LC Operation

Shimadzu High Performance Liquid Chromatography

LC Workstation

[LabSolutions/LCsolution]

Operation Manual

Read the instruction manual thoroughly before you use the product.Keep this instruction manual for future reference.

223-00914BAug. 2006

This page is intentionally left blank.

1

LabSolutions/LCsolution Operation Manual i

Microsoft and Windows are registered trademarks of Microsoft Corporation in the USA and other countries.Acrobat and Adobe are trademarks of Adobe Systems Incorporated.

Other company names and product names mentioned in this manual are trademarks or registered trademarks of their respective companies.

Microsoft Windows 2000 is referred to as Windows 2000.Microsoft Windows XP Professional Edition is referred to as Windows XP.

2003-2006 Shimadzu Corporation. All rights reserved.

>z The copyright of this manual is owned by Shimadzu Corporation. Reproduction and duplication of whole or part of the

content without permission of the company are strictly prohibited.z The content of this manual is subject, without notice, to modifications for the sake of improvement.z Every effort has been made to ensure that the content of this manual was correct at the time of creation. However, in the

event that any mistakes or omissions are discovered, it may not be possible to correct them immediately.z Please be aware that we do not take any responsibility for the consequences of operations described in this manual.z Please be aware that we do not take any responsibility for the consequences of using the device.z Maintenance parts of this product are provided for seven years after production is stopped. Please note that we may not be

able to provide maintenance parts after this period. However, for parts that are not genuine parts of our company, the period of provision is determined by the manufacturer.

z The contents of the hard disk in a PC can be lost due to an accident. To protect your important data from accidents, be sure to backup your data.

To ensure the safe use of this device, be sure to observe the points below.

Follow the procedures in the instruction manuals. Observe all warnings and cautions. Do not disassemble or modify this product without permission. The repair of this product may be ordered at our offices and distributors.

ii LabSolutions/LCsolution Operation Manual

Thank you very much for purchasing the LCsolution software for Shimadzu High Performance Liquid Chromatography Workstation (hereafter referred to as "LCsolution").

LCsolution enables you, from your PC, to control high performance liquid chromatograph (hereafter referred to as "LC"), acquire chromatograms and various types of data, and reanalyze the acquired data under various conditions.

This manual explains the basic operations of LCsolution.

'LCsolution Administration Manual' and 'LCsolution Installation Manual' are provided as separate volumes.

Read this manual thoroughly before using LCsolution. Keep it in a safe place so that it can be referred to whenever you have questions regarding the operation of this product.The details of the functions and the valid range of input parameters for each screen are described in the Online Help. Refer to it for more information.

This manual assumes that readers are knowledgeable of basic operations of Windows. For details on how to operate Windows, see the instruction manual accompanying that product.

Introduction

LabSolutions/LCsolution Operation Manual iii

Our company provides a warranty on this product, as stated below.

Details

* If there is a document such as a warranty attached to the product, or there is a separate contract agreed upon that includes warranty conditions, the rules stated in those documents shall be followed.Warranty periods for products with special specifications and systems are provided separately.

* The license cannot be reissued if you lose the license key disk or the USB dongle provided with the product.

Product Warranty

1. Warranty Period: Please consult your Shimadzu representative for information about the extent of the warranty.

2. Warranty Description:

If failure occurs for reasons attributable to our company during the warranty period, our company will provide repairs or the replacement of parts without charge (including USB dongles). However, we may not be able to provide identical products in the case of products such as PCs, and their peripherals and parts, which have a short lifespan in the market.

3. Warranty Exceptions: The failures caused by the following events are excluded from the warranty, even if they occur during the warranty period.

1) The product is handled in an improper way.

2) Repairs or modifications are performed by companies or people other than our company and our designated companies.

3) This product was used in combination with hardware or software other than those designated by our company.

4) Device failures and damage to data and software, including the basic software, that are caused by computer viruses.

5) Device failures and damage to data and software, including the basic software, that are caused by power failures, including power outages and sudden drops of voltage.

6) Device failures and damage to data and software, including the basic software, that are caused by powering off the device without the proper shutdown procedure.

7) Failures are caused by reasons other than the device itself.

8) Failures are caused by the use in harsh environments, such as in high temperature or humidity, corrosive gas, or vibration.

9) Failures are caused by fires and earthquakes or any other act of providence, contamination by radio active substances and hazardous substances, or any other force majeure event including wars, riots, and crimes.

10)Problems occur because the device is transferred or transported after installation.

11) Expendable items and partsNote: Recording media such as floppy disks and CD-ROMs are considered expendables.

iv LabSolutions/LCsolution Operation Manual

The LCsolution package comes with the following instruction manuals. PDF manuals are included in the installation CD-ROM. There is also an online help that explains the operations and configuration settings. Refer to them as necessary.

The symbols and notations used in the instruction manuals have the following meanings.

Symbols used in the Instruction Manuals

Buttons and menu items on the screen

Instruction Manuals

List of Instruction Manuals

Name Content

Installation Manual (Booklet/PDF)

Explains the basic installation method for LCsolution. Be sure to read this manual first.Information on advanced settings is presented in the 'Operation Manual' and 'Administration Manual'. You can go to the applicable sections in the 'Operation Manual' and 'Administration Manual' from the hyperlinks in this PDF manual.

Quick Manual (PDF)

Explains the operation of LCsolution in a tutorial format. You can learn how to operate the application by actually analyzing data.

Operation Manual (Booklet/ PDF) (This document)

Explains data acquisition and analysis procedures of LCsolution for various purposes. The PDF version is also provided so that you can view it on PCs.

Administration Manual (Booklet/ PDF)

Explains the basic concepts and operating procedures of system and data administration using LCsolution.The PDF version is also provided so that you can view it when necessary.

Help Provides detailed information on parameters and their setting ranges for each LCsolution function.Online Help can be accessed from the LCsolution [Help] menu. For information on how to use the online help, refer to "13.1.1 Using Help" in the 'Operation Manual'.

Notations used in Instruction Manuals

Notation Meaning

NOTE Cautions and supplementary explanations that serve as a reference for operation

Reference Indication of location for related information in the instruction manuals

' ' The names of the instruction manuals

" " The names of programs and products

Notation Meaning

[ ] Texts displayed on the screen, such as the names of buttons, menu items, settings, screens, and icons.Example: Click [OK].

[ ] - [ ] When several operations are performed one after another, hyphens are used to show the sequence.Example: Select [File] - [Print].

Click the [File] menu and select [Print].

LabSolutions/LCsolution Operation Manual v

Contents

1 What is LCsolution?1.1 Features of LCsolution.........................................................................................1

2 Software Overview2.1 LCsolution Fundamentals ....................................................................................5

2.1.1 Starting "LCsolution Launcher" .......................................................................... 52.1.2 Starting "LC Real Time Analysis" ....................................................................... 72.1.3 File Types and File Tabs of the Data Explorer ................................................... 9

3 Starting the Analysis3.1 Before Reading This Chapter ............................................................................11

3.1.1 The Analytical Instruments and Analysis Conditions Described in This Chapter ................................................................................................. 11

3.1.2 The Data Structure of LCsolution..................................................................... 113.1.3 LCsolution Operation Flow............................................................................... 12

3.2 Preparing for Analysis........................................................................................133.2.1 Starting the Analytical Instruments and Logging into "LC Real Time Analysis"133.2.2 Setting the System Configuration of the Analytical Instruments....................... 143.2.3 Checking the Analytical Instruments ................................................................ 163.2.4 Setting the Instrument Parameters................................................................... 173.2.5 Monitoring the analytical instruments ............................................................... 19

3.3 Performing Single Analysis................................................................................213.3.1 Performing Single Analysis .............................................................................. 213.3.2 Setting the Peak Integration Parameters ......................................................... 223.3.3 Setting the Column Performance Parameters.................................................. 25

3.4 Performing Batch Analysis.................................................................................263.4.1 Creating a Batch Table .................................................................................... 263.4.2 Performing Batch Analysis ............................................................................... 27

3.5 Generating Calibration Curves ..........................................................................283.5.1 Creating Data Analysis Parameters ................................................................. 283.5.2 Applying Data Analysis Parameters to the Original Method............................. 313.5.3 Performing Batch Processing........................................................................... 323.5.4 Viewing the Calibration Curve.......................................................................... 343.5.5 Calculating the Concentration of Data Using the Calibration Curve................. 36

3.6 Creating a Report Format ..................................................................................38

vi LabSolutions/LCsolution Operation Manual

Contents

3.7 Creating a Summary Report.............................................................................. 413.8 Viewing Data ..................................................................................................... 42

3.8.1 Viewing and Using [Data Explorer] .................................................................. 423.8.2 Viewing Data .................................................................................................... 443.8.3 Viewing Data with the CLASS-Agent ............................................................... 46

4 Data Acquisition4.1 Using and Viewing the [Data Acquisition] Window............................................ 494.2 Changing the Analysis End Time ...................................................................... 52

4.2.1 Setting the LC Analysis End Time.................................................................... 524.2.2 Stopping the Analysis....................................................................................... 524.2.3 Changing the Analysis End Time during Analysis............................................ 53

4.3 Changing the Data Sampling Interval................................................................ 544.3.1 Types of Parameters Related to Data Sampling.............................................. 544.3.2 Setting the Detector [Base Period]................................................................... 554.3.3 Setting the Sampling Interval of the Detector................................................... 56

4.4 Start/Stop Setup of the LC System.................................................................... 574.5 Registering Column Information........................................................................ 59

5 LC Data Analysis5.1 How to Use and View the [LC Data Analysis] Window...................................... 615.2 Operations Using Data Files.............................................................................. 62

5.2.1 Exporting/Importing Data Analysis Parameters ............................................... 635.2.2 Importing/Exporting a Report Format............................................................... 635.2.3 Exporting Batch Tables .................................................................................... 64

5.3 Setting the Peak Integration Parameters........................................................... 645.3.1 Setting the Peak Integration Parameters to Detect Peaks............................... 645.3.2 Setting the Peak Integration Time Program ..................................................... 665.3.3 Setting Manual Peak Integration (Manipulation) .............................................. 68

5.4 Setting the Peak Identification Parameters ....................................................... 715.5 Setting the Quantitative Parameters.................................................................. 74

5.5.1 Setting the Quantitative Parameters ................................................................ 745.5.2 Setting the Compound Table ........................................................................... 755.5.3 Saving (Exporting) to a Method File................................................................. 76

5.6 Operations in the [LC Calibration Curve] Window............................................. 775.6.1 Displaying the [LC Calibration Curve] Window ................................................ 775.6.2 Generating a Calibration Curve........................................................................ 79

LabSolutions/LCsolution Operation Manual vii

Contents

5.7 Operations in the [LC Data Comparison] Window .............................................795.7.1 Displaying the [LC Data Comparison] Window ................................................ 795.7.2 Calculations Between Chromatograms............................................................ 815.7.3 Moving Chromatograms................................................................................... 81

6 PDA Data Analysis6.1 Using and Viewing the [PDA Data Analysis] Window........................................836.2 Extraction for Different Views ............................................................................86

6.2.1 Extraction in the [Contour View] ....................................................................... 866.2.2 Extraction in the [Chromatogram View] / [Spectrum View]............................... 88

6.3 Registering Chromatograms..............................................................................896.3.1 Registering Chromatograms in the Multi Chromatogram Table ....................... 89

6.4 Leveraging the Multi-Chromatogram for Data Analysis .....................................926.4.1 Leverage for Identification and Quantitative Processing.................................. 92

6.5 Registering the Spectrum ..................................................................................936.5.1 Registering Spectra in the [Spectrum Table].................................................... 93

6.6 Calculating Peak Purity......................................................................................956.6.1 Setting the Purity Parameters .......................................................................... 956.6.2 Displaying the Peak Purity Calculation Results................................................ 96

6.7 Specifying a Compound From a Spectrum........................................................976.7.1 Setting Library Search Criteria ......................................................................... 986.7.2 Performing a Library Search ............................................................................ 98

6.8 Editing the Spectrum Library ...........................................................................1006.8.1 Creating a New UV Library File...................................................................... 1016.8.2 Saving a UV Library File................................................................................. 1016.8.3 Registering a Spectrum to a UV Library File .................................................. 1016.8.4 Editing the UV Library File.............................................................................. 102

6.9 Using a Spectrum for Peak Identification.........................................................1026.10 Performing Other Operations...........................................................................104

6.10.1 Saving (Exporting) to a Method File ............................................................. 1046.10.2 Exporting a Chromatogram / Spectrum ........................................................ 1046.10.3 Updating Snapshots ..................................................................................... 1046.10.4 Performing Chromatogram / Spectrum Calculations .................................... 105

viii LabSolutions/LCsolution Operation Manual

Contents

7 Batch Processing7.1 Performing Continuous Data Acquisition......................................................... 107

7.1.1 Displaying and Editing the [Batch Table] ....................................................... 1077.1.2 Using the Wizard............................................................................................ 1097.1.3 Editing a Batch Table ..................................................................................... 1107.1.4 Performing Batch Processing......................................................................... 111

7.2 Performing Continuous Data Analysis............................................................. 1127.2.1 Displaying the [Batch Table] to Open a Batch File......................................... 1127.2.2 Adding Rows with Selected Data Files .......................................................... 1137.2.3 Performing Batch Processing......................................................................... 1157.2.4 Stopping / Pausing Batch Processing ............................................................ 115

7.3 Batch Processing Parameters ......................................................................... 1157.3.1 Parameters for Batch Processing .................................................................. 1157.3.2 Setting Bracket Calibration............................................................................. 1177.3.3 Configuring the Startup / Shutdown Conditions ............................................. 1187.3.4 Setting the Background Data File .................................................................. 119

7.4 Performing Additional Analysis (The Batch Queue Function) ......................... 120

8 Browser Function8.1 Displaying Quantitative Calculation Results Using the [Quant Browser]......... 121

8.1.1 Displaying the [Quant Browser]...................................................................... 1218.1.2 Displaying the Necessary Information............................................................ 123

8.2 Performing Postrun Analysis on Multiple Data ................................................ 1238.2.1 Changing Data Analysis Conditions Using the [Quant Browser].................... 1238.2.2 Performing Statistical Calculations on Quantitatively Calculated Values....... 124

8.3 Displaying Multiple Data Collectively............................................................... 1248.3.1 Displaying [Data Browser] .............................................................................. 1248.3.2 Extracting Necessary Data............................................................................. 1268.3.3 Removing Unnecessary Display Data............................................................ 1278.3.4 Changing the Display Data Type ................................................................... 1288.3.5 Arranging the Layout...................................................................................... 1298.3.6 Saving the Layout .......................................................................................... 1298.3.7 Performing a Library Search .......................................................................... 129

9 Report Function9.1 Printing the Analysis Results ........................................................................... 131

9.1.1 Report Output in Data Acquisition.................................................................. 1319.1.2 Report Output in Batch Processing................................................................ 1329.1.3 Default Report Format File............................................................................. 133

LabSolutions/LCsolution Operation Manual ix

Contents

9.2 Reprinting the Data Processing Results ..........................................................1339.3 Creating Report Format Files ..........................................................................1379.4 Example of Creating a Report Layout..............................................................1399.5 Adjusting the Report Layout ............................................................................141

9.5.1 Editing Print Items .......................................................................................... 1419.5.2 Placing Print Items on Multiple Pages............................................................ 1429.5.3 Setting the Page Size and Margins................................................................ 1439.5.4 Setting the Header / Footer ............................................................................ 1439.5.5 Viewing the Print Image ................................................................................. 1449.5.6 What Can Be Printed Out............................................................................... 145

10 Common Window Operations10.1 Displaying Windows and Bars .........................................................................147

10.1.1 Window Types of the Program Windows...................................................... 14710.1.2 Displaying Application Windows ................................................................... 148

10.2 Customizing Windows .....................................................................................15010.2.1 Changing the View Size................................................................................ 15110.2.2 Changing the Display Format of the Application Window............................. 15110.2.3 Changing the Display Format of Common Windows .................................... 153

11 Common Table Operations11.1 Operating Table Columns................................................................................155

11.1.1 Selecting Table Columns.............................................................................. 15611.1.2 Changing the Table Column Width............................................................... 15611.1.3 Changing the Display of the Table Columns (Table Style Setting 1)............ 157

11.2 Operating Table Rows .....................................................................................15811.2.1 Selecting Table Rows ................................................................................... 15811.2.2 Adding or Deleting Table Rows .................................................................... 15911.2.3 Changing the Number of Rows Displayed in a Table

(Table Style Setting 2) .................................................................................. 159

11.3 Editing Cells.....................................................................................................16011.3.1 Selecting Multiple Cells................................................................................. 16011.3.2 Editing the Contents of Cells ........................................................................ 160

11.4 Setting Values in Cells.....................................................................................16111.4.1 Setting a Value to a Cell with a Special Input Format................................... 161

x LabSolutions/LCsolution Operation Manual

Contents

12 Common File Operations12.1 File Operations in the Application Windows .................................................... 163

12.1.1 Creating a New File ...................................................................................... 16312.1.2 Opening a File .............................................................................................. 16412.1.3 Saving a File ................................................................................................. 16512.1.4 Viewing File History ...................................................................................... 165

12.2 Efficient File Administration ............................................................................. 16612.2.1 Using [Data Explorer].................................................................................... 16612.2.2 Projects......................................................................................................... 16712.2.3 Creating a New Project (Folder) ................................................................... 16812.2.4 Deleting a Project (Folder)............................................................................ 169

12.3 File Operations in [Data Explorer] ................................................................... 17012.3.1 Opening a File .............................................................................................. 17012.3.2 Copying a File............................................................................................... 17112.3.3 Moving a File ................................................................................................ 17112.3.4 Deleting a File............................................................................................... 17212.3.5 Renaming a File............................................................................................ 17312.3.6 Viewing File History ...................................................................................... 17312.3.7 Converting File Format ................................................................................. 174

13 Appendix13.1 When Having Problems with the Operation..................................................... 177

13.1.1 Using Help .................................................................................................... 17713.1.2 Using Online Manuals................................................................................... 179

13.2 How to Use the LC Maintenance Utility ........................................................... 18013.3 Maintaining the PDA Detectors ....................................................................... 182

13.3.1 Starting the PDA Utility ................................................................................. 18213.3.2 Using "Wavelength Check"........................................................................... 18313.3.3 Using "Wavelength Calibration".................................................................... 18513.3.4 Using "Leak Sensor Check/Adjustment"....................................................... 18813.3.5 Using "Cell Temperature Check" .................................................................. 18913.3.6 Using "Absorbance Accuracy Check/Calibration"......................................... 18913.3.7 Using "Instrument Information" ..................................................................... 192

13.4 Specifications of LCsolution ............................................................................ 192

Index

LabSolutions/LCsolution Operation Manual 1

11

What is LCsolution?

LCsolution is a workstation for Shimadzu High Performance Liquid Chromatography.

With LCsolution, you can control liquid chromatographs from a personal computer and perform various tasks such as chromatographic data acquisition, postrun analysis, report creation, and data management. LCsolution runs on Windows XP Professional, enabling users to perform data acquisition and postrun analysis using similar operations with many general software on Windows.

This chapter explains the features and operability of LCsolution.

1.1 Features of LCsolutionLCsolution supports a networked environment and is equipped with GLP/GMP support and system management functions. Despite its wide range of functions, LCsolution makes work intuitive and easy, displaying just the information necessary for the task at hand.

Enhanced GLP/GMP support functionsLCsolution is equipped with functions that meet the reliability requirements of GLP/GMP and 21 CFR Part 11. The GLP/GMP support functions provide sure, efficient help in achieving compliance with the requirements of 21 CFR Part 11 and GLP/GMP.

Compliance with US FDA's 21 CFR Part 11The electronic record and signature functions of LCsolution comply with the requirements of 21 CFR Part 11.Information such as data measurement methods, schedules, date/time, operator name, and chromatograms can all be saved at once, and human and machine readable data can be saved together as required for compliance with Part 11.

NOTEWhen used in combination with the optional CLASS-Agent Manager, LCsolution meets the Part 11 requirements for electronic records and signatures for review and approval, as well as the long-term storage of data.

Security supportSystem administration functions help fulfill the security needs of a wide variety of work flows.They provide multi-level access management and electronic approval of analytical data stored in databases.

Audit trail supportDetailed user information such as methods, data, and system operations, can be saved to create and manage an appropriate audit history.

Validation supportThe system check and QAQC functions help provide efficient and accurate hardware/software/method validation.

2 LabSolutions/LCsolution Operation Manual

1 What is LCsolution?

Supporting a networked environment

Consolidated management of data You can build a Client/Server data management system when working in combination with the optional database management/registration network tool "CLASS-Agent". This makes it easier to obtain the data found in other PCs on the network, since data from all of the instruments can be consolidated.

All-In-One file structure (Compound structure system)Data files contain all the information about the data, including methods, batch, and report formats, enabling easy management of files. Also, you can analyze the data from any PC on the network.

LAN compatible system administrationThe system administration information can be shared among LCsolution systems on the LAN (Local Area Network).

NOTEUsers are administered using the "SHIMADZU User Authentication Tool". Common user administration for LCsolution and CLASS-Agent is possible, and by setting account policies such as minimum password length, maximum password age, and login attempts, unauthorized access can be effectively prevented.

Ensuring file compatibility

CLASS-LC10 format compatibilityThe method files and data files from CLASS-LC10 can be imported and used as is.

CLASS-VP format compatibilityThe method files and data files from CLASS-VP can be used once they have been converted to the LCsolution format.

AIA ANDI format compatibilityThis software allows users to read and write files in the AIA ANDI format, which is the international standard format for data.

NOTEThis function can be used to transfer data files from LCsolution to CLASS-LC10 and CLASS-VP.

Outputting results in the universal data formatData can be saved in text format so that it can be used in commercially available spreadsheet software. Also, compliance with 21 CFR Part 11 is made easier by functions that allow report images to be saved in PDF format ("Acrobat" link).

NOTEAcrobat must be installed separately in order to use the "Acrobat" link.

Report format with a high degree of flexibility

Report format function with a high degree of flexibility You can efficiently create reports with flexibility in formats using techniques similar to those used in common applications such as Microsoft Word.


1

1.1 Features of LCsolution

Enhanced report itemsReports can be customized in accordance with the intended use, by combining 23 different items such as chromatograms, calibration curves, quantitative results, spectrum indices, and summary reports.

Enhanced identification and quantitative processing functions

Various methods for identification and quantitative calculationsThis software supports a variety of identification methods including Window/Band, spectrum identification, absolute retention time, relative retention time, and six different calculation methods including the external and internal standard methods.

Various calibration curve typesThis software supports seven different types of calibration curves including linear, point-to-point, and exponential.

Flexibility in creating calibration curvesBesides creating a calibration curve from continuous analysis and batch analysis, you can manually add or delete data to produce the most optimum calibration curves.

Acclaimed algorithmThis software uses the peak integration algorithm from the Chromatopac Integrator series.

Abundant functions along with simple operations

Using the latest Windows technologyThis software runs on Windows XP Professional and can be operated using the same techniques commonly used to operate Windows, such as multi-tasking and drag-and-drop. Other functions including advanced security and the support for long file names are also available in this software.

Assistant barEven a beginner can easily perform analysis simply by clicking one of the function icons in the assistant bar.

Data Explorer"Data Explorer" can be used to open files by simply dragging and dropping files to the operation window. It supports file management such as copying, transferring and deleting files, and referring to history information.

Quick instrument settings screenTwo modes, a quick and a detailed one can be employed depending on analysis conditions and use. They support wide range of analysis conditions and add convenience to daily operations.

Batch Table Wizard"The Batch Table Wizard" provides a series of simple on-screen instructions that helps create batch tables for continuous analysis.

Compound Table Wizard"The Compound Table Wizard" provides a series of simple on-screen instructions that helps create a compound table for quantitative calculations.

The functions have been efficiently arranged so that the profusion of functions can be put to effective use.



1 What is LCsolution?


2

2

Software Overview

This software is composed of the following four applications. "LC Real Time Analysis"

Performs single and batch analysis, and controls and adjusts the settings of the instruments. "LC Analysis Editor"

Edits the method files and batch files during analysis. "LC Postrun Analysis"

Creates calibration curves and processes the acquired analysis data. "LC Browser"

Performs analysis of and displays multiple sets of analysis data.

An overview of this software is given in this chapter.

2.1 LCsolution FundamentalsThe four applications of this software can be started from the "LCsolution Launcher".This section provides instructions on how to start the "LCsolution Launcher", the [Administration] menu, and the "LC Real Time Analysis", and explains the types of files used in this software.

2.1.1 Starting "LCsolution Launcher"

To use any applications, you must first start the "LCsolution Launcher" by clicking the (LCsolution)

icon.

Starting the "LCsolution Launcher"

Once this software is installed on your PC, the (LCsolution) icon appears on the desktop.

1 Double-click the (LCsolution) icon.The "LCsolution Launcher" starts.

2 Click the [Operation] tab on the left side of the screen.The [Operation] menu appears.


2 Software Overview

NOTE

The (Database) icon is enabled only when the optional "CLASS-Agent" software is

installed.

To exit "LCsolution Launcher", click (Exit) at the top right corner of the screen.

Icons in the [Operation] menuThe [Operation] menu contains the following icons. Click an icon to start an application specific to that icon.

NOTE The [Analysis] icon and [Offline Editor] icon are enabled as a set for every single LC system connected to

the PC.

When the mouse cursor is pointed to the [Analysis] icon or the [Offline Editor] icon, the name of the instrument registered in the [System Configuration] is shown as a tool tip.

When another program is running in full-screen mode, the "LCsolution Launcher" icons are hidden in the background. In this case, you can either reduce the size or minimize the window of the full screen program to show the icons, or you can start the "LC Launcher" from the [LCsolution] menu in the start-up menu.

Displaying the [Administration] menuClick the [Administration] tab to display the [Administration] menu.

Analysis This icon starts the "LC Real Time Analysis" application, which performs single and batch analysis, and controls and adjusts the settings of the instruments.("LC Real Time Analysis" is available in the online mode.)

Offline Editor This icon starts the "LC Analysis Editor", which allows you to edit method files and batch files that are not used in the current analysis.(This application is similar to "LC Real Time Analysis" but in the edit mode.)

Postrun This icon starts the "LC Postrun Analysis" which imports the acquired analysis data to create calibration curves and to process the data.

Browser This icon starts the "LC Browser" which performs analysis of and displays multiple sets of analysis data.


2

2.1 LCsolution Fundamentals

The icons in the [Administration] menuThe [Administration] menu contains the following icons. Click an icon to start an application specific to that icon.

ReferenceFor details about the [Administration] menu, refer to "2.2 Functions of the System Administration Tools" in 'Administration Manual'.

2.1.2 Starting "LC Real Time Analysis"Start "LC Real Time Analysis" after you have confirmed that the LC system is turned on.

1 Click the [Analysis 1] icon.

The [Login] screen appears.

2 For [User ID], select [Admin] and leave the [Password] blank. Click [OK].

NOTESetting a password for the Admin user or adding new users can be done in [User Administration] of the [Administration] menu.

SystemPolicy

Sets the system administration policies when using system administration functions such as login method and password restriction.

User Administration

Performs user registration and periodical password changes. Also establishes groups according to their operational privileges.

Log Browser Displays the list of search results using the specified search criteria for logs such as operation history.


2 Software Overview

The "LC Real Time Analysis" starts.

NOTE The screen of the [LC Analysis Editor] is the same as the "LC Real Time Analysis" except that it

does not include the layout function of the actual analysis operation. The [LC Postrun Analysis] screen and the [LC Browser] screen are also similar to the "LC Real Time Analysis".

It is unnecessary to turn on the LC system when starting the "LC Analysis Editor", the "LC Postrun Analysis", or the "LC Browser".

No. Name DescriptionTitle bar Displays information such as the name of the application running, the name

of the file used, and the name of the user that is logged in.Menu bar Displays the menus that are enabled for the current window and the

privileges of the currently logged in user.Toolbar Contains frequently used menus and functions that can control analytical

instruments.Applicationwindow

Displays an application window, such as the data acquisition window.

Exit button Click here to exit the application.Output window Displays the message logs and login/ logout history.

When an error occurs, this window pops up automatically and displays the error message.

Data Explorer The following features help provide easy file management; a tab per each file type, easy file import by simple drag-and-drop operation onto the application window, and quick switch between folders.

Assistant bar The icons of the applications are lined up in the same order as the general analysis work flow.The main operations of this software are performed using these icons.


2

2.1 LCsolution Fundamentals

2.1.3 File Types and File Tabs of the Data ExplorerThe following files are used in this software.

File typeFile tab in the Data Explorer

Details

Data file (extension .lcd) The acquired chromatograms and the following information are saved in this file:

Calculation results including area values and concentrations. Operational or error logs of the HPLC instrument at the time of

data acquisition. The content of the method file used in the analysis (including the

system configuration at the time of analysis). The content of the report format file (when the report was

output). The content of the batch file (when the batch process was

performed).Method file(extension .lcm)

The method file contains all of the information about the analysis conditions and the data analysis parameters.The analysis conditions include the setting information of the analytical instrument and its parameters.The data analysis parameters include the chromatogram peak integration parameters, a quantitative calculation method, and calibration curve information.It also records the system configuration at the time the method is edited, and when importing the method file, the software checks to see if it matches the current system configuration.The appropriate method file allows one to easily perform analysis under the same conditions because you can specify the method file in the batch table and import the method parameters into the data file.

Report format file (extension .lcr)

Saves information about the report format, when it is output, including the layout of the report and detailed settings of the items included in the report.The "LC Real Time Analysis" and "LC Postrun Analysis" both have the "Create Report" function.

Batch file (extension .lcb) Saves batch tables for continuous analysis and postrun analysis of the data.The same file can be used in both "LC Real Time Analysis" and "LC Postrun Analysis".



2 Software Overview


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3

Starting the Analysis

This chapter is a tutorial to help you learn how to operate LCsolution. For better understanding, it is recommended that you actually operate LCsolution and work through the analysis procedure of this chapter.

For common operations performed in LCsolution, refer to "10 Common Window Operations" on page 147, "11 Common Table Operations" on page 155, and "12 Common File Operations" on page 163.

3.1 Before Reading This Chapter

3.1.1 The Analytical Instruments and Analysis Conditions Described in This Chapter This chapter explains the analysis procedure using the following instruments under the listed conditions.

Configuration of the analytical instrumentsHigh pressure gradient UV detection systemPump: 2 LC-10ADvp pumpsAutosampler: SIL-10ADvpColumn oven: CTO-10A(C)vpDetector: SPD-10A(V)vp

Analysis conditionsColumn: Shim-pack VP-ODS (150mm x 4.6mm i.d. 5 m)Mobile phase: Pump A: Water

Pump B: AcetonitrileMobile phase ratio: B.Conc = 50%Column temperature: 40 degrees CelsiusDetection wavelength: 254nmSample: Mixtures of parahydroxybenzoate esters* * Consists of methyl, ethyl, and propyl ester compounds. However, the propyl ester is considered to be the internal standard.

3.1.2 The Data Structure of LCsolutionThis section gives a brief explanation about the data structure of LCsolution.Various LCsolution data parameters are stored in the data file, such as instrument and analysis conditions during data acquisition and data analysis (metadata).( "4.1.5 Data File" in 'Administration Manual') This ensures the traceability of data since the analysis conditions and parameters can be referenced from the data file itself. Also, you can reanalyze the data using only the data file.


3 Starting the Analysis

NOTEThe method information stored in the data file is the duplicate of the method file used during the acquisition and analysis of data. Therefore, when modifying the method parameter in the data file, only the "method in data" is modified and no changes are made to the original method file. But the original method file can be updated accordingly with the modified method in the data file by using "Apply to the Method File" (save as a method file) function.

3.1.3 LCsolution Operation FlowIn this chapter, the operation flow of LCsolution is explained in the following order.

NOTE In a batch table, the method file (.lcm) and the report format file (.lcr) are specified for each particular data file

(.lcd). When the batch analysis is completed, it creates a data file (.lcd) containing the same information stored in the batch table (.lcb) and .lcm and .lcr (when a report is output) set for each line in the batch table.

When you open a method file (.lcm) in the [LC Calibration Curve] window, the data files (.lcd) for all the calibration levels included in the calibration curve of the method file (.lcm) are loaded simultaneously. If the method file (.lcm) is edited here, the data can be recalculated using the new analysis conditions for each imported data file (.lcd). You may also optimize the quantitative method and the type of calibration curve in addition to adding and deleting calibration level data.

[Real Time Analysis] - [Data Acquisition]Method file (.lcm)

[Real Time Analysis] - [Data Acquisition]Data file (.lcd) (Created automatically)

[Postrun Analysis] - [LC Data Analysis]Modifying the method in the data file (.lcd) -> Can modify .lcm accordingly

[Postrun Analysis] - [Report Format]Report format file (.lcr)

[Real Time Analysis] - [Batch Table]Batch file (.lcb), data file (.lcd) (Created automatically)

[Postrun Analysis] - [LC Data Analysis]Data file (.lcd)

[Postrun Analysis] - [LC Calibration Curve]Method file (.lcm)

Setting the analysis conditions 3.2.4

Performing single analysis 3.3.1

Setting peak integration parameters 3.3.2~

Viewing results 3.8 Viewing results (calibration curves) 3.5.4

Planning and performing batch analysis 3.4

Creating reports 3.6

[Postrun Analysis] - [LC Data Analysis]Modifying the method in the data file (.lcd) -> Can modify the method file (.lcm) accordingly[Real Time Analysis] - [Batch Table]Batch file (.lcb), data file (.lcd), and method file (.lcm)

Generating calibration curves 3.5


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3.2 Preparing for Analysis

3.2 Preparing for AnalysisPrior to analysis, start the analytical instruments and login to "LC Real Time Analysis", then configure and verify each analytical instrument. When necessary, set instrument parameters that determine the analysis conditions.

3.2.1 Starting the Analytical Instruments and Logging into "LC Real Time Analysis"Prepare the analytical instruments and login to "LC Real Time Analysis".

Preparing the analytical instruments

1 Ensure that all of the units of the analytical instruments (pump, autosampler, column oven, and detector) are connected to the system controller.2 Turn the power on for each of the units.Each unit is initialized.3 Confirm that there are no error messages displayed on any of the units.

Logging into "LC Real Time Analysis"Start "LC Launcher" and login to "LC Real Time Analysis".Ensure that you have started the analytical instruments prior to starting LCsolution. LCsolution does not recognize the analytical instruments if they are started after LCsolution.

1 Double-click (LCsolution)."LCsolution Launcher" starts.

2 Click the [Operation] tab on the left side of the screen.The [Operation] menu appears.

3 Click (Analysis) of the instrument you wish to use.

The [Login] screen appears.



4 Type your user ID and password, then click [OK].

NOTEDuring initial login, use Admin for [User ID], leaving the password space blank.

The "LC Real Time Analysis" starts and the [LC Real Time Analysis] window (initial setting) appears.

3.2.2 Setting the System Configuration of the Analytical InstrumentsSet the system up so that it matches the actual configuration of the instruments.The following settings are necessary only when LCsolution is connected to the analytical instruments for the first time, or when the configuration of the instrument is changed. If the configuration of the instruments has not been changed, go to "3.2.3 Checking the Analytical Instruments" on page 16.

1 Click the (System Configuration) icon in the assistant bar at the left side of the [LC Real Time Analysis] window (initial setting).The [System Configuration] screen appears.

NOTE

If the [Real Time] assistant bar is not shown, click (Top) icon to display it.

2 In the list of [Modules Used for Analysis], double-click [Instrument].


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3 In the [Instrument] screen, select the instrument type and the communication settings, and click [OK].

NOTEWhen using a photodiode array detector, also select the instrument type and the communication settings of the PDA.

4 Check the instrument modules on the [System Configuration] screen.Instrument modules that are connected is automatically added to [Modules Used for Analysis].5 Double-click the icons of each module to view its property screen.6 Once you have checked the properties of each module, click [OK] and exit the [System Configuration] screen.

If the system is configured correctly, the status display shows [Ready].

NOTEWhen controlling the photodiode array detector using a start signal from the system controller, set [Relay Output Terminal] correctly in the [System Controller] properties.



3.2.3 Checking the Analytical InstrumentsCheck the instruments for each module.

1 Click the (System Check) icon in the assistant bar.The [System Check] screen appears.2 Set each item and click [Run].

1 Select the item to be checked.2 Set the output format.3 Click [Run].

NOTE To check the "lamp energy" and "wavelength accuracy" of the detector's light source, set

the [Check Level] to [Complete]. Otherwise, select [Partial].

To output instrument logs (operation logs, maintenance logs, error logs) and various correction factors, select [Detailed] in [Format]. Otherwise, select [Normal].

The system check starts. A screen indicating the progress of the check is displayed. Please wait.Once the system check has been completed, the result (either pass or fail) is displayed halfway down the screen on the left side.

3 When the progress bar indicates that the system check is 100% complete, click [View Results].A result report that includes the most recent check result is displayed.

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4 Make sure there weren't any device failures.The system check results are automatically saved with a filename including the analysis date in the "LCsolution\Log\Syscheck" folder where LCsolution is installed.

NOTETo modify the criteria for the system check, refer to "3.2.2 Setting the System Configuration of the Analytical Instruments" on page 14 and open the property screen of each instrument module. Then click [System Check] and change the criteria.

3.2.4 Setting the Instrument ParametersYou can set the analysis conditions (instrument parameters) in the [Instrument Parameters View] in the [Data Acquisition] window.Save the instrument parameters to a method file, and use it to perform the analysis.

1 Make sure that [Ready] is displayed in the status display area of the [LC Real Time Analysis] window.

NOTEIf [Not Connected] is displayed, refer to "3.2.2 Setting the System Configuration of the Analytical Instruments" on page 14 and adjust the system configuration as necessary.



2 In the [Instrument Parameters View], set the instrument parameters you wish to use for the analysis.Click [Normal] to display a tab where you can enter analysis conditions such as measurement time, pump flow, the wavelength of the detector, and oven temperature. Be sure to set [LC Stop Time] and [End Time] (measurement end time) for each detector.

NOTE [LC Stop Time] is the duration of time the instrument is controlled. When there are no settings for

the time program or the fraction collector time program, leave it at 0.01 min.

Make sure that [Detect Rack] is done in the advanced settings for the autosampler. If the rack type is not set properly, an error occurs and the analysis is not completed.

3 Save the method file under a new name.A screen to save the settings (of the method file) appears.4 Once the instrument parameters have been set, click [Download] to transfer the settings to the instruments.

The parameters are transferred to the instruments.If the system controller is active during transmission, each instrument module begins its operation with

the new parameters. If the system controller is not active, then click the (Instrument On/Off) button to activate the instrument.

5 When the analysis conditions have been changed, click the (Save) button in the toolbar to overwrite the settings of the method file.It is recommend that you save the method file in a location (folder) that is easy to find since it is used for the subsequent analysis.

NOTEYou can specify the default location for saving files by selecting the project folder in [File] - [Select Project (Folder)]. ( "3.8.1 Viewing and Using [Data Explorer]" on page 42)


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How to use the instrument control tool buttons (Instrument Control Bar)You can use the "Instrument Control Tool Buttons" when operating the instrument.

NOTEYou can display/hide the instrument control bar in the [View] menu.

3.2.5 Monitoring the analytical instrumentsWhen the instruments are properly connected and the [System Configuration] is set properly, the current status information for each instrument is shown in both the Instrument Monitor Bar and in the [Chromatogram View] which can be found in the [LC Real Time Analysis] - [Data Acquisition] window.

[Chromatogram View]The signal intensity of the detectors, the pressure, oven temperature, room temperature, and the detector cell temperature can be monitored.

Button Name Description

Instrument On/OffActivates or inactivates the system controller.When using the CBM-20A/20A line, this button is used to control the pump and the column oven.

Controller On/Off Turns on/ off the LCD of the system controller.

Controller Lock/UnlockLocks/ unlocks the buttons on the system controller console.When locked, you can only control using the LCsolution software. This helps prevent operational errors.

Pump On/Off Starts / stops the operation of the pump.

Purge samplerStart purging the autosampler.The purge time for the autosampler can be set in the [Autosampler] tab when selecting [Advanced] in the [Instrument Parameter View].

Rinse sampler Start rinsing an autosampler.This operation washes the sampling needles.

Oven On/Off Turns on/ off the column oven.

Zeros Detector A Click this button to correct the signal intensity of the detector to zero.



Instrument Monitor BarThe parameters for each instrument module are displayed as numeric values.

NOTEYou can modify the parameters by entering values in the instrument monitor bar.

Setting the type and range of the axis for a graph in [Chromatogram View]You can set the axis types and axis ranges for the graph in [Chromatogram View]

1 Right-click on the graph, and select [Display Settings] from the displayed menu.The [Display Settings] screen appears.2 Set each item and click [OK].

1 Check the status items that you wish to display.2 Set the display range of each status item.3 Click [OK].

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3.3 Performing Single Analysis

3.3 Performing Single AnalysisWhen the position of the chromatogram peak and the optimum peak detection conditions are unknown, a sample chromatogram must be measured in advance and used to set the data analysis parameters.This section explains the procedures for setting the optimum analysis parameters by actually performing single analysis.

3.3.1 Performing Single Analysis

1 Make sure that [Ready] is displayed and that the baseline is stable in the [Chromatogram View].

2 Click the (Single Start) icon in the assistant bar of the [LC Real Time Analysis] window.The [Single Run] screen appears.3 Set each item.

No. DescriptionSpecify the method file to be used for the analysis.Enter the name of the data file created.The location in which the data file is created usually is the currently browsed project (folder shown in the [Data Explorer]). If you wish to specify the folder, click the (Browse) button to open the currently specified destination folder. Specify the folder and the name of the data file.Enter the vial number of the sample injected by the autosampler.If no sample is injected, enter "-1".Enter the tray number of the sample injected by the autosampler. Specify the volume of the injected sample.



4 Click [OK].The single analysis starts and the status in the [LC Real Time Analysis] window changes from [Ready] to [Running].

3.3.2 Setting the Peak Integration ParametersYou can set the most appropriate values by adjusting the peak integration parameters.

1 Click the (Data Analysis) icon in the [Acquisition] assistant bar.After the acquired data files are imported, the "LC Data Analysis" of "LC Postrun Analysis" starts.2 Open another data file that has been created as the result of analysis.The data file can be opened from the [File] menu.

Open the data file to show the chromatograms in the [Chromatogram View].

3 Right-click on the [Chromatogram View], and then select [Data Analysis Parameters] in the displayed menu.

The [Data Analysis Parameters (Method in Data File)] screen appears.


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4 Click the [Integration] tab and enter the appropriate values for the peak integration parameters. Then click [Apply].

In the [Integration] tab, you can adjust the peak integration parameters and determine the most appropriate values. The result of the change can be checked in the [Chromatogram View]. Change the values in each field and find the most appropriate peak integration parameters.

NOTEHere is how you can set the peak integration parameters. First set lower values for [Width] and [Slope]. For example, set the values of [Width] to "3" and [Slope] to "1000" (you may want to change these values depending on the target peak). Then double the original values of both and check the peak detection status. As the [Width] becomes larger, the minute noises are not detected as peaks. On the other hand, as the [Slope] becomes larger, the rolling baseline is not detected as peaks. Repeat the process until no unnecessary peaks are detected, and use those peak integration parameters.

No. DescriptionEnter the minimum width of the peak you wish to detect.You can exclude the noise peak by setting the minimum width of the peak to be detected. Peaks with widths at the half peak height that are equal to or greater than approximately 1/4 of the [Width] value are recognized as peaks.Enter values to determine the starting and ending points of the peak. The slope test can be performed to obtain the best slope value from the data.The positions where the absolute value of the slope of a chromatogram becomes the [Slope] value are detected as the starting and ending points of the peak.Enter a value for the [Drift] parameter to adjust the baseline settings when dividing two or more overlapping peaks.Generally in chromatograms, the peaks are wider as the retention time increases. The [T.DBL] is a parameter that automatically changes the values of [Width] and [Slope], allowing the broad peak with the long retention time to be detected. As the time set in [T.DBL] elapses, it automatically doubles the value of [Width] and decreases the value of [Slope] by 1/2.Enter the minimum area or minimum height of the peaks to be detected. Peaks smaller than the minimum value are not considered as peaks.Select whether to specify the minimum value of the peak by area or by height.



5 Once an appropriate peak integration result is obtained, click [OK].

NOTEIf you cannot obtain an appropriate peak integration result at this point, you can use the "Integration Time Program" to perform the peak integration with greater flexibility. The "Integration Time Program"

can be started by clicking the (Time Program) button on the [Integration] tab.

Slope TestBy performing the slope test, the [Slope] value (peak detection sensitivity) is automatically set from the status of the noises and drifts appearing on the chromatogram prior to the analysis.

1 Click the button shown as in the above screen.2 Click the starting point of the slope test on the baseline.3 Click the ending point of the slope test.

The slope test is performed.

4 Check the [Slope] value. To set this value as a parameter, click [Set to Parameter].In the "Integration Time Program", you can use any peak integration command at any time. For a detailed explanation of each command, refer to the help.


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3.3.3 Setting the Column Performance ParametersThe column performance calculation is one of the functions that support the system suitability test.Complete the following settings when calculating column performance.

NOTE LCsolution supports various calculations including USP/JP/JP2/EP/BP/DAB/EMG/EMG(50%)/area/height,

as well as user-defined equations. For details, see "5.1.1 Calculating Column Performance" in 'Administration Manual'.

Column performance is calculated for every identified or detected peak. The calculation results can be viewed in the compound table or in the peak table, both of which can be found in the [LC Data Analysis] window.

1 Displaying the column performance parameter settings screen.1 Open a data file on "LC Postrun Analysis".2 Right-click on the [Chromatogram View], and then select [Data Analysis Parameters] from the

displayed menu.The [Data Analysis Parameters (Method in Data File)] appears.

3 Click the [Performance] tab and you can set the column performance parameters here.

2 Set the column performance parameters and click [OK].

1 Select the column performance calculation method.2 Specify the unretained peak time.

You can either use the first detected peak retention time, or enter the time of your choice.

3 Enter the column length.This value is used for calculating the theoretical plate height.

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4 Specify whether to calculate all peaks or only those identified.When this box is checked, column performance is calculated using detected peaks that match those in the compound table.

5 Click [OK].The column performance settings are completed.

3 Once the column performance is set, click the (Save) button in the toolbar.The data file is overwritten and saved.

3.4 Performing Batch AnalysisThis section explains the procedures for performing batch analysis.

3.4.1 Creating a Batch TableSet the schedule for continuous analysis.

1 Click the (Batch Processing) icon in the assistant bar in "LC Real Time Analysis".The batch table is displayed.2 Select [New Batch File] from the [File] menu.

3 Enter the batch analysis schedule in the batch table.The main items to be entered into the batch table are as follows:

Vial# Enter the vial number of the sample injected by the autosampler. If no sample is injected, enter "-1".

Tray Name Enter the tray number of the sample injected by the autosampler. No entry is necessary when using an autosampler without multiple tray settings.

Inj. Volume Enter the injection volume (unit: L).Sample Type Select the sample type.

Select [Standard] for the sample in order to create the calibration curve, and select [Unknown] to quantify the sample.For the first standard sample for creating a calibration curve, enable [Initialize Calibration Curve].

Analysis Type Specify whether to perform analysis processing.[IT] means to perform peak integration and [QT] means to perform quantitative calculation.

Method File Specify the method file to be used for the analysis.Data File Name the data that is to be saved as the analysis results.

When the file name is entered without a path, the data is created in the currently browsed project folder (as previously specified).

Level# Enter the calibration level number when using the standard sample.


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3.4 Performing Batch Analysis

NOTEWhen the necessary entry items are not shown in the batch table, right-click on the batch table, and select [Table Style] from the pop-up menu. Add the necessary items to [Display Items].

4 Name the batch table and save it.Click the (Save) button in the toolbar and save it under a new name.The content of the batch table is saved.

ReferenceFor details, see "7.1.1 Displaying and Editing the [Batch Table]" on page 107 and "7.1.2 Using the Wizard" on page 109.

3.4.2 Performing Batch AnalysisPerform analysis based on the batch table settings.

1 While the batch table is open in "LC Real Time Analysis", click (Batch Start) in the [Batch] assistant bar.The batch analysis is performed one by one starting from the first line.

NOTE When pausing the batch analysis, subsequent batch tables can be modified while continuing the

current analysis.

To pause the batch analysis at the line that is currently being analyzed or to restart the paused batch

analysis, click the (Pause/Restart) icon in the [Batch] assistant bar.

To stop the batch analysis, click the (Stop) icon in the [Batch] assistant bar.

Viewing the peak integration results during batch analysisUsing the snapshot feature allows you to temporarily display the data in [Postrun Analysis] and to view the peak integration results. This function is useful when you want to make sure the data from batch analysis is integrated in the manner that you intended within the specified method.

1 During the analysis, click the (Snapshot) icon in the [Acquisition] assistant bar while [Data Acquisition] is active.The "Snapshot" of the data is displayed.

Checking the progress of the batch analysis

The progress of the batch analysis can be checked in the output window. Once the batch analysis has been completed, check the messages to see if there were any errors during the analysis. When the batch analysis is unsuccessful, check the content from the log.



3.5 Generating Calibration CurvesLCsolution generates a calibration curve by batch processing.This section explains how to generate a calibration curve by batch processing of postrun analysis using pre-measured data.Following is the procedure for generating a calibration curve:

1 Create a method file.Set the identification/quantitative parameters such as the peak identification method, the quantitative calculation method, and the type of calibration curve, and save them as a method file.

2 Run the batch table.Using the method file, create a batch table and run the batch.NOTE

Whether performing real time or postrun batch analysis, a calibration curve can be generated using the same procedure.

3.5.1 Creating Data Analysis ParametersFrom the pre-measured chromatogram data, set the peak identification method, the quantitative calculation method, and the type of calibration curve.

1 Click the (Postrun) on "LCsolution Launcher".The "LC Postrun Analysis" starts.2 Click the (LC Data Analysis) icon in the [Postrun] assistant bar.The [LC Data Analysis] window appears.

NOTEWhen using data from the photodiode array detector, click the (PDA Data Analysis) icon in the [Postrun] assistant bar to open the [PDA Data Analysis] window.

3 Select [Open Data File] in the [File] menu and select a pre-measured data file.The data file opens.4 Click the (Wizard) icon in the assistant bar.The [Compound Table Wizard] appears.5 Make sure that appropriate peak integration parameters are set, and click [Next].

NOTEYou can also modify the peak integration parameters here. For more details about setting peak integration parameters, refer to "3.3.2 Setting the Peak Integration Parameters" on page 22.

The [Compound Table Wizard 2/5] screen appears.


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3.5 Generating Calibration Curves

6 Register the retention time of the peaks to be identified and click [Next].In the compound table place checks against the peaks to be registered.The peaks that are detected within certain limits during the retention time set here, are quantitated during real time and postrun analysis.

NOTEThe retention time limits can be set in the [Compound Table Wizard 4/5] screen.


7 Specify the quantitative calculation method and the calibration curve type, and click [Next].The settings should be the same as those shown in the screen below.

NOTEWhen [Group Type] is specified, the following group quantitative calculation can be done in addition to the quantitative calculation for each peak. [Group Calibration] ...... Quantifies the data by generating a calibration curve from the total value of

the peak area set in the same group.

[Conc. Summation] ...... Outputs the total concentration of the quantitative calculation concentration set for the peaks in the same group.




8 Set how you want to identify peaks and click [Next].Do not change the settings here. Continue as is.


9 Provide detailed settings for the compound table and click [Finish].The settings specified up to this point are displayed on the screen. Enter or modify the items as necessary.

No. DescriptionEither [Window] or [Band] can be selected.When [Window] is selected, peaks that are within the limits of the "specified retention time ((Input Value)% + 0.02)" are identified.When [Band] is selected, peaks that are within the limits of the "specified retention time (Input Value) min" are identified.A value for either [Window] or [Band] can be entered.The peaks you want to identify can be selected.When [All Peaks] is selected here, if there are two or more peaks within the limits of one retention time, they are identified as one peak.


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NOTEWhen grouping is selected, [Group#] is added in the compound table field. You should assign the same [Group#] for all substances that are to be calculated quantitatively in the same group. Also, after completing the compound table wizard setup, provide the group settings for each group in the [GroupParam's] tab in the [Compound Table View].

The [Compound Table Wizard] is completed.

10 Confirm changes to the compound table.Once you are finished with the [Compound Table Wizard] setup, click the [View] button at the top-right corner of the compound table and confirm all changes.

NOTEIf you wish to cancel your changes, select [Cancel Edit] in the [Edit Table] menu before clicking the [View] button.

3.5.2 Applying Data Analysis Parameters to the Original MethodSave the analysis parameters set in "LC Data Analysis" as a method file.To use the settings in other analysis, the method in data must be saved as a method file.

1 Click the (Apply to Method) icon in the [LC Data] assistant bar.A screen appears prompting you to specify a name for the method file.2 Name the method file and click [Save].The original method file name is automatically entered into the [File name] field. When applying the

contents of a method set in "LC Data Analysis" to the original method file, click [Save] without making any changes and overwrite the method file.

The [Select Method Parameters] screen appears.

No. DescriptionEnter the compound name.Enter the type.When quantifying using the external standard method, specify everything as the [Target].When quantifying using the internal standard method, specify only the internal standard substance as [ISTD].Enter the concentration of the standard substance at each level. The numbers following "Conc." show the levels of the concentration, and are used to specify the target data when generating a calibration curve. In [Conc. 1], enter the level 1 concentration of each compound included in the standard sample. Similarly, enter the level 2 concentration in [Conc. 2], the level 3 concentration in [Conc. 3] and so on.This field is shown when using the internal standard method. Enter the same [ISTD Group] value for the element that is to be calculated using the internal standard element and the internal standard element itself.



3 Click [OK] without making any changes.

The data analysis parameters that have been set or changed are overwritten and saved into the original method file.

3.5.3 Performing Batch ProcessingCreate a batch table and perform batch processing. Specify the method file.

1 Click the (Batch Processing) icon in the assistant bar.The [Batch Table] window appears.2 Click the (Toggle Data Explorer) button in the toolbar.[Data Explorer] appears.3 Set the folder containing the pre-measured data as the project folder.

Click the (Select Browse Folder) button at the top-right corner of [Data Explorer] and select the folder containing the pre-measured data.

4 Select [New Batch File] in the [File] menu.

5 Drag and drop the data files, which are set as the calibration levels, from the [Data Explorer] to the batch table.

Data file names are added to the batch table.


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6 Specify the method file that you just created in the [Method File] field for all the lines of the batch table.You can browse and specify the method file in the screen that can be displayed by clicking the arrow placed to the right of the cell.

7 Make sure that both [IT] and [QT] are displayed in the [Analysis Type] cells.If they are not shown on the table, check both [IT] and [QT] in the [Analysis Type] screen.

8 Set the [Sample Type] cells.Set the first line to [Standard - Initialize Calibration Curve], and set the second and all subsequent lines to [Standard - Add Calibration Level].

1 Click the first line to display the [Sample Type] screen. Select [Standard] and [Initialize Calibration Curve] and then click [OK].

2 Click the second line to display the [Sample Type] screen. Select [Standard] and [Add Calibration Level], then click [OK].

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9 Set the [Level#].Enter and assign the level of the compound table for each line.When the same level is set for multiple data, the mean value of the data is used to calibrate the level.

10 Save the batch table under a new name.

11 Click the (Batch Start) icon in the [Batch] assistant bar.The batch processing is performed and a calibration curve is generated.3.5.4 Viewing the Calibration Curve

View the generated calibration curve in the [Calibration Curve View] of "LC Data Analysis".

1 Open the most recently processed standard sample data in the batch table.You can open a data file by double-clicking the line number.

NOTEWhen you open standard sample data that is in process of generating a calibration curve, an incomplete curve is displayed.

A calibration curve is displayed on the [Calibration Curve View] of "LC Data Analysis".


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2 View the calibration curve for each element.Click the number in the [ID#] field of the [Compound Table View] to display the calibration curve for each element in the [Calibration Curve View].In the [Conc.] field in the [Results] tab of the [Compound Table View], the concentration value, calculated from the area/height of the peaks on the calibration curve saved in the data, is displayed.

Detailed view of the calibration curve

1 Click the (Calibration) icon in the [Postrun] assistant bar.The [LC Calibration Curve] window appears.2 Open the method file used for generating the calibration curve.To view detailed information about the calibration curve, select [Open Method File] in the [File] menu and

then specify the method file name.



The following contents can be viewed on this screen.

Calibration levels can be added or deleted from this screen.To add calibration level data, drag and drop the data file from [Data Explorer] to the area where the file name is displayed.To delete calibration level data, right-click on the data file that you wish to delete and select [Delete] from the displayed menu.

NOTEIf peaks are not identified within the identification conditions set in the method file, the calibration level of the curve is not added. In this case, either select [Data Analysis Parameters] from the [Method] menu and modify the parameters in the [Identification] tab, or optimize the [Ret. Time] settings in the [Param's] tab of [Compound Table View].

3.5.5 Calculating the Concentration of Data Using the Calibration CurveYou can calculate the concentration of an unknown sample using the calibration curve. The concentration can be calculated either by importing the method or by performing the batch processing.

Importing the method

1 Open the data file that you wish to calculate using either "LC Data Analysis" or "PDA Data Analysis".Select [Open Data File] in the [File] menu and specify the data file name.

2 Select the method file used for generating the calibration curve.Select [Load Method Parameters] from the [File] menu and specify the method file name.The [Select Method Parameters] screen appears.

3 Click [OK].The concentration of data is calculated using the calibration curve in the method file. The calculation results can be viewed in the [Results] tab of the [Compound Table View].

No. DescriptionData file names for all levels of calibration dataChromatogramPeak information


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Performing batch processing

1 Create a batch table.Create a batch table by specifying the method file used to generate a calibration curve. Each field should be set as follows:

2 Click the icon in the [Batch] assistant bar.Batch processing is performed and the concentration is calculated quantitatively from the calibration curve in the method file.

No. Field SettingSample Type All [Unknown]Analysis Type Check both [IT] and [QT]Method File The method file used for generating a calibration curve.Data File An unknown sample of data to be calculated.Level# All [0]



3.6 Creating a Report FormatThis section explains the procedures for creating a data file report. In this section, a report displaying a chromatogram and a peak table is created as an example. "9 Report Function" on page 131

1 Click the (Report Format) icon in the assistant bar of "LC Postrun Analysis".The [Report] window appears.2 Click the (LC/PDA Chromatogram) button in the toolbar.

3 Select the area to which you wish to paste the chromatogram.

In the workspace, drag your cursor from the starting point to the ending point of the area where the item is to be pasted. Release the mouse button at the ending point of the area, and the [LC/PDA Chromatogram Properties] screen appears.


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3.6 Creating a Report Format

4 Click [OK].Set the format of the report on the [LC/PDA Chromatogram Properties] screen. In this example, the report is output using the default value, so click [OK] without making any changes.The chromatogram is displayed on the workspace as shown below.

5 Load the file to the report.Drag and drop the (Data File) icon from the [Data Explorer] onto the workspace. The file selected is loaded into the report.

6 Customize the report.1 Right-click on the workspace and select [Properties] from the menu, or double-click the area of the

workspace where the chromatogram is pasted.The [LC/PDA Chromatogram Properties] screen appears.

2 Set the display format of the report, or enable/disable the display of the title.



NOTEYou can view the report format on the workspace, but by using [Preview] (on page 40), you can view it in more detail.

7 Click the (LC/PDA Peak Table) button in the toolbar.

8 Select the area where you wish to paste the peak table.Just like you pasted the chromatogram in Step 3 above, specify the starting point and ending point by dragging the mouse cursor in the workspace.The layout of the report is now complete.

9 Click the (Preview) icon in the [Report] assistant bar.Check the content of the report in the displayed preview screen. For more details, see "9.5.5 Viewing the Print Image" on page 144.

10 To print the report, click [Print] in the toolbar.The report is printed.11 Save the report format.Once the report format is saved, you can use it to create reports for other data with the same format.

Click the (Save) button in the toolbar, and save the report format under a new name.

NOTE

To open the saved report format - click the (Open) button in the toolbar, and specify the file name in the [Open] screen.

Texts and images can also be pasted and positioned onto the report by using (Text) or (Picture) and the same procedure outlined in Step 3.


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3.7 Creating a Summary Report

3.7 Creating a Summary ReportThe function of the summary report is to create a simple report that includes the chromatograms and peak tables of two or more results as shown below.

There are two types of summary reports. [Concentration]: The results of the concentration, area, and height are displayed altogether. [Compound]: Peak information including concentration and column performance are displayed for each

compound peak.

This section explains the procedure for creating a "Summary Report" with postrun batch analysis. A summary report can be created using the same procedures used to create the report format. For more details about procedures that are not explained here, refer to "3.6 Creating a Report Format" on page 38.

1 Create a summary report format.1 Click the (Report Format) icon in the assistant bar of "LC Postrun Analysis" to display the

[Report] window.

2 Click either the (Summary(Compound)) or the (Summary(Conc.)) button in the toolbar.3 In the workspace, drag your cursor from the starting point to the ending point of the area where the

item is to be pasted.

4 Save the report format

2 Open the batch file used for analysis.Click the (Batch Processing) icon in the assistant bar of "LC

Documents

LC Operation