Bio-Rad ChromLab Software User Manual

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3.0

NGC™ Chromatography Systems

and ChromLab™ Software

User Guide

Version 3.0

NGC™ Chromatography Systems and Chr

User Guide

Version 3.0

omLab™ Software

Bio-Rad Technical Support Department

The Bio-Rad Technical Support department in the U.S. is open Monday through Friday, 5:00 AM to 5:00 PM, Pacific Time. Worldwide technical support is available on the Web at

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Web: www.consult.bio-rad.com

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No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage or retrieval system, without permission in writing from Bio-Rad.

Bio-Rad reserves the right to modify its products and services at any time. This user guide is subject to change without notice. Although prepared to ensure accuracy, Bio-Rad assumes no liability for errors or omissions, or for any damage resulting from the application or use of this information.

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Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11

Main NGC Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

NGC Chromatography Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Finding Out More . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

Chapter 2 The Workspace . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17

The Home Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

The System Control Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

The Method Editor Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

The Evaluation Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Instrument Control Touch Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Touch Screen Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Touch Screen Toolbar Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Chapter 3 System Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .29

System Control Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

View Menu Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Manual Menu Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Tools Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Toolbar Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Context Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

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Chromatogram View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

Changing Chromatogram View Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Showing or Hiding Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39

Changing Trace Color . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Zooming In and Out. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Changing the X-Axis Units and Scale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Changing the Y-Scale Values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Autoscaling the UV Trace . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Annotating the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

Showing and Hiding Events, Fractions, Errors, and Annotations . . . . . . . . . 44

Fluidic Scheme Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

Modules. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Fluidic Scheme Configurations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

Working with Fluidic Schemes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .57

Fluidic Scheme Mapping. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Calibrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64

Calibrating a Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .64

Calibrating Flow Rate of Pumps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

Calibrating Pressure Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

Verifying Plumbing with the Point-to-Plumb Feature . . . . . . . . . . . . . . . . . . . . . . 67

System Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .69

Delay Volume Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69

Control Flow Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Remote Access Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .74

Trace Settings Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .74

Device Input Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76

Device Output Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

Air Sensors Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80

Email Notifications Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82

System Name Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85

System Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .86

General Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Devices Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Process Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

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Detector Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87

Single-Wavelength Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87

Multi-Wavelength Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88

Preferences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .89

Email Server Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89

Chapter 4 Performing a Manual Run. . . . . . . . . . . . . . . . . . . . . . . . . . 93

Preparing the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94

Priming and Purging the System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94

Cleaning the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .102

Running an Experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

Changing Module Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Stopping a Manual Run. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Clearing Run Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Saving a Manual Run. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Saving a Manual Run on the Touch Screen . . . . . . . . . . . . . . . . . . . . . . . . . . 110

Viewing Run Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111

Chapter 5 Method Editor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113

Before You Begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .114

Accessing Method Editor Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114

Opening a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115

Method Editor Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117

Edit Menu Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118

View Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .119

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119

Toolbar Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119

Phase Library Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .120

Adding a Phase to a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121

Method Outline Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122

Phase Parameters Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122

Method Steps Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123

Viewing Details of a Step in the Step Table . . . . . . . . . . . . . . . . . . . . . . . . . . 124

Adding, Modifying, and Deleting Steps in the Step Table. . . . . . . . . . . . . . . . 124

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Table of Contents

Method Settings Parameters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125

Fluidic Scheme . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125

Column Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126

Detector Settings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131

Unit Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131

pH Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

Fraction Collection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

Flow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133

Buffer Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133

Phase Controls and Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136

Standard Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138

Sample Application Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138

Column Wash Parameters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143

Elution Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144

System CIP (Clean in Place) — Storage Parameters . . . . . . . . . . . . . . . . .145

Column Preparation/Activation Parameters . . . . . . . . . . . . . . . . . . . . . . . . 152

Column Performance Test Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . .153

System Preparation Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154

Create New Phase Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155

Step Library Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156

Scout Parameters Pane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157

Chapter 6 Creating a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159

Standard Method Templates. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160

Creating a Method from a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .163

Creating a Method from Standard Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165

Adding Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170

Editing Methods and Phases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170

Renaming Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171

Rearranging Phases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171

Deleting Phases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172

Opening a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172

Running a Method and Collecting Fractions. . . . . . . . . . . . . . . . . . . . . . . . . . . . 173

Collecting Fractions Using the BioFrac Fraction Collector . . . . . . . . . . . . .173

Collecting Fractions Using Outlet Valves . . . . . . . . . . . . . . . . . . . . . . . . . .173

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Collecting Fractions in Run Queues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174

Collecting Fractions Using the BioFrac Fraction Collector . . . . . . . . . . . . .174

Collecting Fractions Using Outlet Valves . . . . . . . . . . . . . . . . . . . . . . . . . .174

Saving a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .177

Renaming a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178

Deleting a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179

Scouting on Parameters within a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180

The Scouting Wizard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .181

Page 1: Choose a Scouting Parameter . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181

Page 2: Select Method Steps to Scout . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182

Page 3: Generate Scout Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184

Using the Scouting Wizard to Scout Columns . . . . . . . . . . . . . . . . . . . . . . 185

Using the Scouting Wizard to Scout Samples . . . . . . . . . . . . . . . . . . . . . . 187

Creating a Scouting Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189

Special Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190

Viewing a Summary of the Scouting Method . . . . . . . . . . . . . . . . . . . . . . . . . 192

Running a Scouting Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192

Chapter 7 Evaluating Results. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195

Managing Analysis Projects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196

Evaluation Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 198

File Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199

View Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .201

Analysis Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201

Help Menu Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 202

Tab Toolbar Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 202

Customizing the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203

Displaying Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203

Selecting the Active Trace. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204

Showing or Hiding a Trace . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205

Changing the Axes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .205

Controlling the Range of the Y-Scale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205

Changing Trace Colors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206

Zooming In and Out. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207

Obtaining Statistics of a Selected Region on the Chromatogram . . . . . . . . . 208

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Table of Contents

Annotating the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210

Copying the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211

Customizing the Trace Table Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 212

Changing Table Grouping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 212

Sorting Table Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213

Ordering and Selecting Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213

Showing or Hiding Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213

Copying a Table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214

Showing or Hiding Pre-Injection Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214

Manually Setting the Injection Point (x Alignment) . . . . . . . . . . . . . . . . . . . . . 215

Managing Runs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216

Renaming a Single Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216

Deleting a Single Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216

Viewing Multiple Runs in Separate Tabs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 217

Comparing Traces. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218

Viewing Multiple Runs in One Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219

Stacked View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219

Overlay View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221

Customizing the Trace Comparison Table . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Expanding or Collapsing Groups. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Hiding All Traces in a Group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Showing All Traces in a Group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222

Offsetting Traces — Overlay Mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223

Managing Analyses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .224

Removing Runs from a Trace Comparison . . . . . . . . . . . . . . . . . . . . . . . . . . . 224

Saving a Trace Comparison Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 224

Copying an Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225

Renaming an Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .225

Deleting an Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .226

Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226

Exporting Run Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226

Peak Integration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .227

Automatic Peak Integration Settings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

Baseline Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

viii | NGC Chromatography Systems and ChromLab Software

Table of Contents

Peak Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .229

Peak Filtering . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229

Default Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229

Integrate and Delete . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .230

Starting Peak Integration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230

Displaying Peak Results — the Chromatogram . . . . . . . . . . . . . . . . . . . . . . . 231

Changing Peak View Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232

Displaying Peak Results — the Peaks Table. . . . . . . . . . . . . . . . . . . . . . . . . . 233

Table Display Order and Column Selection . . . . . . . . . . . . . . . . . . . . . . . . 235

Showing or Hiding Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235

Copying the Peaks Table. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236

Grouping or Ungrouping the Peaks Table. . . . . . . . . . . . . . . . . . . . . . . . . . 236

Clearing Extinction Coefficient Values in the Peaks Table . . . . . . . . . . . . . 237

Adjusting Peaks Manually . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 238

Saving Peak Integration Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 240

Evaluating Fractions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241

Displaying Fraction Collection Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241

Displaying Fraction Collection Results — the Fractions Table . . . . . . . . . . . . 243

Calculating Protein Concentration for Fractions . . . . . . . . . . . . . . . . . . . . .245

Viewing Fraction Details . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .246

Pooling Fractions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246

Viewing Fraction Collection Results — the Heat Map. . . . . . . . . . . . . . . . . . . 248

Viewing Heat Map Details . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 249

Column Performance Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251

Column Performance — Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251

Traces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 252

Calculate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 252

Analyzing Column Performance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 253

Resetting the Column Performance Properties. . . . . . . . . . . . . . . . . . . . . . 254

Deleting Column Performance Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254

Column Performance — Manual Adjustment . . . . . . . . . . . . . . . . . . . . . . . . . 255

Displaying Column Performance Results — the Chromatogram . . . . . . . . . . 255

Displaying Column Performance Results — the Column

Performance Table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256

User Guide | i x

Table of Contents

Chapter 8 Importing and Exporting Data . . . . . . . . . . . . . . . . . . . . . 259

Importing an NGC Method or Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 259

Importing Unicorn Data Files. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261

Importing BioLogic DuoFlow Data Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 263

Exporting Data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264

Exporting Data as an NGC File . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 265

Exporting Run Data as a .csv File . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 269

Exporting Diagnostic Logs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .271

Chapter 9 Reports. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275

Producing a Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276

Method Reports. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277

Run Reports. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277

Analysis Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278

Printing a Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278

Saving a Report. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 279

Appendix A Database Management . . . . . . . . . . . . . . . . . . . . . . . 281

Back Up the NGC Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .282

Restore the NGC Database. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 283

Appendix B Multicolumn Purifications . . . . . . . . . . . . . . . . . . . . . 285

Multicolumn Purification Method Templates. . . . . . . . . . . . . . . . . . . . . . . . . . . . 286

Multicolumn Sequential Purification Templates . . . . . . . . . . . . . . . . . . . . . . . 286

Multicolumn Tandem Purification Templates . . . . . . . . . . . . . . . . . . . . . . . . . 289

Performing Multicolumn Purification Chromatography. . . . . . . . . . . . . . . . . . . . 291

Plumbing the NGC System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291

Plumbing NGC Discover Pro Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291

Plumbing the NGC Discover Pro System to Use

Multicolumn Purification Templates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .292

Priming the NGC System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 294

Selecting and Saving a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295

Running Multicolumn Purification Methods . . . . . . . . . . . . . . . . . . . . . . . . . . 296

x | NGC Chromatography Systems and ChromLab Software

1 Introduction

NGC™ chromatography systems provide a general purpose purification platform for purifying all forms of biomolecules using a combination of chromatography techniques. The systems are also useful for developing and optimizing purification protocols. The systems can provide highly purified proteins, peptides, nucleic acids, monoclonal antibodies, and other small molecules.

User Guide | 1 1

1 | Introduction

ChromLab™ software enables you to set up and control an NGC instrument, run protein separations and other operations manually, program methods to automate purification runs, evaluate the results, and generate and print experiment reports. This user guide explains how to perform all these tasks.

Main NGC Features

NGC chromatography systems enable you to do the following:

 Easily create purification and maintenance protocols from predefined

method templates and protocol phases

 Automate multicolumn purification processes using preprogrammed

templates and multiple column switching valves

 Automate multiple sample injections using either the sample inlet valve and

the sample pump or the C-96 autosampler

 Expand sample monitoring using the signal import module (SIM) to export

digital signals to and import digital signals from external detectors

 Collect large-volume fractions using multiple outlet valves while also

collecting small-volume fractions using the BioFrac™ fraction collector

 Automatically prepare buffers using preprogrammed buffer blending

protocols

 Analyze purification results through 1-click peak integration, determine

protein concentration and calculate column performance

 Automate purification protocol optimization using the scouting wizard

 Easily locate fractions containing peaks of interest and view the protein

concentration within each fraction

 Extend the preconfigured systems with additional valves for buffers,

samples, and columns

12 | NGC Chromatography Systems and ChromLab Software

Main NGC Features

 Organize the location of the modules to optimize separation performance

based on method scale and complexity, and to minimize the system swept volume

 Minimize errors when connecting tubing using the Point-to-Plumb™

feature in ChromLab software

User Guide | 1 3

1 | Introduction

NGC Chromatography Systems

All NGC chromatography systems include ChromLab software and the NGC touch screen.

NGC chromatography systems are available in several combinations. Each system is equipped with either two 10 ml system pumps (the 10 series) or two 100 ml system pumps (the 100 series).

The NGC Quest™ chromatography system includes the following:

 Two system pumps

 Mixer

 Sample injection valve

 Conductivity monitor with either a single-wavelength UV detector or a

multi-wavelength UV/Vis detector (available on the Plus systems)

14 | NGC Chromatography Systems and ChromLab Software

NGC Chromatography Systems

The NGC Scout™ chromatography system includes the following:

 All modules on the Quest system

 pH detector valve

 Buffer blending valve

The NGC Discover™ chromatography system includes the following:

 All modules on the Scout system

 Column switching valve

 Two buffer inlet valves

 Sample pump

The NGC Discover Pro chromatography system includes the following:

 All modules on the Discover system

 Fourth expansion tier

 Sample inlet valve

 Outlet valve

User Guide | 1 5

1 | Introduction

Finding Out More

After you install NGC documentation from the NGC Chromatography Systems Documentation DVD, you can access installed NGC guides and tutorials on the Help menu in any ChromLab view.

More information about the NGC chromatography systems and ChromLab software is available fr

 The NGC Chromatography Systems and ChromLab Software Installation

Guide is available on your NGC Chromatography Systems Documentation DVD as a .pdf file. This guide explains how to set up your environment, set up and install the NGC instrument in the lab, install ChromLab software, and connect ChromLab to the NGC system.

 The NGC Chromatography Systems and ChromLab Software Instrument

Guide is available on your NGC Chromatography Systems Documentation DVD as a .pdf file. This illustrated guide details the modules that make up the NGC instrument and includes troubleshooting and maintenance information.

 For ChromLab Help, clicking in dialog boxes accesses relevant

information. Screen-level help is also available on the Help menu.

om the following sour

ces.

 NGC video tutorials are available on the NGC Chromatography Systems

Documentation DVD as .mp4 files.

Tip: You can click the Bio-Rad logo in the upper-left corner of any Chr

omLab window to launch the Bio-Rad website.

16 | NGC Chromatography Systems and ChromLab Software

2 The Workspace

ChromLab™ software provides an intuitive interface for developing chromatography methods, operating an NGC™ instrument, and analyzing data from chromatography runs.

ChromLab software presents four primary workspaces.

 The Home window

 The System Control window

 The Method Editor window

 The Evaluation window

Each workspace is shown and described in this chapter. The NGC instrument touch scr

een is also described.

User Guide | 1 7

2 | The Workspace

The Home Window

ChromLab software opens with the Home window, which displays three panes and the System Control tab.

The three panes provide quick access to the system control, method editor, and evaluation workspaces. The status of connection to the NGC instrument is also displayed. Links to recently completed runs, recently created methods, and recently accessed or analyzed runs and analyses appear listed at the bottom of the window.

Tip: Clicking the Bio-Rad logo in the upper-right corner of any ChromLab window launches the Bio-Rad website.

18 | NGC Chromatography Systems and ChromLab Software

The Home Window

Method Editor and Evaluation tabs become visible in the Home window when you select tasks in the Method Editor and Evaluation panes.

User Guide | 1 9

2 | The Workspace

File Menu Commands

Connect to System — opens a dialog box that enables you to choose an NGC

chromatography system to connect to. ChromLab detects the NGC systems available on the same subnetwork or those systems that are directly connected to the computer. Their system name, network name, and IP address appear in the table. To connect to a system, you can:

 Select a name in the list of detected systems and click Connect.

 Select the appropriate radio button, enter the system’s name or IP

Note: If your system does not appear in the list, click Detect. Chr

omLab searches the network for available NGC systems and

refreshes the list.

address, and click Connect.

Tip: To obtain the system’s name and IP address select System Information on the instrument touch scr

een dropdown menu.

Disconnect System — starting Chr

omLab connects you to the NGC system. This command enables you to disconnect ChromLab software from the system so you can work offline.

Manual Run — opens the System Contr

ol window in manual mode so you can

perform a manual run or set up your system manually.

Calibrate — opens the Calibration dialog box, which displays instructions and options for selecting a module and calibrating it. See Calibrations on page 64 for details.

Open Template — opens the T

emplate dialog box in which you can select a

method template from template folders organized by technique.

Open Method — opens the Method dialog box in which you can select fr your saved methods.

New Method — opens the Method Editor window in which you can cr method using standard method phases and steps.

20 | NGC Chromatography Systems and ChromLab Software

om a list of

eate a

The Home Window

Import — displays links from which you can import the following:

 NGC File — opens a dialog box in which you can import a method, a

method with runs, or a run exported from ChromLab software running on another NGC system. See Importing an NGC Method or Run on page 259 for more information.

 Unicorn Data — opens a dialog box in which you can import a Unicorn

data file into the NGC database. See Importing Unicorn Data Files on

page 261 for more information.

 DuoFlow Data — opens a dialog box in which you can import a BioLogic

DuoFlow™ data file into the NGC database. See Importing BioLogic

DuoFlow Data Files on page 263 for more information.

Export — displays links fr

 Methods/Method Runs — opens a dialog box in which you can export

om which you can export the following:

both single or multiple methods and single or multiple methods with associated runs.

 Runs — opens a dialog box in which you can export single or multiple

runs.

See Exporting Data as an NGC File on page 265 for more information.

Open Run/Analysis — opens a dialog box in which you can select a run or analysis to view or analyze.

Preferences — opens dialog boxes in which you can do the following:

 Select pressure units for all system and software pressure values. This is a

global setting.

 Set up an SMTP server to receive email messages about system

notifications from the ChromLab computer.

Exit — closes Chr

omLab.

User Guide | 2 1

2 | The Workspace

Help Menu Commands

Help — displays screen-level help topics and links to installed manuals.

Export Diagnostic Logs — opens the Export Diagnostic Logs dialog box in which

you can export all critical information that Bio-Rad T diagnose issues. The log files and data are zipped and saved to a location that you choose. See Exporting Diagnostic Logs on page 271 for more information.

echnical Support requires to

About — displays Chr

omLab copyright and version information.

22 | NGC Chromatography Systems and ChromLab Software

The System Control Window

The System Control window enables you to run the instrument manually, monitor method runs while they are running, select fluidic schemes, calibrate pumps and detectors, and verify the accuracy of instrument plumbing. This window displays a chromatogram during a run. A fluidic scheme graphically depicts the flow path of all the modules on the system. A status panel appears above each module displaying its real-time status. In manual mode, clicking a module displays its controls and detailed settings. The Run Log documents each action that occurs during a run. The Run Queue lists all runs ready to be started.

The System Control Window

System Control functionality is detailed in Chapter 3, System Control.

User Guide | 2 3

2 | The Workspace

The Method Editor Window

The Method Editor window enables you to create, open, review, edit, and run a method. You can also open and edit a method template to create a new template. Method Editor functionality is detailed in Chapter 5, Method Editor. See also

Chapter 6, Creating a Method.

24 | NGC Chromatography Systems and ChromLab Software

The Evaluation Window

The Evaluation window enables you to view and compare run data, perform peak integration, and save run data as analyses. Evaluation functionality is detailed in

Chapter 7, Evaluating Results.

The Evaluation Window

User Guide | 2 5

2 | The Workspace

Instrument Control Touch Screen

In addition to ChromLab software running on a computer, the instrument is equipped with a touch screen that accesses system control functionality. You can use this touch screen to run, control, and monitor a run independent of ChromLab. See System Control on page 29 for more information.

Tip: When the NGC system has been inactive for two hours the LED display scr

eens on the instrument tur appears on the touch screen informing you that the system is in standby mode. You can take the system out of standby mode by touching OK in the dialog box, starting the system pumps by initiating a manual or method run, or clicking on a module in the fluidic scheme that has an LED display.

n off, the touch screen dims, and a dialog box

26 | NGC Chromatography Systems and ChromLab Software

Instrument Control Touch Screen

Touch Screen Menu Commands

Calibrate — opens the Calibration dialog box, which displays instructions and

settings for selecting a module and calibrating it. See Calibrating a Module on

page 64 for details.

Point-to-Plumb — starts the Point-to-Plumb™ featur instrument LED lights so you can visually verify or change instrument plumbing. Displays the current fluidic scheme. See Verifying Plumbing with the Point-to-Plumb

Feature on page 67 for details.

Change Fluidic Scheme — opens the Fluidic Scheme Selector dialog box in which you can edit the fluidic scheme or choose another one. See Fluidic Scheme

Configurations on page 51 for details.

Map Fluidic Scheme — opens the Fluidic Scheme Mapping dialog box in which you can map devices on your instrument to their position in the fluidic scheme. See

Fluidic Scheme Mapping on page 60 for details.

System Settings — opens the System Settings dialog box in which you can customize system settings. See System Settings on page 69 for more information about customizing your system.

System Information — opens the System Information dialog box, which lists the serial number and other general information about the NGC device as well as information about the system components, pr detectors.

Service — for Bio-Rad technical service staf

Help — displays detailed information about touch scr

ocesses, and UV and UV/Vis

f use only. Do not select this command.

e and simultaneously turns off

een menu commands.

About — displays Chr

Shut Down — shuts down the NGC system, including the connected computer

omLab version and copyright information.

User Guide | 2 7

2 | The Workspace

Touch Screen Toolbar Commands

Save — saves in a data file the steps executed during a manual run.

Clear — deletes manual run data fr

Autoscale — automatically scales the chr

peak height during the run. Autoscaling is enabled by default.

Reset Zoom — r

Annotate — adds a note to the chr

or after the run completes.

esets the view to show the full chromatogram.

om the touch screen display.

omatogram’s primary y-axis to the tallest

omatogram at points on the x-axis during a run

28 | NGC Chromatography Systems and ChromLab Software

3 System Control

The ChromLab™ System Control window is the main interface to the NGC™ chromatography system instrument. This interface also appears on the instrument touch screen. System Control settings enable you to perform a manual run, monitor and control a method run, verify the device plumbing with the Point-to-Plumb™ feature, control and calibrate the system, and map two or more valves of the same type on your instrument to their position in the fluidic scheme.

In the Home window, you can access the System Control window by selecting the System Contr

System Control Window

The System Control window displays a chromatogram viewer and a graphical fluidic scheme. The chromatogram is a time-, volume-, or column volume-based view of the run data acquired from the instrument. The fluidic scheme is a real-time view of the instrument status and flow.

For enhanced viewing, you can maximize the chromatogram or the fluidic scheme using the Expand buttons on the right side of the window for touch screen viewing.

ol tab.

. This is especially useful

The fluidic scheme graphically depicts the flow between modules and how the system is configur status appears next to its image. For manual runs, a list of executed commands appears in the Run Log pane. In method mode, phases and steps of the method being run appear in their respective tabs, along with controls to stop or pause the run and hold the step. The Run Queue pane lists all the runs that are ready to be started. The Run Queue pane is accessible in both manual and method modes.

ed and plumbed for an experiment. Each module’s real-time

User Guide | 2 9

3 | System Control

LEGEND

1 The menu bar provides quick access to File, View, Manual, Tools, and Help menu

2 Tabs provide quick navigation among open windows (Home, System Control,

3 The tab toolbar pr

4 Expand buttons expand the selected pane to fill the screen.

5 The chromatogram viewer displays data acquired from the instrument as traces

6 The chromatogram legend matches each trace to its trace type and color and

7 In manual mode, the Run Log pane pr







 







commands.

Method Editor, and Evaluation).

ovides commands to save the current run, autoscale the UV trace, change the chromatogram view manual run data from the display.

based on time, volume, or column volume.

displays the wavelength value in nanometers for UV traces. Y traces by clicking them.

and events. In method mode, run data appear in the Method Editor phase and step views. In both modes, the Run Queue pane lists runs that are waiting to start.

, annotate the chromatogram, and delete

ou can view or hide

esents a time-stamped record of run steps

30 | NGC Chromatography Systems and ChromLab Software

System Control Window

LEGEND

8 The Fluidic Scheme pane depicts graphically how modules are configured and

plumbed for an experiment.

File Menu Commands

Connect to System — connects your computer to the NGC system if it is not

already connected. In the dialog box that appears enter the DNS name of the NGC system and click Connect.

Disconnect System — when you start Chr

omLab, you are automatically logged in to the NGC system to which your computer is connected. This command enables you to disconnect ChromLab software from that system so you can work offline.

System Settings — opens the System Settings dialog box in which you can customize system settings. See System Settings on page 69 for more information about customizing your system.

System Information — opens the System Information dialog box, which lists the serial number and other general information about the NGC device as well as information about the system components, pr

ocesses, and UV and UV/Vis

detectors. See System Information on page 86 for more information.

Preferences — opens dialog boxes in which you can do the following:

 Select pressure units for all system and software pressure values. This is a

global setting.

 Set up an SMTP server to send email messages about system notifications

from the ChromLab computer. See Preferences on page 89 for more information about setting up an SMTP server.

Exit — closes Chr

omLab.

User Guide | 3 1

3 | System Control

View Menu Commands

Show Chromatogram — displays a chromatogram of the current run data. Clearing

this command hides the chromatogram from view.

Show Fluidics — displays the fluidic scheme. Clearing this command hides the fluidic scheme fr

Manual Menu Commands

Enter/Exit Manual Mode — toggles ChromLab between manual and automatic

modes.

Save Recorded Manual Run — in manual mode, saves in a data file the steps executed during a manual run.

om view.

Clear Recorded Data — deletes manual run data fr

om the display.

Tools Menu Commands

Calibrate — opens the Calibration dialog box, which displays instructions and

settings for selecting a module and calibrating it. See Calibrating a Module on

page 64 for details.

Point-to-Plumb — starts the Point-to-Plumb featur instrument LED lights so you can visually verify port locations during instrument plumbing. Displays the current fluidic scheme. Gray lines indicate the flow path. Clicking a line in the window turns on LED lights on the instrument corresponding to ports to be connected. See Verifying Plumbing with the Point-to-Plumb Feature on

page 67 for details.

Change Fluidic Scheme — opens the Fluidic Scheme Selector dialog box in which you can edit the fluidic scheme or choose another one. See Fluidic Scheme

Configurations on page 51 for details.

e and simultaneously turns off

32 | NGC Chromatography Systems and ChromLab Software

System Control Window

Map Fluidic Scheme — opens the Fluidic Scheme Mapping dialog box, which displays the location of two or more valves of the same type in the fluidic scheme, for instance two or more inlet valves or column-switching valves. You can use this dialog box to map the device on your instrument to its position in the fluidic scheme. See Fluidic Scheme Mapping on page 60 for details.

Help Menu Commands

Help — displays screen-level help topics and links to installed manuals.

Export Diagnostic Logs — opens the Export Diagnostic Logs dialog box in which

echnical Support requires to

About — displays version and copyright information about Chr

omLab software.

Toolbar Commands

Save — saves in a data file steps executed during a manual run.

Clear — deletes manual run data fr

Autoscale — automatically scales the chr

peak height during the run. Autoscaling is enabled by default.

Reset Zoom — r

Annotate — adds a note to the chr

or after the run completes.

esets the view to show the full chromatogram.

om the display.

omatogram’s primary y-axis to the tallest

omatogram at points on the x-axis during a run

Context Menu Commands

To access context menu commands

 Right-click in the chromatogram and choose a command from the menu that

appears.

Undo Zoom — r

Reset Zoom — r

estores immediately previous zoom level.

esets the view to show the full chromatogram.

User Guide | 3 3

3 | System Control

Autoscale UV Trace — automatically scales the primary y-axis to the tallest peak height during the run. While enabled, UV scale in the legend cannot be manually set.

Copy Chromatogram — copies the chr

omatogram to the clipboard so you can

paste it into another application.

Save Chromatogram As —saves the chr

omatogram in an image format you

choose (.bmp, .gif, .jpeg, .png, or .tiff).

Export as .csv — exports run data as a .csv file, which can be opened in spr

eadsheet applications.

34 | NGC Chromatography Systems and ChromLab Software

Chromatogram View

A chromatogram shows the real-time acquisition of data during the run as well as its outcome. It also depicts the quality of the purification. The chromatogram is recorded only when pumps are running and data are being acquired by the detectors in the fluidic scheme.

The chromatogram legend matches each colored trace to its detector type. When you expand the legend, you can hide traces by clearing the checkbox beside the trace, edit trace colors to dif a trace. See Changing Trace Colors on page 206 and Changing the Axes on

page 205 for more information.

Chromatogram View

entiate among them, and change the y-axis scale for

fer

When a run starts, the chromatogram viewer in the System Control window displays data acquired from the detectors as traces based on time, volume, or column volume. Pausing the pointer on a trace displays a tooltip with the trace x- and primary (left) y-axis values at that location. The legend also displays the wavelength value in nanometers for UV traces.

User Guide | 3 5

3 | System Control

You can expand the chromatogram or the fluidic scheme by clicking the arrow in the upper-right corner of the relevant pane. This is especially useful when viewing a chromatogram on the touch screen. Events that occur during the run, such as valve changes or changes in certain parameters, appear as event markers. Error events such as overpressure appear in red. The view also shows when fractions are collected and their locations in the selected rack.

The chromatogram has two y axes. The left axis is the primary axis. Its default trace is UV absorbance. Conductivity is the default trace for the right axis. The chromatogram also shows the following traces, which are defined in Table 1:

 Single/Multiple UV

 Conductivity

 pH

 Temperature

 Gradient

 Pressure

 Precolumn pressure

 Delta-column pressure

 Flow Rate

36 | NGC Chromatography Systems and ChromLab Software

Chromatogram View

Table 1. Trace Definitions

Trace Module Explanation

UV  1–4 Single-Wavelength UV

and Multi-Wavelength UV/Vis Detectors

Conductivity Single-Wavelength UV

and Multi-W

avelength

UV — with Single-Wavelength UV module, a reading of absorbance

MV — with Multi-Wavelength UV/Vis module, up to four r

eadings ( 1–4)

The conductivity of the fluid, read by the conductivity monitor (mS/cm)

UV/Vis Detectors

%B Theoretical System Pump Programmed by the user in the method or in the

System Pump dialog box

 Gradient, system pump: % of pump B

 Gradient, blending valve: % of valve Q4

 Buffer blending: % of valve Q4

pH Theoretical pH Theoretical pH, as programmed in the method or

System Pump dialog box. A

vailable only with the

buffer blending valve in buffer blending mode

pH pH The measure of pH read from the pH probe,

System Pr

essure

available only when a pH valve is pr

System Pump Measured at the mixer (psi or MPa)

esent

PreCol Pressure Column Switching Valve Available when at least one column switching

valve is pr

Tip: When two or mor

esent

e column switching valves are present, the traces appear in different shades of green on the chromatogram. The trace number corresponds to the valve number in the fluidic scheme. For example, PreCol Pressure 1 in the legend and the chromatogram corresponds to C1 in the fluidic scheme.

User Guide | 3 7

3 | System Control

Table 1. Trace Definitions, continued

Trace Module Explanation

Delta Col Pressure

Column Switching Valve Available when at least one column switching

valve is present

Tip: When two or mor

e column switching valves are present, the traces appear in different shades of blue on the chromatogram. The trace number corresponds to the valve number in the fluidic scheme. For example,

Col Pressure 1 in the

legend and the chromatogram corresponds to C1 in the fluidic scheme.

Sample Pump Pr

essure

Temperature Single-Wavelength UV

Sample Pump Sample pump pressure (psi or MPa)

The temperature of the fluid, read at the Single-

and Multi-W UV/Vis Detectors

avelength

Wavelength UV or Multi-Wavelength UV/Vis module

Flow Rate System Pump Current system flow rate (ml/min)

Sample Pump

Sample Pump Sample pump flow rate (ml/min)

Flow Rate

Changing Chromatogram View Settings

You can change trace display attributes and other run view settings in the System Control window. Changes you make to the following settings are saved and used in the display of subsequent runs:

 Trace Show/Hide state

 Trace color

 Trace y-scale range (Min/Max y values)

 Active x-scale units

 Show/Hide state of events, fractions, errors, and annotations

38 | NGC Chromatography Systems and ChromLab Software

Chromatogram View

When a manual or method run is saved, the new settings are saved with it. When you open the run in the Evaluation window, it displays the new settings. Some chromatogram view settings that are changed on the computer automatically change in the touch screen chromatogram view and vice versa.

The following display attributes are synchronized:

 Trace Show/Hide state

 Trace color

 Trace y-scale range (Min/Max y values)

 Autoscale (on/off)

 Active x-scale units

 Show/Hide state of events, fractions, errors, and annotations

 Active y-scale (selected trace y-scale)

Showing or Hiding Traces

To show or hide a trace

1. Expand the chromatogram legend.

2. Select or clear the trace entry to turn the trace on or off.

Tip: See Trace Settings Tab on page 74 to show all hidden traces.

User Guide | 3 9

3 | System Control

Changing Trace Color

To change trace color

1. Expand the legend and click the trace color in the Edit column.

2. In the Color dialog box, select a color and click OK.

Tip: See Trace Settings Tab on page 74 to revert trace colors to their factory default settings.

Zooming In and Out

To zoom in on a section of the chromatogram

 While clicking in the chromatogram, drag the pointer to mark the zoom region

of interest.

To zoom out to the previous zoom level

 Double-click the chromatogram or right-click the chromatogram and choose

Undo Zoom in the menu that appears.

To zoom out to the full-scale view

 Click Reset Zoom on the tab toolbar or right-click the chromatogram and

choose Reset Zoom in the menu that appears.

40 | NGC Chromatography Systems and ChromLab Software

Chromatogram View

Changing the X-Axis Units and Scale

For runs performed from saved methods, the x-axis unit can be changed to Time (min), Volume (ml), or Column Volume (CV). For runs performed manually, the x-axis unit can be changed to Time (min) or Volume (ml).

To change the x-axis units and scale

 Click the x-axis title to toggle among the available options.

Changing the Y-Scale Values

You can change the maximum and minimum y-scale values in the legend to set the chromatogram scale.

To change the y-scale values

1. Expand the legend in the chromatogram view.

2. Enter maximum and minimum values in the appropriate trace row, ensuring that the maximum value always exceeds the minimum value.

Tip: See Trace Settings Tab on page 74 to revert the y-scale values to their factory default settings.

Autoscaling the UV Trace

Autoscaling the UV trace scales the chromatogram’s primary UV y-axis based on signal intensity. Autoscaling is enabled by default.

When multiple UV absorbance traces are present you can

 Autoscale each UV trace independently .

The y-axis scale of each trace is scaled to the UV signal intensity of the individual trace.

 Autoscale all UV traces to the same scale .

The y-axis scale is based on the signal intensity of the UV trace with the highest intensity and all other traces are normalized to the intensity of that trace.

 Disable autoscaling .

User Guide | 4 1

3 | System Control

When a single UV absorbance trace is monitored with a single-wave UV you can

 Autoscale all UV traces to the same scale .

The y-axis scale is based on the signal intensity of the UV trace with the highest intensity and all other traces are normalized to the intensity of that trace.

 Disable autoscaling .

To autoscale the UV trace

 Do one of the following:

 Click Autoscale on the toolbar to choose a status.

 Right-click the chromatogram, choose Autoscale UV trace, and select

Annotating the Chromatogram

You can add notes to the chromatogram in manual mode at any time during a run or after a run completes to associate observations with data points on the chromatogram. The annotation dialog box contains three fields: Location (in time, volume, or CV), Title, and Description. When the annotation is saved, its title appears at the specified location on the x-axis. The description appears in the run report.

a status.

You can add annotations when a method run is in progress. However, if the run ends while the annotation dialog box is open, the annotation is saved and the dialog box closes automatically to the chr

omatogram in System Control. You can annotate completed runs by

. When a method run is complete, annotations cannot be added

opening them in Evaluation mode.

42 | NGC Chromatography Systems and ChromLab Software

Chromatogram View

To add annotations

1. Click Annotate on the toolbar and drag the icon onto the chromatogram. The green annotation dialog box opens with the Location field automatically filled.

Tip: You can edit the Location field if necessary.

2. Type a title for the annotation.

3. (Optional) Type a description for the annotation.

4. Click Save to save the annotation.

Note: Clicking

closes the dialog box without saving the annotation.

To edit an annotation

1. Double-click the annotation on the chromatogram to open its dialog box.

2. Edit the annotation and click Save to save the changes.

To delete an annotation

 Double-click the annotation to open its dialog box and click Delete.

User Guide | 4 3

3 | System Control

Showing and Hiding Events, Fractions, Errors, and Annotations

You can choose whether to show or hide fractions, events, errors, and annotations by selecting or clearing the appropriate checkboxes at the bottom of the legend.

44 | NGC Chromatography Systems and ChromLab Software

Chromatogram View

When you pause the pointer on a trace, a tooltip displays the real-time reading at the current location in the chromatogram.

User Guide | 4 5

3 | System Control

Fluidic Scheme Pane

The Fluidic Scheme pane graphically displays each module currently in use on the instrument. Real-time status information includes buffer flow rate, sample flow rate, pressure, and valve position through the system. The fluidic scheme reflects how the instrument is plumbed and the flow path through the various modules on the system.

Important: The selected fluidic scheme must match the system’

har

dware modules.

The fluidic scheme shows the real-time flow path (bright green) through the system and, in manual mode, pr scheme components excluded fr

ovides access to device settings. The path to fluidic

om the flow path appears in grey.

s installed

46 | NGC Chromatography Systems and ChromLab Software

Fluidic Scheme Pane

When pumps are not running, the predicted flow path appears in a lighter color corresponding to the pump flow path (light blue for the sample pump or light green for the system pump).

On the NGC Discover system, two flow paths are displayed — one for the system pump (bright green) and another for the sample pump (light blue). In the following screen, the gray path from the column switching valve to the column indicates that the column switching valve is not inline with a pump or injection syringe but is in bypass.

User Guide | 4 7

3 | System Control

The fluidic scheme displays the number of a valve’s active port in bright green. A grey number on a valve indicates that it is not in use. The NGC instrument supports up to four inlet valves (two buffer inlet and two sample inlet valves), three column-switching valves, and two outlet valves. Pausing on a valve displays a tooltip indicating its type.

Tip: If the system includes two sample inlet valves, the fluidic scheme identifies the first sample inlet valve as S1 and displays it as the lower of the two. The second is identified as S2. If the system includes multiple column switching valves or outlet valves, the valves ar ascending or

e identified numerically in

der.

48 | NGC Chromatography Systems and ChromLab Software

Fluidic Scheme Pane

Modules

Fluidic scheme modules are more fully described in Chapter 2, Components, of the Instrument Guide. For ease of reference, Table 2 depicts each module that can appear in a fluidic scheme and describes corresponding dialog box options.

Table 2. Fluidic Scheme Modules

System Pumps and Mixer

This module dialog box controls the NGC system’s two gradient pumps and shows the gradient status. It controls flow rate, gradient type (salt or pH), gradient duration, and system pressure limits.

System Pumps with Buffer Blending Valve

This module dialog box controls the NGC system gradient pumps when connected to a buffer blending module and shows the pump and gradient status. It controls flow rate, buffer blending valve priming, buffer recipe, gradient composition, pH, gradient duration, and system pressure limits.

System Pumps with Buffer Blending Valve and Buffer Inlet A and B

This module dialog box controls the NGC system gradient pumps when connected to inlet valves and a buffer blending valve. It controls gradient mode (two-pump gradient or buffer blending valve), flow rate, buffer blending valve priming, buffer recipe selection, gradient composition, pH, gradient duration, and system pressure limits.

Sample Inject Valve

This module dialog box controls the NGC sample inject valve. It is used to route fluid from the gradient pump, sample pump, and injection port to the loop, column, and waste as required during an experiment.

User Guide | 4 9

3 | System Control

Table 2. Fluidic Scheme Modules, continued

Sample Pump

This module dialog box controls the sample pump and displays the sample pump status. It is used to automatically load samples into a sample loop or to directly inject samples onto a column during an experiment. It is used primarily for large volume samples.

Column Switching Valve

This module dialog box controls the column switching valve and displays the valve status, the precolumn pressure, and the pressure drop across the column ( the five columns that can be connected to it, bypass the columns, or reverse the flow through the columns.

Col). It is used to select one of

UV/Conductivity Monitors

This module dialog box controls the single-wavelength UV or multi-wavelength UV/Vis monitor and displays the UV, temperature, and conductivity monitor status. It is used to turn on the UV monitor and to set the wavelength that will be used for monitoring. It is also used to zero the baseline during a run. The single-wavelength monitor can monitor one wavelength at a time (255 or 280 nm). The multi-wavelength UV/Vis monitor can monitor up to four wavelengths simultaneously (190–800 nm).

Signal Import Module

This module dialog box controls the signal import module (SIM) and displays the output of the attached external detectors. Up to two external devices can be attached to the SIM. This module displays the output from both devices or output from either SIM1 or SIM2 if one device is turned off.

50 | NGC Chromatography Systems and ChromLab Software

Fluidic Scheme Pane

Table 2. Fluidic Scheme Modules, continued

pH Monitor and Valve

This module dialog box controls the pH monitor and displays the pH. It is used to place the pH flow cell inline or to bypass it during an experiment. It also enables you to calibrate the pH probe without taking it offline.

Fraction Collector

This module dialog box controls fraction collection. It is used to select racks and rack location and to start and stop fraction collection. It is also used to determine fraction size and the number of tubes to collect.

Fluidic Scheme Configurations

The fluidic scheme must be customized to match your instrument hardware setup. In manual mode, you can access the Fluidic Scheme Selector dialog box to change the detailed settings of most elements. You can also create custom fluidic schemes. See To create a new fluidic scheme on page 58.

Table 3. Fluidic Scheme Configuration Options

Pumps Configuration Explanation

System pump only Used to start and stop pumps, change

buffer, gradient, and duration, and to set pressure limits.

System pump and inlet A

Select from up to eight different buffers for A.

User Guide | 5 1

3 | System Control

Table 3. Fluidic Scheme Configuration Options, continued

System pump and inlet B

Select from up to eight different buffers for B.

System pump and inlet A and B

System pump and buffer blending valve

System pump and buffer blending valve with inlets A and B

Select from eight buffers each for A and B (16 total).

Start and stop buffer blending valve gradient formation and perform buffer blending.

Start and stop buffer blending valve gradient formation and perform buffer blending. Select from up to seven additional buffers and solutions.

Sample Inject Configuration Explanation

Sample inject valve

Load a specific predetermined sample volume onto a column.

52 | NGC Chromatography Systems and ChromLab Software

Table 3. Fluidic Scheme Configuration Options, continued

Sample inject valve with sample pump

Automatically load a specific predetermined sample volume onto a sample loop, or directly onto the column, using the sample pump.

Fluidic Scheme Pane

Sample inject valve with autosampler

Sample inject valve with sample pump and a single sample inlet valve

Sample inject valve with two sample inlet valves

Automatically load samples onto a column using an autosampler.

For use as a mini autosampler, can select from eight different samples.

For use as a mini autosampler, can select from 15 different samples when two sample inlet valves are daisychained together.

User Guide | 5 3

3 | System Control

Table 3. Fluidic Scheme Configuration Options, continued

Columns Configuration Explanation

Column Self-explanatory when column

switching valve is absent.

Column with column switching valve

Two column switching valves, each with a column attached

Three column switching valves, each with a column attached

Used to select from up to five different columns, bypass the columns, or reverse the flow through the columns.

Used to select from up to 10 different columns, bypass the columns, or reverse the flow through the columns.

Used to select from up to 15 different columns, bypass the columns, or reverse the flow through the columns.

Detectors Configuration Explanation

Single-wavelength UV with conductivity

For selection of a single-wavelength UV detector with conductivity and temperature.

54 | NGC Chromatography Systems and ChromLab Software

Table 3. Fluidic Scheme Configuration Options, continued

Single-wavelength UV with conductivity and

For selection of a single-wavelength UV detector with conductivity, temperature, and pH valve.

pH probe

Fluidic Scheme Pane

Multi-wavelength UV/Vis detector with conductivity and temperature

Multi-wavelength UV/Vis detector with conductivity, temperature, and pH probe

Signal import module (SIM) with single-wavelength UV detector with conductivity and temperature

SIM with single-wavelength UV detector with conductivity, temperature, and pH probe

SIM with multi-wavelength UV/Vis detector with conductivity

For selection of a multi-wavelength UV/Vis detector with conductivity and temperature.

For selection of a multi-wavelength UV/Vis detector with conductivity, temperature, and pH valve.

For selection of the SIM with a single-wavelength UV detector with conductivity and temperatur

For selection of the SIM with a single-wavelength UV detector with conductivity

, temperature, and pH

valve.

For selection of the SIM with a multi-wavelength UV/Vis detector with conductivity and temperature.

SIM with multi-wavelength UV/Vis detector with conductivity and pH probe

For selection of the SIM with a multi-wavelength UV/Vis detector with conductivity, temperature, and pH valve.

User Guide | 5 5

3 | System Control

Table 3. Fluidic Scheme Configuration Options, continued

Fraction Collection Configuration Explanation

BioFrac™ fraction collector

For configuring a BioFrac fraction collector. Used for starting and stopping fraction collection to determine fraction size and number of tubes to collect.

Fraction collector with outlet valve

For collecting a combination of large and small volume fractions. Select outlet ports can be plumbed to divert a fraction eluted from a column onto a sample/storage loop to be subsequently reinjected onto a second column during tandem applications.

Fraction collector with two outlet valves

Outlet valve For use as a fraction collector, can

collect up to 11 large-volume fractions.

Select outlet ports can be plumbed to divert a fraction eluted from a column onto a sample/storage loop to be subsequently r

einjected onto a second

column during tandem applications.

56 | NGC Chromatography Systems and ChromLab Software

Table 3. Fluidic Scheme Configuration Options, continued

Two outlet valves For use as a fraction collector, can

collect up to 22 large-volume fractions when two outlet valves are daisychained together.

Select outlet ports can be plumbed to divert a fraction eluted fr onto a sample/storage loop to be subsequently reinjected onto a second column during tandem applications.

Working with Fluidic Schemes

Five sections of the fluidic scheme can be configured based on the available modules and experiment requirements.

 Pumps

 Sample inject

 Columns

 Detectors

Fluidic Scheme Pane

om a column

 Sample collection

To change a fluidic scheme

1. Do one of the following:

 In the System Control window, select Tools > Change Fluidic Scheme.

 On the touch screen, choose Change Fluidic Scheme on the dropdown

menu.

The Fluidic Scheme Selector dialog box appears. A descriptive tooltip appears when you pause the pointer on a module in the fluidic scheme.

User Guide | 5 7

3 | System Control

2. Click an entry in the Fluidic Scheme Selector pane.

The right pane displays the fluidic scheme you selected.

3. Click Select.

To create a new fluidic scheme

Note: You cannot save a duplicate fluidic scheme.

1. Do one of the following:

 In the System Control window, select Tools > Change Fluidic Scheme.

 On the touch screen, choose Change Fluidic Scheme on the dropdown

menu.

2. Click New.

The current fluidic scheme appears with Edit buttons that enable you to select alter

native modules in spin boxes.

58 | NGC Chromatography Systems and ChromLab Software

3. Click Edit under the group of modules to change.

A spin box displays a number of modules of the type you selected.

4. Scroll up or down to view module choices.

Fluidic Scheme Pane

5. Select a module and click Accept.

6. Repeat steps 3–5 to edit other modules.

7. When you have finished editing modules, click Save and type a name in the New Fluidic Scheme dialog box that appears.

8. Click OK.

To change module settings

1. In the fluidic scheme, double-click a module.

2. In the dialog box that appears, edit module options.

3. Click Apply.

User Guide | 5 9

3 | System Control

Fluidic Scheme Mapping

The NGC instrument supports up to four inlet valves (two buffer and two sample inlet valves), three column-switching valves, and two outlet valves. When more than one of a specific valve type is installed, ChromLab automatically detects the valves and maps the fluidic scheme to their default positions on the NGC instrument. For example, when two or more inlet valves are present in the fluidic scheme, ChromLab automatically detects the position of the valves on the NGC instrument (starting from the left side of the bottom tier) and assigns the first valve Inlet A.

If the configuration of your NGC instrument is different than the default, or if you changed your fluidic scheme, you must manually map the valves on the instrument to their locations on the fluidic scheme. This is done using the Fluidic Scheme Mapping dialog box.

Note: If you have only two inlet valves on your NGC instrument and both valves ar system automatically determines their locations and function on the instrument.

The Fluidic Scheme Mapping dialog box displays the current fluidic scheme in the upper pane and the curr ChromLab automatically detects which valves in the fluidic scheme can be mapped. These valves appear color-coded in the upper pane. The corresponding valves appear in the same color code on the instrument map in the lower pane, in relative location to the mappable modules on the NGC instrument. All other modules appear inactive in the lower pane.

e designated as buffer inlets, you do not need to map them. The

ent NGC instrument configuration in the lower pane.

60 | NGC Chromatography Systems and ChromLab Software

Fluidic Scheme Mapping

The color and labeling scheme for the Fluidic Scheme Mapping dialog box is as follows:

Color Label valve

Blue A and B Buffer inlet valve

Blue S1 and S2 Sample inlet valve

Green C1–C3 Column switching valve

Orange O1 and O2 Outlet valve

To map a fluidic scheme to the instrument, you first unmap the current settings and then map the valves to the new fluidic scheme.

User Guide | 6 1

3 | System Control

To unmap valves

1. Do one of the following:

 In the System Control window, select Tools > Map Fluidic Scheme.

 On the touch screen, choose Map Fluidic Scheme on the dropdown menu.

The Fluidic Scheme Mapping dialog box appears with the valves on the fluidic scheme mapped to their default positions on the instrument map.

2. Select a valve on the instrument map. The border of the valve on the fluidic scheme becomes highlighted, indicating that it is selected.

3. Click Unmap. The border of the valve on the instrument map thins and its label disappears, indicating that the valve is no longer mapped to the fluidic scheme.

4. (Optional) Continue unmapping all valves that need to be remapped.

5. To map the valves, proceed to To map valves on page 63.

To cancel unmapping valves

 Click Cancel to cancel unmapping the valves. The fluidic scheme and

instrument map return to the previous mapping configuration.

62 | NGC Chromatography Systems and ChromLab Software

Fluidic Scheme Mapping

To map valves

1. In the Fluidic Scheme Mapping dialog box, select a valve to map in either the upper or lower pane.

2. Select its position on the corresponding scheme. In the instrument map, a label appears on the selected valve and its bor

der thickens, indicating that it is

mapped.

3. (Optional) Continue mapping the unmapped valves.

4. Click Save to save the new mapping.

User Guide | 6 3

3 | System Control

Calibrations

The NGC instrument arrives factory calibrated. For the most part, the instrument will not require further calibration. The pH probe, however, should be recalibrated each day the instrument is used and when there are large fluctuations in temperature because pH probe settings drift with time.

In addition to the pH probe, ChromLab includes settings for calibrating the following:

 Pump flow rate

 Sample pump pressure

 System pressure

 Column switching valve pressure

 Conductivity monitor

Calibrating these modules is optional.

Calibrating a Module

Note: You cannot run a method during calibration.

On the Tools menu, clicking Calibrate displays a Calibration dialog box in which you can select a module to calibrate. The System Contr title Calibration in Progress in the title bar.

To select a module to calibrate

 Select a module in the Calibrate dropdown list at the top of the Calibration

dialog box.

Instructions for calibrating the module you selected appear in the dialog box.

64 | NGC Chromatography Systems and ChromLab Software

ol window also appears with the

Calibrations

Calibrating Flow Rate of Pumps

If the buffer flow seems to be incorrect, you might want to recalibrate the pumps.

Important: The system must be primed with water befor

e you calibrate the

pumps’ flow rate. Otherwise the calibration will be incorrect.

To calibrate the flow rate of the pumps

 Select Pump Flow Rate in the Calibrate dropdown list at the top of the

Calibration dialog box.

The Calibration dialog box displays instructions and settings for calibrating the pumps.

User Guide | 6 5

3 | System Control

Calibrating Pressure Settings

WARNING! You must remove the tubing from the pressure sensor before you

calibrate it. The sensor must be at atmospheric pressure when you start.

You can calibrate sample pump pressure, system pressure, and column switching valve pr pressure values to zero.

To calibrate a pressure setting

1. Select a pressure module in the Calibrate dropdown list at the top of the Calibration dialog box.

2. Follow the dialog box instructions to reset the pressure setting to zero.

Note: To calibrate pressures at the upper limit, call Bio-Rad Support.

essure to zero. If pressure is not reading zero or close to it, you can reset the

66 | NGC Chromatography Systems and ChromLab Software

Verifying Plumbing with the Point-to-Plumb Feature

The NGC chromatography system instrument arrives plumbed. The Point-to-Plumb feature enables you to change the plumbing or to verify that the instrument is plumbed correctly for the fluidic scheme you want to use.

You can access the Point-to-Plumb feature on the touch screen menu or on the computer running Chr

Tip: Using the touch screen makes it easy to view the Point-to-Plumb window and the fr

ont of the instrument at the same time.

omLab. (Select T

ools > Point-to-Plumb.)

When you select Point-to-Plumb, the curr Point-to-Plumb mode. All LEDs on the instrument tur between two modules, the LEDs corresponding to the two ports to be connected light up.

In the Point-to-Plumb window, the proper plumbing path is indicated by the color gray. Selected flow paths turn green. Path elements that are not plumbed appear in a paler shade of gray and do not change color when you select them.

Note: When the fluidic scheme includes a signal import module (SIM), the SIM is also pr module that connects to the external detector (via SIM) turns on when the fluid

esent in the Point-to-Plumb window

ent fluidic scheme appears in

f. When you select a line

n of

. In this case, the outlet LED on the

User Guide | 6 7

3 | System Control

path between them is selected. If the fluidic path includes an NGC module after the external detector, the inlet LED on that module turns on.

To verify the plumbing path

1. With both the Point-to-Plumb window and the front of the instrument in view, click a segment of the gray path in the Point-to-Plumb window.

The segment you selected turns green to indicate the correct plumbing path.

At the same time, green LED lights flash on the front of the instrument, indicating the ports to be connected to each other

2. Check the instrument plumbing and the Point-to-Plumb display to verify that the selected segment of the plumbing is corr

3. Continue selecting segments of the plumbing path and verifying that they are corr

ect.

4. When you have finished verifying the plumbing path, click Exit Point-to-Plumb.

The LEDs on the instrument turn on. Depending on the module, LEDs indicate

 Flow from the system pumps (green LEDs)

 Flow from the sample pump (blue LEDs)

68 | NGC Chromatography Systems and ChromLab Software

ect. If it is not correct, replumb it.

System Settings

The System Settings dialog box enables you to customize your system. From this dialog box you can

 Set the delay volume

 Control the flow rate to prevent overpressure

 Enable remote access to ChromLab from an iPad, Android device, or

another PC

 Reset the trace settings, the minimum and maximum y-scale values, and

the trace colors in the chromatogram to their factory default settings

 Enable or disable external detectors that are connected to the NGC

instrument via a SIM device

 Enable or disable the ability to send UV signals to external devices that are

connected to the NGC instrument via a SIM device

 Set air sensors to detect either end of sample or end of buffer

System Settings

 Enable the system to send email messages about system events to a list of

users

 Set a unique name for the NGC system

Note: This dialog box is accessible in manual mode. The settings are saved and the fraction collector is also synchr

onized in method runs.

Delay Volume Tab

Delay volume is the volume of plumbing between the UV detector and the fraction collector drophead and/or the outlet valve. It indicates the delay between the detection of the fraction’s UV peak and the time it takes for this peak to traverse the volume of tubing and flow cells to the fraction tube into which it is collected.

User Guide | 6 9

3 | System Control

When the delay volume is set, the fraction collector or outlet valves wait for the fraction peak to travel from the detector to the drophead (or port) before collection begins. Fraction collection on the instrument can be delayed relative to the collection information shown in the chromatogram and status panels. When the run reaches the end of rack or the last port on the outlet valve, the chromatogram and status panel might display the start tube and rack or new outlet port before collection actually starts.

Note: If an outlet valve is included in the fluidic scheme and you change the collection port or fraction size during the run, the first container in the new scheme collects the designated fraction size as well as the indicated delay volume. All succeeding fraction containers collect only the designated fraction size.

To set the delay volume

1. Select File > System Settings to open the System Settings dialog box.

2. Choose the Delay Volume tab.

70 | NGC Chromatography Systems and ChromLab Software

System Settings

3. Select Synchronize with detector.

4. Specify the inner diameter of the tubing in use.

Tubing Color Inner Diameter

Orange 0.02" (0.5 mm)

Green 0.03" (0.75 mm)

Clear 0.062" (1.6 mm)

5. If your system includes the BioFrac fraction collector, select the checkbox Include BioFrac – 12 l and specify the tubing length between the UV outlet port and the fraction collector drophead.

6. If your system includes an outlet valve, select the checkbox Include Outlet valve – 50 l and specify the tubing length between the UV outlet port and the outlet valve.

User Guide | 7 1

3 | System Control

7. If your system includes both the BioFrac fraction collector and an outlet valve, select both checkboxes. In the Include Outlet valve – 50 l section, specify the tubing length between the UV outlet port and the outlet valve as well as the tubing length between the outlet valve and the fraction collector drophead.

8. Select or clear the next four checkboxes, depending on whether these modules ar

Note: If a pH detector module is present on your system and you select the checkbox to include its cell volume, determine whether the detector is set to Inline or Bypass mode and select the appr

9. If one or more of the options is not applicable, or if you change the length of the tubing, enter the appr

The system calculates the delay volume based on your specifications and displays the r

10. Click OK.

Control Flow Tab

Use the option Control Flow to avoid overpressure to prevent the system from exceeding the maximum pressure limit. Control Flow reduces the flow rate by half whenever the system pressure gets within 80% (the default) of the maximum system pressure limit. Control Flow lowers the flow rate multiple times if necessary. However, it will not reduce the flow rate below the minimum user-specified flow rate. For methods, Control Flow resets the flow rate at the start of each phase and then readjusts it if necessary. The pumps are immediately stopped if the pressure exceeds the maximum pressure limit at any time.

e present on your system.

opriate radio button.

opriate values in the Additional volume box.

esults in l at the bottom of the dialog box.

The option Control Flow to avoid overpressure is available in the System Pump and Sample Pump dialog boxes in manual mode as well as in the Method Editor window a method to control the flow rate within the pressure limit range using settings in the Control Flow tab of the System Settings dialog box.

72 | NGC Chromatography Systems and ChromLab Software

. You can choose either of these settings before performing a run or running

System Settings

When Control Flow to avoid overpressure is selected, the system lowers the flow rate to avoid exceeding the maximum pressure. Typically, Control Flow activates when the pressure reaches 80% of the maximum set pressure from the column. This limit can be changed. The flow rate is lowered to half the set flow rate. Flow rate returns to normal when the phase changes and drops only if the pressure in the next phase continues to exceed the upper pressure limit of the column.

Note: If the pressure increases rapidly and exceeds the specified limits, or the flow rate dr

ops below the specified flow rate, the system shuts down.

To control the flow rate

1. Select File > System Settings to open the System Settings dialog box.

2. Choose the Control Flow tab.

3. Specify a percentage of maximum pressure at which you want Control Flow to activate (the default is 80%).

The system reduces the flow rate by 50% when pressure reaches the specified per

centage of the maximum pressure.

User Guide | 7 3

3 | System Control

4. (Optional) Specify a minimum flow rate for the system pump and for the sample pump if the sample pump is available.

Tip: When you change the pump heads on the system pumps, ChromLab softwar setting in the System Settings dialog box Control Flow tab is compatible with the range that the new pump heads allow. If it is not, the flow rate is set to the minimum that the new pump heads allow.

Remote Access Tab

You can access ChromLab software remotely with an iPad, Android device, or another personal computer utilizing a virtual network computing (VNC) service.

To enable remote access to ChromLab

1. Select File > System Settings to open the System Settings dialog box.

2. Choose the Remote Access tab.

3. Select Enable VNC service for remote screen sharing.

Trace Settings Tab

e checks at instrument startup whether the most recently used flow rate

ChromLab saves changes to trace display settings and applies them to subsequent manual or method runs. You can revert the following display settings to their factory defaults:

 Trace Show/Hide state

 Trace y-scale range (Min/Max y values)

 Trace color

Note: When you revert a display setting to its factory default, the change applies to the curr does not affect previously saved or evaluated runs.

74 | NGC Chromatography Systems and ChromLab Software

ent and all subsequent manual or method runs. The change

System Settings

To revert trace display settings to their factory defaults

1. Select File > System Settings to open the System Settings dialog box.

2. Choose the Trace Settings tab.

3. Do one or more of the following:

 To show all traces in the System Control legend, click Show All.

 To reset the y-scale values to their factory default settings, click Reset.

 To reset all trace colors to their factory default settings, click Reset.

4. Click OK.

User Guide | 7 5

3 | System Control

Device Input Tab

You can import analog (voltage) signal from up to two external devices and convert it to digital NGC data via the signal import module (SIM).The SIM imports and converts the signal to the appropriate trace units through its SIM channels. When enabled, the details appear in the chromatogram’s legend, the trace appears on the chromatogram, and the data can be analyzed in the Evaluation window. For each device you can define its

 Trace name

 Minimum and maximum output range (in volts)

 Trace unit type (displayed in the chromatogram’s legend)

 Minimum and maximum unit range

To enable trace measurement from external detectors

1. Ensure that the SIM is connected to the NGC instrument.

2. On the back of the SIM, determine which external devices are connected to the SIM 1 and SIM 2 ports.

3. Select File > System Settings to open the System Settings dialog box.

76 | NGC Chromatography Systems and ChromLab Software

4. Choose the Device Input tab.

System Settings

5. By default, the fields in this dialog box are disabled. Select the Enabled checkbox for SIM 1 and provide the following details to enable connection to the first detector:

a. Type a trace name for the detector in the Trace Name field.

Note: This field has a 20-character limit.

b. Range (Volt) is the output from the detector. Set the minimum and

maximum volt range.

c. Type a unit type for the trace in the Trace Unit field.

Note: This field has a 5-character limit.

d. Scale converts the output to the trace’s units. Set the minimum and

maximum unit range (y-axis scale).

User Guide | 7 7

3 | System Control

6. (Optional) Select the Enabled checkbox for SIM 2 and repeat steps 5a—5d for a second external detector.

7. Click OK.

To disable the connection

 On the Device Input tab, clear the Enabled checkbox for the device and click

OK.

Note: Disabling the SIM connection does not turn off the detector.

Device Output Tab

You can convert digital NGC data to analog voltage via the SIM and output the voltage to an external recording device, such as a light scattering detector. The SIM converts and outputs NGC trace information as voltage through its digital-to-analog converter (DAC) channels. When enabled, the external device can receive the converted trace information. For the device you can define its

 Signal type

 Minimum and maximum output range (from 0–10 volts)

 Minimum and maximum unit range

To enable output to external devices

1. Ensure that the SIM is connected to the NGC instrument.

2. On the back of the SIM, ensure that the device is connected to DAC A.

3. Select File > System Settings to open the System Settings dialog box.

4. Choose the Device Output tab.

78 | NGC Chromatography Systems and ChromLab Software

System Settings

5. By default, the fields in this dialog box are disabled. Select the Enabled checkbox for Signal Output A and provide the following details:

a. Choose a signal type from the Signal dropdown list.

b. Scale converts the trace units to the device’s input type. Set the minimum

and maximum unit range (y-axis scale).

c. Range is the output to the device. The SIM’s DAC channels output 0—10 V.

Recommended settings within this range ar

e specific to the external

device. Refer to the device’s user guide for more information.

6. Click OK.

To disable the connection

 On the Device Output tab, clear the Enabled checkbox and click OK.

Tip: Disabling the SIM connection does not turn off the external device.

User Guide | 7 9

3 | System Control

Air Sensors Tab

The system supports up to two air sensor modules and up to eight air sensors. Each air sensor module can contain up to four air sensors. The system detects the number of air sensors that are attached to the instrument.

Air sensors can be set to detect air, indicating end of buffer, in lines connected to the system pumps. When the air sensor detects end of buf system pumps and the run. You can continue the run after replenishing the buffer and purging the lines.

Air sensors can also be set to detect air, indicating end of sample, in lines connected to the sample pump. When the sample pump is used to load sample in manual mode and the air sensor detects end of sample, the system stops the sample pump to keep it fr detects end of sample during the sample application phase, the sample pump stops. In this case the run proceeds to the next phase.

To activate air sensors

1. Select File > System Settings to open the System Settings dialog box.

2. Choose the Air Sensors tab.

fer, the system stops the

om pumping air onto the column. When the air sensor

By default, only the air sensors that the system detects are enabled. The other sensors ar

80 | NGC Chromatography Systems and ChromLab Software

e disabled.

System Settings

3. To activate one or more air sensors, select its checkbox and from the dropdown list choose either Sample (to detect end of sample) or Buffer (to detect end of buffer).

4. Click OK.

To deactivate an air sensor

 On the Air Sensors tab, clear its checkbox and click OK.

User Guide | 8 1

3 | System Control

Email Notifications Tab

You can connect ChromLab to your outgoing email server to send email notifications to a list of users. When this option is enabled, ChromLab sends messages informing users of specific event types such as

 Pumps stopped

 Method run completed successfully

 Method run stopped

 Connection to the NGC system is lost

The email identifies

 Type of event

 Time of the event

 Name of the method that was running when the event occurred

 Elapsed time of the method before the event occurred

To set up the connection between ChromLab and your SMTP server see To connect

ChromLab to an email server on page 90.

To set up email notifications

1. Select File > System Settings to open the System Settings dialog box.

2. Choose the Email Notifications tab.

By default, email notification is not enabled.

82 | NGC Chromatography Systems and ChromLab Software

System Settings

3. Select Enable Email Notification.

Note: If you have not set up a valid SMTP connection for ChromLab, an error message appears informing you that Chr

omLab cannot connect to the email server. Click OK. The Email Server dialog box appears. To set up the connection between ChromLab and your SMTP server see To connect

ChromLab to an email server on page 90.

4. Below the Email Recipients box, click Add. The Add Email Address dialog box appears.

5. Type an email address in the text box and click OK.

Perform this step for each user who wants to receive email alerts from Chr

omLab. Y

ou can also enter a user group email address to email a group of

users.

User Guide | 8 3

3 | System Control

All added users receive an email from DoNotReply_NGC notifying them that they are registered to receive email about NGC system events.

6. In the Event Types section, clear the checkboxes for the events about which you do not to want to r

Note: All email recipients will receive email about all selected event types.

7. Click OK to save the changes and exit the dialog box.

To edit a name in the Email Recipients section

1. Double-click the name in the Email Recipients list. The Edit Email Address dialog box appears.

2. Modify the email address and click OK.

To remove a name from the Email Recipients list

 In the Email Recipients section, select the email recipient and click Remove.

To stop all email notifications

 On the Email Notifications tab, clear the Enable Email Notifications and click

OK.

eceive email notifications.

84 | NGC Chromatography Systems and ChromLab Software

System Settings

System Name Tab

You can specify a unique name for the NGC instrument. ChromLab uses this name to identify the system to which the computer is connected. The system name appears in the Home window when ChromLab connects to the NGC system. It also appears in the formatted run reports to identify the system on which the run was generated.

Note: The system name is limited to any combination of 10 alphanumeric and special characters.

User Guide | 8 5

3 | System Control

System Information

The System Information dialog box enables you to view general information about your system as well as information about the system components, processes, and UV and UV/Vis detectors.

Note: This dialog box is accessible in manual mode and from the menu on the touch scr

General Tab

This tab displays details about the NGC instrument, including

 Type of configuration (for example, NGC Discover 10)

 System’s serial number

 Name of your NGC system

 Name of the ChromLab computer

 Version of ChromLab software running on the system

 System’s IP address

een.

 System’s network name

 System’s available disk space

 System’s BIOS information (including the BIOS version and build date)

Devices Tab

This tab displays details about each module on the NGC instrument, including its version number and serial number. This information is useful when you need to order replacement modules.

Process Tab

This tab displays details about the processes running on the NGC instrument, as well as on the ChromLab computer and the touch screen.

86 | NGC Chromatography Systems and ChromLab Software

System Information

Detector Tab

This tab displays details about UV or UV/Vis lamp usage, from which you can determine whether the lamps need to be replaced. The system determines which detector module is installed and displays information specific to that module.

Single-Wavelength Detector

The system retrieves the lamp status for both lamps (255 nm and 280 nm) and displays the reference voltage, pulse-width modulation (PWM) percentage, and total lamp hours for each lamp on the Detector tab.

The system must have stopped running before lamp status can be determined. If the pumps ar Status, informing you that the system is unable to acquire the lamp status because the system is busy.

The system turns off the lamp if it is on when the pumps stop running. The system then tur the wavelength to 255 nm, waits for the reference signal to stabilize, and reads the reference signal data. The system then sets the wavelength to 280 nm and follows the same protocol. The values appear on the Detector tab after the data for both wavelengths have been determined.

e running, a warning message appears when you click Get Lamp

ns the lamp on, which updates the PWM percentage data. The system sets

You can use the lamp usage hours or the PWM percentage to determine how close the lamp is to its end of life. Higher per

After you change the lamps, reset the lamp hours to reflect the new lamps’ usefulness.

centage values indicate lower usefulness.

To display the signal and lamp usage time

1. Select File > System Settings to open the System Information dialog box.

2. Choose the Detector tab. The empty detector screen appears.

3. Click Get Lamp Status.

4. Click Close to close the System Information dialog box.

User Guide | 8 7

3 | System Control

To reset the lamp hours

 On the Detector tab, click Reset Lamp Time.

Note: A dialog box appears warning you that this cannot be undone. Click No if you do not want to r

Multi-Wavelength Detector

The tab displays the measured signal counts at specific wavelengths. It also displays the lamp usage time (in hours) for both the deuterium and tungsten lamps.

The system must have stopped running before lamp status can be determined. If the pumps ar Status, informing you that the system is unable to acquire the lamp status because the system is busy.

Note: The lamps must be turned on and in Standby mode. If the lamps are off, a war that the lamps must be turned on and the detector must be in Standby mode. In Manual mode, turn the lamps on and wait for the lamps to display Standby. This might take some time.

When the pumps are idle and the lamps’ signal is stable, the system sets the deuterium lamp to 240 nm and the tungsten lamp to 600 nm. After the measur lamp usage data (in hours). After you close the dialog box, the system restores the detector to the wavelength that was set before the procedure started. You can use the lamp usage hours to determine how close the lamps are to their end of life.

eset lamp hours.

e running, a warning message appears when you click Get Lamp

ning message appears when you click Get Lamp Status, informing you

ements are complete, the tab displays the measured reference counts and

To display the reference counts and lamp usage time

1. Select File > System Settings to open the System Information dialog box.

2. Choose the Detector tab. The empty detector screen appears.

3. Click Get Lamp Status.

4. Click Close to close the System Information dialog box.

88 | NGC Chromatography Systems and ChromLab Software

Preferences

The Preference dialog box enables you to select pressure units for all system and software pressure values. This is a global setting.

This dialog box also enables you to connect ChromLab to your internal email server. Chr

omLab can then send email alerts about specific system events.

Email Server Tab

Note: See your system administrator to connect ChromLab to the internal

email server.

ChromLab can connect to your internal email server and send email notifications about system events to a list of users.

The email identifies

 Type of event

 Time of the event

 Name of the method that was running when the event occurred

Preferences

 Elapsed time of the method before the event occurred

To enable email notification see To set up email notifications on page 82.

User Guide | 8 9

3 | System Control

To connect ChromLab to an email server

1. In ChromLab select File > Preferences.

2. Choose the Email Server Setup tab.

3. Provide the following information for your company:

 SMTP server name — the name of the outgoing email server at your

company.

 Port — the port number for the SMTP server. The default is 25.

 Use SSL — by default Secure Socket Layer (SSL) is disabled. If you use

SSL at your company, select this checkbox.

 From address — the name of the email server at your company.

90 | NGC Chromatography Systems and ChromLab Software

Preferences

 Authentication — by default, Authentication is disabled. If your site

requires account authentication, select this checkbox. The Authenticated Account and Password fields become active.

 Authenticated Account — the name of the authenticated account.

 Account Password — the password for the authenticated account.

4. To verify that the SMTP server settings are correct, enter a valid email address in the T

est Email Address field and click Send Test Email.

5. Click OK to close the dialog box.

User Guide | 9 1

3 | System Control

92 | NGC Chromatography Systems and ChromLab Software

4 Performing a Manual Run

The NGC™ chromatography system instrument operates in two modes: manual mode and automated, programmed method mode. In manual mode you have full control of each NGC pump, valve, detector, and fraction collector. This mode is used primarily for nonautomated processes such as priming and cleaning or purging the system, but it can also be used to load samples, optimize chromatography parameters, and run simple experiments.

Method mode enables you to execute preprogrammed steps automatically. See

Chapter 5, Method Editor, and Chapter 6, Creating a Method, for information about

working in method mode.

When the NGC system is turned on, the instrument touch screen displays the Home window in manual mode. By default the system r calibration is in progress or a method is running.

You can enter the required instrument settings by double-clicking the individual module dialog boxes in the fluidic scheme. This chapter describes these settings. See also Chapter 3, Pr Systems and ChromLab Software Instrument Guide for more detailed information on tasks described in this chapter.

eparing the Instrument, in the NGC Chromatography

emains in manual mode unless

Manual tasks can be performed in the System Control window using the computer connected to the NGC instrument or on the touch scr can be saved to the ChromLab™ database located on the computer.

Note: The NGC system can be operated in manual mode without a connected computer

een. Runs performed manually

User Guide | 9 3

4 | Performing a Manual Run

Preparing the System

System preparation consists of priming the system to remove air from the pumps, tubing, and valves followed by flushing the fluidics system to remove storage buffer, cleaning solutions, or old buffer. Priming is performed in manual mode, while both purging and cleaning tasks can be performed either manually or through preprogrammed method phases. See Chapter 5, Method Editor for more information about working with methods.

Priming and Purging the System

Perform the following procedures consecutively to prime and purge your NGC systems.

Important: Whenever you add or r

NGC system hardware configuration, you must replumb and reprime the system.

emove a module or change or upgrade your

To prime the system

1. Power on the NGC system and its connected computer.

2. To enter manual mode, do one of the following:

 On the computer, click Manual Run in the ChromLab Home window or

click the System Control tab and then click in the fluidic scheme.

 On the touch screen, click Enter Manual Mode if the application is not

already in manual mode.

3. (Optional) If your hardware configuration has changed, select a fluidic scheme that matches your curr

a. Select Tools > Change Fluidic Scheme.

b. Select the appropriate fluidic scheme in the Fluidic Scheme Selector pane.

c. Click Select to choose the manual mode fluidic scheme.

4. Insert appropriate inlet lines into the buffer, storage solution, and/or cleaning solutions to be primed and flushed thr

ent system configuration.

ough the system.

94 | NGC Chromatography Systems and ChromLab Software

Preparing the System

5. Prime the system. (See Priming and Purging the Systems in Chapter 3 of the NGC Chromatography Systems and ChromLab Software Instrument Guide for detailed instructions.)

To purge the NGC Quest system pumps

1. In the fluidic scheme, touch or click the Sample Inject valve module to open its dialog box.

2. Set the sample inject valve to System Pump Waste.

3. In the fluidic scheme, double-click the System Pump module to open its dialog box.

User Guide | 9 5

4 | Performing a Manual Run

4. In the System Pump dialog box:

a. Set Flow Rate to 10 ml/min.

b. (Optional) Set Duration to at least 1 min.

c. If selected, clear the Enable Air Sensor checkbox.

d. Click Start.

96 | NGC Chromatography Systems and ChromLab Software

Preparing the System

To purge the buffer blending valve

1. In the fluidic scheme, double-click the System Pump module to open its dialog box.

2. Select Priming in the Mode dropdown list.

 To purge an individual line of air bubbles, proceed to step 3 on page 98.

 To purge all lines of air bubbles sequentially, proceed to step 4 on page 98.

Tip: The available buffer blending valve modes are defined as follows:

Mode Description

Buffer Blending Blends stock acid, base, water, and salt solutions to form

isocratic or linear buffer gradients at a user-specified buffer concentration and pH using the Buffer Blending module.

Priming Opens the buffer blending inlet ports for priming and turns

the pumps on to flush the inlet line and pumps with the buf

fer at all four ports.

Gradient via Inlet V

alves

Buffers are selected at the inlet valve. Gradients are made by pumps A and B using the selected buffers.

User Guide | 9 7

4 | Performing a Manual Run

Mode Description

Gradient via Blending Valve (High Flow)

3. To purge an individual line of air bubbles and fill it with buffer:

a. Select a port to open.

b. (Optional) Change the default flow rate. The default flow rate is set to

20 ml/min.

c. Click Start.

d. Repeat steps 3a–c for any other port to purge.

4. To purge all lines of air bubbles and fill them with buffer:

a. Click All.

b. (Optional) Change the default flow rate. The default flow rate is set to

20 ml/min.

c. Click Start.

Buffer blending valve ports Q1 and Q4 are used as inlets for buffers A and B. Gradients are formed by the buffer blending valve rather than by the pumps. This enables each pump to run at their full flow rate capacity which doubles the total flow rate, for example to 20 ml/min for F10 pumps.

5. Monitor the pump pressure and outlet flow to ensure that the fluid is flowing pr

operly.

The system pumps and the buffer blending valve start operating at the specified flow rate, exchanging the solution in the inlet lines and expelling any trapped bubbles, cycling sequentially thr specified duration, the pumps stop momentarily while the blending valve switches to port Q3 in order to flush the lines with water before closing the valve. No change is allowed during this process for the specified duration.

Note: You can stop the cycle before it completes by clicking Stop. Before stopping the pur

ge cycle, ensure that all buffer lines are filled with the

appropriate buffers before starting a run.

98 | NGC Chromatography Systems and ChromLab Software

ough the ports Q1 to Q4. After the

Bio-Rad ChromLab Software User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual

Table of Contents

1 Introduction

Main NGC Features

NGC Chromatography Systems

Finding Out More

2 The Workspace

The Home Window

File Menu Commands

Help Menu Commands

The System Control Window

The Method Editor Window

The Evaluation Window

Instrument Control Touch Screen

Touch Screen Menu Commands

Touch Screen Toolbar Commands

3 System Control

System Control Window

File Menu Commands

View Menu Commands

Manual Menu Commands

Tools Menu Commands

Help Menu Commands

Toolbar Commands

Context Menu Commands

Chromatogram View

Changing Chromatogram View Settings

Fluidic Scheme Pane

Modules

Fluidic Scheme Configurations

Fluidic Scheme Mapping

Calibrations

Calibrating a Module

Verifying Plumbing with the Point-to-Plumb Feature

System Settings

Delay Volume Tab

Control Flow Tab

Remote Access Tab

Trace Settings Tab

Device Input Tab

Device Output Tab

Air Sensors Tab

Email Notifications Tab

System Name Tab

System Information

General Tab

Devices Tab

Process Tab

Detector Tab

Preferences

Email Server Tab

4 Performing a Manual Run

Preparing the System

Priming and Purging the System