Thermo Fisher Scientific Xcalibur LCquan Quantitative Analysis User guide

Xcalibur

Thermo LCquan

Quantitative Analysis

Version 2.6.1

User Guide

XCALI-97236 Revision B April 2011

LCQ, BioWorks, LCquan, Quantum, and Web Access are trademarks; and Xcalibur, Surveyor, LTQ, and

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document is strictly prohibited, except with the written authorization of Thermo Fisher Scientific Inc.

The contents of this document are subject to change without notice. All technical information in this

document is for reference purposes only. System configurations and specifications in this document supersede

all previous information received by the purchaser.

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Release history: Revision A - January 2009; Revision B - April 2011.

Software version: LCquan 2.6.1; Xcalibur 2.1 SP1; Foundation 1.0.2 SP2; TSQ Quantum 2.3 SP2; LC

Devices 2.5; Microsoft .Net 3.5 SP1; Microsoft Windows XP Professional with Service Pack 3 or Microsoft

Windows 7 Professional 32-bit; Microsoft Office 2007 or 2010.

For Research Use Only. Not for use in diagnostic procedures.

Thermo Scientific LCquan User Guide iii

C

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix

Related Documentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .ix

Special Notices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x

System Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x

Contacting Us . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x

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

Quantitative Analysis with LCquan. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

Overview of LCquan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

LCquan Folder Structure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

LCquan Applications Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Acquiring and Processing Data with LCquan . . . . . . . . . . . . . . . . . . . . . . . . 10

LCquan for Web Access . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

LCquan for Citrix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

Chapter 2 Preparing to Perform Quantitative Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13

Creating a New Study and Workbook. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Create a New Workbook. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Saving a Workbook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

File Tracking Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Workbook Summary Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Introduction to the Instrument Setup Window. . . . . . . . . . . . . . . . . . . . . . . . . 26

Creating Instrument Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Importing an Existing Instrument Method . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Introduction to the Acquisition Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Using the Setup Sequence View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

Acquisition Sequence Header . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Acquisition Sequence History . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Acquisition Sequence Grid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

Using the Run Sequence Dialog Box. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68

Change Instruments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75

Using the Status View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77

Using the Status View Control Buttons. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79

Acquisition – Status – Chromatogram Pane . . . . . . . . . . . . . . . . . . . . . . . . . 80

Acquisition – Status – Spectrum Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Acquisition – Status – Run Status . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90

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Acquisition – Status – Acquisition Queue . . . . . . . . . . . . . . . . . . . . . . . . . . . 93

Acquisition – Status – Acquisition History . . . . . . . . . . . . . . . . . . . . . . . . . . 97

Processing Data While Continuing to Acquire Data . . . . . . . . . . . . . . . . . . . 99

Verifying Disk Space . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101

Save Settings Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

Emergency Shutdown . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

Shutting Down LCquan While Acquiring. . . . . . . . . . . . . . . . . . . . . . . . . . . . 104

Remote Acquisition. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105

Chapter 3 Exploring the Data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .109

Introduction to the Explore Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

Creating Explore Methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110

Defining Chromatographic Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111

Defining Peak Integration Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113

Acquisition Sequence History . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128

Preview Panes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130

Exploring the Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131

Navigation Features. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

Opening a Raw File and Displaying the Chromatogram . . . . . . . . . . . . . . . 138

Generating a Peak List. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139

Identifying Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143

Exporting the Active Raw File to Qual Browser. . . . . . . . . . . . . . . . . . . . . . . . 144

Data Views . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145

Toolbar for Selectable Views . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145

Chromatogram View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148

Chromatogram List View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152

Error Report View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153

Filter List View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154

General Parameters Plot View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155

Instrument Method View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157

Mass List View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158

Peak List View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159

Sample Info View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163

Spectrum View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164

Status Plot View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166

Status Report View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168

Tune Method View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170

Multi Peak Plot View for Review Only . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171

Chapter 4 Creating a Processing Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175

Creating a Processing Method with the New Method Wizard . . . . . . . . . . . . . 176

Start the Wizard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 176

Create or Import a Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177

Import a Processing Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178

Specify Calibration Standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178

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Thermo Scientific LCquan User Guide v

Select a Raw file. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178

Complete the Wizard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179

Specifying Method Identification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179

Chromatogram Definition Area. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180

Retention Time Area . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183

Peak Integration Area . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184

Peak Identification Area . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185

Ion Ratio Confirmation Area. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185

Summing Ions Dialog Box . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187

Chromatogram Normalization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189

Specifying Component ID Parameters and Integrating the Peaks . . . . . . . . 191

Defining Peak Integration Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194

IRC Detection Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208

Identification Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223

Ion Ratio Confirmation of Analytes Example . . . . . . . . . . . . . . . . . . . . . . . 225

Specifying Method Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226

Component Type Area . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227

Target Compound Area. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

Component List Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232

Calibration Shortcut Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233

Correction for Isotope Contribution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236

Specifying Calibration Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237

Specifying Internal Standards and Target Compounds . . . . . . . . . . . . . . . . 237

Reviewing the Integrated Peaks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 240

Saving the Processing Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241

Importing a Processing Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241

Exporting Components and Levels to an Acquisition Sequence. . . . . . . . . . . . 241

Data Views . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 242

Toolbar for Selectable Views . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 242

Calibration Curve View for Survey or Review All Results Only . . . . . . . . . . 245

Chromatogram View for Create Method Only . . . . . . . . . . . . . . . . . . . . . . 247

Chromatogram View for Survey or Review All Results Only . . . . . . . . . . . . 251

Spectrum View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256

IRC Chromatogram View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 258

Chromatogram List View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 262

Error Report View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 263

Filter List View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264

General Parameters Plot View for Survey or Review All Results Only . . . . . 265

Instrument Method List View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 267

Mass List View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 268

Peak List View for Survey or Review All Results Only . . . . . . . . . . . . . . . . . 270

Sample Info View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 271

Status Plot View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 273

Status Report View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275

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Tune Method View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277

Multi Peak Plot View for Survey or Review All Results Only. . . . . . . . . . . . 278

Chapter 5 Creating a Processing Sequence. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .283

Defining a Processing Sequence. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 284

Using the New Processing Sequence Wizard . . . . . . . . . . . . . . . . . . . . . . . . 286

Copying an Acquisition Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 288

Importing a Processing Sequence. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289

Copying the Last Acquired Sequence. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 290

Editing a Processing Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 290

Using the Create Sequence (edit) View . . . . . . . . . . . . . . . . . . . . . . . . . . . . 290

Using the Create Sequence (create) View. . . . . . . . . . . . . . . . . . . . . . . . . . . 304

Resolving Discrepancies in Level Names . . . . . . . . . . . . . . . . . . . . . . . . . . . 307

Exporting a Processing Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 309

Chapter 6 Processing the Raw Files and Reviewing the Analytical Results. . . . . . . . . . .311

Processing the Raw Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 311

Reviewing the Calibration Curve and QCs . . . . . . . . . . . . . . . . . . . . . . . . . . . 312

Opening the Survey View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 312

Reviewing the Calibration Standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 319

Modifying Calibration Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 322

Excluding a Calibration Standard from the Calibration Curve . . . . . . . . . . 328

Modifying the Peak Detection and Integration Settings. . . . . . . . . . . . . . . . 331

Specifying Additional Peak Detection Criteria. . . . . . . . . . . . . . . . . . . . . . . 332

Manually Integrating Peaks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 349

Reviewing All Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 350

Opening the Review All Results View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 351

Reviewing the Unknowns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 351

Reviewing Peak Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352

Adding Comments to the Chromatogram Display . . . . . . . . . . . . . . . . . . . 359

Creating and Reviewing Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360

Generating Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 362

Review Reports Warning. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368

Report Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368

Exporting Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 381

Locking the Workbook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 382

Saving the Workbook and Exiting the LCquan Application . . . . . . . . . . . . . . 385

Appendix A Quantitative Analysis Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .387

About Quantitative Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 387

Considering the Variables of Quantitative Analysis by LC/MS/MS. . . . . . . . . 388

Using LC for Analyte Separation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 388

Using MS/MS for Analyte Detection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389

Quantitative Analysis Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 390

Using External Standards for Quantitative Analysis. . . . . . . . . . . . . . . . . . . 392

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Using Internal Standards for Quantitative Analysis . . . . . . . . . . . . . . . . . . . 392

Sample Types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 394

Standards. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 394

Unknowns. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 394

QCs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395

Blanks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395

Appendix B LCquan Flow Diagrams . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .397

LCquan Flow Diagram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 398

LCquan for Web Access Flow Diagram. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399

LCquan for Citrix Flow Diagram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 400

LCquan Acquisition Flow Diagram. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 401

LCquan Quantitate Flow Diagram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 402

Appendix C LCquan Menu and Toolbar Reference. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .403

Using the Menus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 403

Actions Menu for Acquisition Window Only . . . . . . . . . . . . . . . . . . . . . . . . . 404

Apps Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406

Change Menu for Acquisition Window Only . . . . . . . . . . . . . . . . . . . . . . . . . 406

File Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406

Help Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 411

Options Menu. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 412

View Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 415

Zoom Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 418

Icon Bars . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 419

Toolbar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 420

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .425

Thermo Scientific LCquan User Guide ix

P

Preface

The LCquan™ application is part of the Thermo Xcalibur™ mass spectrometry data system.

This user guide describes how to use LCquan to perform quantitative analysis of compounds.

The LCquan application is a secure data quantitation package. It provides all the tools you

need to perform quantitation tasks, including importing sequence information from external

systems, acquiring data, developing processing methods, reviewing and processing data,

generating reports, and exporting results to external systems.

Documentation

The following LCquan manuals are available on the LCquan software CD as PDF files:

•LCquan Administrator Guide describes how to configure the software for compliance and

security.

•LCquan User Guide describes how to use LCquan to perform quantitative analysis of

compounds.

•LCquan Tutorial describes step-by-step procedures to perform quantitative analysis with

sample data.

To view the LCquan manuals

Go to Start > Programs > Thermo Xcalibur > Manuals > LCquan.

To open the LCquan Help

From the LCquan window, choose Help > LCquan Help.

To find a particular topic, use the Help Contents, Index, or Search panes.

Note The LCquan for Web Access or Citrix application does not include the Acquisition

feature.

Preface

xLCquan User Guide Thermo Scientific

Special Notices

This guide includes the following types of special notices:

System Requirements

In addition, your system must meet these minimum requirements.

IMPORTANT Highlights information necessary to prevent damage to software, loss of

data, or invalid test results; or might contain information that is critical for optimal

performance of the system.

Note Highlights information of general interest.

Tip Highlights helpful information that can make a task easier.

System Requirements

Computer • 2.4 GHz processor dual core with 4 GB RAM (use 4 GB for

enhanced speed)

•DVD/CD-ROM drive

• Video card and monitor capable of 1280 × 1024 resolution

(XGA)

• 250 GB available on the C: drive

•NTFS format

Software • Microsoft™ Windows™ XP Professional SP3 (U.S. version) or

Microsoft Windows 7 Professional 32-bit

• Microsoft Office Pro 2007 SP2 or 2010 (required for

Windows 7)

• Microsoft .NET Foundation 3.5 SP1

• Thermo Scientific software:

−Thermo Xcalibur™ 2.1 SP1

−Thermo Foundation™ 1.0.2 SP2

Preface

Thermo Scientific LCquan User Guide xi

Contacting Us

There are several ways to contact Thermo Fisher Scientific for the information you need.

To contact Technical Support

Find software updates and utilities to download at mssupport.thermo.com.

To contact Customer Service for ordering information

To get local contact information for sales or service

Go to www.thermoscientific.com/wps/portal/ts/contactus.

To copy manuals from the Internet

Go to mssupport.thermo.com, agree to the Terms and Conditions, and then click

Customer Manuals in the left margin of the window.

To suggest changes to documentation or to Help

• Fill out a reader survey online at www.surveymonkey.com/s/PQM6P62.

• Send an e-mail message to the Technical Publications Editor at

techpubs-lcms@thermofisher.com.

Phone 800-532-4752

Fax 561-688-8736

E-mail us.techsupport.analyze@thermofisher.com

Knowledge base www.thermokb.com

Phone 800-532-4752

Fax 561-688-8731

E-mail us.customer-support.analyze@thermofisher.com

Web site www.thermo.com/ms

Thermo Scientific LCquan User Guide 1

1

Introduction

This chapter provides a brief overview of quantitative analysis with the LCquan application

and an overview of using the LCquan application with Thermo Scientific Web Access Suite™

or Citrix Presentation Server™.

Quantitative Analysis with LCquan

The LCquan application is a complete quantitative analysis software package that lets you

acquire data specifically for an analyte of interest, to quantitate the results, and to produce

reports. For more information about quantitative analysis concepts, see Appendix A,

“Quantitative Analysis Overview.”

This section contains the following topics:

•Overview of LCquan

•LCquan Folder Structure

•LCquan Applications Menu

•Acquiring and Processing Data with LCquan

•LCquan for Citrix

Contents

•Quantitative Analysis with LCquan

•LCquan for Web Access

•LCquan for Citrix

1 Introduction

Quantitative Analysis with LCquan

2LCquan User Guide Thermo Scientific

Overview of LCquan

The LCquan application is divided into four main windows: Instrument Setup, Acquisition,

Explore, and Quantitate. By using these windows, you can develop instrument methods,

acquire data, explore data, and process data files.

Instrument Setup Window

Use the LCquan Instrument Setup window to develop instrument methods for the

instruments you selected in the Xcalibur Instrument Configuration window. An instrument

method is a set of experimental parameters and operating settings for a specific instrument,

such as an autosampler, LC pump or MS pump, mass spectrometer, divert valve, or syringe

pump. Within the Instrument Setup window, you can create new methods or modify existing

methods. You can also import previously collected data files to help you set up time segments

and scan events based on the acquired data. The Instrument Setup window within LCquan

functions similar to the Instrument Setup window of the Xcalibur Home Page.

The Instrument Setup window contains separate pages for each instrument that you selected

in the Xcalibur Instrument Configuration window (see Figure 1).

Note The Instrument Setup window is not available in the LCquan for Web Access or

Citrix application.

1 Introduction

Quantitative Analysis with LCquan

Thermo Scientific LCquan User Guide 3

Figure 1. Instrument Setup window

1 Introduction

Quantitative Analysis with LCquan

4LCquan User Guide Thermo Scientific

Acquisition Window

In the LCquan Acquisition window, you can set up a sequence of samples for acquisition (see

Figure 2). The sequence can contain unknown samples, calibration standard samples, quality

control samples, and blank samples. After you have defined a sequence, you can specify

acquisition options and processing action options for the sequence and start running it. The

Acquisition window also contains a real-time display so you can watch updates of the

acquisition status and the data being acquired.

The Acquisition window contains the Setup Sequence view, the Run Sequence dialog box,

and the Status view.

Figure 2. Acquisition window

Note The Acquisition window is not available in the LCquan for Web Access or Citrix

application.

1 Introduction

Quantitative Analysis with LCquan

Thermo Scientific LCquan User Guide 5

Explore Window

In the Explore window, you can display a multi-peak chromatogram for a single sample and

experiment with peak detection and integration parameters to see how they affect the

chromatogram (see Figure 3). Use the Explore window to determine the optimum parameters

used in the processing method. The LCquan application can also generate a peak list that you

use to generate the quan components in the processing method. The Create Explore Methods

view of the Explore window opens.

Figure 3. Explore window

1 Introduction

Quantitative Analysis with LCquan

6LCquan User Guide Thermo Scientific

Quantitate Window

In the Quantitate window, the LCquan application steps you through a procedure for batch

processing raw files during quantitative analysis (see Figure 4). Because the LCquan window

incorporates an integrated calibration curve, peak integration, and results view, you can

interactively review and rework standards, QCs, and unknowns in one window. You can also

enter the Quantitate window to process the acquired data as you acquire it.

Figure 4. Quantitate window

The LCquan application uses a processing method to automatically determine the

concentration (or amount) of the sample being analyzed. The processing method measures

the response of the detection system to the compounds in your sample by integrating the area

under each peak.

1 Introduction

Quantitative Analysis with LCquan

Thermo Scientific LCquan User Guide 7

After measuring several known standards, the LCquan application constructs a calibration

curve that it uses to accomplish the quantitative analysis of unknown samples. It provides a

number of options for fitting the calibration data to generate the calibration curve. You can

have it fit the calibration data to the following curve types:

•Linear

•Quadratic

•Linear log-log

•Quadratic log-log

• Average response factor (RF)

• Point-to-point

•Cubic spline

•Locally weighted

You can also have the LCquan application weight the calibration data with the following

weighting functions when performing the least squares fit to the calibration data:

•Equal

•1/X

2

•1/Y

2

•1/S

2, where S2 = X2 + Y2

In addition, you can have it ignore the origin, use the origin as a data point, or force the

calibration curve to include the origin.

The Quantitate window contains these views: Create Method, Create Sequence, Survey,

Review All Results, and Review Reports.

1 Introduction

Quantitative Analysis with LCquan

8LCquan User Guide Thermo Scientific

LCquan Folder Structure

When you start a new quantitative analysis project, the LCquan application creates a

hierarchical folder structure for the project on the hard drive.

The top-most level in the file structure is the study folder. This folder is used to organize

one or more workbook folders. Each workbook folder holds all the information used by the

LCquan application for an individual quantitative analysis project. The workbook folder

contains the LCquan file (.lqn), the instrument method files (.meth), and the audit log files.

In addition, the workbook folder contains these folder names: Exports, Imports, raw files, and

Temp.

The Exports folder stores copies of all files that are exported from the LCquan application,

such as report files. The Imports folder stores a copy of legacy files that you import into the

workbook, such as instrument method files or sequence files. The raw files folder contains

acquired data files (.raw), and any imported raw files. The Temp folder contains backup and

temporary files that it uses.

When you create a new project, the LCquan application requires you to name the study folder

and the workbook folder. You can have as many different workbooks in the same study folder

as you want, but each workbook must have a unique name. This is convenient for organizing

your quantitative analysis projects. For example, you can use one study folder to organize all

the separate quantitative analysis projects that you are doing for a particular client. You cannot

rename the folders that the LCquan application creates in the workbook folder.

The LCquan application manages all the files associated with the project. You do not have to

remember what individual files are named or where they reside on your hard drive. All

necessary files are contained within the workbook. In addition, it uses a file tracking feature to

maintain a secure file system. Check with your administrator if you have problems with the

security configuration.

IMPORTANT Always use the LCquan application to open and modify the workbook

and its associated files. To maintain secure file operations, it tracks the files in the

workbook to ensure that none has been modified by an external process. If you modify a

file in the workbook by using another software application (including Xcalibur), it

prevents you from reopening the workbook.

Thermo Fisher Scientific Xcalibur LCquan Quantitative Analysis User guide

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