Dionex As Operator's Manual

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AS Autosampler

Operator's Manual

Document No. 065051

Revision 03

January 2008

This publication is protected by federal copyright law. No part of this publication may be copied or distributed, transmitted, transcribed, stored in a retrieval system, or transmitted into any human or computer language, in any form or by any means, electronic, mechanical, magnetic, manual, or otherwise, or disclosed to third parties without the express written permission of Dionex Corporation, 1228 Titan Way, Sunnyvale, California 94088-3603 U.S.A.

DISCLAIMER OF WARRANTY AND LIMITED WARRANTY

THIS PUBLICATION IS PROVIDED “AS IS” WITHOUT WARRANTY OF ANY KIND. DIONEX CORPORATION DOES NOT WARRANT, GUARANTEE, OR MAKE ANY EXPRESS OR IMPLIED REPRESENTATIONS REGARDING THE USE, OR THE RESULTS OF THE USE, OF THIS PUBLICATION IN TERMS OF CORRECTNESS, ACCURACY, RELIABILITY, CURRENTNESS, OR OTHERWISE. FURTHER, DIONEX CORPORATION RESERVES THE RIGHT TO REVISE THIS PUBLICATION AND TO MAKE CHANGES FROM TIME TO TIME IN THE CONTENT HEREINOF WITHOUT OBLIGATION OF DIONEX CORPORATION TO NOTIFY ANY PERSON OR ORGANIZATION OF SUCH REVISION OR CHANGES.

TRADEMARKS

Chromeleon is a registered trademark and PolyVial is a trademark of Dionex Corporation. PEEK is a trademark of Victrex plc. Teflon and Tefzel are registered trademarks of E.I. duPont de Nemours & Co. Microsoft, Windows XP, and Windows 2000 are registered trademarks of Microsoft Corporation.

PRINTING HISTORY

Revision 01, May 2005 Revision 02, August 2006 Revision 03, January 2008

1 • Introduction

1.1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

1.2 About This Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

1.2.1 Safety Messages and Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

1.3 Safety and Regulatory Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

1.3.1 Safety Labels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

2•Description

2.1 Overview of Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

2.2 Front Panel Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

2.2.1 Keypad Button Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

2.2.2 Display Screens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

2.3 Autosampler Compartment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

2.3.1 Syringe and Reservoir Organizer . . . . . . . . . . . . . . . . . . . . . . . 15

2.3.2 Sampling Needle Arm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

2.3.3 Sampling Needle Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

2.3.4 Flush, Waste, and Inject Ports . . . . . . . . . . . . . . . . . . . . . . . . . 19

2.3.5 Sample Vials and Well Plates . . . . . . . . . . . . . . . . . . . . . . . . . 19

2.3.6 Injection Valve Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

2.3.7 Sample Temperature Control Option . . . . . . . . . . . . . . . . . . . . 22

2.3.8 Simultaneous Injection Option . . . . . . . . . . . . . . . . . . . . . . . . . 23

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2.3.9 Sequential Injection Option . . . . . . . . . . . . . . . . . . . . . . . . . . . .23

2.3.10 Concentrator Injection Option . . . . . . . . . . . . . . . . . . . . . . . . . .24

2.4 Rear Panel Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25

2.4.1 USB Receptacle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25

2.4.2 TTL/Relay Connectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26

2.4.3 Fuse Holder and Power Receptacle . . . . . . . . . . . . . . . . . . . . . .26

2.5 Autosampler Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27

2.5.1 Local Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27

2.5.2 Computer Control (Locked Remote Mode) . . . . . . . . . . . . . . .30

2.6 System Wellness and Predictive Performance . . . . . . . . . . . . . . . . . . . .31

2.7 Theory of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33

2.7.1 Understanding the Wait Operation . . . . . . . . . . . . . . . . . . . . . .33

2.7.2 Overlapping Sample Preparation . . . . . . . . . . . . . . . . . . . . . . . .34

2.7.3 Running Samples Without Overlap . . . . . . . . . . . . . . . . . . . . . .35

2.7.4 Operating the AS as the System Master (Disabling the

Wait Operation) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .35

2.7.5 Understanding the Cycle Time . . . . . . . . . . . . . . . . . . . . . . . . .36

2.7.6 Operating Events During a Schedule . . . . . . . . . . . . . . . . . . . .37

2.7.7 Understanding the Status Display During a Run . . . . . . . . . . . .40

2.7.8 Operating Events During Flushing and Priming . . . . . . . . . . . .41

2.7.9 Sample and Prep Syringe Fluid Schematics . . . . . . . . . . . . . . .42

3 • Operation and Maintenance

3.1 Getting Ready to Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45

3.1.1 Turn On the Power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45

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3.1.2 Fill the Vials or Wells and Load the Sample Tray . . . . . . . . . . 46

3.2 Selecting Computer or Front Panel (Local) Control . . . . . . . . . . . . . . . 51

3.2.1 Connecting to Chromeleon or Chromeleon Xpress . . . . . . . . . 52

3.2.2 Running a Sequence of Injections in Chromeleon or

Chromeleon Xpress . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

3.3 Default Operating Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55

3.4 Specifying Sample Positions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56

3.4.1 Vial Tray Sample Positions . . . . . . . . . . . . . . . . . . . . . . . . . . . 56

3.4.2 Well Plate Tray Sample Positions . . . . . . . . . . . . . . . . . . . . . . 57

3.5 Specifying Injection Volumes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58

3.6 Sample Injection Types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

3.6.1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

3.6.2 Determining the Injection Type . . . . . . . . . . . . . . . . . . . . . . . . 62

3.6.3 Determining the Sample Loop Injection Type . . . . . . . . . . . . . 63

3.6.4 Guidelines for Sample Volumes Used, Loop Sizes, and

Injection Volumes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

3.6.5 Normal, Full-Loop Injections . . . . . . . . . . . . . . . . . . . . . . . . . . 70

3.6.6 Normal, Large-Loop Injections . . . . . . . . . . . . . . . . . . . . . . . . 72

3.6.7 Normal, Partial-Loop Injections . . . . . . . . . . . . . . . . . . . . . . . . 73

3.6.8 Normal, Partial-Loop, Limited-Sample Injections . . . . . . . . . . 76

3.6.9 Simultaneous Injections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

3.6.10 Concentrate Injections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82

3.6.11 Sequential Injections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

3.6.12 Sequential Concentrate Injections . . . . . . . . . . . . . . . . . . . . . . 90

3.7 Selecting Syringe Speed Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92

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3.7.1 Selecting Syringe Speeds on the Control Panel . . . . . . . . . . . .93

3.7.2 Entering Syringe Speeds in the Program . . . . . . . . . . . . . . . . . .94

3.8 Syringe Speed and Tubing Size Guidelines . . . . . . . . . . . . . . . . . . . . . .97

3.9 Opening the Autosampler Door During Operation . . . . . . . . . . . . . . .100

3.10 Routine Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .102

3.10.1 Daily . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .102

3.10.2 Periodically . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .103

3.10.3 Annually . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .103

3.10.4 Manually Flushing the Inject Port . . . . . . . . . . . . . . . . . . . . . .103

3.11 System Shutdown . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .104

4 • Troubleshooting

4.1 Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .105

4.2 Liquid Leaks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .124

4.2.1 Leaking Syringe or Syringe Valve Port . . . . . . . . . . . . . . . . . .124

4.2.2 Leaking Drain Line Connection . . . . . . . . . . . . . . . . . . . . . . .125

4.2.3 Leaking Inject Port . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .125

4.2.4 Leaking Fitting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .125

4.2.5 Broken Liquid Line . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .125

4.3 Condensation on Well Plate Covers . . . . . . . . . . . . . . . . . . . . . . . . . . .126

5•Service

5.1 Replacement Tubing and Fittings . . . . . . . . . . . . . . . . . . . . . . . . . . . . .127

5.1.1 Syringe and Reservoir Connections . . . . . . . . . . . . . . . . . . . .127

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5.1.2 Assemblies, Tubing, and Fittings . . . . . . . . . . . . . . . . . . . . . . 128

5.2 Resetting the Usage Counters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129

5.2.1 Resetting a Usage Counter on the AS Wellness Panel . . . . . . 130

5.2.2 Resetting a Usage Counter on the Usage Log Screen . . . . . . 131

5.3 Replacing the Sample or Prep Syringe . . . . . . . . . . . . . . . . . . . . . . . . 132

5.3.1 Removing the Existing Syringe . . . . . . . . . . . . . . . . . . . . . . . 133

5.3.2 Filling the New Syringe and Removing Bubbles . . . . . . . . . . 134

5.3.3 Connecting the New Syringe and Flushing . . . . . . . . . . . . . . 134

5.3.4 Initializing the Syringe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135

5.3.5 Resetting the Syringe Usage Counter . . . . . . . . . . . . . . . . . . . 136

5.4 Replacing the Sampling Needle Assembly . . . . . . . . . . . . . . . . . . . . . 136

5.4.1 Removing the Old Sampling Needle Assembly . . . . . . . . . . . 138

5.4.2 Installing the New Sampling Needle Assembly . . . . . . . . . . . 142

5.4.3 Resetting the Sampling Needle Assembly Usage Counter . . 145

5.5 Removing the Drip Tray . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145

5.6 Installing the Drip Tray . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146

5.7 Replacing the Needle Seal Assembly . . . . . . . . . . . . . . . . . . . . . . . . . 148

5.8 Aligning the Sampling Needle in the Inject Port . . . . . . . . . . . . . . . . . 151

5.9 Calibrating the Inject Port Volume . . . . . . . . . . . . . . . . . . . . . . . . . . . 155

5.10 Replacing the Leak Sensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161

5.11 Calibrating the Leak Sensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162

5.12 Removing the Sample or Prep Syringe Valve . . . . . . . . . . . . . . . . . . . 164

5.13 Replacing the Sample Syringe Valve . . . . . . . . . . . . . . . . . . . . . . . . . 165

5.14 Replacing the Prep Syringe Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167

5.15 Replacing a Sample or Prep Syringe Pump Assembly . . . . . . . . . . . . 169

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5.16 Rebuilding the Injection or Diverter Valve . . . . . . . . . . . . . . . . . . . . .170

5.17 Replacing the Well Plate Tray Thermal Pads . . . . . . . . . . . . . . . . . . . .172

5.18 Changing the Main Power Fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . .173

A • Specifications

A.1 Autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175

A.1.1 Electrical. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175

A.1.2 Environmental/Physical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175

A.1.3 Valves (Optional) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .176

A.1.4 Injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .176

A.2 Sample Temperature Control (Optional) . . . . . . . . . . . . . . . . . . . . . . .177

B • Installation

B.1 Facility Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .179

B.2 Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .180

B.3 Autosampler Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .186

B.3.1 Before You Begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .186

B.3.2 Installation Checklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .186

B.3.3 Connecting the Syringes and Drain Line . . . . . . . . . . . . . . . . .187

B.3.4 Connecting a Single Injection Valve . . . . . . . . . . . . . . . . . . . .191

B.3.5 Connecting Two Injection Valves (for Simultaneous

Injections) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .193

B.3.6 Connecting a Diverter Valve (for Sequential Injections) . . . . .197

B.3.7 Connecting a Second AS to a Dual ICS-3000 System . . . . . . .200

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B.3.8 Connecting the AS to Chromeleon or Chromeleon

Xpress (Optional) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203

B.3.9 Connecting the Power Cord . . . . . . . . . . . . . . . . . . . . . . . . . . 206

B.3.10 Turning On the Power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207

B.3.11 Selecting Module Setup Options . . . . . . . . . . . . . . . . . . . . . . 208

B.3.12 Configuring the AS in Chromeleon or Chromeleon Xpress . . 212

B.3.13 Priming the Liquid Lines. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214

B.3.14 Removing Bubbles from the Syringe . . . . . . . . . . . . . . . . . . . 218

B.4 Injection Valve Plumbing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219

B.5 Setting Up Chromeleon or Chromeleon Xpress for Simultaneous Injections 220

B.5.1 Assigning Unique Names to Devices . . . . . . . . . . . . . . . . . . . 220

B.5.2 Creating Sequences, Programs, and Quantification Methods . 224

C • Display Screens

C.1 Menu of Screens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

C.2 Main Status Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229

C.3 Detail Status Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232

C.4 Method Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236

C.4.1 Sample Prep Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237

C.4.2 Method Setup Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 242

C.4.3 Timed Events Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243

C.5 Schedule Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 245

C.6 Module Setup Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247

C.6.1 Installed Options Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247

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C.6.2 Front Panel Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .248

C.6.3 Plumbing Configuration Screen . . . . . . . . . . . . . . . . . . . . . . . .249

C.6.4 Time/Date Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .251

C.6.5 System Parameters Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . .252

C.6.6 Inject Port Alignment Screen . . . . . . . . . . . . . . . . . . . . . . . . . .255

C.6.7 Door Interlock Bypass Screen . . . . . . . . . . . . . . . . . . . . . . . . .256

C.7 Flush/Prime Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .257

C.8 Time Function In Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .259

C.9 Diagnostic Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .260

C.9.1 Power-Up Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .260

C.9.2 Diagnostic Tests Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .261

C.9.3 XYZ Test Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .262

C.9.4 Temperature Statistics Screen . . . . . . . . . . . . . . . . . . . . . . . . .263

C.9.5 Liquid Control Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .264

C.9.6 Logs Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .267

C.9.7 Leak Sensor Calibration and Status Screen . . . . . . . . . . . . . . .270

C.9.8 Keyboard Test Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .271

C.9.9 Code Versions Screen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .272

C.10 Time Function Out Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .272

D • Front Panel Control

D.1 Accessing AS Screens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .273

D.2 Entering or Changing Values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .273

D.3 Running a Schedule of Injections from the Front Panel . . . . . . . . . . . .274

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D.3.1 Selecting and Starting the Schedule . . . . . . . . . . . . . . . . . . . . 275

D.3.2 Placing a Running Schedule on Hold . . . . . . . . . . . . . . . . . . . 276

D.3.3 Stopping a Running Schedule . . . . . . . . . . . . . . . . . . . . . . . . . 276

D.4 Running Under Direct Control from the Front Panel . . . . . . . . . . . . . 277

D.5 Creating Methods from the AS Front Panel . . . . . . . . . . . . . . . . . . . . 279

D.5.1 Creating a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 279

D.5.2 Saving a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 280

D.5.3 Defining Sample Prep Steps . . . . . . . . . . . . . . . . . . . . . . . . . . 281

D.5.4 Selecting Method Setup Parameters . . . . . . . . . . . . . . . . . . . . 291

D.5.5 Defining Timed Events Steps . . . . . . . . . . . . . . . . . . . . . . . . . 294

D.5.6 Editing a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 297

D.5.7 Deleting a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 297

D.5.8 Copying a Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 297

D.5.9 Example Method for Dilution . . . . . . . . . . . . . . . . . . . . . . . . . 298

D.6 Creating Schedules from the AS Front Panel . . . . . . . . . . . . . . . . . . . 298

D.6.1 Creating a New Schedule . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299

D.6.2 Saving a Schedule . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 300

D.6.3 Selecting an Action if a Scheduled Vial Is Missing . . . . . . . . 300

D.6.4 Defining Schedule Lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 301

D.6.5 Editing a Schedule. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303

D.6.6 Deleting a Schedule. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303

D.6.7 Copying a Schedule. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303

D.7 Selecting Syringe Speeds for Front Panel Operation . . . . . . . . . . . . . 304

D.8 Aligning the Sampling Needle from the AS Front Panel . . . . . . . . . . 306

D.9 Calibrating the Inject Port Volume from the AS Front Panel . . . . . . . 311

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AS Autosampler Operator’s Manual

E • TTL and Relay Control

E.1 TTL and Relay Connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .317

E.1.1 Connecting a TTL or Relay . . . . . . . . . . . . . . . . . . . . . . . . . . .318

E.1.2 Selecting TTL Input Control Types . . . . . . . . . . . . . . . . . . . . .319

E.2 Controlling TTL and Relay Outputs . . . . . . . . . . . . . . . . . . . . . . . . . . .321

E.2.1 Example TTL/Relay Connections . . . . . . . . . . . . . . . . . . . . . .322

F • Reordering Information

x Doc. 065051-03 1/08

1.1 Overview

The Dionex AS Autosampler precisely delivers from 1 to 8,000 μL of sample to an injection valve. The injection volume range delivered by a particular AS depends on the size of the sample syringe and the sampling mode.

Modes

Several sampling modes are available:

• In normal mode, sample is delivered to a single system and a sample loop is

installed on the injection valve.

• In concentrate mode, the AS delivers sample to a Dionex ultra-low pressure

concentrator column.

• In simultaneous mode, the AS can deliver sample to two independent ion

chromatography (IC) systems. Sample is delivered equally to the two IC systems (50% of the volume to each). This mode requires the simultaneous injection option.

1 • Introduction

• In sequential mode, a diverter valve allows sample to be delivered to two

independent channels or IC systems through sequential injections. This mode requires the sequential injection option.

• In sequential concentrate mode, a diverter valve allows sample to be delivered

to two independent channels or IC systems on which Dionex ultra-low pressure concentrator columns are installed.

Sample Temperature Control

The sample temperature control option provides uniform heating and cooling of the vial tray.

Sample Preparation

The sample preparation option is used for automated dilutions and pipet functions.

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AS Autosampler Operator’s Manual

Injection Valve Options

The AS can be configured either without an injection valve, or with one or two injection valves installed in the autosampler compartment.

Vials, Well Plates, and Sample Trays

The AS can sample from either vials or well plates. Several types of sample trays are available to accommodate the various vial and well plate sizes. Insulated trays are available for use with the sample temperature control option.

During sampling, the tray remains stationary and a sampling needle arm moves from position to position. Up to 99 injections can be taken from each vial or well.

Control

The AS can be controlled locally, from the front panel keypad and display, or remotely, with a personal computer running Windows Chromeleon or later), or Chromeleon Xpress (Release 1.0 or later). Limited remote control is also available, using TTL and relay signals.

For communication between the AS and the software, the AS is connected to a USB (Universal Serial Bus) port on the computer, USB hub, or other Dionex module equipped with a USB hub.

® Chromatography Data Management System software (Release 6.7

® 2000 or Windows XP and

2 Doc. 065051-03 1/08

1.2 About This Manual

1 • Introduction

Chapter 1 Introduction

Chapter 2 Description

Chapter 3 Operation and Maintenance

Chapter 4 Troubleshooting

Chapter 5 Service

Appendix A Specifications

Appendix B Installation

Appendix C Display Screens

Appendix D Front Panel Control

Introduces the AS; explains the conventions used in this manual, including safety-related information.

Describes the AS operating features and system components; provides the theory of operation.

Provides operating instructions for the AS and describes routine preventive maintenance procedures.

Lists problems and presents step-by-step procedures for how to isolate and eliminate the cause of each problem.

Provides step-by-step instructions for routine service and parts replacement procedures that the user can perform.

Lists the AS specifications and installation site requirements.

Describes how to install the AS.

Describes the front panel operational and diagnostic display screens.

Provides step-by-step instructions for operating the AS from the front panel.

Appendix E TTL and Relay Control

Appendix F Reordering Information

NOTE For details about using Chromeleon or Chromeleon

Xpress to operate the AS, refer to the Chromeleon or Chromeleon Xpress Help. If you are using Chromeleon Xpress, also refer to the Chromeleon Xpress Quick Reference Guide (P/N 065070), provided on the Dionex Reference Library CD-ROM (P/N 053891).

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Describes the AS TTL and relay control features.

Lists spare parts for the AS.

AS Autosampler Operator’s Manual

1.2.1 Safety Messages and Notes

This manual contains warnings and precautionary statements that can prevent personal injury and/or damage to the AS when properly followed. Safety messages appear in bold type and are accompanied by icons, as shown below.

Indicates an imminently hazardous situation which, if not avoided, will result in death or serious injury.

Indicates a potentially hazardous situation which, if not avoided, could result in death or serious injury.

Indicates a potentially hazardous situation which, if not avoided, may result in minor or moderate injury. Also used to identify a situation or practice that may seriously damage the instrument, but will not cause injury.

Indicates that the function or process of the instrument may be impaired. Operation does not constitute a hazard.

Messages d'avertissement en français

Signale une situation de danger immédiat qui, si elle n'est pas évitée, entraînera des blessures graves à mortelles.

Signale une situation de danger potentiel qui, si elle n'est pas évitée, pourrait entraîner des blessures graves à mortelles.

Signale une situation de danger potentiel qui, si elle n'est pas évitée, pourrait entraîner des blessures mineures à modérées. Également utilisé pour signaler une situation ou une pratique qui pourrait gravement endommager l'instrument mais qui n'entraînera pas de blessures.

4 Doc. 065051-03 1/08

1 • Introduction

Warnhinweise in Deutsch

Bedeutet unmittelbare Gefahr. Mißachtung kann zum Tod oder schwerwiegenden Verletzungen führen.

Bedeutet eine mögliche Gefährdung. Mißachtung kann zum Tod oder schwerwiegenden Verletzungen führen.

Bedeutet eine mögliche Gefährdung. Mißachtung kann zu kleineren oder mittelschweren Verletzungen führen. Wird auch verwendet, wenn eine Situation zu schweren Schäden am Gerät führen kann, jedoch keine Verletzungsgefahr besteht.

Notes

Informational messages also appear throughout this manual. These are labeled NOTE and are in bold type:

NOTE NOTES call attention to certain information. They alert

you to an unexpected result of an action, suggest how to optimize instrument performance, etc.

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AS Autosampler Operator’s Manual

1.3 Safety and Regulatory Information

The AS is designed for IC (ion chromatography) and HPLC (high-performance liquid chromatography) applications and should not be used for any other purpose. Operation of an AS in a manner not specified by Dionex may result in personal injury.

1.3.1 Safety Labels

The TUV GS and cTUVus Mark safety labels and the CE Mark label on the AS indicate that the AS is in compliance with the following standards: EN 61010-1:2001 (safety), CAN/CSA-C22.2 No. 1010.1-92 + A2:97 (safety), UL 61010C-1:2002 R8.02 (safety), and EN 61326:1997, including A1:1998 and A2:2001 (EMC susceptibility and immunity).

The symbols below appear on the AS or on AS labels.

Alternating current

Protective conductor terminal (earth ground)

Power supply is on

Power supply is off

Indicates a potential hazard. Refer to the operator’s manual for an explanation of the hazard and how to proceed.

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2.1 Overview of Features

Figure 2-1 illustrates the main features of the AS Autosampler.

Flush

Reservoir

2 • Description

Sample and Prep Syringes

Front Panel

Autosampler Compartment

Power

Switch

Figure 2-1. AS Operating Features

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AS Autosampler Operator’s Manual

Front Panel

The front panel contains a liquid crystal display (LCD) and membrane keypad (see Section 2.2

Power Switch

The power switch is in the lower left corner of the AS. The switch functions only when the door is fully closed.

Autosampler Compartment

The autosampler compartment contains a stationary sample tray and an XYZ sampling needle arm that moves from vial to vial during operation (see

Section 2.3

port. Valves for sample injection or for diverting sample flow to different systems can optionally be included.

Syringe and Reservoir Organizer

The organizer on top of the autosampler holds the sample syringe, the flush liquid reservoir, and the optional prep syringe and reagent reservoirs. See Section 2.3.1 for details.

). The front panel permits manual control of AS functions.

). The compartment also contains an inject port, flush port, and waste

Options

• Sample Preparation: When this option is installed, the AS can perform

sample preparation functions such as diluting standards or dispensing reagents. See Section 2.3.1

for details.

• Sample Temperature Control (not pictured): When this option is installed,

the AS provides uniform heating and cooling of the sample tray. See

Section 2.3.7

for details.

• Simultaneous Injection (not pictured): When this option is installed, the AS

can deliver sample to two independent ion chromatography (IC) systems simultaneously. See Section 3.6.9

for details.

• Sequential Injection (not pictured): When this option is installed, the AS can

deliver sample to two independent ion chromatography (IC) systems alternately.

• Concentrator Injection (not pictured): When this option is installed, the AS

can load sample to a Dionex ultra-low pressure concentrator column. See

Section 2.3.10 for details.

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Valve Options

One or two injection valves can be installed in the AS autosampler compartment. Or, the AS can be configured without an injection valve and instead be connected to one or two valves installed in other chromatography system modules. Two injection valves are required for simultaneous injections.

For the sequential injection option, a diverter valve is installed in the AS.

2.2 Front Panel Features

2 • Description

Adjust screen contrast

Figure 2-2. AS Front Panel

Information is displayed on the LCD, or screen. To adjust the screen contrast, use the knurled knob in the recess below the keypad (see Figure 2-2

From the keypad, you can:

• Directly control AS operation

• Create and modify programmed series of operating steps, called methods

• Create and modify programmed series of injections, called schedules

Refer to Section 2.2.2

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for a summary of how to select screens and edit parameters.

AS Autosampler Operator’s Manual

(

)

2.2.1 Keypad Button Functions

Button Function

Stop

Stops a schedule if the AS is in Local mode, Schedule control (see

Section 2.5.1

). A confirmation message appears; press Stop again to

abort the schedule or Hold/Run to resume. If the AS is in Direct Control (see Section 2.5.1 the current syringe and/or sampling arm movement.

Note: Pressing Stop while the AS is in Direct Control, Locked Remote mode (software control) (see Section 2.5.2 recommended. Use the controls on the software Control panel (see

Section 2.5.2

) to stop a sequence.

), pressing Stop stops

) is not

Home

Insert

Delete

(Hold)

Run

Sends the needle arm to its home position. This button functions only when the AS is in Local mode. If a schedule is in progress, it must be on hold. If you press Home when a schedule is on hold, and then resume the schedule, the needle arm returns to the location it was in when you pressed Home.

Inserts a new step into a method or schedule. Move the cursor to the leftmost field on the

SAMPLE PREP, TIMED EVENTS, or SCHEDULE

screen and press Insert. The new step is added above the cursor position. See Appendix D

for details about front panel operation.

Delete performs several functions:

• Cancels an entry that is in progress and restores the previous value.

• Returns a field to its default value (if an entry is not in progress).

• Deletes a line in a method or schedule. To do this, position the

cursor in the leftmost field of the line that you want to delete and press Delete twice.

• Deletes an entire method or schedule. To do this, position the

cursor in the method or schedule edit field, or in the time field of the

INIT step, and press Delete twice.

At power-up, the AS is on hold (the left LED is illuminated). After you select a schedule to run, pressing Hold/Run starts the schedule (the right LED is illuminated). When the schedule is complete, the AS returns to hold. When running, press Hold/Run to pause the schedule. This button functions only when the AS is in Local mode.

Note: If the wait operation is enabled (see Section 2.7.1 pauses when it reaches the wait step in the method. Press Hold/Run to continue. You can disable the wait (see Section 2.7.4

), the AS

10 Doc. 065051-03 1/08

Button Function

If you press the Door button when a sequence (or front panel

Door

schedule) is running, a message screen indicates whether it is currently safe to open the door. If it is, the screen displays for how much longer it is safe to open the door. Opening the door without first pressing this button, or when the message indicates it is not safe, ends the currently running sequence (or schedule).

Select

The Select buttons scroll through predetermined options in entry fields. To confirm the selected value, press Enter or move out of the field by pressing a cursor arrow button. In fields that have

Select

predetermined numeric choices, Select Δ increases the value by one unit and Select ∇ decreases the value by one. Holding down a Select button increases (or decreases) the value continuously.

The arrow buttons move the cursor in the direction of the arrow to the next entry field (if one exists). At the end of a line, the left arrow wraps the cursor around to the next entry field on the line above; the right arrow wraps the cursor to the next entry field on the line below. The up and down arrows do not wrap around.

2 • Description

The autosampler door must remain closed during operation. If the door is opened during operation, the sampling arm stops immediately. If a sequence (or schedule) is running, it is ended. If you want to open the door during a sequence (or schedule), first press the Door button, and then follow the onscreen instructions.

After entering a new value in an entry field, pressing an arrow button saves and/or executes the change.

Displays a help screen specific to the current entry field.

Help

Displays a list of the available screens.

The numeric buttons enter the selected number into the current entry field. On a menu, pressing a numeric button opens the corresponding screen.

Enter

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Saves and/or executes changes made in entry fields. If a menu screen is displayed, pressing Enter opens the highlighted screen.

AS Autosampler Operator’s Manual

2.2.2 Display Screens

The LCD displays status information and allows access to all AS operations.

When the power is turned on, the AS performs an initialization sequence (see Section 3.1.1 is an example of the

) and then displays the MAIN STATUS screen. Figure 2-3

MAIN STATUS screen during AS operation.

VOL

8 9

20.013

METHOD: 01 SCH LINE

Time since last injection

Volume to be injected

Current method

Control mode

Current s chedule

Schedule line currently in progress

121113

min

INJ#

10 10.6

°C

↑

status

m ode

VIAL#

TRAY:

LOCAL

Help Prompt

Operating status

Current vial or well

2 3

Current injection Total injections s cheduled

for this vial Current tray temperature

Tray temperature

(increasing) Operatin

LOADING LOOP

TIME

Figure 2-3. Main Status Screen

NOTE TRAY temperature is displayed only when the

sample temperature control option is installed.

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2 • Description

To access other AS screens:

Press the Menu button. The MENU of SCREENS appears (see Figure 2-4).

MENU of SCREENS

FLUSH/PRIME

MAIN STATUS

DETAIL STATUS

METHOD MENU

SCHEDULE

MODULE SETUP MENU

Help Prompt

Figure 2-4. Menu of Screens

To select a screen on the menu, use one of the following methods:

TIME FUNCTION IN

DIAGNOSTIC MENU

8 9

TIME FUNCTION OUT

• Press the keypad numeric button corresponding to the screen’s

number on the menu.

• Move the cursor to highlight the screen number and press Enter.

To display a brief description of each screen:

Press the Help button. See Appendix C for details about each screen.

To edit a field on the screen:

1. Press a cursor arrow button to position the cursor in the field to be

edited. Fields that are in reverse video (blue letters on white background) can be edited. Other fields display information only.

2. If the field accepts numeric values, press the desired numeric buttons

to enter the value.

If the field has predetermined options, press the

Select Δ or Select ∇

button to choose the desired option.

3. To confirm the selected value, press

Enter or move the cursor out of

the field by pressing an arrow button.

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AS Autosampler Operator’s Manual

2.3 Autosampler Compartment

Prep Syringe

(Optional)

Sample

Syringe

Reagent

Reservoir (Optional)

Flush

Reservoir

XYZ Sampling

Needle Arm

Flush, Waste,

and Inject Ports

Figure 2-5. Autosampler Compartment Features

Injection and/or Diverter Valves

(Optional)

Sample Tray

(for Vials or Well Plates)

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2.3.1 Syringe and Reservoir Organizer

The organizer on top of the autosampler holds the sample syringe, 1 L flush reservoir, and the optional prep syringe and reagent reservoirs (see

Figure 2-5

Sample Syringe Functions

The sample syringe performs these operations:

• Pipettes liquid from one vial to another or to waste.

• Mixes a vial by repeatedly drawing and expelling the vial contents. If

AS is equipped with the sample preparation option, mixing is done

the with the sample prep syringe instead of the sample syringe.

• Delivers sample to the injection valve.

The following sample syringe volumes are available:

2 • Description

Sample Syringe Vol ume

100 μL 055064 250 μL (standard) 053916 500 μL 055065 1,000 μL 055066

5 mL* 053915

10 mL* 055068

*A 5 or 10 mL syringe is required for simultaneous injections and optional for all other sampling modes. If a 5 or 10 mL syringe is installed, an 8.5 mL sampling needle assembly (P/N 061267) is required.

Part Number

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AS Autosampler Operator’s Manual

Sample Preparation Option Functions

The AS can be equipped with a sample preparation option that includes a sample prep syringe, a sample prep valve, and one 250 mL reagent reservoir. Up to three additional reagent reservoirs can be configured.

The sample prep syringe performs these operations:

• Dispenses reagents from external reservoirs to any vial

• Mixes a vial by repeatedly drawing and expelling the vial contents

• Performs dilutions, allowing preparation of multilevel calibration

standards

• Performs liquid-liquid extractions by adjusting the height of the

needle in the vial

The following sample prep syringe volumes are available:

Prep Syringe Volume Part Number

250 μL 053916 500 μL 055065 1,000 μL 055066

2.5 mL 055067

5 mL (standard) 053915

10 mL 055068

The reagent reservoir connects to port A on the sample prep valve. Additional reservoirs can be connected to ports B, C, and D. Commands from the AS front panel screens or from Chromeleon or Chromeleon Xpress let you select the reservoirs.

NOTE The 5 mL and 10 mL syringes are intended for most

sample prep functions. Use the smaller syringes only for applications that require low flow rates.

NOTE The 2.5 mL syringe (P/N 055067) can only be used

with the sample preparation option. It cannot be used as a sample syringe.

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2.3.2 Sampling Needle Arm

The autosampler door must remain closed during operation. If you open the door during operation, the sampling arm stops immediately. If a schedule is running, it is ended. To safely open the door during a schedule, press the Door button. See Section 3.9 for details.

During operation, the sampling needle arm moves the sample needle along X-, Y-, and Z-axes. The sample tray remains stationary throughout the sampling process.

To sample from a vial or well plate, the arm moves to the specified position in the tray and lowers the needle into the vial or well. The sampling needle pierces the vial cap or well plate cover to allow fluid to be drawn or delivered. When sampling is complete, the arm raises the needle out of the vial or well. When running a schedule of injections, vials or wells can be sampled in any order.

The needle height (the distance from the tip of the needle to the bottom of the vial or well) can be adjusted, allowing operations such as liquid-liquid extraction.You can set the needle height in a Chromeleon or Chromeleon Xpress program (see Section 3.2.2

page 292

2 • Description

) or in a front panel program (see

NOTE Changing the needle height is not recommended

when sampling from a micro-well well plate, because the well depth is shallow. You can adjust the needle height in a deep-well well plate as required, provided the needle remains submerged in sample. This ensures accurate sample aspiration.

During a flush operation, the sampling needle arm delivers flush fluid to the flush port (see Figure 2-7

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AS Autosampler Operator’s Manual

eedle

2.3.3 Sampling Needle Assembly

The sampling needle assembly consists of a PEEK™ sampling needle and a line, which connects the sample syringe and the sampling needle (see

Figure 2-6

ample Syringe

Valve

Sample Syringe

Sampling Needle

ssembl

Flush Reservoir

Sampling N

Figure 2-6. Sample Syringe with Sampling Needle Assembly

Two assemblies are available (see the table below).

Sampling Needle Assembly

PEEK Needle with

1.2 mL line

PEEK Needle with

8.5 mL line

a. The 8.5 mL sampling needle assembly is standard when the AS is configured with the simultaneous injection option.

18 Doc. 065051-03 1/08

Used With Part Number

Sample syringe volumes

054271

1mL or less

Sample syringe volumes

061267

greater than 1 mL

2.3.4 Flush, Waste, and Inject Ports

The flush port flushes the outside of the sampling needle with fluid from the flush reservoir. Excess fluid flows to the waste port. The waste port accepts waste fluid from the sampling needle and the flush port, and directs it out the drain line. The inject port (see Figure 2-7 sample from the sampling needle and delivers it to the injection valve.

Waste Port

Flush Port

Figure 2-7. Flush, Waste, and Inject Ports

2.3.5 Sample Vials and Well Plates

2 • Description

) accepts the

Inject Port

Vial Trays and Vials

The following vial trays are available. See the following section for well plate information.

Tray Material Vial Size Held Tray Capacity Part Number

Plastic 10 mL 49 062374

Aluminum 0.3 mL, 1.5 mL 100 062481

Insulated Aluminum

a. Required for the sample temperature control option. Includes tray covers for empty tray rows.

0.3 mL, 1.5 mL 100 062377

The following vials are available:

Vial Size and Material Quantity Part Number

0.3 mL polymer Package of 100 055428

1.5 mL glass Package of 100 055427

10 mL PolyVial™ Package of 100 055058

1.5 mL plastic Package of 100 061696

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AS Autosampler Operator’s Manual

Dionex does not recommend using the 10 mL plastic tray with the sample temperature control option because the plastic tray is inefficient at heating and cooling the vials. In addition, the tray is not designed for temperatures above 40 °C (104 °F) and may deform at

temperatures above 40

Well Plates and Trays

°C.

Dionex recommends using well plates in the AS if you have limited amounts of sample or if you require more than 100 sample positions. Follow these guidelines when using well plates:

• 2 mL deep-well plates are suitable for ion chromatography (IC)

applications, where concentration of ions is above 1 ppm.

• 0.5 mL and 0.45 mL micro-well plates are not recommended for IC

applications.

• Where concentration of ions is below 1 ppm, Dionex recommends

using 10 mL PolyVials (P/N 055058, package of 100).

The following well plate tray packages are available:

Well Plate Tray Type Includes Part Number

Micro-well plate tray One micro-well plate tray, two micro-well

plates with 0.5 mL round (U-shaped) wells, and two covers (not suitable for IC applications)

Deep-well plate tray One deep-well plate tray, two deep-well plates

with 2 mL wells, and two covers (suitable for

IC applications, where concentration of ions is above 1 ppm)

066324

066325

Additional well plates and covers can be ordered separately. See

Appendix F

Well Plate Tray Notes

for reordering information.

• All well plate trays can be used with the sample temperature control

option.

• Each well plate tray also holds ten, 1.5 mL vials.

• The micro-well plate tray can hold either type of micro-well plate (V-

shaped wells or U-shaped wells).

20 Doc. 065051-03 1/08

• The AS automatically detects the type of tray installed.

2.3.6 Injection Valve Options

The AS can optionally have one or two injection valves installed inside the autosampler compartment. The injection valves are six-port, electrically-activated Rheodyne valves. The valves have two operating positions: Load and Inject (see Figure 2-8

2 • Description

Concentrator

Column

(optional—

replaces loop)

Sample In

Sample Loop

To Wa st e

LOAD POSITION

(5)

(6)

(1)

(4)

(3)

(2)

From Pump

Sample In

To Co l u m n

Sample Loop

(6)

To Wa st e

INJECT POSITION

(5)

(1)

From Pump

(4)

(2)

To C o l u m

(3)

Figure 2-8. Injection Valve Flow Schematics

Eluent flows through either the Load or Inject path, depending on the valve position:

• In the Load position, sample flows from the AS inject port line, through the

valve, and into the sample loop, where it is held until injection. Eluent flows from the pump, through the valve, and to the column, bypassing the sample loop. Excess sample flows out to waste.

• In the Inject position, sample is swept to the column for analysis. Eluent flows

from the pump, through the sample loop, and on to the column, carrying the contents of the sample loop with it.

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AS Autosampler Operator’s Manual

2.3.7 Sample Temperature Control Option

The sample temperature control option provides heating and cooling of the sample tray. You can program the temperature set point from 4 60

°C. Samples in the tray are uniformly heated or cooled to the

programmed set point (to a minimum of 20 maximum of 40

The sample temperature control option sits on the workbench below the autosampler compartment (see Figure 2-9

°C above ambient). The absolute range is 4 °C to 60 °C.

°C to

°C below ambient and a

Sample Temperature Control Option

Figure 2-9. AS with Sample Temperature Control Option

NOTE The temperature inside the autosampler compartment

may exceed ambient by up to 2 °C. The sample temperature control option is recommended for samples that are temperature sensitive.

22 Doc. 065051-03 1/08

2.3.8 Simultaneous Injection Option

When the AS simultaneous injection option is installed, a single AS can deliver sample to two independent chromatography systems. The sample is injected simultaneously to both systems (two injection valves are required). Dual analyses can be performed with only one sample, thus increasing sample throughput and eliminating the need to label, fill, and track two sample vials.

In Chromeleon or Chromeleon Xpress, the two chromatography systems and the AS are configured into a single timebase and each system is assigned a unique device name and channel. This lets you monitor and control both systems from one Control panel and run all samples in one sequence.

2 • Description

The simultaneous injection option requires a 5 or 10 mL and an 8.5 mL sampling needle assembly (see Section 2.3.3 injections are required with this option.

See Section 3.6.9

for details about simultaneous injections.

2.3.9 Sequential Injection Option

When the simultaneous option is installed, a single AS can deliver sample to two independent chromatography systems. With the sequential option, sample is delivered to the first system, flow is rerouted (diverted), and sample is then delivered to the second system. Sequential injection allows different samples to be injected into each system.

The sequential injection option requires two injection valves and a diverter valve (for switching sample flow between the two systems). The diverter valve is installed in the AS autosampler compartment. The injection valves must be installed in the IC system’s chromatography compartment.

See Section 3.6.11

NOTE The diverter valve can only be controlled by

for details about sequential injections.

Chromeleon or Chromeleon Xpress. It is not controllable from the AS front panel.

sample syringe

). Full-loop

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AS Autosampler Operator’s Manual

2.3.10 Concentrator Injection Option

The AS concentrator injection option lets an AS deliver sample and reagent to a Dionex ultra-low pressure concentrator column. With this option, a concentrator column is installed on the injection valve instead of a sample loop. A 5 or 10 mL larger sample loading volumes. If the 5 or 10 mL installed, an 8.5 mL sampling needle assembly must also be installed (see

Section 2.3.3

The AS is capable of delivering at a maximum pressure of 690 kPa (100 psi). When setting up concentrator injections, make sure to select syringe speeds that will keep the backpressure below 690 kPa (100 psi), taking into account the pressure from the installed tubing and the concentrator column. A syringe speed of 1 or 2 is required. See

Section 3.8

for details.

sample syringe can be installed to allow

sample syringe is

See Section 3.6.10

for details about concentrator injections.

24 Doc. 065051-03 1/08

2.4 Rear Panel Features

1 2 3

2 • Description

USB

Receptacle

TTL/Relay

Connectors

Syringe

Connectors

Auxiliary

Power Connector

Main Power

Receptacle and Fuse Holder

Figure 2-10. AS Rear Panel

2.4.1 USB Receptacle

The USB receptacle provides a connection to the Chromeleon or Chromeleon Xpress computer. To enable computer control of the autosampler, connect a USB cable between the USB receptacle on the AS rear panel and a USB port on the computer or USB hub. See Section B.3.8 for USB connection instructions.

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AS Autosampler Operator’s Manual

2.4.2 TTL/Relay Connectors

The eight TTL and relay connectors interface with Dionex and nonDionex modules. The input connectors allow for relay and TTL control of the autosampler by a connected module(s). The output connectors allow the AS to control other module(s). Appendix E TTL functions and the connections between the AS and other modules.

2.4.3 Fuse Holder and Power Receptacle

The fuse holder contains two 3.15 amp fast-blow fuses (P/N 954745). For instructions on how to change the fuses, see Section 5.18

The power cord plugs into the IEC 320 three-prong receptacle.

The power supply cord is used as the main disconnect device. Make sure the socket-outlet is located near the AS and is easily accessible.

Le cordon d'alimentation principal est utilisé comme dispositif principal de débranchement. Veillez à ce que la prise de base soit située/installée près du module et facilement accessible.

Das Netzkabel ist das wichtigste Mittel zur Stromunterbrechung. Stellen Sie sicher, daß sich die Steckdose nahe am Gerät befindet und leicht zugänglich ist.

describes the relay and

26 Doc. 065051-03 1/08

2.5 Autosampler Control

The AS can be controlled locally, from the front panel keypad and display (see

Section 2.5.1

Chromeleon Xpress software is installed (see Section 2.5.2 from an integrator or other remote controller can also be used to control the AS. See Appendix E

2.5.1 Local Control

), or remotely, with a personal computer on which Chromeleon or

for details about TTL control.

In Local mode, the AS receives commands from the front panel buttons and screens. When the AS is powered up, it is always in Local mode.

When the AS is in Local mode, two types of control are available: Direct and Schedule.

• In Direct control, you enter control commands and parameters on the

AS screens, and the AS executes the commands immediately.

• In Schedule control, you define a list of injections (a schedule) to be

run in sequence. The schedule can be defined, saved, and then run at a later time.

2 • Description

). TTL input signals

The table below summarizes the AS Local control configurations:

Operating/Control Mode Autosampler Operation

Local/Direct Control Commands are entered from the AS front panel

and executed immediately after being entered.

Local/Schedule Control Commands are entered from the AS front panel

and executed by running a schedule programmed from the front panel.

Local/Direct Control Mode

You can directly control the following AS functions from the front panel:

• TTL and relay outputs

• Injection valve position

• Second injection valve position or diverter valve (if installed)

• Tray set point (if installed)

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AS Autosampler Operator’s Manual

Set these functions from the DETAIL STATUS screen (see Figure 2-11).

DETAIL

VIAL#:

INJ#:

METHOD:

TRAY:

LOCAL

Help Prompt

Local/Schedule Control Mode

20 °C25

Figure 2-11. Detail Status Screen Example

IDLE

TIME:

VOL:

TRAY:

LOOP: NORMAL FULL

DIRECT CONTROL

100

min uL

2mL

TTL1

TTL2

RLY1

RLY2

INJ VLV L

The AS runs all injections according to a programmed schedule. The schedule defines a series of vials to be sampled and the operating parameters to run on each vial. A schedule consists of the following parameters:

• The vials to sample

• The number of injections to draw from each vial

• The volume of each injection

• The method to run on each injection

• The action to take when a scheduled vial is missing

1 0 0 0

You program, save, and edit schedules from the

Figure 2-12

). The AS can store up to nine schedules in memory. The

SCHEDULE screen (see

stored schedules are retained in memory when the AS is turned off.

Section D.6

MISSING VIAL ACTION:

LINE ENDSTART

Help Prompt

28 Doc. 065051-03 1/08

describes how to create schedules.

EDIT

VIAL#

Figure 2-12. Schedule Screen

SAVE TO

STOP INJ/ INJ VIAL METHODVOL(uL)

RUNSCHEDULE

25.0 111

2 • Description

Each injection in a schedule is assigned a method (a series of operating instructions that tells the AS how to perform a single injection). A method consists of the following three phases:

Method Phase Used To Examples

Sample prep Prepare the sample for injection Pipetting, mixing, diluting

Method setup Set parameters that remain

constant throughout the method

Timed events Perform functions at a specific

time in the method

When the method runs, the

AS performs the sample prep steps first,

followed by the method setup, and then the timed events steps.

Section D.5

Notes

describes how to create methods.

• See Appendix D for details about front panel operation.

• Some autosampler operations are not available from the AS front

panel and can only be accessed from Chromeleon or Chromeleon Xpress. These include setting different aspirate and dispense syringe speeds and sampling from well plates.

Sample needle height, sample temperature

Loading the sample loop, injecting

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2.5.2 Computer Control (Locked Remote Mode)

In Locked Remote mode, Chromeleon or Chromeleon Xpress software sends commands from the host computer via the USB interface. Connecting the AS to a Control panel (see Figure 2-13 selects the Locked Remote mode. In this mode, you cannot change any operating parameters from the front panel keyboard. To return the AS to Local mode, clear the Connect check box on the AS Control panel.

) immediately

Figure 2-13. Example AS Control Panel

There are two modes for controlling the AS using Chromeleon or Chromeleon Xpress:

• In Direct mode, you select operating parameters and commands

directly from the Control panel. The AS executes the commands immediately.

30 Doc. 065051-03 1/08

2 • Description

• In Automatic mode, you create a list of samples (a sequence) to be

processed automatically. The sequence defines a series of vials to be sampled and the AS operating parameters to run on each vial. The sequence is similar to a front panel schedule (see Section 2.5.1

For instructions on how to connect the AS to Chromeleon or Chromeleon Xpress, see Section 3.2.1 autosampler using Chromeleon or Chromeleon Xpress, see Section 3.2.2 For detailed instructions, refer to the Chromeleon Help or Chromeleon Xpress.

. For an overview of how to operate the

2.6 System Wellness and Predictive Performance

The Chromeleon System Wellness panels provide easy access to the autosampler’s built-in diagnostic and calibration features. You can use the Wellness panels to perform the following tasks:

• Calibrate the inject port tubing volume

• Align the sampling needle in the inject port

• Calibrate and test the leak sensor

• Monitor usage of replaceable parts (see the description of Predictive

Performance below)

Wellness panels for calibrating the inject port tubing, aligning the sampling needle, and monitoring usage of parts are available on the Control panel (see

Figure 2-13

sensor (see Section 5.11

The Chromeleon Predictive Performance feature provides functions for monitoring the lifetime of replaceable parts and for planning and recording service procedures. You can use the Chromeleon Predictive Performance feature to perform the following tasks:

). A separate panel is available for calibrating and testing the leak

NOTE You can also perform System Wellness tasks from the

front panel screens. See Appendix D

for details.

• Monitor how many times a part has been used since it was last replaced.

Chromeleon monitors usage of the following AS parts: injection valve, diverter valve, XYZ sampling needle arm, sampling needle assembly, needle seal assembly, sample syringe, and prep syringe.

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• Display a warning message in the Chromeleon Audit Trail when a monitored

part is approaching its usage limit and when the limit is reached.

• Set the usage limits for monitored parts to suit your needs.

• Record when a part is serviced and reset the usage counter.

All Predictive Performance commands and parameters are available in the Chromeleon Commands dialog box (press the F8 key or select Command on the Control menu to open the Commands dialog box).

You can view and reset the usage counters for the various replaceable parts on either the AS Wellness panel (see Figure 2-14

Section C.9.6

above, are available only in Chromeleon.

). The remaining Predictive Performance functions, described

) or the USAGE LOG SCREEN (see

Figure 2-14. Example AS Wellness Panel

(Default Warning Values for All Parameters Shown)

32 Doc. 065051-03 1/08

2.7 Theory of Operation

2.7.1 Understanding the Wait Operation

NOTE This section pertains to AS operation in

Local/Schedule Control mode (see Section 2.5.1 only.

The wait operation is a step in the sample prep phase of a method. After performing all other sample prep steps, the AS waits for a continue command before performing the method’s timed events. The continue command can be sent via the front panel signal.

The wait’s position in the sample prep steps determines whether samples are overlapped or run sequentially:

• When the wait operation is the last step in the sample prep phase (the

default), samples are overlapped (see Section 2.7.2

• When the wait operation is at the beginning of the sample prep phase,

samples are run sequentially (see Section 2.7.3

2 • Description

)

Hold/Run button or a TTL input

NOTE The wait operation can be disabled (see

Section 2.7.4

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AS Autosampler Operator’s Manual

2.7.2 Overlapping Sample Preparation

When the wait operation is at the end of the sample prep phase (the default), the collection is occurring for one injection, the AS performs the sample prep steps for the next injection (see Figure 2-15 the schedule run flow of operations, see Section 2.7.6

AS overlaps sample preparation. This means that while data

). For a detailed description of

Start First Method

SAMPLE PREPWASH /

FLUSH

NOTE: The WAIT is ignored for the first injection in a Schedule.

Sample Overlap with Chromeleon or Chromeleon Xpress

If Chromeleon or Chromeleon Xpress is controlling the AS, front panel settings for the wait operation are ignored. Instead, the Sample Overlap option determines whether sample preparation is overlapped. Sample overlap, which is selected in the Chromeleon Server Configuration program, is enabled by default. For instructions on how to configure the AS in Chromeleon, see Section B.3.12

Injection 1

SETUP TIMED EVENTSINIT

0.0

Overlap

Start Next Method

WAS H/

SAMPLE PREP SETUP TIMED EVENTSINIT

FLUSH

Wait for Continue Command

WAI T

CONTINUE

Injection 2

0.0

Overlap

Start Next Method

WASH /

SAMPLE PREP

Figure 2-15. Sample Prep Overlap: Wait Operation at

End of Sample Prep Phase (Default)

WAI T

34 Doc. 065051-03 1/08

2.7.3 Running Samples Without Overlap

Placing the wait operation at the start of the sample prep steps allows samples to be run sequentially. The timed events for an injection before starting the next injection (see

Figure 2-16

AS completes the sample prep and

2 • Description

Start First Method Start Next Method

Run

FLUSH

SAMPLE PREPWAS H/

SETUP

TIMED EVENTS

INIT

Injection 1

Injection 2

WASH / FLUSH

Continue Continue

WAIT

SAMPLE PREP

SETUP

TIMED EVENTS

INIT

Figure 2-16. No Sample Prep Overlap: Wait Operation at

Beginning of Sample Prep Phase

2.7.4 Operating the AS as the System Master (Disabling the Wait Operation)

When the wait operation is disabled, a single run command (for example, pressing the performs the schedule lines without requiring additional commands. This allows the sending TTL signals to start the pump and detector methods).

To disable the wait operation, press

SETUP MENU and press 5 to go to the SYSTEM PARAMETERS screen. Set

WAIT field to DISABLED (see Figure 2-17).

the

METHOD SYRINGE SPEED:

CUT SEGMENT VOLUME:

FLUSH SYRINGE SPEED:

Help Prompt

Hold/Run button) starts the schedule. Thereafter, the AS

AS to act as the system master (the AS controls the system by

Menu and 5 to go to the MODULE

SYSTEM PARAMETERS

WAIT:

DISABLED 4

SAMPLE MODE:

INJECT PORT VOLUME:

NORMAL

WASH / FLUSH

Figure 2-17. System Parameters Screen

For details about the order of events during a schedule when the wait operation is disabled, see Figure 2-20

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AS Autosampler Operator’s Manual

0.0

2.7.5 Understanding the Cycle Time

Cycle time is an optional feature that can be used to control the time between injections. When a method is assigned a cycle time, the AS delays sample injection until the specified time has elapsed since the previous injection. Cycle time is measured from the start of the previous method's timed events (see Figure 2-18 time zero for each method (the default), specifying a cycle time allows a consistent time interval between injections, regardless of differences in sample prep and timed events among methods.

). Assuming injection occurs at

WASH / FLUSH

SAMPLE PREP

SETUP TIMED EVENTSINIT

Injection 1Start First Method

0.0

Start Next Method

WAS H/

SAMPLE PREP SETUP

FLUSH

Cycle Time

Wait fo r end of cycle time

INIT

Injection 2

TIMED EVENTS

0.0

Start Next Method

WASH / FLUSH

Cycle Time

SAMPLE PREP SETUP INIT

Figure 2-18. Cycle Time Example

A cycle's duration is never shorter than the specified time. However, cycle time will be ignored, thus delaying injection, if any of the following events occurs:

• WAIT FOR TEMP STABLE is enabled in the METHOD SETUP screen

(see Section C.6 the cycle time.

) and the temperature did not stabilize by the end of

• The sample prep wait operation is enabled and a run command did

not occur by the end of the cycle time.

• The previous method’s timed events extend beyond the cycle time.

If the cycle time expires before the injection occurs, a warning message is displayed and the message is logged on the

Section C.9.6

36 Doc. 065051-03 1/08

MESSAGE LOG screen (see

2.7.6 Operating Events During a Schedule

• Figure 2-19 shows the order of events during a schedule when the

wait operation is enabled.

• Figure 2-20 shows the order of events during a schedule when the

wait operation is disabled.

2 • Description

See Section 2.7.1

for details about the wait operation.

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AS Autosampler Operator’s Manual

Schedule Started

Preparing to Run:

Flush Inject Port and

Wash Needle

See Note #1

Wait for

Temperature

to Stabilize

See Note #2

Timed

Events

See Note #4

Method Starts for

Injection #1

Flush Inject Port and

Wash Needle

Vial Present?

Yes

Set

Temperature(s)

Do Sample Prep

(Pipetting, Mixing, etc.)

Method Setup

Execute INIT Step

Load Loop

Method Time 0.0

Inject

Start Cycle Timing

Method Times x.x to x.x

Set TTLs/Relays, etc.

Method Finished

See Note #3

End of Cycle Time

See Note #1

Sample Prep

Overlap

Continue

See Note #2

Method Starts for

Injection #2

Flush Inject Port and

Wash Needle

Vial Present?

Yes

Set

Temperature(s)

Do Sample Prep

(Pipetting, Mixing, etc.)

Wait for Previous

Method to Finish

Wait for Continue

Command

Method Setup

Execute INIT Step

Load Loop

Wait for Cycle Time

Method Time 0.0

Inject

Method Times x.x to x.x

Set TTLs/Relays, etc.

Method Finished

Sample Prep

Overlap

Method Starts for

Injection #3

Continue as Shown for Method #2 Until

Last Scheduled

Injection Completed

Flush Inject Port and

Wash Needle

Schedule Ends

Notes:

1. The schedule either stops and waits or skips to the next vial, depending on the action selected in the schedule.

2. If multiple injections from the same vial are specified in the schedule, the sample prep steps are perfomed once only (before the first injection).

3. The continue command can be sent via the front panel Hold/Run button or a TTL input signal.

4. Use of cycle time is optional.

Figure 2-19. Order of Events During a Schedule Run (Wait Enabled)

38 Doc. 065051-03 1/08

Schedule Started

Preparing to Run: Flush Inject Port and Wash Needle

2 • Description

See Note #1

Wait for

Temperature

to Stabilize

See Note #2

Timed Events

Method Starts for

Injection #1

Flush Inject Port and

Wash Needle

Vial Present?

Yes

Set

Temperature(s)

Do Sample Prep

(Pipetting, Mixing, etc.)

Set Up Method

Execute INIT Step

Load Loop

Method Time 0.0

Inject

Start Cycle Timing

Method Times x.x to x.x

Set TTLs/Relays, etc.

Method Finished

End of Cycle TimeSee Note #3

See Note #1

Sample Prep

Overlap

See Note #2

Yes

Method Starts for

Injection #2

Flush Inject Port and

Wash Needle

Vial Present?

Yes

Set

Temperature(s)

Do Sample Prep

(Pipetting, Mixing, etc.)

Wait for Previous Method to Finish

Setup Method

Execute INIT Step

Load Loop

Wait for Cycle Time

Method Time 0.0

Inject

Method Times x.x to x.x

Set TTLs/Relays, etc.

Method Finished

Sample Prep

Overlap

Method Starts for

Injection #3

Continue as Shown for Method #2 Until

Last Scheduled

Injection Completed

Flush Inject Port and

Wash Needle

Schedule Ends

Notes:

1. The schedule either stops and waits or skips to the next vial, depending on the action selected in the schedule.

2. If multiple injections from the same vial are specified in the schecule, the sample prep steps are perfomed once only (before the first injection).

3. Use of cycle time is optional.

Figure 2-20. Order of Events During a Schedule Run (Wait Disabled)

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AS Autosampler Operator’s Manual

2.7.7 Understanding the Status Display During a Run

During runs, the MAIN STATUS and DETAIL STATUS screens display the operations currently being performed. Because of sample prep overlap, information on multiple injections may be displayed simultaneously at certain points in the schedule. For example, in Figure 2-21 injecting the sample and performing the timed events for the first injection from vial 1 while preparing for the second injection.

, the AS is

Method Timed Events Clock for Injection #1

DETAIL

VIAL#:

INJ#:

METHOD: TRAY:

TRAY:

LOCAL

Help Prompt

1 1/3

12.53

12mL

20 °C

Figure 2-21. Detail Status Screen Example

Preparing for Injection #2

FLUSHING

TIME:

VOL:

LOOP: NORMAL PARTIAL

SCH 1

12.5 25

100

LINE 10

Elapsed Time Since Injection #1

min uL

INJ VLV

TTL1 TTL2 RLY1 RLY2

0 1 0 0 L

Notes

• The INJ# field is updated when the AS loads the loop for the next

injection.

• The TIME field and the METHOD timed events clock are reset to zero at

injection.

• The TIME field counts up from the injection until the next injection

occurs.

• The METHOD timed events clock counts up until all timed events have

been performed for an injection. The clock then stops and is no longer displayed.

40 Doc. 065051-03 1/08

2 • Description

2.7.8 Operating Events During Flushing and Priming

The AS automatically flushes the inject port and washes the outside of the needle at the following times:

• At the beginning and end of a Chromeleon or Chromeleon Xpress

sequence or a front panel schedule

• Before each injection, while the schedule or sequence is running

Flushing can also be initiated manually when the AS is idle, or as a step in the sample prep phase of a Chromeleon or Chromeleon Xpress program or front panel method.

Manual priming of the flush and reagent lines must be initiated after refilling a reservoir, or after the AS has been idle for a period of time. See

Section B.3.13

Flushing

The AS performs the following operations during flushing:

for instructions on priming the liquid lines.

• The sampling needle arm moves to the waste port and expels any

fluid from the needle.

• The sampling needle arm moves to the inject port and delivers a

volume of fluid for flushing the inject port.

• The sampling needle arm moves to the flush port and delivers a

volume of flush fluid. This washes the outside of the needle.

The flush reservoir and sample syringe are always used to perform the flush.

The needle is washed with a factory-set volume. Select the volume of liquid for flushing or priming on the Chromeleon or Chromeleon Xpress Control panel (see Figure 2-13

Section C.7

syringe volume, the volume is reached.

If you are using Chromeleon or Chromeleon Xpress to control the autosampler, you can specify separate syringe speeds for aspirating and dispensing liquid during flushing or priming. Specify the speeds on the Control panel or in the program (see Section 3.7

Doc. 065051-03 1/08 41

). If you specify a flush volume that is greater than the sample

AS performs multiple syringe strokes until the

) or on the FLUSH/PRIME screen (see

AS Autosampler Operator’s Manual

eedle

Priming

The AS performs the following operations during priming:

• The sampling needle arm moves to the waste port and expels any

fluid from the needle.

• The sampling needle arm moves to the flush port and delivers a

volume of flush fluid. This primes the lines from the flush reservoir to the syringe and through the sampling needle assembly (see

Figure 2-22

When the sample preparation option is installed, additional priming is required to prime the lines from the reagent reservoir(s) to the sample syringe (see Figure 2-24

). See Section B.3.13 for instructions on priming

the liquid lines.

2.7.9 Sample and Prep Syringe Fluid Schematics

Figure 2-22 shows the fluid connections to the sample syringe and flush

reservoir when no sample prep option is installed.

Sample Syringe

Valve

Sampling Needle

ssembly

Sample Syringe

Flush Reservoir

Sampling N

Figure 2-22. Fluid Schematic for Sample Syringe

42 Doc. 065051-03 1/08

2 • Description

Needle Positi

Flush Positi

Figure 2-23 shows the flow paths through the sample syringe valve ports

for the two valve positions:

• In the needle position, flow is between ports A–B, C–D, and E–F.

• In the flush position, flow is between ports A–F, B–C, and D–E.

To Flush

Reservoir

To Sample

ringe

To Sampling

Needle

To Flush

Reservoir

To Sample

ringe

To S amp lin g

Needle

Figure 2-23. Sample Syringe Valve Port Flow Path Schematics

Figure 2-24

shows the fluid connections to the sample syringe, flush reservoir, sample prep syringe, and reagent reservoirs when the sample prep option is installed.

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AS Autosampler Operator’s Manual

(

)

ldi

To R

Reagent B (Optional)

Reagent C (Optional)

(*Port E not connected)

(Back Fitting)

Prep

Syringe

Valve

Reagent D

Optional

(Behind

Port S*)

(Front Fitting)

Prep Syringe

(Front Fitting)

(Back Fitting)

Reagent A

eagent Ho

ng Loop

(To/From Needle)

Sample

Syringe

Valve

Flush Reservoir

Sample Syringe

Sampling Needle Line

Figure 2-24. Fluid Schematic for Sample Syringe with Optional Prep Syringe

The prep syringe valve has two rows of port fittings. Figure 2-25 shows a detailed view of the front and back port connections.

Samplin Needle

To P ort A on

Sample Syringe

Front

Fittings

To P rep S

ringe

To Reservoir C

To R eser voir D

To Port F on Sample Syringe

eservoir

Back

Fittings

Not

Connected

To P ort B on Sample Syringe

To Reservoir A

Notes on Valve Port Flow Paths:

• Ports S and F are

always connected.

• Port L connects to

the port selected on the back fittings.

Figure 2-25. Prep Syringe Valve Port Connections

44 Doc. 065051-03 1/08

3 • Operation and Maintenance

3.1 Getting Ready to Run

3.1.1 Turn On the Power

Press the power switch in the lower left corner of the front door to turn on the AS power. When the power is turned on, a series of screens are displayed and the autosampler performs a power-up sequence.

• The COPYRIGHT NOTICE screen is displayed briefly, and then the

POWER-UP screen appears.

• While the POWER-UP screen is displayed, the AS performs

initialization procedures and a series of diagnostic tests. If one of the tests fails, testing stops and the displayed. See Section C.9.2

• When the initialization procedures are complete and the AS has

passed the diagnostics, the a few seconds (or if you press a button), the The power-up sequence is then complete.

DIAGNOSTIC TEST screen is

if this occurs.

INSTALLED OPTIONS screen appears. After

MAIN screen is displayed.

After power-up, you can display the

INSTALLED OPTIONS screens by selecting these options on the MENU of SCREENS

Doc. 065051-03 1/08 45

. See (see Section C).

POWER-UP, DIAGNOSTIC TEST, and

AS Autosampler Operator’s Manual

)

3.1.2 Fill the Vials or Wells and Load the Sample Tray

Filling Vials

1. Fill the vials to the vial shoulder or lower (see

Figure 3-1

). Do not fill to the

Never fill above this point

top.

During sampling, make sure the needle’s vent is not immersed in liquid.

All vial sizes

For 10 mL vials only: If necessary, raise the needle height inside the vial to position the vent above the liquid. You can set the needle height in a Chromeleon or Chromeleon Xpress program (see Section 3.2.2

) or in a

Needle vent (Do not immerse

front panel program (see

page 292

10 mL vial

Figure 3-1. Maximum Level to Fill Vials

2. Screw a cap onto each vial and tighten. To prevent septa from falling into the vials when they are pierced by the sampling needle, make sure the cap septa are pushed fully into the caps and the caps are securely tightened.

NOTE For 0.3 mL and 1.5 ml vials (any type), Dionex

recommends using vial caps with slit septa only (see

Section 2.3.5

3. Load the vials in the tray in the order required for the Chromeleon or Chromeleon Xpress sequence (see Section 3.2.2 panel schedule (see Section D.6

) or for the front

4. If the sample temperature control option is installed, position tray covers (P/N 061010) over any empty positions on the tray (see

46 Doc. 065051-03 1/08

Figure 3-2). This ensures temperature stability and minimizes

condensation, especially at low temperature set points.

Tray Covers

(for empty

positions)

Sample Vials

3 • Operation and Maintenance

Figure 3-2. Installing Tray Covers (Sample Temperature Control Option Only)

5. Install the tray in the autosampler compartment.

6. Close the autosampler compartment door. Make sure the door remains closed during operation.

If the door is opened during operation, the sampling arm stops immediately. If a schedule is running, it is aborted. To safely open the door during operation, press the Door button (see Section 3.9 details).

Doc. 065051-03 1/08 47

for

AS Autosampler Operator’s Manual

Filling Well Plates

1. Put on powder-free latex gloves before handling well plates and covers to minimize sample contamination.

2. Rinse the well plates and covers thoroughly with deionized water before use to remove any contaminants.

3. Dry the well plates completely to prevent any changes in sample concentration. You can use 690 kPa (100 psi) nitrogen gas to assist in drying.

4. Use a pipet to fill the wells in the well plate. In general, pipet the amount of sample to be aspirated plus 70 details about the sample volumes aspirated for each type of injection.

The table below lists the maximum fluid volumes for each well size and the maximum sample volumes available when the well is filled:

μL. See Section 3.6.4 for

Well Plate Type Maximum Volume

per well (

0.45 mL micro-well with conical (V-shaped) wells

0.5 mL micro-well with round (U-shaped) wells

2.0 mL deep-well 1800 1700

a. Assumes the well contains the maximum volume and the needle height is 2 mm (the default).

NOTE Changing the needle height is not recommended

320 250

μL)

Sample Available

μL)

(

5. Place a cover over the well plate and firmly press the indentations on the cover into each well. This creates a tight seal and prevents sample evaporation.

NOTE Replace covers after each use to prevent

contamination and sample evaporation. Well plates can be reused.

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3 • Operation and Maintenance

6. Orient the well plate with well A1 at the front-left corner and set the well plate in the well plate tray (see Figure 3-3

7. Fill another well plate and install it on the right side of the tray. The two plates must be of the same type. You can install either two deepwell plates or two micro-well plates (0.50 mL or 0.45 mL).

8. Position the clamp in the well plate tray with the thumbscrews in the center of the tray and the arms across both well plates.

Always install two well plates in the tray to ensure even pressure from the clamp. Make sure that the arms on the clamp are positioned over the small “lip” on the front and back sides of both well plates. This ensures that the clamp presses down on each well plate evenly and the well plate doesn’t move during sampling.

9. Tighten the thumbscrews on the clamp snugly.

Clamp Thumbscrews

A1 Wells (in left and right well plates)

Figure 3-3. Well Plate Tray with Two Well Plates Installed

10. Install the tray in the autosampler compartment.

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11. Close the autosampler compartment door. Make sure the door remains closed during operation.

If you open the door during operation, the sampling arm stops immediately. If a schedule is running, it is ended. To safely open the door during operation, press the Door button (see Section 3.9 details).

Guidelines for improving precision when sampling from well plates:

• To avoid contaminants, rinse well plates and covers thoroughly with

deionized water before use.

• Dry well plates completely to prevent any changes in sample

concentration. You can use 690 kPa (100 psi) nitrogen gas to assist in drying.

• Follow the sample volume guidelines (see Step 4) when pipetting

sample into the wells.

• Check the orientation of the cover to make sure the notches in the

corners of the cover and well plate are aligned.

for

• Press the well plate cover tightly into each well.

• To avoid contaminants and evaporation, do not reuse covers. The

small hole created when the sampling needle pierces the cover allows slight, but noticeable, evaporation of sample. This is especially true when samples must wait for several hours before being run.

• If sample evaporation does occur, try reducing the tray temperature.

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3.2 Selecting Computer or Front Panel (Local) Control

When the AS is powered up, it is always in front panel (Local) control. The MAIN and

DETAIL screens indicate this by displaying LOCAL in the operating mode field.

See the example in Figure 3-4

IDLE

TIME

VOL

TIME

VOL

25.011

METHOD: 0

DIRECT CONTROL

25.011

METHOD: 0 DIRECT CONTROL

min

MAIN and DETAIL screens

min

VIAL#

INJ#

TRAY:

LOCAL

Help Prompt

To select computer control, follow the steps in Section 3.2.1 a Chromeleon or Chromeleon Xpress Control panel.

When the AS is under computer control, the front panel indicate this by displaying and control mode fields. See the example in Figure 3-5

VIAL#

INJ#

TRAY:

LOCKED RMT

Help Prompt

5 99.9

°C

- -

Figure 3-4. Main Screen—Local Control

LOCKED RMT and DIRECT CONTROL in the operating

5 99.9

°C

- -

Figure 3-5. Main Screen—Computer Control

to connect the AS to

To return the AS to Local mode, clear the Connect check box on the Chromeleon or Chromeleon Xpress Control panel (see Figure 3-6

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3.2.1 Connecting to Chromeleon or Chromeleon Xpress

1. Start the Chromeleon Server, if it is not already running:

• If the Chromeleon Server icon on the taskbar is crossed out in red

, the Server is not running. Start the Server by right-clicking

the icon and selecting Start Server. When the server is running

(and data acquisition is not occurring), the icon is gray

• If the Server Monitor icon is not on the taskbar, click Start on the

taskbar and select All Programs (or Programs, depending on the operating system) > Chromeleon > Server Monitor. Click Start to start the server.

2. Start the Chromeleon or Chromeleon Xpress client:

a. Click Start on the Windows taskbar and select All Programs (or

Programs) > Chromeleon > Chromeleon to start the Chromeleon client.

b. If Chromeleon is installed, select View > Default Panel Tabset

or click the corresponding toolbar button to display the panel tabset.

NOTE If Chromeleon Xpress is installed, starting the

client automatically displays the panel tabset.

c. To display the AS Control panel, select the Autosampler tab on

the panel tabset (see Figure 3-6

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d. Verify that the Connect check box is selected. If it is not, click

the box to connect the AS to Chromeleon or Chromeleon Xpress.

Figure 3-6. AS Control Panel

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3.2.2 Running a Sequence of Injections in Chromeleon or Chromeleon Xpress

This section provides a brief description of the steps required to set up and run the autosampler with Chromeleon or Chromeleon Xpress. For detailed instructions, refer to the Chromeleon or Chromeleon Xpress Help.

1. Create a program that specifies the functions the autosampler (and

other system instruments) should perform at a specific time. Programs can be created automatically (with the help of a software wizard) or manually (by editing an existing program).

In addition to commands for loading and injecting the sample, a program for the AS can include commands for setting up the autosampler (syringe speed, sample needle height, etc.) and for preparing the sample before injection (pipetting, mixing, flushing, etc.).

2. Create a sequence (a list of injections). For each injection, specify the

vial or well position (see Section 3.4 program to run, and other parameters. Sequences can be created automatically (with the help of a software wizard) or manually (by editing an existing sequence).

), the injection volume, the

3. Load the sequence into a batch.

4. Start the batch.

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3.3 Default Operating Parameters

The Table 3- 1 lists the factory-set values for AS operating parameters:

Operating Screen Parameter Default Value

Method Sample Prep Step 1 wait step only

Method Setup Cycle Time

Sample Needle Height Sample Temperature Wait for Temp Stable

Method Timed Events

Schedule Missing Vial Action

System Parameters Wait

Flush/Prime Flush Volume

Time=Init

Inject Valve

TTL/Relays

Time=0.00

Inject Valve

TTL/Relays

Vial # Start/End Injections per Vial Injection Volume Method Number

Syringe Speeds

Cut Volume Sample Mode Inject Port Volume

Air Bubble Volume (not user changeable)

Flush Reservoir Flush Syringe Prime Volume Prime Reservoir Prime Syringe

Off 2 mm 20 °C No

Load (for simultaneous or sequential injection, both valves at Load position) All off (0)

Inject (for simultaneous or sequential injection, both valves at Inject position) All off (0)

Stop 1/1 1

25.0 μL; 1 mL for simultaneous injection 1

Enabled Refer to Section 3.7 syringe speeds. Refer to Section D.7 front panel control syringe speeds. 10 μL (see Section 3.6.3 Normal varies (each inject port is individually calibrated) 8 μL if the sample syringe is <5 mL; 60 μL if a 5 or 10 mL syringe is installed

250 μL Flush (always) Sample (always) 400 μL Flush Sample

for Chromeleon control

for

for details)

Table 3-1. Default Operating Parameters

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3.4 Specifying Sample Positions

When creating a schedule or sequence of injections, use the following guidelines to specify sample positions in the vial or well plate sample tray.

3.4.1 Vial Tray Sample Positions

For the 49-position vial tray, vial position 1 is in the lower-left corner of the tray and vial position 49 is in the upper-right corner (see

Figure 3-7

43 49

1234

For the 100-position vial tray, vial position 1 is in the lower-left corner of the tray and vial position 100 is in the upper-right corner (see

Figure 3-8

Figure 3-7. 49-Position Vial Tray

1234

567

100

8910

Figure 3-8. 100-Position Vial Tray

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3 • Operation and Maintenance

3.4.2 Well Plate Tray Sample Positions

NOTE Chromeleon or Chromeleon Xpress is required

to sample from a well plate.

Sample positions in the well plates are specified with a two letter and one number identifier (for example, LB8):

• The first letter (either L or R), specifies the left or right well plate (see

Figure 3-9

• The second letter (A through H) and the number (1 through 12)

specify the well position in the well plate. Well letters and numbers are printed on well plates and well plate covers.

For example, well plate position LA1 identifies the well located at position A1, in the well plate installed on the left side of the AS well plate tray. When the well plate is installed in the tray, position LA1 is in the lower-left corner of the tray (see Figure 3-9

RH12

V10

Position

LA1

LH12

LA1

12 3456789101112

ABCDEFG

1234567 8910

RA1

12 3456789101112

ABCDEFG

Figure 3-9. Well Plate Tray: Well Plate and Vial Positions

NOTE To specify the position of a 1.5 mL vial installed in the

well plate tray, precede the vial position number with a V (V1, V2,…V10) (see Figure 3-9

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3.5 Specifying Injection Volumes

The range of injection volumes the AS can deliver to the injection valve depends on the size of the sample syringe installed on the AS and on the sampling mode selected on the

SYSTEM PARAMETERS screen (see Section 3.6.2).

Table 3- 3

Syringe Size(s) Sampling Mode Injection Volume

100–1,000 μL Normal and Sequential 1.0–1,000 μL

5 mL Normal and Sequential 1.0–4,700 μL

10 mL Normal and Sequential 1.0–8,000 μL

lists the volume range for each syringe size and sampling mode.

Concentrate and Sequential Concentrate

Simultaneous 1,000–4,700 μL

Concentrate and Sequential Concentrate

Simultaneous 1,000–8,000 μL

Table 3-2. Summary of Injection Volume Ranges

100–1,000 μL

100–4,700 μL

100–8,000 μL

You can enter the injection volume in the Chromeleon or Chromeleon Xpress sequence (see Figure 3-10

Section D.3

). For volumes less than 100 μL, specify the value in 0.1 μL

and Figure 3-11) or in the front panel schedule (see

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increments. For volumes of 100 μL or greater, specify the value in 1 μL increments.

Figure 3-10. Sequence Wizard: Specifying the Injection Volume

Figure 3-11. Sequence Control Panel: Specifying the Injection Volume

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3.6 Sample Injection Types

3.6.1 Overview

The type of injection the AS performs for a particular sample depends on several factors, including the number of systems the AS is connected to and whether a sample loop or concentrator column is installed on the injection valve. Table 3- 3

Injection Type Description

Full Full-loop injection to one system

Partial Partial-loop injection to one system

Partial LS Partial-loop, limited-sample injection to one

Large Loop Large-loop injection to one system

Simultaneous Full-loop injections to two systems simultaneously

Concentrate Concentrate injection to one system

Sequential Full Full-loop injections to two systems alternately

Sequential Partial Partial-loop injections to two systems alternately

Sequential Partial LS Partial-loop, limited-sample injections to two

Sequential Large Loop Large-loop injection to two systems alternately

Sequential Concentrate Concentrate injections to two systems alternately

summarizes all of the available injection types.

system (no extra sample is aspirated)

systems alternately

Table 3-3. Summary of Available Injection Types

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During an injection, the injection type is displayed on the Chromeleon or Chromeleon Xpress Control panel (see Figure 3-12

STATUS screen (see Figure 3-13).

Figure 3-12. Injection Type Shown on Control Panel

) and on the DETAIL

DETAIL

VIAL#:

INJ#:

METHOD:

TRAY:

1 1/3 01 P10 mL

20 °C

LOCAL

Help prompt

Figure 3-13. Injection Type Shown on Detail Status Screen

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INJECTING

TIME:

VOL:

0.0

TRAY:

LOOP:

25.0

NORMAL FULL

SCH 1

LINE 1

min uL

TTL1

TTL2

RLY1

RLY2

INJ VLV

0 1 0 0 L

AS Autosampler Operator’s Manual

3.6.2 Determining the Injection Type

To determine the injection type performed by the AS, first select the sample mode on the

Menu, 5, and 5 to go to the SYSTEM PARAMETERS screen.

Press

METHOD SYRINGE SPEED:

CUT SEGMENT VOLUME:

INJECT PORT VOLUME:

FLUSH SYRINGE SPEED:

Help Prompt

SYSTEM PARAMETERS screen (see Figure 3-14).

SYSTEM PARAMETERS

SAMPLE MODE:

Figure 3-14. System Parameters Screen

The sample mode corresponds to the AS configuration. The following sample modes are available:

• Normal—Sample is delivered to a single system; a sample loop is

installed on the injection valve.

WAIT:

ENABLED 3 10

NORMAL S1 43 4

uLS2

• Concentrate—Sample is delivered to a single system; a concentrator

column is installed on the injection valve.

• Simultaneous—Sample is delivered to two systems simultaneously;

a sample loop is installed on both injection valves.

• Sequential—Sample is delivered to two systems alternately; a

sample loop is installed on both injection valves.

• Sequential concentrate—Sample is delivered to two systems

alternately; a concentrator column is installed on both injection valves.

NOTE The SAMPLE MODE must be selected on the

SYSTEM PARAMETERS screen. It cannot be selected

from Chromeleon or Chromeleon Xpress.

If normal or sequential mode is selected, you must also enter additional parameters to specify the type of sample loop injection performed (see

Section 3.6.3

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3.6.3 Determining the Sample Loop Injection Type

When the AS is in normal or sequential mode, it can perform the following types of sample loop injections:

• Full-loop—A full loop volume is injected.

• Large-loop—A full (large) loop volume is injected. Large loops are

200.0 to 7000.0

μL in volume.

• Partial-loop—A partial loop volume is injected.

• Partial-loop, limited-sample—A partial loop volume is injected; no

extra sample is aspirated from the vial or well.

NOTE When the AS is in simultaneous mode, it performs

only full-loop injections.

The following three parameters are used to determine the type of sample loop injection the AS performs: cut volume, injection volume, and loop size (see Table 3-4)

Criteria for Determining the Sample Loop Injection Type

Injection volume ≥ loop size (loop size = 2.5 to 199.9 μL)

Injection volume ≥ loop size (loop size = 200.0 to 7000.0 μL)

Injection volume < loop size (cut volume > 0)

Injection volume < loop size (cut volume = 0)

Table 3-4. Determining the Sample Loop Injection Type

Type of Sample Loop Injection Performed

Full-loop

Large-loop

Partial-loop

Partial-loop, limited-sample

The cut volume, which is used only for partial-loop and partial-loop, limited-sample injections, specifies a volume of sample to be “cut” (or omitted) from each side of the sample during the injection process. Using a smaller cut volume minimizes the amount of total sample aspirated from the vial or well. However, better precision is achieved with a larger cut volume. If you enter a cut volume of 0, the AS draws only the injection volume from the vial or well.

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1. Enter the size of the installed loop on the PLUMBING CONFIGURATION

screen (see Figure 3-15

CONFIGURATION screen.

SAMPLE SYRINGE VOLUME:

SAMPLE SYRINGE CONFIG:

PREP SYRINGE VOLUME:

PREP SYRINGE CONFIG:

Help Prompt

Figure 3-15. Plumbing Configuration Screen

2. Enter the injection volume in the Chromeleon or Chromeleon Xpress

sequence (see Figure 3-16

Section D.3

). Press Menu, 5, and 3 to go to the PLUMBING

PLUMBING CONFIGURATION

LOOP SIZE V1: LOOP SIZE V2:

250

READY

) or in the front panel schedule (see

Figure 3-16. Sequence Wizard: Specifying the Injection Volume

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3 • Operation and Maintenance

• If you enter an injection volume equal to or larger than the

installed loop size, the AS performs a full-loop (or large-loop) injection.

• If you enter an injection volume smaller than the loop size, the

AS performs a partial-loop or partial-loop, limited-sample injection.

3. Enter the cut volume program (see Figure 3-17 (see Section C.6.5

NOTE The cut volume is used only for partial-loop or

partial-loop, limited-sample injections. For full-loop or large-loop injections, the cut volume is ignored. See page 63

in the Chromeleon or Chromeleon Xpress

) or on the SYSTEM PARAMETERS screen

for details about cut volumes.

Figure 3-17. Program Wizard: Specifying the Cut Volume

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3.6.4 Guidelines for Sample Volumes Used, Loop Sizes, and Injection Volumes

Sample Volumes Used for Each Type of Sample Loop Injection

Table 3- 5 shows the volume of sample the AS aspirates from the sample

vial or well plate well and the volume of sample injected onto the column.

Injection Type Sample Volume Aspirated

from Vial or Well

Full-loop (2.5–16.9 μL loop)

Full-loop (17.0–199.9 μL loop)

Large-loop (200.0–7000.0 μL loop)

Partial-loop Injection volume + 2 × cut

Partial-loop, limitedsample

Table 3-5. Sample Volumes Used for Each Injection Type

See Section 3.6.3

for descriptions of sample loop injection types and the

Injection volume or

2.5 × loop size + 25 μL (whichever is greater)

Injection volume or 4 × loop size (whichever is greater)

Injection volume or 1 × loop size + 500 μL (whichever is greater)

volume

Injection volume Injection volume

Sample Volume Injected

Loop size

Injection volume

criteria used to determine which type of injection is performed.

The following tables give several examples for full-loop, large-loop, and partial-loop injections. The examples show the amount of sample used and injected for various loop sizes and injection volumes.

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3 • Operation and Maintenance

Full-Loop Sample Volume Examples

For full-loop injections, if the injection volume is more than 4 times the loop size, the full injection volume is aspirated. The excess aspirated sample is flushed through the loop, but the amount injected is equal to the loop size.

Installed Loop Size (

μL)

Injection Volume from Chromeleon Sequence (μL)

Sample Aspirated from Vial or Well

μL)

(

Sample Volume Injected onto Column (μL)

10 10 40 10

10 35 40 10

10 60 60 10

25 25 100 25

25 75 100 25

25 150 150 25

Table 3-6. Full-Loop Sample Volume Examples

a. Excess aspirated sample is used to flush the loop before and after injection.

Large-Loop Sample Volume Examples

For large-loop injections, if the injection volume exceeds the loop size +

μl, the full injection volume is aspirated. The excess sample aspirated

500 is flushed through the loop, but the amount actually injected is equal to the loop size.

Installed Loop Size (

μL)

Injection Vol ume from Chromeleon Sequence (μL)

Sample Aspirated from Vial or Well

μL)

(

Sample Volume Injected onto Column (μL)

500 500 1,000 500

500 700 1,000 500

500 1,100 1,100 500

Table 3-7. Large-Loop Sample Volume Examples

a. Excess aspirated sample is used to flush the loop before and after injection.

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Partial-Loop and Partial-Loop, Limited-Sample Volume Examples

For partial-loop injections, the amount of sample aspirated is equal to the injection volume + 2 to the injection volume.

For partial-loop, limited-sample injections, the amount of sample aspirated and injected is equal to the injection volume.

times the cut volume. The amount injected is equal

Installed Loop Size (

μL)

25 10

25 20

25 10

25 20

Table 3-8. Partial-Loop and Partial-Loop, Limited-Sample

a. The cut volume specifies a volume of sample to be “cut” (or omitted) from each side of the sample during the injection process. See Section 3.6.3 details.

Maximum Sample Loop Sizes and Injection Volumes

Injection Vol ume from Chromeleon Sequence (μL)

Sample Volume Examples

Cut Volu me

μL)

(

Sample Aspirated from Vial or Well (μL)

Sample Volume Injected onto Column (μL)

20 10

40 20

10 10

20 20

for

The maximum loop size allowed depends on which sample syringe and sampling needle assembly are installed. If the sample loop is too large, the sample volume drawn exceeds the syringe/sampling needle assembly volume, causing sample to enter the flush bottle.To prevent sample from contaminating the flush bottle, the sample loop size must not exceed the values shown in Tab le 3-9

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Table 3- 9 also lists the maximum injection volume that can be specified in

the Chromeleon or Chromeleon Xpress sequence or in the front panel schedule.

Sample Syringe

≤1,000 μL1.2mL 500μL 5mL 8.5mL 4,000μL 10 mL 8.5 mL 7,000 μL

Table 3-9. Maximum Sample Loop Sizes and Injection Volumes

a. Requires a 1.5 mL vial or a 2 mL deep well plate. b. Requires a 10 mL vial.

Notes

Sampling Needle Assembly

Maximum Sample Loop Size

Maximum Injection Volume

1,100 μL 4,700 μL 8,000 μL

• To minimize the backpressure applied to the sample syringe pump,

use the largest ID tubing possible to make the injection loop.

• If a 1.5 mL vial is used, the maximum loop size is 500 μL and the

maximum injection volume is 1100

μL, regardless of the syringe size.

• For syringes that are 1,000 μL or smaller, the maximum loop size and

injection volume requires the syringe to aspirate from the sample vial multiple times. This can result in reduced precision. For maximum precision, use a loop size and injection volume that require only a single sample aspiration.

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)

3.6.5 Normal, Full-Loop Injections

For a normal, full-loop injection, the AS draws a volume of sample from the sample vial or well plate well. The volume of sample drawn depends on the loop size and the specified injection volume (see Section 3.6.4 details). A portion of the sample is flushed through the loop, the loop is filled, and an additional portion remains before the loop. The sample in the loop is then injected onto the column (see Figure 3-18

Flush

Air

Sample Syringe

Sample

for

1. Draw air

2. Draw sample

3. Draw air again to isolate the sample

Waste Waste Waste

Pump

INJECT

C Column

4. Prepare to load sample (excess sample used)

Pump

LOAD

C Column

5. Deliver sample to the loop (excess sample sent to waste)

Figure 3-18. Full-Loop Injection Sequence

Pump

INJECT

Column

6. Inject sample onto column. Empty syringe (remaining sample + air bubble

KEY

Flush Air Eluent Sample

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Setting Up a Full-Loop Injection

1. On the SYSTEM PARAMETERS screen (see Section C.6.5), select

NORMAL in the SAMPLE MODE field.

2. In the Chromeleon or Chromeleon Xpress sequence (see Figure 3-19 or in the front panel schedule (see Section D.3

), enter an injection volume equal to or greater than the volume of the loop installed on the injection valve.

)

Figure 3-19. Sequence Wizard: Specifying the Injection Volume

Full-Loop Injection Notes

• Entering an injection volume equal to or greater than the sample loop

size sets up the AS for a full-loop injection. The actual volume of sample injected equals the volume of the sample loop installed.

• If the entered injection volume is more than four times the loop size,

the entire injection volume is delivered during loading of the loop. For example, if a 100 600

μL, then 350 μL is flushed through the loop, 100 μL is loaded

into the loop, and 150

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μL loop is installed and the injection volume is

μL remains before the loop.

AS Autosampler Operator’s Manual

Example Program for Normal Mode

The following example Chromeleon or Chromeleon Xpress program shows the AS commands for sample loading and injecting when the autosampler is in normal mode (full-loop, partial-loop, or partial-loop, limited sample).

0.000 Load ;Switch the inject valve to Load

and load sample onto the loop

Wait CycleTimeState ;Wait for the end of the specified

Cycle Time

Inject ;Switch the inject valve to Inject

and sweep the sample onto the column

Wait InjectState ;Wait for the injection to finish

3.6.6 Normal, Large-Loop Injections

The injection sequence for a normal, large-loop injection is the same as described for a normal, full-loop injection (see Section 3.6.5 when a large loop (200.0 to 7000.0 μL) amount (500

μL) of extra sample from the vial for flushing the loop,

instead of drawing a factor of the loop volume (see Section 3.6.4 details).

is installed, the AS draws a set

). However,

for

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3.6.7 Normal, Partial-Loop Injections

For a partial-loop injection, the AS draws the volume of sample to be injected from the sample vial plus two times the cut volume.The cut volume specifies a volume of sample to be “cut” (or omitted) from each side of the sample during the injection process. details. The middle portion of the sample is positioned in the loop and

3. Draw air again to isolate the sample.

Flush Air

Sample

1. Draw air

injected (see Figure 3-20

Sample Syringe

2. Draw sample (injection volume + 2X cut volume)

See Section 3.6.3 for

4. Draw flush fluid to position sample in loop.

Waste Waste Waste

Pump

INJECT

Column

5. Prepare to load sample (flush fluid + air bubble + cut volume #1 is used)

Pump

LOAD

C Column

6. Deliver sample (inject volume) the loop

Pump

INJECT

C Column

7. Inject sample onto column. Empty sample syringe (cut volume #2 + air bubble)

KEY

Flush Air

Eluent Sample

Figure 3-20. Partial-Loop Injection Sequence

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Setting Up a Partial-Loop Injection

1. On the SYSTEM PARAMETERS screen (see Section C.6.5), select

NORMAL in the SAMPLE MODE field.

2. In the Chromeleon or Chromeleon Xpress sequence (see Figure 3-19 or in the front panel schedule (see Section D.3

), enter an injection volume that is less than the volume of the sample loop installed on the injection valve.

3. In the Chromeleon or Chromeleon Xpress program (see Figure 3-21 or on the

SYSTEM PARAMETERS screen, enter a cut volume greater

than zero.

)

Figure 3-21. Program Wizard: Specifying the Cut Volume

NOTE For the best accuracy when performing partial-loop

injections, specify an injection volume equal to half the loop size or less.

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3 • Operation and Maintenance

Example Partial-Loop Injection Parameters:

Sample loop size: 100 Injection volume: 50 Cut volume: 20

To perform the injection, the AS aspirates 90

μL

μL from the sample vial

(the inject volume plus double the cut volume) and delivers it to the injection valve. Then, enough flush fluid is drawn into the syringe to position the sample. One cut volume (20 next 50

μL is loaded into the loop. At injection, 50 μL is injected.

After injection, the remaining cut volume (20

μL) flows to waste and the

μL) flows to waste.

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)

3.6.8 Normal, Partial-Loop, Limited-Sample Injections

For a partial-loop, limited-sample injection, the AS draws only the volume to be injected from the sample vial. The AS positions the sample in the loop and injects it onto the column (see Figure 3-22

Flush Air

Sample Syringe

Sample

1. Draw air

2. Draw sample (injection volume)

Waste Waste Waste

Pump

INJECT

Column

5. Prepare to load sample (flush fluid + air bubble is used)

Pump

LOAD

Column

6. Deliver sample (injection volume) the loop

3. Draw air again to isolate the sample.

7. Inject sample onto column. Empty sample syringe (air bubble + an

Figure 3-22. Partial-Loop, Limited-Sample Injection Sequence

Pump

INJECT

C Column

excess fluid

4. Draw flush fluid for cleaning inject port.

KEY

Flush Air

Eluent Sample

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3 • Operation and Maintenance

Setting Up a Partial-Loop, Limited-Sample Injection

1. On the SYSTEM PARAMETERS screen (see Section C.6.5), select

NORMAL in the SAMPLE MODE field.

2. In the Chromeleon or Chromeleon Xpress sequence (see Figure 3-19 or in the front panel schedule (see Section D.3

), enter an injection volume that is less than the volume of the sample loop installed on the injection valve.

3. In the Chromeleon or Chromeleon Xpress program (see Figure 3-21 or on the

SYSTEM PARAMETERS screen, enter a cut volume of zero.

Example Partial-Loop, Limited-Sample Injection Parameters:

Sample loop size: 100 Injection volume: 50 Cut volume: 0

μL

To perform the injection, the AS aspirates 50

μL

μL from the sample vial

(the inject volume) and then enough flush fluid is drawn into the syringe to position the sample. The 50

μL of sample is positioned just

before the loop and is then loaded onto the loop and injected.

)

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)

3.6.9 Simultaneous Injections

The AS must be in simultaneous mode to perform a simultaneous injection (see Section 3.6.2 volumes of sample to two injection valves (see Figure 3-23 injection is a full-loop size.

Flush

Air

Sample Syringe

Sample

1. Draw air 3. Draw air again to

2. Draw sample (inject volume)

). In simultaneous mode, the AS delivers equal

). Each

isolate the sample

Waste

Pump

Inject Valve #1

LOAD

Column

Waste

Pump

Inject Valve #2

4. Deliver 1/2 of sample to the loop of each inject valve

LOAD

Column

Waste Waste

Pump

INJECT

Inject Valve #1

5. Inject sample simultaneously onto both system’s columns. Empty sample syringe (remaining

C Column

Column

Pump

INJECT

Inject Valve #2

KEY

Flush Air Eluent Sample

sample + air bubble

Figure 3-23. Simultaneous Injection Sequence

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3 • Operation and Maintenance

Setting Up a Simultaneous Injection

The AS must be equipped with the simultaneous injection option in order to perform simultaneous injections. The following setup steps are required:

• Follow the instructions in Section B.3.5 to connect the AS to two

injection valves (either in the autosampler compartment or two IC systems). In this configuration, the inject port tubing is connected to a Y-connector and the legs of the Y are connected to the inject valves in both systems (see Figure 3-23

). The tubing from the Y to each valve

must be the same length.

• Install a sample syringe of 5,000 or 10,000 μL.

• Enter the sample syringe size of 5,000 or 10,000 μL on the

PLUMBING CONFIGURATION screen (see Section C.6.3).

• On the SYSTEM PARAMETERS screen (see Section C.6.5), select

SIMULTANEOUS in the SAMPLE MODE field.

• In the Chromeleon or Chromeleon Xpress sequence (see Figure 3-19)

or in the front panel schedule (see Section D.3 volume that is large enough to allow flushing the inject port, flushing the tubing from the inject port to each valve (25 tubing), plus enough to flush and fill both injection loops (this is about 5 times the combined loop sizes).

), enter an injection

μL per inch of

Example Simultaneous Injection Parameters:

μL = inject port volume

μL = 20 inches of connector tubing (combined from both

500 systems) x 25

625

μL = 5 x (100 μL loop #1 + 25 μL loop #2)

total = 1170

μL

The specified injection volume is loaded into the sample syringe and then delivered equally to the two sample loops.

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Example Program for Simultaneous Mode

The following example Chromeleon or Chromeleon Xpress programs show the AS commands for sample loading and injecting when the autosampler is in simultaneous mode. Commands differ slightly, depending on where the valves are installed.

Example Simultaneous Injection Program: Two Valves Installed in the AS

0.000 Sampler_InjectValve.LoadPosition ;Switch the first inject

valve to Load

Sampler_InjectValve_2.LoadPosition ;Switch the second

inject valve to Load

Load ;Load sample onto the

loop

Wait CycleTimeState ;Wait for the end of the

specified Cycle Time

Sampler_InjectValve.InjectPosition ;Switch the first inject

valve to Inject

Sampler_InjectValve_2.InjectPosition ;Switch the second

inject valve to Inject

Inject ;Sweep the sample onto

the column

Wait InjectState ;Wait for the injection to

finish

Example Simultaneous Injection Program: Two Valves Installed in an ICS-3000 Detector/Chromatography Module

0.000 InjectValve_1.LoadPosition ;Switch the first inject

valve to Load

InjectValve_2.LoadPosition ;Switch the second

inject valve to Load

Load ;Load sample onto the

loop

Wait CycleTimeState ;Wait for the end of the

specified Cycle Time

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3 • Operation and Maintenance

InjectValve_1.InjectPosition ;Switch the first inject

valve to Inject

InjectValve_2.InjectPosition ;Switch the second

inject valve to Inject

Inject ;Sweep the sample onto

the column

Wait InjectState ;Wait for the injection to

finish

Example Simultaneous Injection Program: Two Valves Installed in an ICS-1000/1500/2000

0.000 Pump_InjectValve.LoadPosition ;Switch the first inject

valve to Load

Pump_InjectValve_2.LoadPosition ;Switch the second

inject valve to Load

Load ;Load sample onto the

loop

Wait CycleTimeState ;Wait for the end of the

specified Cycle Time

Pump_InjectValve.InjectPosition ;Switch the first inject

valve to Inject

Pump_InjectValve_2.InjectPosition ;Switch the second

inject valve to Inject

Inject ;Sweep the sample onto

the column

Wait InjectState ;Wait for the injection to

finish

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3.6.10 Concentrate Injections

The AS must be in concentrate mode to perform a concentrate injection

). In concentrate mode, the AS draws the volume to be

). The example in Figure 3-24 also shows the reagent prime

Flush

Sample

Air

(see Section 3.6.2 injected from the sample vial and delivers it to a concentrator column (see

Figure 3-24

and reagent flush steps for matrix elimination.

Sample Syringe

3. Draw air again to isolate the sample

Reagent

Waste

Pump

Concentrator

LOAD

6. Flush concentrator with reagent (matrix

elimination)

Sample

Eluent

Column

Waste

Pump

7. Inject sample onto column

ReagentAir

Waste Waste

1. Draw air

Pump

Concentrator

LOAD

Column

2. Draw sample (inject volume)

Reagent

Waste

Port

Pump

Concentrator

LOAD

. Prime sampling

Column

needle assembly line

with reagent

KEY

DI Water

Figure 3-24. Concentrate Injection Sequence (Example for Matrix Elimination)

C Column

Concentrator

INJECT

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3 • Operation and Maintenance

Setting Up a Concentrate Mode Injection

1. Install a concentrator column on the injection valve.

2. If the sample syringe volume is greater than 1 mL, install an 8.5 mL sampling needle assembly (P/N 061267). See Section 5.4 installation instructions.

for

3. On the

SAMPLE MODE field.

4. For front panel control, select

SYRINGE

SYSTEM PARAMETERS screen, select CONCENTRATE in the

METHOD SYRINGE and FLUSH

speeds on the SYSTEM PARAMETERS screen.

5. For Chromeleon or Chromeleon Xpress control, enter the syringe speed in the Program Wizard. 1 is the slowest speed and 5 is the fastest. You can enter separate aspirate and dispense speeds for the Concentrate, Reagent Flush, and Reagent Prime functions. In general, use a faster speed for aspirating and a slower speed for dispensing fluid to the concentrator. See Section 3.7

for details about selecting

syringe speeds.

NOTE If a 5 or 10 mL syringe is installed, the dispense

speed is limited to speed 1 or 2 only.

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Figure 3-25 shows the Program Wizard Sampler Options tab page

with the Concentrate function selected.

Figure 3-25. Program Wizard Sampler Options for Concentrate

NOTE Be sure to select syringe speeds that will keep the

backpressure below 690 kPa (100 psi), taking into account the pressure from the tubing and the concentrator column. See Section 3.8

for details.

6. In the Chromeleon or Chromeleon Xpress sequence (see Figure 3-19) or in the front panel schedule (see Section D.3

), enter an injection volume equal to the amount of sample to be loaded onto the concentrator.

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Example Program for Concentrate Mode (Matrix Elimination)

The following example Chromeleon or Chromeleon Xpress program shows the AS commands when the autosampler is in concentrate mode. Commands similar to these could be used for a matrix elimination application.

Flush Volume = 100 ;AS options commands

Wait FlushState

NeedleHeight = 2

CutSegmentVolume = 0

SyringeSpeed = 3

CycleTime = 0

Pipet Volume = 20.0, SourceVial = CurrentVial, DestinationVial = CurrentVial

Mix SourceVial = CurrentVial, NumberOfTimes = 5, Volume = 250.0

FlushSP Volume = 250.0

DelaySP Time = 0.0

Dispense Volume = 1000.0, SourceReservoir = Reservoir_A, DestinationVial = CurrentVial

WaitForTemperature = False

Concentrate ValvePosition = LoadPosition, AspirateSpeed = 3, DispenseSpeed = 1

ReagentPrime Volume = 10000.0, SourceVial = Reservoir_A, ValvePosition = NoChange

ReagentFlush Volume = 5000.0, SourceVial = Reservoir_A, ValvePosition = NoChange

Wait SampleReady

0.000 Wait CycleTimeState

3 • Operation and Maintenance

;AS sample prep commands

;Switch the inject valve to Load and load sample onto the concentrator

;Prime sampling needle assembly with reagent

;Flush concentrator with reagent; this is the matrix elimination step

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AS Autosampler Operator’s Manual

Sampler_InjectValve.InjectPosition ;Switch the inject valve

to Inject and sweep the sample onto the column

5.000 BeginOverlap ;Start preparing the next

sample in the sequence (see note below)

30.000 End

NOTE If the AS is in concentrate mode, sample preparation

overlap is available only if the autosampler is controlled by Chromeleon or Chromeleon Xpress. If the AS is under front panel control, sample overlap is not available.

3.6.11 Sequential Injections

The AS must be in sequential mode to perform sequential injections (see

Section 3.6.2

separate chromatography systems. Each system is operated independently and can have a different sample and sample volume injected.

). In sequential mode, the AS delivers sample to two

NOTE Precision for partial-loop injections may be slightly

affected by the longer tubing length from the inject port to the injection valve.

NOTE Figure 3-26

When the diverter valve is switched to position 2, the injection sequence is identical, except that flow is diverted to inject valve #2.

86 Doc. 065051-03 1/08

shows an injection to inject valve #1.

Flush

Sample

Air

Sample Syringe

3 • Operation and Maintenance

1. Draw air

Diverter Valve

Waste

Pump

Inject Valve #1

INJECT

Column

Waste

Pump

4. Prepare to load sample (excess sample used). Diverter valve selects inject valve #1 or #2.

Column

INJECT

Inject Valve #2

2. Draw sample (at least 4X loop volume)

Waste Waste

Pump

Inject Valve #1

LOAD

Column

To Inject Valve #2

5. Deliver sample to the loop (excess sample sent to waste)

3. Draw air again to isolate the sample

Pump

Inject Valve #1

INJECT

Column

To Inject Valve #2

6. Inject sample onto column Empty syringe (remaining sample + air bubble)

KEY

Flush

Eluent

Sample

Unselected ValveAir

Figure 3-26. Sequential, Full-Loop Injection Sequence

NOTE Partial-loop and partial-loop, limited-sample injections

can also be performed in sequential mode. The sequences of events for these injection types are the same as for normal mode, except for selection of the diverter valve position.

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Setting Up a Sequential Mode Injection

The AS must be equipped with the sequential injection option (P/N 063294) in order to perform sequential injections. The following setup steps are required:

1. Follow the instructions in Section B.3.6

to install the sequential injection diverter valve, connect the AS inject port tubing to the diverter valve, and connect the diverter valve to the injection valves on two systems.

2. Calibrate the inject port volume of each system. This is the volume from the inject port through the diverter valve to the injection valve (see Section 5.9

3. On the

SAMPLE MODE field (see Figure 3-27).

METHOD SYRINGE SPEED:

Help Prompt

4. On the

SYSTEM PARAMETERS screen, select SEQUENTIAL in the

CUT SEGMENT VOLUME:

INJECT PORT VOLUME:

FLUSH SYRINGE SPEED:

Figure 3-27. System Parameters Screen: Sequential Mode

PLUMBING CONFIGURATION screen, enter the sample loop size

installed on each valve (see Figure 3-28

SYSTEM PARAMETERS

WAIT:

SAMPLE MODE:

ENABLED 3 10

SEQUENTIAL S1 43 4

S2 47

PLUMBING CONFIGURATION

LOOP SIZE V1: LOOP SIZE V2:

SAMPLE SYRINGE VOLUME:

SAMPLE SYRINGE CONFIG:

PREP SYRINGE VOLUME:

PREP SYRINGE CONFIG:

Help Prompt

Figure 3-28. Plumbing Configuration Screen: Sequential Mode

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100

uL 250 READY

READY

Dionex As Operator's Manual

Specifications and Main Features

Frequently Asked Questions

User Manual

Contents

1.1 Overview

1.2 About This Manual

1.2.1 Safety Messages and Notes

1.3 Safety and Regulatory Information

1.3.1 Safety Labels

2.1 Overview of Features

2.2 Front Panel Features

2.2.1 Keypad Button Functions

2.2.2 Display Screens

2.3 Autosampler Compartment

2.3.1 Syringe and Reservoir Organizer

2.3.2 Sampling Needle Arm

2.3.3 Sampling Needle Assembly

2.3.4 Flush, Waste, and Inject Ports

2.3.5 Sample Vials and Well Plates

2.3.6 Injection Valve Options

2.3.7 Sample Temperature Control Option

2.3.8 Simultaneous Injection Option

2.3.9 Sequential Injection Option

2.3.10 Concentrator Injection Option

2.4 Rear Panel Features

2.4.1 USB Receptacle

2.4.2 TTL/Relay Connectors

2.4.3 Fuse Holder and Power Receptacle

2.5 Autosampler Control

2.5.1 Local Control

2.5.2 Computer Control (Locked Remote Mode)

2.6 System Wellness and Predictive Performance

2.7 Theory of Operation

2.7.1 Understanding the Wait Operation

2.7.2 Overlapping Sample Preparation

2.7.3 Running Samples Without Overlap

2.7.4 Operating the AS as the System Master (Disabling the Wait Operation)

2.7.5 Understanding the Cycle Time

2.7.6 Operating Events During a Schedule

2.7.7 Understanding the Status Display During a Run

2.7.8 Operating Events During Flushing and Priming

2.7.9 Sample and Prep Syringe Fluid Schematics

3.1 Getting Ready to Run

3.1.1 Turn On the Power

3.1.2 Fill the Vials or Wells and Load the Sample Tray

3.2 Selecting Computer or Front Panel (Local) Control

3.2.1 Connecting to Chromeleon or Chromeleon Xpress

3.2.2 Running a Sequence of Injections in Chromeleon or Chromeleon Xpress

3.3 Default Operating Parameters

3.4 Specifying Sample Positions

3.4.1 Vial Tray Sample Positions

3.4.2 Well Plate Tray Sample Positions

3.5 Specifying Injection Volumes

3.6 Sample Injection Types

3.6.1 Overview

3.6.2 Determining the Injection Type

3.6.3 Determining the Sample Loop Injection Type

3.6.4 Guidelines for Sample Volumes Used, Loop Sizes, and Injection Volumes

3.6.5 Normal, Full-Loop Injections

3.6.6 Normal, Large-Loop Injections

3.6.7 Normal, Partial-Loop Injections

3.6.8 Normal, Partial-Loop, Limited-Sample Injections

3.6.9 Simultaneous Injections

3.6.10 Concentrate Injections

3.6.11 Sequential Injections