Thermo Fisher Scientific Xcalibur LCquan Quantitative Analysis User guide

  • Hello! I've reviewed the Thermo Fisher Scientific Xcalibur LCquan User Guide. This document provides detailed instructions on how to use the LCquan software for quantitative analysis, including data acquisition, processing methods, and report generation. It describes features like peak integration, calibration curve generation, and data review. I'm ready to answer questions about the software and its functions.
  • What is LCquan used for?
    Can LCquan be used for data acquisition?
    What are the main windows in LCquan?
    What can be done in the Quantitate window?
Xcalibur
Thermo LCquan
Quantitative Analysis
Version 2.6.1
User Guide
XCALI-97236 Revision B April 2011
© 2011 Thermo Fisher Scientific Inc. All rights reserved.
LCQ, BioWorks, LCquan, Quantum, and Web Access are trademarks; and Xcalibur, Surveyor, LTQ, and
Accela are registered trademarks of Thermo Fisher Scientific Inc. in the United States.
The following are registered trademarks in the United States and other countries: Microsoft, Windows, Excel,
and Vista are registered trademarks of Microsoft Corporation. Adobe, Acrobat, Reader, and Flash are registered
trademarks of Adobe Systems Incorporated. Oracle is a registered trademark of Oracle Corporation and/or its
affiliates. Waters is a registered trademark of Waters Corporation.
The following are registered trademarks in the United States and possibly other countries: Agilent is a registered
trademark of Agilent Technologies, Inc. Citrix is a registered trademark of Citrix Systems Incorporated.
Citrix Presentation Server is a trademark of Citrix Systems.
All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries.
Thermo Fisher Scientific Inc. provides this document to its customers with a product purchase to use in the
product operation. This document is copyright protected and any reproduction of the whole or any part of this
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.
Thermo Fisher Scientific Inc. makes no representations that this document is complete, accurate or error-
free and assumes no responsibility and will not be liable for any errors, omissions, damage or loss that might
result from any use of this document, even if the information in the document is followed properly.
This document is not part of any sales contract between Thermo Fisher Scientific Inc. and a purchaser. This
document shall in no way govern or modify any Terms and Conditions of Sale, which Terms and Conditions of
Sale shall govern all conflicting information between the two documents.
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
Contents
Contents
iv LCquan User Guide Thermo Scientific
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
Contents
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
Contents
vi LCquan User Guide Thermo Scientific
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
Contents
Thermo Scientific LCquan User Guide vii
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
•1/X
2
•1/Y
•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.
/