BioLogic HR CHROMATOGRAPHY SYSTEM Instruction Manual Rev D

SAFETY

SAFETY

!

Caution/Warning Disconnect power to any BioLogic HR system component before servicing. No user-serviceable parts are inside any component. Refer servicing to Bio-Rad service personnel.

This instrument is intended for laboratory use only. The BioLogic HR system is certified to meet the I.E.C. 1010* standard for safety of laboratory equipment. Certified products are safe to use when operated in accordance with the instruction manual. This safety certification does not extend to other chromatography equipment or accessories not I.E.C. 1010 certified, even when connected to this BioLogic HR system. The BioLogic HR system conforms to the class A standards for Electromagnetic Emissions, intended for laboratory equipment applications. It is possible that emissions from this product may interfere with some sensitive appliances when placed nearby or on the same circuit as those appliances. The user should be aware of this potential and take appropriate measures to avoid interference. This instrument should not be modified or altered in any way. Alteration of this instrument will void the manufacturer’s warranty, void the I.E.C. 1010 certification, and create a potential safety hazard for the user. Bio-Rad is not responsible for any injury or damage caused by the use of this instrument for purposes other than for which it is intended or by modifications of the instrument not performed by Bio-Rad or an authorized agent.

*I.E.C. 1010 is an internationally accepted electrical safety standard for laboratory instruments.

TABLE OF CONTENTS

TABLE OF CONTENTS

Safety

SECTION 1. Chapter 1.0 1.1 1.2 1.3 1.4

SYSTEM OVERVIEW Introduction ...1 Overview ...1 Features ...2 Unpacking ...3 Description of System Components...3

Chapter 2.0 2.1 2.2 2.3

Description of the BioLogic HR System Components...7 Controller...7 Workstation ...9 UV and Conductivity Detectors ...13 2.3.1 UV Detector...14 2.3.2 Conductivity Monitor...15 2.4 Bio-Rad System Options...16 2.4.1 Signal Import Module - HR (SIM-HR) ...16 2.4.2 pH Monitor...17 2.4.3 Dyna Loop Large Sample Volume Loop ...17 2.4.4 Model EP-1 Econo Pump...18 2.4.5 Model 2110 Fraction Collector ...19 2.4.6 Model 2128 Fraction Collector ...20 2.4.7 Model 1327 Chart Recorder...21 2.4.8 BioLogic HR Enviro-Case ...22 2.4.9 Uninterruptible Power Supply...22 2.4.10 AS-100 Auto Sampler...22 2.5 Non-Bio-Rad System Options ...23 2.5.1 Generic Fraction Collectors...23 2.5.2 Generic Chart Recorders ...23 2.5.3 Printers ...23 2.6 Columns and Column Fittings...24 2.6.1 Anion Exchange: Q-Strong Anion Exchange ...24 2.6.2 Anion Exchange: S-Strong Cation Exchange ...24 2.6.3 Anion Exchange: DEAE Weak Anion Exchange...25 2.6.4 Anion Exchange: Carboxy Methyl (CM) Weak Cation Exchange ...25 2.6.5 Ceramic Hydroxyapatite (CHT) ...25 2.6.6 Size Exclusion Chromatography (SEC) ...26 2.6.7 High Pressure Reversed Phase Columns ...26 2.6.8 Hydrophobic Interaction Chromatography (HIC)...26 2.6.9 Affinity Chromatography...27 2.6.10 Empty Columns...27 2.6.11 Column Fittings ...28

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TABLE OF CONTENTS

SECTION 2. Chapter 3.0 3.1 3.2 3.3

SYSTEM INSTALLATION AND SETUP System Configurations...31 System Configurations ...31 Example 1 of a System Configuration: Manual BioLogic HR System ...32 Example 2 of a System Configuration: Automated BioLogic HR System...37

Chapter 4.0 4.1 4.2 4.3

System Plumbing ...39 General Guidelines for Plumbing the System...39 Example of a Manual Plumbing Arrangement ...40 Plumbing Bio-Rad’s Low Pressure valves ...43 4.3.1 Plumbing the SV5-4 Buffer Select Valve...43 4.3.2 Plumbing the SV3-2 Diverter Valve...45 4.4 Plumbing Bio-Rad’s High Pressure Valves ...46 4.4.1 High Pressure Inject Valves (V7-3 and AV7-3) ...46 4.4.2 High Pressure Stream Select Valves (V9-8 and AV9-8) ...48 4.5 Priming the System ...49

SECTION 3. Chapter 5.0 5.1 5.2 5.3

SYSTEM OPERATION Introduction to the System Software ...51 Using the System Interface...51 Standard Mouse and Keyboard Functions...52 System Menus ...53 5.3.1 System Toolbar Menu Buttons ...53 5.3.2 System Drop-down Menus...55

Chapter 6.0 Introduction To The Browser Screen ...63 Chapter 7.0 7.1 7.2 7.3 7.4

Modes Of Operation...69 Manual Control Screen ...70 Setup Screen ...72 Protocol Screen...75 Run Screen ...87 7.4.1 Pausing/Stopping a Method in Progress...89 7.4.2 Working Offline...90 7.4.3 Editing a Protocol While a Run is in Progress ...91 7.4.4 Run Notebook Screen...93 7.4.5 Run Log Screen ...93 7.5 Post Run Screen ...94 7.5.1 Resizing...94 7.5.2 Chromatogram Information ...94 7.5.3 Annotating (“tagging”) the Chromatogram ...95 7.5.4 Transferring Data into other Programs ...95 7.5.5 Exporting Chromatogram Images ...96

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TABLE OF CONTENTS

SECTION 4.

ADVANCED SYSTEM APPLICATIONS

Chapter 8.0 Sample Loading ...97 8.1 Sample Loading through the Gradient Pump ...97 8.1.1 System Setup...97 8.1.2 Writing the Protocol...98 8.2 Sample Loading using the DynaLoop Sliding-Piston Loop...99 8.2.1 System Setup...99 8.2.2 Writing the Protocol...100 8.3 Sample Loading onto a Low Pressure Column Using the Econo Pump ...103 8.3.1 System Setup...103 8.3.2 Writing the Protocol...104 8.4 Loading Multiple Samples ...106 8.4.1 System Setup...106 8.4.2 Writing the Protocol...107 8.5 Multiple Injections of a Single Sample ...108 8.5.1 System Setup...108 8.5.2 Writing the Protocol...108 8.6 Using the AS-100 Autosampler with the BioLogic HR System ...109 8.6.1 System Setup...109 8.6.2 Writing the Protocol...110 Chapter 9.0 Multiple Columns ...113 9.1 Column switching ...113 9.1.1 System Setup...113 9.1.2 Writing the Protocol...113 9.2 Running One Sample Sequentially over Several Different Columns...115 9.2.1 System Setup...115 9.2.2 Writing the Protocol...116 Chapter 10.0 Reverse Flow Chromatography ...117 10.1 System Setup...117 10.2 Writing the Protocol...118

SECTION 5.

MAINTENANCE AND TROUBLESHOOTING

Chapter 11.0 Maintenance...119 11.1 Cleaning the Outer Surfaces of the Instruments...119 11.2 Storage of the BioLogic HR System ...119 11.3 Care and Maintenance of the Gradient Pump ...120 11.3.1 Priming the Gradient Pump and Removing Trapped Air Bubbles...120 11.3.2 Daily Maintenance...120 11.3.3 Routine Maintenance of the Gradient Pump...121 11.4 Maintenance of the UV Optics Module and the Conductivity Flow Cell ...124 11.4.1 Cleaning the UV Optics Module and the Conductivity Flow Cell...124 11.4.2 Replacing the Lamp in the UV Optics Module...125 Chapter 12.0 Troubleshooting the BioLogic HR System ...127 12.1 Troubleshooting the BioLogic HR Controller...127 12.2 Troubleshooting the BioLogic HR Gradient Pump ...129 12.3 Troubleshooting the UV Optics Module and UV Trace ...133 12.4 Troubleshooting the Conductivity Flow Cell and Trace...135 12.5 Troubleshooting the Model 2128 Fraction Collector ...136

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TABLE OF CONTENTS

SECTION 6.

APPENDICES

Appendix A. Specifications ...137 Appendix B. Pressure Conversion Table...139 Appendix C. Warranty Statement ...140 Appendix D. Ordering Information ...141 Appendix E. Column Information ...141 Appendix F. Declaration of Conformity and ISO Certification ...145

LIST OF FIGURES 1. BioLogic HR System ...1 2. Model OM-II UV Optics Module and Conductivity Flow Cell ...13 3. Signal Import Module (front and rear views)...16 4. Cable Connections to the Signal Import Module-HR ...16 5. pH Electrode and Flow Cell ...17 6. Dynaloop ...17 7. Model EP-1 Econo Pump...18 8. Model 2110 Fraction Collector ...19 9. Model 2128 Fraction Collector ...20 10. Model 1327 Chart Recorder...21 11. AS-100 Autosampler ...22 12. BioLogic HR System as used with a Condensing Cold Room ...31 13. Manual BioLogic HR System ...32 14. Rack Assembly...34 15. Manual System Cable Connections...35 16. Automated BioLogic HR System...37 17. Automated BioLogic HR System Cable Connections ...38 18. Making 1/4-28 Flat Bottom Fittings ...39 19. Plumbing Connections to the Gradient Pump...40 20. System Plumbing ...41 21. Inject Valve Plumbing for V7-3 or AV7-3...42 22. Backpressure Device ...42 23. SV5-4 Buffer Select Valve...43 24. Configuration of the SV5-4 as a Gradient Pump Inlet Buffer Select Valve...44 25. SV3-2 Diverter Valve...45 26. Configuration of the SV3-2 as a Fraction Collector Diverter Valve...45 27. V7-3 and AV7-3 Inject Valves...46 28. V9-8 and AV9-8 High Pressure Stream-Select Valves ...48 29. Layout of the Screen Display ...51 30. The Browser Screen ...63 31. The Browser Screen’s Select CopyIn Window ...67 32. CopyIn with Information in Browser Tab Window ...68 33. Relationships between Modes of Operation ...69 34. Manual Mode Screen, showing a Model 2128 Fraction Collector and Four Valves...70 35. Setup Screen ...72 36. Protocol Screen...76 37. Run Screen showing a Run in Progress...87 38. Run Screen’s Abort, Pause, and Hold Buttons ...89 39. Editing during a Run ...91 40. Protocol Screen during a Run...92 41. Run Notebook Screen...93 42. Run Log Screen ...93 43. Post-Run Screen...94 44. Post-Run Tags...95

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TABLE OF CONTENTS

45. 46. 47. 48. 49. 50. 51. 52. 53. 54. 55. 56 57. 58. 59. 60. 61. 62. 63. 64. 65. 66. 67. 68. 69. 70.

Export Data Setup Screen ...96 Exporting a Chromatograph Image...96 Sample Loading Through the Gradient Pump ...97 Writing a Protocol for Sample Injection Through the Gradient Pump...98 Plumbing the DynaLoop for use with an Inject Valve ...99 Load/Inject Sample Window for a Manual Load/Inject of the Sample ...100 Load/Inject Sample Window for an Automatic Load/Inject of the Sample ...101 Valve Positions During a Run using the DynaLoop ...102 Connecting an EP-1 Econo Pump to the BioLogic HR Workstation...103 Plumbing an AV7-3 Inject Valve with an EP-1 Econo Pump as an Auxiliary Load Pump ...104 Writing the Protocol: Direct Sample Loading ...105 Valve Positions during a Run with Direct Sample Loading and Injection ...105 Using a Peristaltic Pump and an AV9-8 or V9-8 Valve to Load Multiple Samples ...106 Load/Inject Sample Window for Loading Multiple Samples...107 Rear View of AS-100...109 Column Switching using Two AV7-3 Valves...113 Protocol for using an AV7-3 Valve Inject Valve for Column Switching...114 Column Switching using two AV9-8 Valves...115 Programming a Sequential Run...116 Injection Valve Plumbing for Use in Affinity Chromatography...117 Programming for Reverse Flow Chromatography ...118 BioLogic HR Workstation Pump Mechanism Parts...121 Removing the Piston Seals (view showing the back of the Pump Head Module)...122 Pump Head Module Assembly...123 Replacing a Mercury Lamp in a Model OM-II Optics Module...125 Replacing a Zinc Lamp in a Model OM-II Optics Module ...125

LIST OF TABLES 1. Description of System Components...3 2. Controller’s Front Panel Features ...7 3. Controller’s Rear Panel Connectors ...8 4. Workstation’s Front Panel Controls...9 5. Workstation’s Rear Panel Connectors ...11 6. Columns and Column Fittings...28 7. Special Function Keys ...52 8. Toolbar Buttons ...53 9. File Drop-Down Menu ...55 10. Edit Drop-Down Menu...57 11. View Drop-Down Menu ...59 12. Utilities Drop-Down Menu ...60 13. Options Drop-Down Menu ...61 14. Valve Setup Information ...74 15. Isocratic Flow Button...77 16. Load/Inject Sample Button ...78 17. Linear Gradient Button ...79 18. Change Valve Button ...79 19. Hold Button ...80 20. Miscellaneous Buttons ...81 21. Fraction Collection Button ...82 22. Protocol Screen’s Editing Toolbar Buttons...86 23. Run Screen’s Control Buttons...88

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SYSTEM OVERVIEW

INTRODUCTION

1.0

INTRODUCTION

1.1

OVERVIEW

The BioLogic HR is a full-featured, gradient chromatography system specifically designed for the high resolution purification of proteins, peptides and other biomolecules where recovery of biological activity is of primary concern. The BioLogic HR system is microprocessor controlled, with an easy-to-use graphic interface and menudriven software for manual operation, system setup, method editing and run operations. The flexible control architecture allows the seamless integration of a wide variety of configurations with other Bio-Rad and non-Bio-Rad components to meet your purification requirements.

UNO™ Q1 COLUMN

MODEL 1327 CHART RECORDER

FITTING TIGHTENER UV DETECTOR

AV7-3 INJECT VALVE

CONDUCTIVITY MONITOR

COLOR MONITOR

CONTROLLER

WORKSTATION

MODEL 2128 FRACTION COLLECTOR

Figure 1. BioLogic HR System

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INTRODUCTION

1.2

SYSTEM OVERVIEW

FEATURES

The BioLogic HR system provides the following features:

2

•

A space-saving modular and stackable design which minimizes the System footprint on the bench. The BioLogic HR Controller, Workstation and optional peripherals are linked by a single “instrument bus” cable in any configuration, making electrical connections simple and fast.

•

Control software specifically designed for the bio-chromatographer. The pre-loaded, dedicated software is based on the easy-to-use Windows® 95 platform and is designed for multiple users in the laboratory. Programming separation methods is menu-driven, easy and intuitive using either Time-based or Volume-based steps. Method storage capacity is virtually unlimited.

•

On-screen help function.

•

Single-point control of chart recorders, fraction collectors, signal import modules and auxiliary pumps (e.g., Bio-Rad’s Econo Pump or a peristaltic pump) for sample loading.

•

Binary gradient formation by two dual piston, low-pulsation pump heads which operate up to a maximum pressure of 1000 psi at flow-rates up to 10 ml/min. This permits the use of a wide range of both preparative and analytical columns.

•

Dynamic mixing for accurate and reproducible gradient formation.

•

The use of biocompatible materials throughout the flowpath to ensure maximum recovery of biological activity.

•

Standard high resolution optics unit (mercury lamp) for fixed wavelength (280 nm and 254 nm) detection and a choice of preparative or analytical flow-cells. A zinc lamp and filter combination for 214 nm detection is optional. Other filters are available from Bio-Rad.

•

Standard conductivity cell to monitor gradient formation.

•

Simple-to-sophisticated Fraction Collection schemes are standard. Collection is volume-based with Collect All, Collection Windows, Threshold and Threshold + Collection Window modes available with a range of collector types.

•

3 ports to accommodate a range of low pressure solenoid divert and buffer select valves.

•

3 ports to accommodate a range of high pressure, automated stream select and sample injection valves.

•

Sufficient tray space on the basic System Rack to hold a Model 2110 Fraction Collector or a Model 1327 Chart Recorder. Additional rack components are available to make a three-tray configuration.

•

Standard column clamping arrangement to eliminate rack clutter.

•

Available manual inject valve with position-sensing switch to satisfy Hold conditions programmed in a Method.

•

Water-resistant integrated mini-keyboard designed for the laboratory.

SYSTEM OVERVIEW

INTRODUCTION

1.3 UNPACKING When you receive the BioLogic HR system, carefully inspect the shipping containers for any damage which may have occurred in shipping. Severe damage to a container may indicate damage to its contents. If you suspect damage to the contents may have occurred, immediately file a claim with the carrier in accordance with their instructions before contacting Bio-Rad Laboratories.

!

Caution Lift items from the bottom as you remove them from their containers!

Open each of the shipping cartons and lift the contents out of its packing. Check the contents of each box against the supplied packing list. Remove the plastic bag from each unit and inspect the unit for external damage. If any part is missing or damaged, contact Bio-Rad Laboratories immediately.

1.4

DESCRIPTION OF SYSTEM COMPONENTS

The following sections identify the key features of the BioLogic HR system and available options.

Table 1. Description of System Components

Component

Function

Controller

This is the “brain” of the system. It provides the graphic user-interface from which the system must be set up and run. From the controller, you can set up and run methods, perform simple data analysis, and store method and run data. Control of the Workstation (which includes the Gradient Pump, Valves, UV/Conductivity Detectors) and peripheral instruments (Model 2128 fraction collector, the Model EP-1 Econo Pump, and Signal Import Modules) is via the instrument bus connection.

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INTRODUCTION

SYSTEM OVERVIEW

Table 1. (continued) Description of System Components

Component Workstation

Function This consists of the following: • Gradient Pump. This consists of two bio-compatible dual-piston pumpheads, labeled A and B. It is controlled from the Controller. Purge and Pause buttons (Purge A, Purge B, and Pause) are present on the front of the Workstation. The Gradient Pump has a maximum delivery of 10 ml/min at a maximum pressure of 1000 psi. The pump system includes a built-in, integrated pressure transducer and dynamic mixing chamber. • Control circuitry and connectors for the UV and Conductivity Detector signals. Signal export connectors for UV and Conductivity data at 1 V and chart recorder control (pen up/down, start/stop). • Control circuitry and connectors for the system valves (low pressure solenoid and automated high pressure inject and select valves). • An Auxiliary (AUX) connector for the Model 2110 fraction collector, generic fraction collectors, the position-sensing cable from the manual inject valve, and/or the Model EP-1 Econo Pump for sample loading. • An output power connector to the UV lamp. • Two instrument bus (phone-type) connectors.

System Rack

This is the system organizer for columns and cartridges, buffer containers, sample inject and stream select valves, UV and Conductivity Detectors, and other devices used with the system. The optional BioLogic HR Rack Expansion kit extends the rack to 3 trays.

UV Detector

The UV Detector with a mercury lamp and filters for fixed wavelength UV detection at 280 nm and 254 nm is provided with all BioLogic HR systems. An optional zinc lamp with a 214 nm filter is available, as are filters for other wavelengths. Two flow cells are available: • An analytical “Z” cell for high resolution protein chromatography applications is provided with all BioLogic HR systems. It has a path length of 5 mm for maximum sensitivity and a swept volume of only 8 µl for maximum sensitivity at flow rates from 0.1 to 10 ml/min. • A cuvette cell is available as an option. It is for preparative work where high sensitivity is not an absolute requirement. It has a path length of 2 mm and a swept volume of 30 µl.

Conductivity Detector

A Conductivity flow cell, which rests in a specially designed holder on the front of the UV Optics unit, provides instantaneous, on-line readings of conductance for monitoring a salt gradient and optimization of column chromatographic conditions. It has a nominal swept volume of 2 µl.

pH Monitor

The pH Monitor, which is available as an option, provides real-time monitoring of buffer pH. It is designed for flow-through pH readings.

Valves

The following valves are available as options to the BioLogic HR system. • High pressure (to 1000 psi) valves include: AV7-3 Automated Inject Valve, V7-3 Manual Inject Valve, AV9-8 Automated Stream Select Valve, V9-8 Manual Stream Select Valve. • Automated, low pressure (to 30 psi) solenoid valves for applications such as buffer selection and flow diversion include: SV5-4 Select Valve and SV3-2 Diverter Valve.

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SYSTEM OVERVIEW

INTRODUCTION

Table 1. (continued) Description of System Components

Component

Function

Sample Loops

The following sample loops are available for use with the inject valves: 25 µl, 50 µl, 100 µl, 250 µl and 500 µl, 1.0 ml, 2.0 ml and 5.0 ml. The 1.0 ml loop is provided with the System. In addition, the DynaLoop 25 and DynaLoop 90 Sample Loops are available for automated introduction of large sample volumes.

Columns

Bio-Rad makes a number of different columns. A description of each type of column is located in Chapter 2, section 2.6, Columns and Column Fittings.

Auxiliary Load Pump

The Model EP-1 Econo Pump may be used as an auxiliary pump for loading large sample volumes onto low pressure columns. This is available as a system option. For loading large volumes onto high pressure columns, load directly through the Gradient Pump via a select valve, or use the Bio-Rad DynaLoop sliding-piston sample loop.

Fraction

Bio-Rad offers two fraction collectors as system options.

Collector

• The Model 2128 provides X-Y motion drop dispensing over any of 5 types of racks. The racks accommodate a wide range of tube diameters and lengths, microtiter plates, microtubes, and “bottle size” fractions. The Model 2128 is suited for both analytical and preparative applications. It is controlled by the BioLogic HR Controller via the instrument bus. • The Model 2110 uses a stationary drop-dispensing head to collect up to 80 fractions in a carousel. To divert non-peak material to waste, use the SV3-2 Diverter valve. The Model 2110 is controlled by the BioLogic HR Controller via the AUX connector on the Workstation. • Other fraction collectors may be used with the BioLogic HR system as long as tube advances can be initiated by an active low pulse (>100 ms). (Refer to the fraction collector’s documentation for discussion of its active low pulse.) The SV3-2 Diverter valve is required for full operation. Fraction advance marks (event marks) for a generic collector are embedded in the UV signal sent from the Workstation to a chart recorder. Generic collectors are controlled by the BioLogic HR Controller via the AUX connector on the Workstation. Regardless of which fraction collector is chosen, the fraction volume and collection parameters such as Collect All, Collection Windows, Threshold, and Threshold + Collection Windows can be controlled from the BioLogic HR Controller.

Chart Recorder Bio-Rad offers the Model 1327 Chart Recorder as a system option. This is a dual pen chart recorder for Conductivity and UV Detector readings. Stop/Start and Pen Up/Down functions are controlled by the BioLogic HR Controller via the BioLogic HR Workstation. Chart speed is set at the recorder itself (the BioLogic HR Controller does not control this function). Event marks including fraction advance marks are embedded in the UV signal sent from the Workstation to the chart recorder. The BioLogic HR system may be used with any generic single or dual-pen chart recorder, provided Stop/Start and Pen Up/Down pin-out information is available.

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INTRODUCTION

SYSTEM OVERVIEW

Table 1. (continued) Description of System Components

Component Signal Import Module

Function Up to two Signal Import Modules (SIMs-HR) may be connected to the BioLogic HR using the instrument bus. This module allows you to connect the pH Detector to the BioLogic HR. There is also a 3-pin connector which may be used to connect other detectors (such as variable wavelength, UV, fluorescence, and RI detectors) which transmit an analog signal of up to 2.5 V. The SIM-HR converts the detector’s analog signal to a digital signal and communicates the information to the BioLogic HR system.

Tubing

The following tubing is supplied with the unit: System Tubing: 1/16” OD, 0.020” ID Tefzel® tubing. The approximate volume of 1 cm of tubing is 2 µl. Pump Inlet Tubing: 1/8” OD, 0.062 “ ID Teflon® tubing. The approximate volume of 1 cm of tubing is 20 µl.

Fittings

Flangeless 1/4-28 flat bottom fittings.

Auxiliary Pump

Bio-Rad’s Model EP-1 Econo Pump is available as a system option. It is a two-channel bidirectional variable speed peristaltic pump for low-pressure chromatography. It offers a full range of features to facilitate ease of use as a stand-alone pump or as an accessory to the Biologic HR system to load large volumes, especially onto low-pressure columns.

Teflon and Tefzel are registered trademarks of E.I. du Pont de Nemours & Co.

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SYSTEM OVERVIEW

2.0 2.1

DESCRIPTION OF BIOLOGIC HR SYSTEM COMPONENTS

DESCRIPTION OF THE BIOLOGIC HR SYSTEM COMPONENTS CONTROLLER

The Controller is the “brain” of the BioLogic HR system. It consists of the Color Display Monitor, the Controller unit, the keyboard and mouse devices, and a floppy disk drive. Note: The sound capabilities of the CD-ROM drive and the audio and mic connectors on the back of the controller are not for use with the BioLogic HR system.

Table 2. Controller’s Front Panel Features

FLOPPY DISK DRIVE BioLogic HR Controller COMPACT

POWER SWITCH CD-ROM DRIVE

Feature

Description

CD ROM Drive

Use this drive to load updates of the BioLogic HR operating software. To open the drive, press the button on the front of the drive.

Keyboard & Mouse

These two interface devices (not shown) are used to control the system. The keyboard can withstand the minor spills associated with use in a laboratory.

Floppy Disk Drive

Use it to backup and restore methods onto floppy disk. Press the button next to the drive slot to manually eject diskettes from the drive.

Power Switch

Turns on/off the Controller and Monitor.

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DESCRIPTION OF BIOLOGIC HR SYSTEM COMPONENTS

SYSTEM OVERVIEW

Table 3. Controller’s Rear Panel Connectors

INSTRUMENT BUS

CONTROLLER'S POWER CONNECTOR

KEYBOARD PORT

COLOR MONITOR PORT

MOUSE PORT

Connector

PRINTER PORT

Description

Instrument bus (phone jack connector): The Instrument Bus allows all components in the BioLogic HR System to communicate with the BioLogic HR Controller. The components may be cabled together in any order via their respective Instrument Bus connectors. This is termed “daisy-chaining.” The presence of a component on the Instrument Bus is recognized when it is switched on. Even when one component is switched off, another component “daisy-chained” to it can be controlled by the Controller. Note that the Controller has only one phone jack connector.

Video connector: Use this connector to connect the Color Monitor to the Controller.

Keyboard connector: Use this connector to connect the Keyboard to the Controller. It supports any standard PC compatible keyboard.

Mouse connector: Use this connector to connect the mouse to the Controller. It supports any standard PC compatible mouse or trackball.

Parallel connector: Use this connector when connecting any parallel printer. Laser and Dot Matrix printers are examples of parallel printers. Refer to your Windows® 95 and/or your printer documentation for instructions on how to set up the printer for use with the BioLogic HR System. Some printer drivers are pre-installed on the Controller.

Power Connector: This is the power connector for the Controller.

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SYSTEM OVERVIEW

2.2

DESCRIPTION OF BIOLOGIC HR SYSTEM COMPONENTS

Workstation

The Workstation is the “heart” of the BioLogic HR system. It contains the following: •

Gradient Pump. This consists of two bio-compatible dual-piston pumpheads, labeled A and B. The Gradient Pump is rated to 1,000 psi (maximum), with a flow rate range of 0.1 to 10 ml/min. This module also contains an integrated pressure transducer and dynamic mixer.

•

The control circuitry for the Gradient Pump, the UV and Conductivity Detectors, and the Valves (solenoid and automated high pressure valves). Connectors for all valves and Detector inputs are on the rear panel along with output connectors for UV and Conductivity signals to a chart recorder.

•

An AUX combicon connector used for (a) controlling fraction advances of the Model 2110 and generic fraction collectors, (b) receiving a signal from the position sensing switch of the V7-3 Manual Inject Valve, and (c) starting and stopping the Model EP-1 Econo Pump for sample loading.

•

Power supply for the Gradient Pump, the UV optics unit, the Conductivity cell, and the Workstation electronics.

Table 4. Workstation’s Front Panel Controls

TUBING FROM PUMP OUTLET TO INJECT VALVE'S PORT #2 MIXER AND TRANSDUCER

BioLogic HR Workstation

PRIMING PORT A POWER ON/OFF, PAUSE, PURGE A & B

Feature

POWER

PAUSE

PURGE A

A

B

B

PRIMING PORT B WASH PORT OUTLET INLET PORTS A & B

Description

Power Button

Press this switch to turn on power to the Workstation.

Pause Button

Stops the Gradient Pump and pauses a running Method. When the Pause button is pressed, the status LEDs for pumps A and B change from green to flashing red. This button is useful in the event of a leak in the plumbing (the fittings or tubing) or if there are not enough tubes in the Fraction Collector (Panic stop!). To resume the run, • Press the button again, or • Press the Continue button in the Run screen, or • Press the START button in the Manual screen.

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DESCRIPTION OF BIOLOGIC HR SYSTEM COMPONENTS

SYSTEM OVERVIEW

Table 4. (continued) Workstation Front Panel Controls

Feature

Purge A & B Buttons

Description

These buttons are used to purge the tubing lines during setup of the system. When the purge buttons are pressed, the pump heads run at their (default) maximum flow of 7.0 ml/min. (To alter the purge flow rate, select Edit User Preferences from the Options pull-down menu.) When the buttons are pressed, the green LEDs for pumps A and B flash. To stop the flow, press the purge buttons again and note the LEDs go off. These buttons do not operate when the system is running a Method. Note: It is important to move any in-line inject valve to its purge position to avoid exposing a column to high flow rates when the purge function is used.

10

Alert Light

This LED serves two functions: a. Constant red: Indicates that the pump has shut down due to a mechanical or electrical problem. This includes a shutdown due to the pressure limit being exceeded. b. Flashing red: Indicates that a pre-programmed ALARM step has been reached in a running Method (e.g., to indicate that a manual valve position needs changing). This LED will flash simultaneously with an audible tone from the Controller.

Plumbing Connections

Ports on the front of the Workstation: a. Pump Head Inlet ports: Buffer inlet lines attach to the ports at the bottom of each pump head. These ports accept standard 1/4-28 flat-bottom fittings. The pump head inlet tubing is 1/8” OD, 0.062” ID Teflon tubing with flat bottom fittings which are supplied in the fittings kit. b. Pump Head Output and Pressure Transducer Mixer Input ports: Output from the two pump heads is to the integral Pressure Transducer/Mixer, located between the pump heads. These ports are pre-plumbed using standard 1/4-28 flat-bottom fittings and 1/16” OD, 0.020” ID Tefzel tubing. c. Mixer Output port: The output from the mixer accepts standard 1/4-28 flatbottom fittings. Use the 1/16” OD, 0.020” ID Tefzel tubing with Super Flangeless flat bottom fittings supplied in the fittings kit. d. Pump Head Priming port: There is a port on each pump head which is used when priming the pump. The port accepts any size syringe with a luer (not luer lock) fitting. This draws buffer along the inlet line and to the pump head. Twist the port counter-clockwise to open it. e. Pump head washout port: The angled port at the top of each pump head is used for rinsing the piston behind the piston seals to remove crystallized salts. It accepts any syringe with a luer fitting. The rinse output is at the lower front of the pump housing.

SYSTEM OVERVIEW

DESCRIPTION OF BIOLOGIC HR SYSTEM COMPONENTS

4

UV CHART COND CHART UV OPTICS

TEST PORT

AUTOMATED VALVES

INSTR BUS

5

6

COND FLOWCELL UV LAMP

Connector

AUX 1. Inject 2. n/c 3. n/c 4. n/c 5. FC Adv 6. AUX Pump 7. n/c 8. n/c 9. Gnd

SOLONOID VALVES

Table 5. Workstation’s Rear Panel Connectors

1 2 3 AUX 1. Inject 2. n/c 3. n/c 4. n/c 5. FC Adv 6. AUX Pump 7. n/c 8. n/c 9. Gnd

Description

Aux connector: This 9-pin connector is used to connect a variety of peripheral modules which do not communicate with the BioLogic HR Controller over the Instrument Bus. Pin # 1

2 3 4 5 6 7 8 9

Description Inject. The V7-3 manual inject valve reports its position (either LOAD or INJECT). This information may be used to satisfy a Hold command which has been programmed in a Method Protocol. n/c. No connection n/c. No connection n/c. No connection FC Adv. Model 2110 and generic Fraction Collector Advance output. AUX Pump. A Stop-Start command is sent to a pump (e.g. Bio-Rad EP-1). n/c. No connection n/c. No connection GND. Ground

Solenoid Valves: For connecting Bio-Rad’s low pressure solenoid valves (SV5-4 and SV3-2) to the system.

Automated Valves: For connecting Bio-Rad’s high pressure automated valves (AV7-3 Inject and AV9-8 Stream Select) to the system.

Cond Flowcell: For connecting the Conductivity flow cell to the system.

UV Lamp: This specialized 6-pin square connector provides electrical power to the mercury or zinc lamp in the UV Optics module’s lamp housing.

UV Optics: For connecting the UV Optics module to the system.

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DESCRIPTION OF BIOLOGIC HR SYSTEM COMPONENTS

SYSTEM OVERVIEW

Table 5. (continued) Workstation Rear Panel Connectors Key

Description

Cond Chart: For conductivity signal output to a single or dual pen chart recorder. An 8-pin mini-DIN to banana plug cable (System Cable 4) for connection to the Model 1327 Chart Recorder is available. Connect the red line to the positive (+) terminal and the black line to the negative (–) or ground terminal of channel 2 (CH2). The chart recorder should be set to 1 V full scale. If you are using the Model 1327 Chart Recorder, move all buttons to the “green” settings.

UV Chart: For UV signal output to a single or dual pen chart recorder. In addition, when the Bio-Rad Model 1327 is used, chart recorder Pen Up/Down, Stop/Start commands, and event marks are sent from this connector. The Bio-Rad Model 1327 dual pen recorder needs an 8-pin mini-DIN to standard DIN cable (System Cable 2) available from Bio-Rad. Generic chart recorders require an 8-pin mini-DIN to breakout cable (System Cable 7) available from Bio-Rad. When a Signal Import Module signal replaces the standard BioLogic HR UV Detector, use System Cable 20 to control a Bio-Rad Model 1327 dual pen recorder. The chart recorder should be set to 1V.

Power Cord Connector: This grounded 3-prong connector is for input power to the Workstation and output power to any unit connected to the Workstation. The Workstation’s input power cord should be plugged into a 3-prong grounded power outlet.

Instr Bus: These RJ-45 modular phone connectors and their cables connect the Workstation to the other components in the system. The Instrument Bus handles all communications between the Controller and each of the components in the system. For example, the Instrument Bus may connect the Workstation to the Model 2128 Fraction Collector. Components can be connected in any order in the system.

Test connector: This connector is reserved for internal Bio-Rad use.

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SYSTEM OVERVIEW

2.3

DESCRIPTION OF BIOLOGIC HR SYSTEM COMPONENTS

UV AND CONDUCTIVITY DETECTORS

The UV detector consists of the Optics bench, a filter tray, a flow cell, and a lamp. It is designed to hold the Conductivity Monitor flow cell if required.

Figure 2. Model OM-II UV Optics Module and Conductivity Flow Cell

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