. If using the Permeate Scale accessory, the data will be automatically entered into
the worksheet and all the calculations will be completed. Please refer to the Permeate
Scale section of this manual for the installation and use of the scale (see Section 11.1).
9.3.4 Compliancy
SpectrumLabs.com offers an additional software program that enables the KF COMM program to comply
with 21 CFR Part 11 regulations. Please contact SpectrumLabs.com for further information.
30
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
BASIC CONCEPTS OF TANGENTIAL FLOW FILTRATION
10. Basic Concepts of Tangential Flow Filtration (TFF)
Membranes use the principle of barrier separations to differentiate components based on size.
Components larger than the membrane pore are quantitatively held back by the membrane while
smaller components pass through the membrane structure along with the permeate. Although there
are other methods for driving the separation process — such as electric charge (e.g. caustic-chlorine
cells) and diffusion (e.g. dialysis and oxygenation devices) — SpectrumLabs.com hollow fiber membrane modules are designed for pressure-driven applications.
10.2 Dead-end Filtration and Tangential Filtration
Traditional sieve (or dead-end) filtrations consist of forcing a solution containing suspended solids
directly through the membrane structure. Solids retained by the membrane collect on the surface of
the membrane media, continually reducing the permeation rate and eventually plugging the device.
See illustration below.
The driving force for permeation is the pressure difference between the feed and the permeate, called
the transmembrane pressure (TMP). For dead end filtration, this is given by the following equation:
PTMP = Pfeed – Ppermeate
TFF is an efficient way to separate streams that would quickly become plugged if processed by deadend techniques. When using tangential flow techniques, most of the process fluid flows along the
membrane surface rather than passing through the membrane structure. Fluid is pumped at a relatively high velocity parallel to the membrane surface. Except for water treatment applications, only a
small percentage of the tangential flow along the membrane surface ends up as permeate. In most
cell and particle separations, only 1 – 5% of the inlet flow to the membrane device becomes permeate. The remaining 95 – 99% exits the membrane device as “retentate”. The retentate is recirculated
back to the process reservoir and the module inlet such that another 1% to 5% can be removed as
permeate. This recirculation process continues in rapid succession, generating a significant and continuous permeation rate. See the illustration below.
31
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
BASIC CONCEPTS OF TANGENTIAL FLOW FILTRATION
Some examples of tangential flow membrane geometries include: stacked plate and spiral devices
that utilize flat sheet membranes, tubular devices, along with shell and tube devices that use hollow
fiber membranes. In the case of tangential flow separations, the driving force is the TMP, the difference between the average of the module feed and retentate pressures and the permeate pressure:
PTMP = (Pfeed + Pretentate) / 2 – Ppermeate
Filtrate flow results in a build up of retained components on the membrane inner lumen surface.
Generally, these components are carried down the length of the hollow fiber and out the end of the
module by the sweeping action of the recirculating fluid. Occasionally, a cake layer may accumulate
on the surface of the membrane. This boundary layer is composed of solids and/or solute macromolecules which are retained by the membrane from filtration. This phenomenon, often erroneously
referred to as “concentration polarization,” can affect module performance by reducing the functional
size of the membrane pore. In other words, the cake layer becomes the membrane barrier, a “dynamic membrane.”
Caking is influenced by fluid variables: degree of solvation, concentration and nature of the solids and
solutes, fluid temperature, along with operating variables — such as solution velocity along the membrane — and TMP. Controlling this phenomenon is the key to maximizing flux, solute passage, and
optimizing the process parameters. Caking can usually be controlled by ensuring adequate fluid velocity at the liquid-membrane wall interface. Fluid velocity is controlled by the pumping rate. Recirculation
rates depend on the quantity of fibers in a module and shear rate considerations. Typically, a shear
rate of 12,000 s-1 is used for filtration applications and up to 4,000 s-1 is used for perfusion applications.
These rates are guidelines only and should be optimized for each process application.
Certain applications may work well at reduced rates while others may require rates that are significantly
higher. When protein passage through the membrane structure is important, particular attention should
be paid to feed or recirculation rate. In general, high feed rates allow more efficient protein passage.
Depending on the characteristics of the retained components (cells, cell debris, diagnostic particles,
etc.), a caking layer can form on the membrane wall that is actually tighter than the membrane pores.
In these instances, high recirculation rate and low transmembrane pressure often help. Variables
such as viscosity or shear sensitivity of the solution components may prevent the user from generating enough velocity to reduce membrane caking. Often in these cases, applying a determined amount
of back pressure on the permeate helps to prevent caking. Cell perfusion applications frequently use
permeate back pressure to prevent caking.
32
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
ASSEMBLING THE FLOWPATH
10.2 Assembling the Flowpath
The following are the instructions for assembling the KrosFlo® Research IIi TFF System flowpaths.
Figure 11 is given as a representative layout of the flowpath. For a description of the difference
between Constant Volume Mode and Batch Mode, refer to Section 10.4.
10.2.1 Constant Volume Mode
Refer to Figures 12, 13 and 14 for assembling either one of the three different size flowpaths
(MiniKros® Sampler, MidiKros® and MicroKros®, respectively) for constant volume mode.
1.
Attach one Reservoir Trilobite (holder) to the bottom end of the right post on either side
of the KrosFlo® Research IIi Pump Drive. The Reservoir Trilobites should be oriented
such that the front (wide) groove faces forward and its respective thumb screw is directed away from the pump. Attach both Module Trilobites to the left post above the
Reservoir Trilobite. Reservoir and Module Trilobites can be attached to the posts by
hooking the posts into the groove on the back (narrow) end of the Trilobite and securing
the Trilobites in place with the back thumb screw (Figures 11A and 11B).
2.
Attach the HF filtration module by inserting it into the front groove on both the Module
Trilobites. Slide the Trilobites away from each other until the module is held firmly in place.
Secure the Trilobites by tightening the back thumb screws (Figure 11B).
3.
Attach the smaller processing reservoir by inserting it into the front groove of the
Reservoir Trilobite on the right or left side of the pump. Secure in place by tightening the
front corresponding thumb screw (Figure 11B).
4.
A secondary Reservoir Trilobite or standard lab clamp can be used to secure the buffer
reservoir (Figure 11B).
NOTE:
The reservoir Trilobites are designed with a thumb screw to hold the smaller reservoirs
by either the bottle or cap, and to hold the large reservoirs by the cap only. The smaller
reservoirs can be held suspended above the surface of the bench top, while the larger
reservoirs need to rest on the bench top.
NOTE:
SpectrumLabs.com’s conical bottom process reservoirs are designed to facilitate mixing
— however, in applications utilizing low velocites and TMP’s, a flat bottom reservoir may
be used to avoid turbulent mixing in stratified fluids.
5.
Assemble the flowpath as shown in Figures 12, 13 and 14 for the MiniKros Sampler (or
MiniKros), MidiKros and the MicroKros, respectively. The Universal Flowpath kit comes
with 3 different sizes of plastic fittings to make the necessary connections. These include
1
⁄16” Hose Barb fittings for tubing sizes 13 and 14 used for the MicroKros or MidiKros, 1⁄8”
Hose Barb fittings for tubing size 16 used for MidiKros and 1⁄4” Hose Barb fittings for tubing size 17 used for the MiniKros Sampler or MiniKros filters.
The reservoirs that come with the system include the 15 ml, 50 ml and 250 ml. All have
three ports. The middle port is used as the inlet (flowing into the filter), the other diptube
is used as the retentate (returning from the filter), and the third port without any diptube
is used as the vent/buffer addition line. The 15 ml reservoir is typically used with #13 tub-
33
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
ASSEMBLING THE FLOWPATH
ing and MicroKros filters for small volume applications (1-50 mls). The 50 ml reservoir is
typically used with #14 tubing and MidiKros modules for 5-1000 mls. The 250 ml reservoir is typically used with #16 tubing or #17 tubing using hose barb adaptors for
MiniKros filters (30 ml -10 L). A 500 ml reservoir (ACBT-500-F1N) is available with ⅛”
diptubes if larger flow rates are needed.
6.
Connect the appropriate size HB x HB connector to the tubing coming out of the buffer
reservoir (#6 in Figures 12, 13 and 14) and connect it to the vent of the process reservoir
(#5 in Figures 12, 13 and 14).
NOTE:
The buffer line can also be connected to a Tee in the retentate line (#4 in Figures 12,
13 and 14) for adding in the wash buffer. This can be done to increase mixing efficiency.
During the filtration, make sure the vent is closed and open the permeate line. Ensure
that a vacuum has been created in the closed system before opening the pinch clamp
to the buffer reservoir. The inlet pressure will decrease if the vacuum is created. If a vacuum is not created, the sample has the potential of flowing into the buffer reservoir
instead of the recirculation reservoir.
7.
Assemble the rest of the flowpath — including pressure transducers and HF filtration
module for MiniKros® Sampler, MidiKros® and MicroKros® — according to Figures 12,
13, and 14 respectively, and the following steps:
MiniKros® Sampler Flowpath (Figure 12, page 35)
34
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
OVERALL FLOWPATH: FRONT AND TOP VIEW
MODULE
TRILOBITE
(HOLDER)
RESERVOIR
TRILOBITE
(HOLDER)
1000
Figure 11A. System Schematic - Top View
35
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
ASSEMBLING THE FLOWPATH
i.
NOTE:
Cut an appropriate length of #17, ¼’’ ID tubing (ACTU-P17-25N) to run from the process
reservoir (#5 in Figure 12) through the pump head (#7) and to the inlet of the filter. Attach
one end of tubing to a ⅛’’ HB x ¼’’ HB connector and attach the connector to the tubing
on the 500 ml process reservoir that corresponds to the lowest diptube.
The 250 ml process reservoir can be used in place of the 500 ml.
Place tubing through the pump head and then connect other end of tubing to the ProConnex fitting (HB x TC x Female Luer™).
ii.
Attach Tee (⅛” HB x ¼” HB x ¼” HB) to retentate line on the 500 ml reservoir. On one
¼” HB Tee port, attach a buffer reservoir (#6 in Figure 12). On the other ¼” HB Tee
Port, use another length of tubing (#4 in Figure 12) for the outlet of the filter through the
Automatic Backpressure valve (if used).
iii. Close off the lower permeate side port of the HF module with a ¾” TC endcap with
tab.
iv. Connect a short piece of tubing from the upper ¾” TC permeate side port (use ¾” x
HB fitting provided with module if necessary) to a Tee (¼” HB x ¼” HB x Female luer)
and connect permeate pressure transducer. Slide a pinch clamp (#9 in figure 12) over
the permeate tubing. Connect an appropriate length of tubing from the free end of the
Tee to the desired permeate collection vessel (not provided).
v.
Attach remaining transducers (#1 in Figure 12) to the inlet and retentate pressure ProConnex or Tee connectors. Plug the pressure transducer cables into the appropriate
labeled jacks on the Octopus cable at the rear of the pump.
vi. Attach a buffer reservoir (#6) to the buffer addition port (1⁄8” ID tubing) on the 500 ml
reservoir for constant volume diafiltration mode.
vii. Attach a vent line stopcock or pinch clamp to open/close vent.
viii. If preferred, suspended tubing can be secured in the tubing notches on the sides of
the module and reservoir Trilobites.
ix. All HB connections may be secured with tie wraps and a tie wrap gun.
36
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
MINIKROS® SAMPLER FLOWPATH
10
11
LEGEND:
1. Polysulfone Pressure Sensor (ACPM-799-01N)
2. Pro-Connex® TC x HB x Female Luer
3. Manual Backpressure clamp
4. Retentate Line (#16 or #17 tubing)
5. 500 ml 3 Port Process Reservoir (ACBT-500-F1N)
6. Buffer Reservoir
7. Feed Pump
8. Permeate Scale (ACR2-SC4-01N)
9. Permeate Clamp
10. ¼” HB x ¼” HB x Female Luer Tee
11. ⅛” HB x ¼” HB x ¼” HB Tee
Figure 12. MiniKros® Sampler Flowpath
37
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
MIDIKROS® FLOWPATH
LEGEND:
1. Inline Polysulfone Pressure Sensor (ACPM-799-01N)
2. Male Luer x ⅛” Hose Barb connector
3. Manual Backpressure clamp
4. Retentate Line (#14 or #16 tubing)
5. 250 ml 3 port Process Reservoir (ACBT-250-F1N)
6. Buffer Reservoir
7. Feed Pump
8. Permeate Scale (ACR2-SC4-01N)
9. Permeate Clamp
38
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
MICROKROS® FLOWPATH
LEGEND:
1.
Inline Polysulfone Pressure
Sensor (ACPM-799-01N)
2.
Female Luer x 1/16” Hose Barb
3.
Manual Backpressure clamp
4.
Retentate Line
(#13 or #14 tubing)
5.
15 ml 3 Port Process Reservoir
(ACBT-015-C1N)
6.
Buffer Reservoir
7.
Feed Pump
8.
Permeate Scale
(ACR2-SC4-01N)
9.
Permeate Clamp
Figure 14. MicroKros® Flowpath
39
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
ASSEMBLING THE FLOWPATH
MidiKros® Flowpath (Figure 13, page 36)
NOTE:
i.
Attach the pressure transducers (#1 in Figure 13) inline to the inlet, retentate and permeate for MidiKros filters with luer connectors.
ii.
Cut an appropriate length of #16 tubing to connect from the 1⁄8” HB x HB connector
on the middle port of the 250 ml process reservoir (#5 in Figure 13) through the pump
head and then to the luer x 1⁄8” HB fitting (#2 in Figure 13, attached to the inline pressure transducer).
The 50 ml reservoir (ACBT-050-F1N) can be used with #14 tubing for smaller volume applications.
iii.
Cut another length of #16 tubing (#4) from the outlet of the filter through the Automatic
Backpressure valve (if used) to the other diptube on the process reservoir.
iii.
Close off the lower permeate side port of the HF module with a male Luer cap (provided with module).
iv.
Connect an appropriate length of tubing from the Luer x HB connector on the pressure sensor to the desired permeate collection vessel (not provided).
v.
Plug the pressure transducer cables into the appropriate labeled jacks on the Octopus
cable at the rear of the pump.
vi.
Attach a buffer reservoir (#6 in Figure 13) to the vent line (⅛” ID tubing) on the 250 ml
reservoir for constant volume diafiltration mode.
vii.
If preferred, suspended tubing can be secured in the tubing notches on the sides of
the module and reservoir Trilobites.
viii.
All HB connections may be secured with tie wraps and a tie wrap gun.
MicroKros® Flowpath (Figure 14, page 37)
NOTE:
40
i.
Attach the pressure transducers (#1 in Figure 14) inline to the inlet, retentate and permeate luer ports. For small volume applications (~1ml holdup volume), the pressure
transducers can be put on tees using the 1⁄16" Tees, cut tubing and 1⁄16" HB x Female
Luer connectors
ii.
Cut an appropriate length of #14 or #13 tubing to connect from the 1⁄16” HB x HB connector on the middle port of the 15 ml process reservoir (#5 in Figure 14) through the
pump head and then to the luer x 1⁄16” HB fitting (#2 in Figure 14, attached to the inline
pressure transducer).
The 50 ml reservoir (ACBT-050-F1N) can be used with #14 tubing for large volume applications.
iii.
Cut another length of #14 or #13 tubing (#4 in Figure 14) from the outlet of the filter
through the Automatic Backpressure valve (if used) to the other diptube on the process
reservoir.
iii.
Close off the lower permeate side port of the HF module with a male Luer cap
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
INTEGRITY TESTING
(provided with module).
iv. Connect an appropriate length of tubing from the luer x HB connector on the pressure
sensor to the desired permeate collection vessel (not provided).
v.
Plug the pressure transducer cables into the appropriate labeled jacks on the Octopus
cable at the rear of the pump.
vi. Attach a buffer reservoir (#6 in Figure 14) to the vent line on the 15 ml reservoir for constant volume diafiltration mode.
vii. If preferred, suspended tubing can be secured in the tubing notches on the sides of the
module and reservoir Trilobites.
viii. All HB connections may be secured with tie wraps and a tie wrap gun.
10.2.2 Batch Mode
For batch mode, there is no buffer reservoir connected to the vent line. Batch mode is typically used
for concentration or clarification of a sample that is able to fit in the selected reservoir.
10.3 Integrity Testing
All SpectrumLabs.com HF membrane modules are integrity tested in our class 10,000 cleanroom
prior to packaging and shipping. Membrane integrity tests generally measure the capillary pressure of
wetted pores. It is strongly recommended to integrity test all HF membrane modules again prior to
use. Dry modules must be fully primed to ensure complete membrane wetting, otherwise a false
integrity failure will be indicated due to non-wetted or incompletely wetted pores. For wetting and
integrity testing procedures, please refer to the “Hollow Fiber Module – Preparation and
Instruction Guide” (Document 400-12058-000) provided with each hollow fiber module. For
specific instructions on how to use the KrosFlo® Research IIi System to complete a system leak test,
see below.
10.3.1 Leak Test
The flowpath connections should first be pressure tested with air prior to filling the systems with fluids.
1.
2.
41
Close the pinch clamp on the permeate line. Close the backpressure control valve on the
retentate line.
Turn on the KRIIi and ensure that it is tared to zero.
3.
Open the vent on the processing reservoir and run the pump until a feed pressure of ~5
psi is reached and maintained.
4.
If there is no significant pressure drop (<0.5 psi/min), the system has no leaks and is
ready for use.
5.
If the pressure decay is >0.5 psi/min, then check to make sure all the seals and fittings
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KRIIi AS A SYSTEM
are secure. Adjust connections as necessary to eliminate pressure decay.
6.
After assuring system is free of air leaks, relieve the pressure by opening the backpressure clamp.
10.4 KRIIi as a System
The basic application methods for the KrosFlo® Research IIi System are for:
•
Batch Concentration of cells, virus, precipitates, proteins or diagnostic particles
•
Batch Clarification of cells, cell debris, virus, precipitates or proteins
•
Topped-off Batch for reduced volume operations
•
Diafiltration (washing) of cells, cell debris, virus, precipitates, proteins or
diagnostic particles
10.4.1 Batch Concentration (Product in the Retentate)
Generally, the term “concentration” is applied to applications where the material retained by the membrane is (or contains) the desired product. As the process fluid is recirculated through the membrane
and back to the feed reservoir, the feed reservoir becomes the process reservoir — the volume diminishes due to the removal of permeate and the product is concentrated. This mode of operation is
used, for example, with fermentation recoveries in which the desired product is the cell itself or as an
initial processing step in which the product is intracellular.
10.4.2 Batch Clarification (Product in the Permeate)
The term clarification is generally used for applications in which the desired product is in the permeate, such as soluble proteins. This mode of operation is used, for example, to harvest animal cell cultures in which the desired product is secreted by the cells, or microbial fermentation in which the
desired product has been released into solution by cell lysis.
When operated for concentration or clarification applications, the membrane quantitatively removes
solids larger than the pores of the membrane and allows the passage of soluble materials that are
smaller than the membrane pores. Set-up is the same for both batch concentration and clarification
processes. The retentate is returned to the feed reservoir, acting as a process reservoir, and more
clarified product permeates through the membrane. The degree of concentration, called the concentration factor (CF) or volume reduction factor (VRF), is given by the following equation where Vi is the
initial volume and Vf is the final volume:
CF = VRF = Vi / Vf.
10.4.3 Constant Volume
A disadvantage of cross-flow batch operations is the relatively high flow rate required for efficient tangential flow — which is 20 – 100 times the permeate rate. As a result, it is difficult to achieve high
concentration factors without foam or vortex formation in the process reservoir. A way of avoiding this
problem is to set up the system to work with a constant volume, where a smaller intermediate reservoir is utilized as the process reservoir to maintain a constant working volume. While the pump creates a positive pressure that drives the filtration, it also creates an equal negative pressure (vacuum)
42
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KRIIi AS A SYSTEM
that pulls the feed in at the same rate. So there is an overall volume movement from the buffer addition
reservoir through the process reservoir and out the permeate line. The module simply inhibits the
passing of any material larger than the membrane pore size and concentrates it in the working volume
in the process reservoir.
In the example of harvesting from a fermenter or stirred-tank bioreactor, attempts are often made to
use the bioreactor vessel as the process reservoir with poor results. The fermenter exit and entrance
ports are frequently too small for adequate recirculation flow. The return line must be modified so that
it is submerged at high concentrations (low volumes).
10.4.4 Diafiltration
Materials that pass through the membrane can be washed away from materials that are retained by
the membrane (cells, particles, etc.). This technique, called diafiltration, is used to recover additional
product in clarification applications with the goal of achieving better product purity in concentration
applications. For best efficiency, the wash buffer should be free of the solute that is being recovered
or removed. Diafiltration may be accomplished either by adding buffer at the same rate as the permeation rate (constant volume diafiltration) or by reducing the volume in the feed reservoir and adding
more buffer to regain the original volume (discontinuous diafiltration). The amount of diafiltration performed can be expressed by the volume of wash buffer added divided by the batch volume, i.e. the
number of wash volumes. During a constant volume diafiltration, in which soluble components pass
freely through the membrane, each wash volume of permeate removed reduces the solute concentration by a factor of e (2.718…). For example, a four time wash volume diafiltration will reduce the
concentration of solute by a factor of e 4, i.e. 50-fold or over 98%. Using this technique, the concentration of solute can be monitored in the permeate until the desired level of purification of product
recovery is achieved. Components that are partially retained by the membrane cannot be diafiltered
or washed with the same efficiency. The membrane rejection (or retention) is defined as:
Rejection = 1 – Concentration in the permeate
Concentration in the retentate
As the membrane rejection of an undesired component increases, diafiltration becomes less effective
as a technique for removing it.
43
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KRIIi AS A SYSTEM
Figure 15. The Effect of Solute Rejection on Diafiltration Efficiency
The system set-up for diafiltration is the same as for topped-off batch applications, except that
wash buffer enters the process reservoir instead of more product. For more detailed information on
diafiltration, please refer to the diafiltration literature on SpectrumLabs.com.
44
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
OPERATING THE SYSTEM
10.5 Operating the System
There are two basic system set-ups and operating instructions for the different application methods.
“Batch” operating instructions are used when the KrosFlo® Research IIi TFF System is used for batch
clarification and concentration. “Constant Volume” operating instructions are used when the system
is used for topped-off batch and diafiltration modes.
10.5.1 Instructions for Batch Concentration / Clarification
1.
Assemble the system according to the instructions in Section 10.2.2.
2.
Integrity Test: Test the integrity of the system and the membrane module according to
the instructions in Section 10.3.
3.
Introduce the sample to be filtered into the reservoir and seal the lid.
4.
Priming: Close the permeate pinch clamp and make sure the second permeate port
(drain) is closed. Turn the pump on in the forward direction and slowly increase the rate
just high enough to prime the system, but low enough to avoid foaming and vortexing
the solution. Continue until a clear, bubble-free stream is coming out through the retentate line back to the feed reservoir. It is important to eliminate all air from the recirculation
loop to avoid product shear.
5.
Setting Operating Parameters: Adjust the pump rate to the desired velocity after the
air is eliminated.
NOTE:
Refer to KFComm as a guide by typing in the model number of the filter or selecting the
filter from the pull down menu. The recommended feed flow rates at specific shear rates
are given on the right side of the header. For fouling (eg. protein) applications or cell applications, shear rates of 12,000 s-1 or a maximum shear rate of 4,000 s-1 are recommended respectively.
6.
If using the KF Comm data acquisition software, open the ‘Trial Data’ worksheet and
begin collecting data. The permeate flow rate (Qpermeate) or total volume of permeate
(Vpermeate) will need to be manually entered (unless using the optional Permeate Scale) at
their respective times to get graphs of the Flux Rate decay and the Pressures vs.
Volumetric Throughput (VT). OR, equivalently, the mass of the permeate (Mpermeate) can be
manually entered at the respective times using an appropriate scale.
7.
Check for system leaks and tighten fittings and connections if necessary.
8.
Open the permeate pinch clamp. Filtrate should start coming out the permeate line.
9.
Adjust the pressure if necessary by using the Automatic Backpressure Valve or the manual backpressure clamp on the retentate line to induce backpressure.
10. Concentration / Clarification: Allow the system to run while monitoring the filtrate (flux)
rate until the desired concentration factor has been achieved, or the desired volume has
been filtered through the membrane module. The flux rate may diminish as the retentate
material concentrates in the process reservoir.
11. If the retentate starts to vortex or foam, stop processing the batch. For clarification appli-
45
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
OPERATING THE SYSTEM
cations, the system can run dry. When batch processing is complete, close the permeate pinch clamp and turn off the pump.
12. Product Recovery: To recover retentate, lift the lid above the feed reservoir with the diptubes above the solution level and turn the pump on at a low rate — this will drive the
retentate in the recirculation loop into the feed reservoir using air.
13. To recover permeate in the extracapillary space of the membrane module, open the permeate pinch clamp as well as the drain pinch clamp on the module lower side-port.
Drain permeate into the permeate reservoir used during the filtration process.
14. Post-process Integrity Test: Since SpectrumLabs.com hollow fiber modules are disposable, assurance of membrane integrity can be achieved by performing a postprocess integrity test.
NOTE:
A post-process integrity test renders the module non-reusable.
10.5.2 Instructions for Constant Volume & Diafiltration
1.
Assemble the system according to the instructions in Section 10.2.1.
2.
Integrity Test: Test the integrity of the system and the membrane module according to
the instructions in Section 10.3.
3.
Fill the processing reservoir with the process fluid to be washed.
4.
Establish a pump velocity (corresponding to suggested recirculation rates based on a
desired shear).
5.
The membrane inlet pressure and velocity of process fluid should be adjusted for each
specific application. Transmembrane pressure can be increased by increasing the recirculation rate, pressurizing the buffer addition tank, or pulling a vacuum on the filtrate line.
6.
Re-adjust the processing reservoir volume to a desired level by introducing more sample
from the buffer addition reservoir, as described in Step 3.
NOTE:
The level of fluid in the processing reservoir plus the volume in the tubing in the processing loop will determine the final volume of the sample after diafiltration. Below are the procedures to adjust the fluid level of the processing reservoir:
Lowering Processing Reservior Fluid Level
•
During the filtration run, close the buffer addition pinch clamp
•
Make sure the permeate pinch clamp is open
•
Open the vent on the processing reservoir by removing the buffer addition line
•
When desired reservoir volume is reached, reattach the buffer addition line and open the
shut-off clamp; to avoid vortexing and bubbling, keep the fluid level in the processing
reservoir above the dip tubes that extend toward the bottom of the reservoir
Raising Processing Reservoir Fluid Level
46
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PRE-STRAINING
•
During the filtration run, make sure the buffer addition pinch clamp is open and the
permeate line is closed
•
Open the cap on the recirculation reservoir and gravity feed in the buffer/sample from
the buffer reservoir
•
When desired reservoir volume is reached, close the cap on the recirculation reservoir
and open the permeate line
If the processing reservoir volume drains with the vent closed or the buffer line attached,
then there is a leak in the system.
7. Open the pinch clamps for both the buffer addition and permeate lines
8. Use the KF Comm data acquisition software to maintain a record of the operating conditions
9. After the desired number of washings has been achieved, the system can be drained
10.6 Pre-Straining
Some process streams may require a strainer to prevent the lumen of the hollow fibers from becoming
occluded by solution components larger than the diameter of the hollow fibers. If the fiber lumens
become blocked, the pressure between the pump and the module will increase and the permeate
rate will decline. In extreme cases, the resulting pressure build-up between the pump and the inlet to
the module can result in membrane module failure. A strainer should be used on the fluid prior to
entering the process reservoir.
For applications where large particles or agglomerates are a concern, a test should be performed
prior to processing. Install a 50 mesh screen in place of the membrane module. Circulate the process
fluid at a high rate. If the feed pressure begins to increase while the pumping rate is constant, the
screen is being blocked. If this is the case, the process fluid must be strained prior to entering the
process reservoir.
Strain the process fluid using a 50 mesh stainless steel screen. If the pressure remains constant after
pre-straining the feed, the pre-strainer worked. If there is difficulty in pre-straining the process fluid, a
larger mesh strainer can be used in tandem with modules having 1 mm fiber diameters. There are,
however, some fluids that plug screens quickly but do not adversely affect the membrane.
10.7 Process Optimization
As a pressure-driven process, the two most important variables for TFF are recirculation velocity and
TMP. In terms of flux and passage efficiency, these two variables are at odds with one another. Higher
recirculation rates, which generate the convective forces parallel to the membrane surface, act to
cleanse the membrane. Conversely, TMP, which generates the driving force for permeation, acts to
form a cake layer. Optimum operation of the module depends on establishing an equilibrium between
these two parameters. Optimization requires that the pressure drop through the module be monitored. This pressure drop (∆P) is generated by the fluid resistance of the membrane channels and is
defined by the equation:
47
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PROCESS OPTIMIZATION
∆P = Pin - Pout
Pressure drop is generated by the recirculation rate through the module and is related to the characteristics of the fluid, such as viscosity and density, as well as the characteristics of the module, such
as membrane diameter and length. The TMP is the average pressure differential generated across the
membranes and is defined by the equation:
TMP = (Pin - Pout) / 2 – Ppermeate
TMP is generated by the recirculation rate though the module plus any downstream restrictions. The
following procedure produces a graph of Steady State Flux vs. TMP that generally resembles Figure
16 (see next page):
Figure 16. Typical Flux and Solute Passage Curve
The first section of the curve (1) is the pressure dependent region and indicates that a cake layer has
not yet been formed. In the pressure dependent range, solute passage is generally unrestricted —
raising the TMP increases the flux. The top section (2) of the curve is the pressure independent range
and indicates that a cake layer has been formed. In the pressure independent range, solute passage
is controlled by the cake layer. Raising the TMP does not increase the flux. The mid-section of the
curve (3) shows the onset of cake layer formation. The optimum TMP is the highest flux where solute
passage is close to 100%. The following is a step-wise approach to optimizing the operating conditions:
48
1.
With the permeate line clamped off and no downstream restriction, begin the run with the
highest possible recirculation rate. This may be dictated by one or more considerations:
inlet pressure to the module, pump capacity, solution shear sensitivity, etc.
2.
Open the permeate line. Allow the permeate rate to reach a steady state, which normally
takes 5 – 10 minutes. Measure the flux and take a permeate sample to determine solute
passage. Collect the data (recirculation rate, transmembrane pressure, and flux) using KF
Comm on a computer. Collect the solute passage data separately.
3
Increase the TMP of the module by adding backpressure. This can be done using a
retentate backpressure valve, or by applying fluid or head pressure to the process vessel.
Generally this increase should be in 7.25 psi (0.5 bar) increments. If the module was
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PROCESS OPTIMIZATION
already being run at the maximum inlet pressure of 30 psi (2 bar), then the initial recirculation rate will have to be decreased.
4.
Establish a new equilibrium, measure the flux, take another sample of the permeate, and
record the recirculation rate and transmembrane pressure.
5.
Repeat Steps 3 and 4 until either flux does not increase, or until the passage of desired
solutes begins to decline. Optimum is just prior to the point of flux decline and solute
passage decline.
In certain instances, even the highest recirculation rates and lowest TMP’s may generate caking. In
these cases, restricting or metering the permeate will reduce TMP by increasing the permeate pressure (refer to the TMP equation). When permeate pressure is other than atmospheric, the pressure
transducer must be connected to the permeate line. In Step 2 above, partially open the permeate line,
then proceed to Step 3.
TMP is increased in this case by opening the permeate line or speeding the metering pump incrementally and checking the permeate for solute passage. Continue to increase TMP in this manner
until solute passage declines. Optimum is just prior to the point of flux decline.
10.8 In-Process Module Cleaning
Even under optimum conditions, a slow decrease in the permeate rate may occur. There are three
techniques for restoring permeate flux. These include: pump off, forward flushing, and reverse flushing. All three, however, stop or reverse permeation.
10.8.1 Pump Off Cleaning
Turning the pump off will allow the cake layer to loosen. Restarting the pump will flush loosened material from the membrane lumen surface. Although allowing one minute before restarting the pump is
usually sufficient, the optimal time will vary with the solution being processed and should be investigated for each individual case.
10.8.2 Forward Flushing
Closing the permeate pinch clamp while recirculating will clean the membrane in the downstream-half
of the module. Closing the permeate line causes the permeate pressure to rise and exceed the retentate pressure in the downstream-half of the module. In this region, the permeate will flow from the
exterior of the fiber to the interior — loosening and carrying away caked material. Normally, forward
flushing for one minute is sufficient to clean the downstream-half of the membrane. Open the permeate line again and continue processing.
The principle of forward flushing can be explained as follows: when the permeate line is closed, the
net filtration rate in the module is zero — however permeation still occurs internally. The inlet-half of
the membrane module (the high pressure end) generates permeate that back flushes the downstream-half of the membrane module (the low pressure end). This phenomenon is called Sterling
Flow.
Reversing the pump with the permeate line closed will back flush the other half of the module and is
an effective cleaning technique.
49
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
MODULE SELECTION AND SCALE-UP
10.8.3 Reverse Flushing
To clean both halves of the module simultaneously, operate the pump in the reverse direction such
that the recirculation flow enters the module from the retentate end of the module and exits the inlet
end. Open the permeate line and allow the permeate to be pulled back into the module through the
permeate port for one minute. Then switch the pump to the forward direction again and continue
processing.
Reversing the pump induces a negative pressure along the entire length of the membrane fibers and
causes permeate to be pulled back through the walls of the membrane and into the retentate. This
flow reversal will flush caked material back into the bulk process fluid. Using caution, the membrane
can also be back flushed by pumping permeate directly into the extracapillary space of the module.
This must be done with caution at 10 psi (0.7 bar) maximum, or the membrane hollow fibers can be
permanently damaged.
Note that reverse flushing will cause the bulk process fluid volume to increase in the processing reservoir. Precautions must be taken to accommodate for this volume increase.
10.9 Module Selection and Scale-up
The KrosFlo® Research IIi TFF System can successfully be used to determine the membrane surface
area needed for scale-up. The performance of the membrane module depends on many processing
variables — such as the fluid type being processed, the conditions of the fluid (temperature, % solids,
viscosity, etc.), fluid volume, recirculation rate, extent of caking, etc. For scale-up purposes, the
amount of surface area and module size needed to process a batch depends on the following:
•
Steady state flux
•
Batch size to be processed
•
Desired process duration
•
Time required for equipment turn-around (cleaning, rinsing, etc.)
Steady state flux can usually be determined by a trial run with the fluid to be processed. The batch
size and the desired time to process the batch are determined by the user. SpectrumLabs.com
KrosFlo® modules are especially useful for small volume scale-up studies.
It is generally prudent to include a safety factor in designing the process. Oversizing the membrane
surface area by 10 – 30% is generally sufficient to account for problems associated with fluid variability.
In order to determine the amount of membrane surface area required for scale-up, the steady state
flux must be determined. Accuracy of the scale-up procedure is dependent on several variables
being constant from the test runs to the final process. These variables include: solution composition,
recirculation rate, processing temperature and operating pressures. Probably the most important
consideration is that the solution being tested is representative of the scale up fluid to be processed.
Steady state flux occurs when the permeate rate remains essentially constant over a period of 15 to
30 minutes.
50
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
MODULE SELECTION AND SCALE-UP
By knowing the steady state flux, the batch size to be processed and the desired processing duration, a simple calculation can be made to determine the necessary membrane surface area:
Required surface area =
Filtrate volume desired
Required time x Steady state flux
Units are:
Required surface area
= square meters (1 m2 = 10,000 cm2)
Filtrate volume desired
= liters
Required time
= hours
Steady state flux
= liters per square meter hour (L/m2hr)
Example 1:
Clarify animals cells from 1.5 liters of bioreactor broth
The steady state flux is 90 L/m2hr.
The desired processing time is 45 minutes or 0.75 hours.
Required Surface Area =
1.5 liters
0.75 hours x 90 L/m2hr
= 0.022 m2 or 220 cm2
Referring to SpectrumLabs.com’s product catalog, part no. D06-P20U-05-N, A MidiKros module
with 460 cm2 is sufficient for this application and will actually process 1.5 liters of permeate in 0.36
hours (22 minutes).
11. Optional Equipment
51
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
OPTIONAL EQUIPMENT
11.1 Permeate Scales Compatible with the KrosFlo® Research IIi System
The KrosFlo® Research IIi TFF System is the ideal tool for performing small scale Tangential Flow
Filtration processes. Permeate scales compatable with the KrosFlo® Research IIi System, such as the
KrosFlo® Research IIi Permeate Scale and the Ohaus® Navigator XL Scale, are accessory compo-
nents that automate the data collection by measuring the mass of the collected permeate over time.
Both scales connect to the KrosFlo® Research IIi System via a serial connection. The system sends
the data to an Excel® Spreadsheet through a USB connection. The KF COMM software then automatically tabulates and graphs the data.
NOTE:
All editions of KF COMM prior to 1.14 do not come with the Ohaus® Nevigator scale driver
preloaded.
11.1.2 Installation
1. Connect Permeate Scale serial cable to “SCALE” serial port on Pump Drive’s Octopus cable,
52
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM DATA COLLECTION SOFTWARE
then connect Permeate Scale to power source
2. Attach USB extension cable to the USB connector labeled “COMPUTER” on the Octopus
Cable, and plug in the other end of the USB extension cable to the PC
3. Turn on Pump Drive, but leave Permeate Scale off
NOTE:
If on, Permeate Scale will not be recognized by PC when booting.
4. Boot PC, and allow Windows® to search for the Pump Drive’s driver — if not found, locate
the driver on the included CD
5. Turn on Permeate Scale
6. Open Excel® Workbook “KR2i with Scale.xlt”
7. Enable Workbook’s macros (this may not be possible if user’s security settings are not high
enough), if successful:
a.
Older versions of Excel® will display four icons for data collection control on the standard
toolbar
b.
Newer versions of Excel will display data collection controls in Add-Ins tab
8. Establish and verify Pump Drive’s built-in Pressure Monitor connection
a.
53
Click on “Configure Collection” to set up Pressure Monitor
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
KF COMM DATA COLLECTION SOFTWARE
Figure 17: Configure Scale Menu on Windows XP
!
i.
Locate Pressure Monitor by clicking “Find COMM,” which will prompt the PC to continuously cycle through available COM ports until it connects to Pressure Monitor, or until
user clicks “Cancel”
ii.
If the Pressure Monitor’s COM is unavailable due to another program’s interference,
close interfering program
b.
NOTE:
Verify Pressure Monitor’s live connection to PC by adding pressure to Inline Pressure
Sensor — the “Trial Data” spreadsheet will update its corresponding yellow Live cell,
depending on which port Pressure Sensor is connected to in Octopus Cable, at top of
the sheet to reflect the pressure
“Override.xls” spreadsheet may be used to force Pressure Monitor’s COM port to the correct setting.
9. Establish and verify Permeate Scale connection
a. Click on “Configure Scale” to set up Permeate Scale
Figure 18: Configure Permeate Scale Menu on Windows 8
i.
54
Locate Permeate Scale by clicking “Find Comm,” which will prompt the PC to continu-
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
PERMEATE SCALE
ously cycle through available COM ports until it connects to Permeate Scale, or until
user clicks “Cancel”
ii.
iii.
If Permeate Scale’s COM is unavailable due to another progran’s interference, close
interfering program
Select “Scout Pro” under “Scale Accessory for KrosFlo® Research IIi TFF System” if
using KrosFlo® Research IIi Permeate Scale, or “Navigator” if using Ohaus®
Navigator XL Scale in the drop-down menu within “Configure Scale” dialog
iv. Click “OK.”
b.
Verify Permeate Scale’s live connection to PC by placing a small object on the scale —
the “Trial Data” spreadsheet will update its yellow Live cells at top of the sheet to reflect
the small object’s mass.
NOTE:
“Scale Override.xls” Spreadsheet my be used to force Permeate Scale’s COM port to the
correct setting
10. Permeate Scale is now ready to use in conjunction with KrosFlo® Research IIi TFF System
and will zero automatically when the “Start Collection” button is pressed
11.1.3 Troubleshooting
a.
COM Port Setting
i.
b.
Driver Recognition
i.
55
If the COM port cannot be detected for the Pressure Monitor or the Permeate Scale, the
Pressure Monitor Override or the Scale Override Excel® spreadsheets can be used
respectively. These are provided on the CD and force the COM port to the correct setting. To do this, open the file and change the COM port to the correct setting (seen in
the Device Manager), then save and close the file. Reopen the spreadsheet and select
yes when asked to force the COM setting.
If Windows® does not recognize the Permeate Scale USB signal or thinks that it is a different device, unplug the USB connector from the computer, turn off the scale and plug
in the USB connector again. If the driver is still not recognized, try loading the driver
again and/or rebooting to reset the drivers.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
AUTOMATIC BACKPRESSURE VALVE
11.2
Automatic Backpressure Valve for KrosFlo Research IIi System
The KrosFlo® Automatic Backpressure Valve controls TMP or permeate pressure during
tangential flow filtration processes when used in conjunction with the KrosFlo® Research
IIi TFF System. The valve is designed to pinch flexible tubing on the retentate or permeate line based on digital pressure readings to maintain the user-set pressure.
11.2.1 Installation
1.
56
The cable on the KrosFlo® Automatic Backpressure Valve has both an RS232 port
(female) and a power supply. Connect the RS232 port to the terminal labeled “Valve” on
the Octopus cable at the back of the Digital Pressure Monitor. Ignore the power supply
connection; it is used to power a stand-alone pressure monitor.
2.
Connect the second RS232 port on the opposite side
(male plug) to the serial cable for sending data to a
PC. The data can also be sent to the PC using the
“Computer” USB connection on the Octopus cable.
3.
Plug in the transformer and connect to the power
supply on the Automatic Backpressure Valve.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
AUTOMATIC BACKPRESSURE VALVE
KRIIi’s Port
for ABV
ABV’s Port
for KRIIi
Power Requirements
ACPC-F16-01N, ¼” (max) OD tubing valve 0.5A @ 12V
ACPC-F17-01N, ⅝” (max) OD tubing valve 1.0A @ 12V
ACPC-F82-01N, 1” (max) OD tubing valve 1.0A @ 12V
11.2.2 Operation
Automatic TMP Pressure Control
1.
To operate the backpressure valve in the auto TMP mode, switch the toggle on the top
of the valve to the “TMP” setting. Press the OPEN button to load the tubing and reset
the pressure controls.
2.
Set the desired TMP using the INC and DEC buttons
3.
With the pump running at the desired speed for the TTF process, press the GO button
!
57
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
AUTOMATIC BACKPRESSURE VALVE
and the valve will start to pinch the tubing until the desired TMP is reached (this may take
a few minutes until it equilibrates).
4.
To pause the valve, press the GO button (the light will turn off). To resume, press the GO button (the light will turn back on).
NOTE:
Under manual control, the Automatic Backpressure Valve does not need to equilibrate.
NOTE:
If the valve is not in the fully open position, it will adjust the pinch slowly to avoid the
potential pressure spikes.
a.
If the KrosFlo Research IIi Pump is turned to off or to 1/2 the rpm when the GO button
is pressed, the valve will pause and will maintain the same pinch distance. The valve will
flash “SLO” while in this paused state. The valve will resume the normal pinch control
when the pump is turned on or recovers at least 1/2 the original rpm.
Automatic Permeate Pressure Control
1.
To operate the backpressure valve in the auto PERMEATE mode, switch the toggle on
the top of the valve to the “Permeate” setting. Press the OPEN button to load the tubing
and reset the pressure controls.
2.
Set the desired Permeate pressure using the INC and DEC buttons
3.
Press the GO button before starting the recirculation pump — the valve will pinch shut
(when using the maximum tubing size rated for the valve). Begin the process and then
the valve will start to open/pinch the tubing until the desired Permeate pressure is
reached (this may take a few minutes to equilibrate).
Manual
To control the valve manually, hold down the GO button for >3 seconds. The light will begin
to flash and then the INC and DEC buttons can be used to adjust the pinch.
Updating Automatic Backpressure Valve
58
1.
Connect the KrosFlo® Automatic Backpressure Valve to the computer via the RS232
port
2.
Turn on the valve and wait 60 seconds
3.
Ensure the appropriate update for the specific valve has been obtained
4.
Press and hold the Open button on the valve for 6 seconds, blanking the display
5.
Start the .exe file by double-clicking on it
6.
Click on the Start button to begin the process
7.
Either click the Find Comm button, or select the Comm port used by the valve
8.
Click on the OK button and the valve display will go dark — the valve will begin to update
9.
To verify a successful update, toggle the TMP/Permeate to restart the valve. The valve
should now display the most current version
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
COMMUNICATIONS
Communications
Serial Communication
The Valve has a serial input cable for communication and control.
The communication parameters are 19200 baud, 1 stop bit, no parity, and no flow control.
When communicating with an external PLC, the valve should receive about 1 line of data
per second. The lines should end with a return character and a line feed character.
Each line should have several values, each separated from the next by a single space:
<InletPressure><space><RetentatePressure><space> <PermeatePressure><space>
<TMP><space><RPM><space><return>
1.
Inlet Pressure: This should contain a decimal and may have a leading - sign for negative
pressures. There should be no sign provided for positive values. If the - sign is included,
there should not be a space after the - sign. The number will not be less than 2 characters,
nor more than 5 characters in length. The number will contain a decimal point. If in units
of millibars, there will be no digits to the right of the decimal. If in psi, there will be 1 or 2
digits to the right of the decimal. The pressure must be no more than 50 psi nor less than
-9.99 psi.
2.
Retentate Pressure: Formatted same as the inlet pressure.
3.
Permeate Pressure: Formatted same as the inlet pressure.
4.
Transmembrane Pressure (TMP): Formatted same as the inlet pressure.
5.
Pump speed in RPM: This is a number with no decimal point and no possible leading sign. It is the unsigned pump speed in RPM. It may be 0. When the GO button is pressed,
the pump speed is remembered. If the speed drops by 50% or more, the valve stops
adjusting position until the pump speed recovers past the 50% mark. If the RPM is 0 when
the GO button is pressed, then the pump speed will never fall below the 50% limit and the
Pump Slow mode will never be entered.
Note: The pump speed should be followed by a space. This space may be followed by
other characters, but they are ignored by the valve.
The line should be no more than 78 characters in length.
59
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
VALVE EXTERNAL CONTROL
Valve External Control
If the input line is “G0,” then the valve will stop and the green light will turn off.
If the input line is “G1,” then the valve will go and the green light will turn on.
If the input line is “G2,” then the valve will open for loading or unloading tubing.
Please note that while the valve is moving a long distance (as it usually does
immediately following a go or load command), additional commands are silently ignored.
Also, the set point may be adjusted from the data stream. Use "S # " the letter S followed
by a space, followed by a number with a decimal point, followed by a space, which may
be followed by any additional characters desired. The number should conform to the standards used for the pressure readings (no decimal digits is mb, 1 or 2 decimal digits is psi,
and 3 decimal digits is bar).
The S and G lines count toward the valve timing. Wait at least 100 ms between sending
lines.
Fine Tuning
The valve positioning and timing can be modified using terminal emulators, such as
HyperTerminal (available on Mircrosoft® Windows 95 up to XP) or PuTTY (a free and opensource program for multipul platforms).
To set the start position, connect the valve to the computer using a serial cable (or USB to
serial adapter if no serial connector is available on the computer). Do not plug the serial
connector into the pressure monitor. Turn on the valve and wait 60 seconds. Press and
hold the OPEN button for about 6 seconds until the display is all - signs. Use
the terminal emulator to communicate with the valve, using the usual settings — 19200
baud, 8 data bits, no parity, 1 stop bit, and no flow control. Select the correct COM port
and make sure that the KF Comm software is not running.
Once connected, press the enter key and then type:
<Start><space><start position number>
Example: Start 98 and then the enter key to set the start position. The default
number is usually 80. Larger numbers are more closed. Numbers bigger than 255
are reduced to numbers less than 256.
To display the current start position, type <Start> and press enter. When the correct
start position number has been entered, type <Lock> and press enter to go back to
normal valve operation. The number will be stored, and the valve will close to the
new position when GO is pressed.
There are 4 other parameters that can be tuned if needed:
1.
60
“Change” is the minimum number of seconds before a change is made in the position
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
VALVE EXTERNAL CONTROL
after a change in the sign of the pressure error. The default is 3.
61
2.
“Interval” is the minimum number of seconds between changes in position. The default
is 4.
3.
“Step” is the number of steps to take in the same direction before the movements get
larger. The default is 5.
4.
“Reverse” is the minimum number of seconds to wait after the first movement in a new
direction. The default is 9.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
ORDERING INFORMATION
12. Ordering Information
KrosFlo® Research IIi Tangential Flow Filtration System
Part Number
Description
SYR2-U20-01N
KrosFlo Research IIi System with pump drive, pump head and
Integrated Digital Pressure Monitor.
KF Comm software, Octopus cable, Flowpath kit and Starter kit also included.
KrosFlo Research IIi System IQ/OQ Paperwork
KrosFlo Research IIi System IQ/OQ Sales Rep Service
ACR2-IQOQ-PW
ACR2-IQOQ-SR
System Components & Accessories
Pressure Transducer ACPM-499-03N
Part Number
ACPM-799-01N
ACPM-799-01S
ACPM-899-01N
ACPM-05TC-01N
ACPM-05TC-C1N
ACPM-10TC-01N
ACPM-10TC-C1N
Fitting Kit ACPX-CD-01N
ACPX-T-01N
ACPX-S-01N
ACPX-N-01N
ACPX-K-01N
ACPX-KE-01N
ACR2-SKT-01N
ACR2-UFP-01N
Pump Head ACR2-H3I-01N
ACR2-H4I-01N
ABV ACPC-F16-01N
ACPC-F17-01N
ACPC-F82-01N
Scale ACR2-SC5-01N
ACR2-SC3-01N
ACR2-SC4-01N
ACR2-SC2-01N
ACM3-SC7-01N
ACM3-SC8-01N
ACM3-SC9-01N
ACM3-SC10-01N
ACM3-SC3-01N
ACM3-SC4-01N
ACM3-SC1-01N
ACM3-SC2-01N
ACM3-SC5-01N
ACM3-SC6-01N
62
Description
Disposable Pressure Transducer, Sterile, 3/Pkg
Polysulfone Pressure Transducer 1/Pkg
Polysulfone Pressure Transducer, Irradiated 1/Pkg
Pressure Transducer, FLL x MLL, Polysulfone, Non-Sterile, Calibrated
Pressure Transducer, 1/2” TC x 1/2” TC, Polysulfone, Non-Sterile
Pressure Transducer, 1/2” TC x 1/2” TC, Polysulfone, Non-Sterile, Calibrated
Pressure Transducer, 1” TC x 1” TC, Polysulfone, Non-Sterile
Pressure Transducer, 1” TC x 1” TC, Polysulfone, Non-Sterile, Calibrated
MicroKros/MidiKros Fitting Kit
MidiKros TC Fitting Kit
MiniKros Sampler Fitting Kit
MiniKros Fitting Kit
KrosFlow Fitting Kit
KrosFlo Essentials Fitting Kit
KrosFlo Research IIi Starter Kit
Universal Disposable Flowpath Kit for use with L/S 13 To L/S 17 Tubing
KrosFlo Research II Pump Head for use with Precision Tubing
KrosFlo Research II Pump Head for use with High Performance Tubing
KrosFlo Automatic Backpressure Valve for use with 1/8" To 1/4" OD Tubing, 110V
(L/S # 13, 14, 16 Tubing)
KrosFlo Automatic Backpressure Valve for use with 3/16" To 5/8" OD Tubing, 110V
(L/S # 25, 17, 18, 15, 24, 35, 36 and IP # 26, 73)
KrosFlow Automatic Backpressure Valve for use with 3/4” to 1” OD Tubing, 110V
(L/S # 73 and #82 Tubing)
400g (0,01 g), 120V
400g (0,01 g), 220V
4000g (0,01 g), 120V
4000g (0,01 g), 220V
10kg (0,001 kg), 120V
10kg (0,001 kg), 220V
25kg (0,002 kg), 120V
25kg (0,002 kg), 220V
50kg (0,01 kg), 120V
50kg (0,01 kg), 220V
100kg (0,01 kg), 120V
100kg (0,01 kg), 220V
250kg (0,02 kg), 120V
250kg (0,02 kg), 200V
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
ORDERING INFORMATION
Reservoir ACBT-015-C1N
ACBT-050-C1N
ACBT-250-F1N
ACBT-1TC-01N
ACBT-500-F1N
ACBT-10TC-01N
ACBT-20TC-01N
ACBT-2TC-01N
ACBT-4TC-01N
ACBT-50TC-01N
Tubing ACTU-P13-25N
ACTU-P14-25N
ACTU-P16-25N
ACTU-P25-25N
ACTU-P17-25N
ACTU-P18-25N
ACTU-E13-25N
ACTU-E14-25N
ACTU-E16-25N
ACTU-E17-25N
ACTU-E18-25N
NOTE:
63
Process Reservoir - Conical Bottom, 15 ml, 3 dip tubes
Process Reservoir - Conical Bottom, 50 ml, 3 dip tubes
Process Reservoir - Conical Bottom, 250 ml, 3 dip tubes, bent inlet
Process Reservoir - Bottle, 1 L, 3/4” TC
Process Reservoir - Conical Bottom, 500ml, 3 dip tubes, bent inlet
Process Reservoir - Carboy, 10 L, 3/4” TC
Process Reservoir - Carboy, 20 L, 3/4” TC
Process Reservoir - Bottle, 2 L, 3/4” TC
Process Reservoir - Bottle, 4 L, 3.4” TC
Process Reservoir - Carboy, 50 L, 3/4” TC
Pharmapure® Tubing, L/S 13, (1/32" ID), 25 ft/pkg
Pharmapure® Tubing, L/S 14, (1/16" ID), 25 ft/pkg
Pharmapure® Tubing, L/S 16, (1/8" ID), 25 ft/pkg
Pharmapure® Tubing, L/S 25, (3/16" ID), 25 ft/pkg
Pharmapure® Tubing, L/S 17, (1/4" ID), 25 ft/pkg
Pharmapure® Tubing, L/S 18, (3/8" ID), 25 ft/pkg
Extended Life Silicone Tubing Size 13, 0.03" (0.8mm) ID, 1/16" Hose Barb
Extended Life Silicone Tubing Size 14, 0.06" (1.6mm) ID, 1/16" Hose Barb
Extended Life Silicone Tubing Size 16, 0.12" (3.1mm) ID, 1/8" Hose Barb
Extended Life Silicone Tubing Size 17, 0.25" (6.4mm) ID, 1/4" Hose Barb
Extended Life Silicone Tubing Size 18, 0.31" (7.9mm) ID, 3/8" Hose Barb
Contact SpectrumLabs.com for assistance selecting the appropriate H F Module, Flowpath,
Reservoir Bottles and Customized Sterile Flowpaths (MBT's) for your application.
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS
CONTACT INFORMATION - WORLD-WIDE
Contact Information:
THE AMERICAS
World-wide Headquarters
voice
fax
e-mail
web
310-885-4600 (world-wide) • 800-634-3300 (toll-free US and Canada)
310-885-4666 (world-wide) • 800-445-7330 (toll-free US and Canada)
[email protected]
www.spectrumlabs.com
EUROPE
European Headquarters
voice +31 (0) 76 5719 419
fax +31 (0) 76 5719 772
e-mail [email protected]
web www.spectrumlabs.eu
Southern Europe
voice
fax
e-mail
web
+33 (0) 6 80 99 41 58
+31 (0) 76 5719 772
[email protected]
www.spectrumlabs.fr
UK and Ireland
voice
fax
e-mail
web
+44 (0) 7508 201 001
+31 (0) 76 5719 772
[email protected]
www.spectrumlabs.co.uk
Central Europe
voice
fax
e-mail
web
+49 (0) 151 1269 2738
+31 (0) 76 5719 772
[email protected]
www.spectrumlabs.de
Benelux
voice
fax
e-mail
web
CHINA
JAPAN
voice (+86) 21 68810228
400-6284448 (toll-free Mainland China)
fax (+86) 21 60919246
e-mail [email protected]
web www.spectrumlabs.cn
voice
fax
e-mail
web
+32 (0) 484 677 532
+31 (0) 76 5719 772
[email protected]
www.spectrumlabs.eu
+81-77-552-7820
+81-77-552-7826
[email protected]
www.spectrumlabs.jp
400-11545-000 Rev. 05 • 151408-jm
64
KrosFlo® RESEARCH IIi TFF SYSTEM • PRODUCT INFORMATION AND OPERATING INSTRUCTIONS