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Empower Light Scattering Software

Getting Started Guide

34 Maple StreetMilford, MA 01757

71500043903, Revision A

NOTICE

The information in this document is subject to change without notice and should not be construed as a commitment by Waters Corporation. Waters Corporation assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with, or arising from, the use of this document.

© 2002 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA. ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.

Alliance, Millennium, and Waters are registered trademarks, and Empower, LAC/E, and SAT/IN are trademarks of Waters Corporation.

Microsoft and Windows are registered trademarks of Microsoft Corporation.

All other trademarks or registered trademarks are the sole property of their respective owners.

Table of Contents

Preface ......................................................................................... 9

Chapter 1 Light Scattering Software Overview ................................................. 14

1.1 What Is Light Scattering (LS) Data? ..................................... 14

1.2 What Is Light Scattering Software?....................................... 14

1.3 Tutorial Overview................................................................... 18

1.4 Restoring the Light Scattering Default Project ...................... 19

Chapter 2 Setting Up Your Software ................................................................. 22


2.2 Enabling Light Scattering ...................................................... 23

2.3 Obtaining Light Scattering Data ............................................ 24

2.3.1 Importing Light Scattering Data ................................. 24

2.3.2 Setting Up Imported Narrow Standard Information ... 26

2.3.3 Acquiring Light Scattering Data ................................. 28

2.4 Creating an Instrument Method for Acquiring Data............... 28

2.5 Creating an Instrument Method for Imported Data ............... 32

2.6 Performing Light Scattering Calibration and Normalization .. 38

2.7 Tutorial Summary .................................................................. 48

Table of Contents 3

Chapter 3 Processing GPCV-LS Data .............................................................. 49


3.2 Preparing to Process Data.................................................... 51

3.3 Processing GPCV-LS Data ................................................... 52

3.4 Checking the Integration for All Channels and Quantitating Unknowns............................................................................. 55

3.5 Reviewing Results................................................................. 57

3.5.1 Checking the Molecular Weight Distribution .............. 57

3.5.2 Checking the LS Data Plot......................................... 58

3.5.3 Checking the Conformation Plot ................................ 59

3.5.4 Checking the Calibration Plot .................................... 60

3.5.5 Customizing Plot Views ............................................. 61

3.6 Saving Your Results .............................................................. 62


Chapter 4 Automating Your Processing Procedures ........................................ 63


4.2 Using Background Processing.............................................. 63

4.3 Viewing and Checking Results.............................................. 65

4.4 Examining and Printing Reports Using Preview ................... 67


Table of Contents 4

Chapter 5 Reporting and Backing Up Project Data .......................................... 70


5.2 Generating Reports .............................................................. 70

5.3 Backing Up Data ................................................................... 71


Index ....................................................................................... 73

Table of Contents 5

List of Figures

1-1 Empower Login Dialog Boxes ........................................................ 201-2 Empower Pro Window ................................................................... 201-3 Configuration Manager (After Restoring the GPCVLS_Default

Project) .......................................................................................... 21

2-1 Steps in Calibrating Your Light Scattering Detector-Solvent System........................................................................................... 23

2-2 Enabling the Light Scattering Option ............................................. 242-3 Browse Project Dialog Box ............................................................ 252-4 Importing Data ............................................................................... 252-5 Project Window with Imported Data .............................................. 262-6 Alter Sample Window .................................................................... 272-7 Entering Molecular Weight Values for Narrow Standards.............. 272-8 Instrument Method Editor Instruments Tab.................................... 292-9 Add Instrument Dialog Box ............................................................ 292-10 Instrument Method Editor General Tab for Acquiring LS Data....... 302-11 Instrument Method Editor Channels Tab for the LS Detector ........ 312-12 Saving the Instrument Method for Acquiring LS Data.................... 322-13 Project Window.............................................................................. 332-14 Review Main Window..................................................................... 342-15 Instrument Method Editor GPCV Tab ............................................ 352-16 Instrument Method Editor General Tab for the LS Detector........... 362-17 Instrument Method Editor Channels Tab for the LS Detector ........ 372-18 Selecting a Processing Method Type............................................. 382-19 Setting the Peak Width .................................................................. 392-20 Setting the Peak Threshold............................................................ 392-21 Setting the Integration Region ....................................................... 402-22 Setting Minimum Area and Height ................................................. 412-23 Setting GPC Calibration Parameters ............................................. 41

List of Figures 6

2-24 Setting V0 and Vt Values................................................................ 422-25 Naming the Processing Method..................................................... 422-26 Saving the Method Set .................................................................. 432-27 Initial LS Calibration Page.............................................................. 442-28 Selecting LS Calibration Procedures ............................................. 452-29 Detector and Solvent Page ............................................................ 452-30 Normalization Standard Page........................................................ 462-31 Normalization Constants Page ...................................................... 472-32 Calibration Standard Page............................................................. 472-33 Calibration Constants Page ........................................................... 48

3-1 Steps in Processing Broad Unknowns........................................... 503-2 Selecting the Broad Unknown Injection ......................................... 513-3 Broad Unknown Displayed in Review ............................................ 523-4 Selecting the Broad Unknown Name............................................. 533-5 Saving the Processing Method ...................................................... 543-6 Checking the Integration of Each Channel .................................... 553-7 GPC Processing Method Wizard Dialog Box................................. 563-8 Quantitated Broad PS Sample....................................................... 573-9 Molecular Weight Distribution Plot ................................................. 583-10 LS Data Plot................................................................................... 593-11 Conformation Plot .......................................................................... 603-12 Calibration Plot for Processed Standards ...................................... 613-13 Editing Plot Properties ................................................................... 62

4-1 Injections View in the Project Window ........................................... 644-2 Background Processing and Reporting Dialog Box....................... 654-3 Results View of the Project Window .............................................. 664-4 Refractometer Channel of the First Processed Injection ............... 674-5 Open Report Method Dialog Box................................................... 684-6 Result Shown in GPCVLS Results Format.................................... 68

5-1 Background Processing and Reporting Dialog Box....................... 71

List of Figures 7

Preface

The Empower Light Scattering Software Getting Started Guide is intended for a wide variety of users whose familiarity with computers and software ranges from novice to expert. This guide describes the basics of how to use the Empower™ software. Using a standard set of data, this guide takes you through the steps of logging in, calibrating your system, acquiring data, developing methods and method sets, reviewing processed data, and printing reports.

Organization

This guide contains the following:

Chapter 1 describes light scattering software features and procedures for logging in to Empower software and restoring the default project.

Chapter 2 describes calibrating and normalizing your system.

Chapter 3 describes processing unknown samples using dual (GPC-LS) and triple (GPCV-LS) detection.

Chapter 4 describes automating your processing procedures.

Chapter 5 describes generating reports of your results and backing up your projects.

Related Documentation

Waters Licenses, Warranties, and Support: Provides software license and warranty information, describes training and extended support, and tells how Waters handles shipments, damages, claims, and returns.

Online Documentation

Empower Help: Describes all Empower windows, menus, menu selections, and dialog boxes for the base software and software options. Also includes reference information and procedures for performing all tasks required to use Empower software. Included as part of the Empower software.

Empower Read Me File: Describes product features and enhancements, helpful tips, installation and/or configuration considerations, and changes since the previous version.

Empower LIMS Help: Describes how to use the Empower LIMS Interface to export results and import worklists.

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Empower Toolkit Professional Help: Describes how to use the common-object- model, message-based protocol to communicate with the Empower software from a third-party application.

Printed Documentation for Base Product

Empower Software Getting Started Guide: Provides an introduction to the Empower software. Describes the basics of how to use Empower software to acquire data, develop a processing method, review results, and print a report. Also covers basic information for managing projects and configuring systems.

Empower Software Data Acquisition and Processing Theory Guide: Provides theories pertaining to data acquisition, peak detection and integration, and quantitation of sample components.

Empower System Installation and Configuration Guide: Describes Empower software installation, including the stand-alone Personal workstation, Workgroup configuration, and the Enterprise client/server system. Discusses how to configure the computer and chromatographic instruments as part of the Empower System. Also covers the installation, configuration, and use of acquisition servers such as the LAC/E32 module, the busLAC/E™ card, and interface cards used to communicate with serial instruments.

Empower System Upgrade and Configuration Guide: Describes how to add hardware and upgrade the Empower software using an import-and-export upgrade method.

Empower Software System Administrator’s Guide: Describes how to administer the Empower Enterprise client/server system and Workgroup configuration.

Empower Software Release Notes: Contains last-minute information about the product. Also provides supplementary information about specific Empower software releases.

Printed Documentation for Software Options

Empower System Suitability Quick Reference Guide: Describes the basics of the Empower System Suitability option and describes the equations used by the System Suitability software.

Empower PDA Software Getting Started Guide: Describes the basics of how to use the Empower PDA option to develop a PDA processing method and to review PDA results.

Empower GC Software Getting Started Guide: Describes how to use the Empower GC option to develop a GC processing method and to review GC results.

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Empower GPC Software Getting Started Guide: Describes how to use the Empower GPC option to develop a GPC processing method and to review GPC results.

Empower GPCV Software Getting Started Guide: Describes how to use the Empower GPCV option to develop a GPCV processing method and to review GPCV results.

Empower Light Scattering Software Getting Started Guide: Describes how to use the Empower Light Scattering option to develop a light scattering processing method and to review light scattering results.

Empower ZQ Mass Detector Software Getting Started Guide: Describes installation, configuration, calibration, and tuning methods, as well as how to operate the ZQ Mass Detector with Empower software.

Empower Chromatographic Pattern Matching Software Getting Started Guide: Describes how to use the Chromatographic Pattern Matching option to develop a pattern matching processing method and to review pattern matching results.

Empower Dissolution System Software Quick Start Guide: Describes how to operate the Alliance® Dissolution System using Empower software.

Empower Toolkit Programmer’s Reference Guide: Describes how to use the common-object-model, message-based protocol to communicate with Empower software from a third-party application.

Waters Integrity System Getting Started Guide: Describes features of the Waters Integrity® System and provides step-by-step tutorials that guide a user through the use of the Empower Mass Spectrometry (MS) option.

Empower AutoArchive Software Installation and Configuration Guide: Describes how to install and configure the Empower AutoArchive option.

Documentation on the Web

Related product information and documentation can be found on the World Wide Web. Our address is http://www.waters.com.

Related Adobe Acrobat Reader Documentation

For detailed information about using Adobe® Acrobat® Reader, see the Adobe Acrobat Reader Online Guide. This guide covers procedures such as viewing, navigating, and printing electronic documentation from Adobe Acrobat Reader.

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http://www.waters.com

Printing This Electronic Document

Adobe Acrobat Reader lets you easily print pages, page ranges, or the entire document by selecting File > Print. For optimum print quantity, Waters recommends that you specify a PostScript® printer driver for your printer. Ideally, use a printer that supports 600 dpi print resolution.

Documentation Conventions

The following conventions can be used in this guide:

Convention Usage

Purple Purple text indicates user action such as keys to press, menu selec-tions, and commands. For example, “Click Next to go to the next page.”

Italic Italic indicates information that you supply such as variables. It also indicates emphasis and document titles. For example, “Replace file_name with the actual name of your file.”

Courier Courier indicates examples of source code and system output. For example, “The SVRMGR> prompt appears.”

Courier Bold Courier bold indicates characters that you type or keys you press in examples of source code. For example, “At the LSNRCTL> prompt, enter set password oracle to access Oracle.”

Underlined Blue Indicates hypertext cross-references to a specific chapter, section, subsection, or sidehead. Clicking this topic using the hand symbol brings you to this topic within the document. Right-clicking and selecting Go Back from the shortcut menu returns you to the origi-nating topic. For example, “Light Scattering Calibration is described in Section 2.6, Performing Light Scattering Calibration and Normalization.”

Keys The word key refers to a computer key on the keypad or keyboard. Screen keys refer to the keys on the instrument located immediately below the screen. For example, “The A/B screen key on the 2414 Detector displays the selected channel.”

… Three periods indicate that more of the same type of item can optionally follow. For example, “You can store filename1, filename2, … in each folder.”

> A right arrow between menu options indicates you should choose each option in sequence. For example, “Select File > Exit” means you should select File from the menu bar, then select Exit from the File menu.

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Notes

Notes call out information that is helpful to the operator. For example:

Note: Record your result before you proceed to the next step.

Attentions

Attentions provide information about preventing damage to the system or equipment. For example:

Cautions

Cautions provide information essential to the safety of the operator. For example:

STOPAttention: To avoid damaging the detector flow cell, do not touch the flow cell window.

Caution: To avoid burns, turn off the lamp at least 30 minutes before removing it for replacement or adjustment.

Caution: To avoid electrical shock and injury, unplug the power cord before performing maintenance procedures.

Caution: To avoid chemical or electrical hazards, observe safe laboratory practices when operating the system.

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Chapter 1Light Scattering Software Overview
This chapter defines GPC-LS and GPCV-LS data, describes the basic features of the Empower™ GPC-LS software, lists the goals of the tutorials included in this book, and explains how to obtain sample GPC-LS and GPCV-LS data on your workstation. After loading the sample data, you can view and manipulate it in a variety of ways, as described in subsequent chapters. Throughout this guide:

• GPC indicates gel permeation chromatography.

• GPCV indicates gel permeation chromatography with viscometry.

• GPC/V indicates gel permeation chromatography with or without viscometry.

• GPC-LS indicates gel permeation chromatography with light scattering.

• GPCV-LS indicates gel permeation chromatography with viscometry and light scattering.

• GPC/V-LS indicates gel permeation chromatography with or without viscometry and with light scattering.

• GPC/V/LS indicates gel permeation chromatography with or without viscometry and with or without light scattering (GPC, GPC-LS, GPCV, and GPCV-LS).

1.1 What Is Light Scattering (LS) Data?

GPC/V-LS data is collected from a system that has a column set that separates samples by the size of the molecules in solution. In the case of GPC-LS (dual detection), the data comes from two detectors attached to the system: a concentration detector and a light scattering detector. In the case of GPCV-LS (triple detection), the data comes from three detectors attached to the system: a concentration detector, a viscometer detector, and a light scattering detector.

1.2 What Is Light Scattering Software?

The Empower Light Scattering software enables you to process data collected using your light scattering detector along with your gel permeation chromatography (GPC) or gel permeation chromatography with viscometry (GPCV) data acquired using Empower to yield one series of results.

What Is Light Scattering (LS) Data? 13

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The Empower software is a chromatography and results management system you can adapt to your individual requirements. It consists of:

• Empower computer in one of the following configurations:

– Personal workstation configuration

– Workgroup configuration

– Enterprise client/server system

• Empower software running one of four user interfaces:

– Pro

– QuickStart

– Open Access

– Web

• Empower database

Empower Light Scattering software is an integrated part of the Empower software. Data acquisition, processing, and reporting with the Light Scattering software requires use of the base Empower software and database.

For GPC-LS (dual detection) you must enable both the GPC and GPC/LS options in the Project Properties dialog box of Configuration Manager.

For GPCV-LS (triple detection) you must enable both the GPC/V and GPC/LS options in the Project Properties dialog box of Configuration Manager.

Features of the Base Empower Software

Empower software provides an icon-based graphical user interface to acquire, process, and manage chromatographic data. All user actions are performed by pointing and clicking with the mouse (keyboard shortcuts are also supported).

Empower software supports operation in both Windows® 2000 Professional and Windows XP Professional environments, allowing you to have multiple windows open at once. You can view a real-time data acquisition run while simultaneously producing summary results of previously acquired data, or fine-tuning integration parameters for the last injection performed.

The base Empower software provides tools for:

• Creating projects

• Configuring chromatographic systems

• Developing instrument methods to control chromatographic systems

• Acquiring data from samples and standards with SAT/IN™ modules or importing data

• Developing processing methods to perform integration, calibration, and quantitation

• Processing data and obtaining results

Light Scattering Software Overview 14

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• Creating report methods to generate custom reports

• Viewing and printing reports

• Backing up, deleting, restoring, and copying the contents of an individual project

• Enabling Full Audit Trail for system and project actions

Empower Software Reference Information

You can perform additional procedures for data acquisition, processing, and report generation depending on the complexity of your application. Refer to the Empower Help for more information. For information on using Empower Help, refer to the Empower Software Getting Started Guide.

Light Scattering Software Functions

The Empower Light Scattering software supports the following functions:

• Sample loading – Defines narrow and broad standards and narrow and broad unknowns.

• Component loading – Defines the following parameters:

– Molecular weights for narrow standards used to calibrate the light scattering detector-solvent system

– Concentrations for standards and unknowns

– dn/dc values for standards and unknowns

– Solvent for light scattering samples

– Scattering function

– A2 second virial coefficient

– Component names for identifying peaks

– Refractive Index (RI) sensitivity

– Mark-Houwink (K, alpha, K(LS), and alpha(LS)) values for standards and unknowns

– Polydispersity

• Calibration standard choices for calibration curves – Include the following types:

– Narrow

– Broad

– Combined narrow and broad

Note: Light scattering processing calculates the molecular weight values for the standards and builds a calibration curve based on the calculated molecular weights.

What Is Light Scattering Software? 15

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• Calibration technique for calibration curves

– Relative

– Modified universal calibration

– Universal calibration using Benoit’s protocol (with the GPCV option)

• Calibration curve fit types – These fit types are not usually needed for light scattering data but are available for GPC/V-LS data. Include the following fit types:

– First-order

– Second-order

– Third-order

– Fourth-order

– Fifth-order

– Bounded

– Cubic-spline

– Point-to-point

• Review – Displays chromatograms, results (including molecular weight distributions, Viscosity Law, LS Data, Calibration, and Conformation plots), and calibration curves.

• Reporting – Includes custom report groups as well as the following plots, tables, and curves:

– GPCV Viscosity Law plot

– GPC Distribution plot

– GPC General Distribution plot

– GPC Distribution table

– GPC/V Calibration Distribution plot

– GPC Calibration curves

– GPC Calibration Point table

– GPCLS Conformation plot

– Viscosity Law Calibration curves

– LS Instrument Calibration table

– LS Data plot

• GPC/V-LS channel set – Combines up to three types of data (RI detector, viscometer, and light scattering) into one channel set, which enables the software to create a single result from up to three types of data.

• GPC/V-LS processing options – Consist of the Fitting Model options Structural and Calibration for the dual or triple detection data, including a polynomial fit, Random fit, first- and second-order step function fits, and Branching Model options of Star, 3 Branch Points, or 4 Branch Points.


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• Viscosity Law plot – Calculates a Viscosity Law plot (log [η] versus log MW) for GPCV data. Calculates the slope (alpha) and y-intercept (log K) of Viscosity Law plot.

• Conformation plot – Calculates a Conformation plot (log Fitted Rg versus log MW) for LS data. Analogous to the Viscosity Law plot for GPCV data except that log Rg and g are plotted instead of log [η] and g′.

• LS Data plot – Displays the data collected using the light scattering detector.

• Branching index – Calculates the long-chain branching index gLCB and g′LCB (for GPCV), and short-chain branching index gSCB and g′SCB (for GPCV), of a sample. Displays the g and g′ value at each point in the distribution and at each molecular weight average, as well as the overall g and g′ for the polymer. Plots the theoretical linear conformation plot and fitted Rg used to calculate g and the theoretical linear Viscosity Law plot and the fitted viscosity used to calculate g′. For details about calculating g and g′, see “GPC/V/LS Results in the Distribution Table” in the Empower Help.

• Axial dispersion correction – Corrects the broadening of broad standard and broad unknown peaks caused by large-particle (20-µ) columns in the chromatographic system. Empower provides two types of axial dispersion correction:

– Iterative deconvolution

– Peak compression

• Compare calibration – Overlays Calibration Curve plots and their corresponding Viscosity Law plots.

• Compare distribution – Overlays Distribution plots and their corresponding Viscosity Law, Calibration, and Conformation plots.

1.3 Tutorial Overview

The tutorials in this book show you how to:

• Restore the GPCVLS_Default project from the Empower CD-ROM. This project is used in all the examples in this guide.

• Enable the LS option.

• Import or acquire data.

• Create an instrument method for either imported or acquired data.

• Normalize and calibrate the LS detector-solvent system.

• Bring samples into Review.

• Process broad unknowns using triple (GPCV-LS) detection.

• Integrate and quantitate unknowns.

• Review Molecular Weight Distribution, Conformation, LS Data, and Calibration plots.

• Save results.

Tutorial Overview 17

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• Automate processing procedures using background processing.

• Report GPC/V-LS results.

• Use the Preview function to examine and print results.

• Back up your project data.

This guide does not explain how to acquire GPCV data. The data acquisition tutorial is in the Empower Software Getting Started Guide along with other basic Empower tutorials.

Before You Begin

Note: This guide uses the Empower Pro interface. If you do not have access to this interface, see your system administrator.

Before you proceed with the tutorial, ensure that:

• You have installed Empower software as described in the Empower System Installation and Configuration Guide.

• You have installed the GPC or GPCV and LS software options as described in the Empower System Installation and Configuration Guide.

• You have reviewed and followed the basic operating procedures in the Empower Software Getting Started Guide.

• Your printer is properly configured in the Windows 2000 or Windows XP software as described in the appropriate Microsoft documentation.

1.4 Restoring the Light Scattering Default Project

The GPCVLS_Default project contains GPC-LS (dual detection) and GPCV-LS (triple detection) example data used in all the tutorials in this guide. Before you can run the tutorials, you must restore the GPCVLS_Default project to your workstation. Restoring this default project also copies the GPC/V-LS example data to your workstation. The GPCVLS_Default project is on the Empower CD-ROM.

To restore the GPCVLS_Default project:

1. Start Empower software by selecting Start > Programs (for Windows XP, All Programs) > Empower > Empower Login. One of the following Empower Login dialog boxes appears (Figure 1-1).


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Figure 1-1 Empower Login Dialog Boxes

2. Enter your user name and password. If you do not know your user name or password, see your system administrator.

3. If you are using an Empower Enterprise system, select the correct database from the Database list. This list appears only when you are connected to a client/server system.

4. Click Advanced and verify that the Requested Interface field is set to Pro. If you cannot select the Empower Pro interface, see your system administrator.

5. Click OK. The Empower Pro window appears (Figure 1-2).

Figure 1-2 Empower Pro Window

6. Click Configure System. Configuration Manager appears (Figure 1-3).

Workgroup or Enterprise SystemPersonal System

Restoring the Light Scattering Default Project 19

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Figure 1-3 Configuration Manager (After Restoring the GPCVLS_Default Project)

7. Click (Restore Project(s)) to start the Restore Projects Wizard. Follow the instructions on each wizard page. For details, see “Restoring a Project Using the Wizard” in the Empower Help.

Restore Project(s)


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Chapter 2Setting Up Your Software

This chapter provides procedures required to calibrate and normalize your system for operation using narrow standards.


This tutorial familiarizes you with Empower software tools and procedures used to calibrate and normalize your system from narrow standard data, and shows you how to:

• Enable the Empower Light Scattering option

• Create an instrument method for acquiring light scattering data

• Create an instrument method for importing light scattering data

• Create a method set using the wizard

• Edit the processing method for LS channels

• Calibrate and normalize your LS system

For details on acquiring GPCV data using Run Samples, refer to the Empower Software Getting Started Guide, Chapter 3, Starting Up and Making Your First Injection.

Figure 2-1 outlines the steps to calibrate your system using GPC/V-LS narrow standards.


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Figure 2-1 Steps in Calibrating Your Light Scattering Detector-Solvent System

2.2 Enabling Light Scattering

The GPCVLS_Default project has the Light Scattering option enabled. To enable it in another project:

1. In the Empower Pro window (Figure 1-2 on page 20), click Configure System. Configuration Manager appears (Figure 1-3 on page 21).

2. Right-click the project, then select Properties. The Project Properties dialog box appears (Figure 2-2).

Start

Enable the Light Scattering Option (Section 2.2)

Create an Instrument Method for Imported Data

(Section 2.5)

Create an Instrument Method for Acquiring Data

(Section 2.4)

Perform Light Scattering Calibration and Normalization

(Section 2.6)

End

Import Light Scattering Data (Section 2.3.1)

Acquire Light Scattering Data (Section 2.3.3)

Set Up Imported Narrow Standard Information

(Section 2.3.2)

Setting Up Your Software 22

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Figure 2-2 Enabling the Light Scattering Option

3. For dual detection data processing, ensure that the GPC and GPC/LS options are selected. For triple detection data processing, ensure that the GPC, GPC/V, and GPC/LS options are selected.

Note: The Light Scattering (GPC/LS) option is enabled only if the GPC option is selected.

2.3 Obtaining Light Scattering Data

You can acquire light scattering data using SAT/IN modules or you can import data from AIA or ASCII files into Empower (Section 2.3.1). For information on configuring SAT/IN modules, refer to the Empower System Installation and Configuration Guide. When you import narrow standard data, you must set the sample type and enter the molecular weight values (Section 2.3.2).

2.3.1 Importing Light Scattering Data

This example uses data acquired with the Precision Detectors’ light scattering detector. The example project, Import, is not on the Empower CD. You can also import data acquired using other manufacturers’ light scattering detectors.

Note: The information imported into the Empower software varies according to each manufacturer’s software for the light scattering detector.

Obtaining Light Scattering Data 23

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To import light scattering data:

1. Click Browse Project in the Empower Pro window, select the project into which you will import data, then click OK (Figure 2-3).

Figure 2-3 Browse Project Dialog Box

2. Select Database > Import Data from the Project window. The Import Data dialog box appears.

3. Select PD Ascii Data Files from the Files of type field. Browse to the location of the ASCII data files with an extension of .A00 (or equivalent from another manufacturer), select files to import, then click Open (Figure 2-4).

Figure 2-4 Importing Data

4. Click the Injections tab in the Project window to display the imported data (Figure 2-5).


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Figure 2-5 Project Window with Imported Data

When you import ASCII data, each file contains the data for all channels of one injection. The last two digits of the file extension represent the injection number imported into the Empower database.

2.3.2 Setting Up Imported Narrow Standard Information

If your data includes narrow standards used to perform either LS calibration and normalization or GPC calibration, you must set the sample type and enter the molecular weight values. Use a narrow standard to calibrate the LS system. By default, the Empower sample type is set to Narrow Unknown.

Note: The information imported into the Component Editor varies according to the manufacturer’s software for the light scattering detector.

To set up narrow standard information:

1. Select the narrow standard samples from the Project window, then click (Alter Sample). The Alter Sample icon is enabled only when a sample is highlighted. The Alter Sample window appears (Figure 2-6).

Alter Sample

Obtaining Light Scattering Data 25

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Figure 2-6 Alter Sample Window

2. Select each standard, click in the Sample Type column, then select Narrow Standard or the appropriate sample type from the list.

3. Scroll to the right and ensure the injection volume appears in the Inj Vol column.

4. Enter 1 in the RI Sensitivity field.

5. Click (Amount) to open the Component Editor.

Figure 2-7 Entering Molecular Weight Values for Narrow Standards


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6. Click the bottom tab All Samples and the top tab Mol Weights (Figure 2-7). If they do not appear, enter the following information for your narrow standards:

• Molecular Weight

• Concentration

• dn/dc

7. Click Next to move the next standard into the Component Editor, then repeat step 6 for each standard you will use to calibrate the light scattering detector-solvent system.

8. Click the top tab Components and accept the default values for the A2 and Scattering Function columns or enter appropriate values.

9. Click OK to close the Component Editor window.

10. Click (Close) to close the Alter Sample window. When prompted to save the changes to the samples, click Yes. The Project window reappears (Figure 2-5 on page 26).

11. Select View > Update to refresh the Project window.

2.3.3 Acquiring Light Scattering Data

You can configure a Waters® busSAT/IN module (SAT/IN) to acquire up to two channels of LS data. Connect the busSAT/IN module to an Empower workstation by a busLAC/E™ interface. For more information on configuring the busSAT/IN module, see the Empower System Installation and Configuration Guide and the Waters busSAT/IN Module Installation Guide.

2.4 Creating an Instrument Method for Acquiring Data

An instrument method specifies the instrument parameters that you use to acquire LS data. You must set up the instrument method from the Project window before acquiring data.

To create an instrument method:

1. In the Empower Pro window (Figure 1-2 on page 20), click Browse Project. The Browse Project dialog box appears (Figure 2-3 on page 25).

2. Select GPCVLS_Default, then click OK. The Project window appears (Figure 2-13 on page 33).

3. Select File > New Method > Instrument Method. The Instrument Method Editor appears (Figure 2-8).

Creating an Instrument Method for Acquiring Data 27

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Figure 2-8 Instrument Method Editor Instruments Tab

4. Click (Add Instruments) or select Edit > Add Instrument. The Add Instrument dialog box appears (Figure 2-9).

Figure 2-9 Add Instrument Dialog Box

5. Select W410 or your concentration detector from the list, then click OK. The Instrument Method Editor appears. Fill in the fields as appropriate (see the Empower System Installation and Configuration Guide).

6. Click (Add Instruments). The Add Instrument dialog box reappears (Figure 2-9).


2

7. Select SAT/IN 2 from the list, then click OK. The Instrument Method Editor reappears. Fill in the fields as appropriate (see the Waters busSAT/IN Module Installation Guide).

8. Click (Add Instruments). The Add Instrument dialog box reappears (Figure 2-9).

9. Select LS, then click OK. The Instrument Method Editor reappears (Figure 2-10).

Figure 2-10 Instrument Method Editor General Tab for Acquiring LS Data

10. In the Concentration Detector area, select 410 or your RI detector from the Channel list, then enter 1.000 (or the flow rate for your system) in the Flow Rate field.

11. Select the Connected to RI Detector check box.

12. In the Light Scattering Detector area:

• Enter the manufacturer of your LS detector in the Name field.

• Select PD as the detector type from the list.

• Enter 0.170 (or the RI/LS interdetector offset for your system) in the Interdetector Time field.

• Enter 680.0 (or the wavelength of your laser) in the Laser Wavelength field.

Creating an Instrument Method for Acquiring Data 29

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13. Click the Channels tab (Figure 2-11).

Figure 2-11 Instrument Method Editor Channels Tab for the LS Detector

14. In the first line of the table, select Satin Ch 1 from the Channel column, then enter 90.0 in the Geometric Angle column.

15. In the second line of the table, select Satin Ch 2 from the Channel column, then enter 15.0 in the Geometric Angle column.

16. Select File > Save As to save the instrument method. The Save current Instrument Method dialog box appears (Figure 2-12).


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Figure 2-12 Saving the Instrument Method for Acquiring LS Data

17. Save the method as Acquiring LS Data, then click OK. Click (Close) or select File > Exit to close the Instrument Method Editor.

2.5 Creating an Instrument Method for Imported Data

An instrument method specifies the instrument parameters that you use to acquire LS data. When importing data, the instrument method is assigned an ID of 1 in the project database and named Import by default.

To create an instrument method:


2. Select GPCVLS_Default, then click OK. The Project window appears (Figure 2-13).

Creating an Instrument Method for Imported Data 31

2

Figure 2-13 Project Window

3. Click the Injections tab, select PS 6.2K and PS 43.9K, then click (Review). The Review Main window appears (Figure 2-14).

Review


2

Figure 2-14 Review Main Window

4. Select View > View Instrument Method. The Instrument Method Editor appears (Figure 2-15).


2

Figure 2-15 Instrument Method Editor GPCV Tab

5. In the Concentration Detector area, select Ch1 from the Channel list, then enter 1.000 (or the flow rate for your system) in the Flow Rate field.


7. In the Viscometry Detector area:

• Select Ch4 from the Channel list.

• Enter –0.076 (or the interdetector offset for your system) in the Interdetector Time field.

If you are processing data from the Waters Alliance® GPCV 2000 System, divide the interdetector offset value from the RI to Viscometer Volume parameter (mL) in the Detector tab of the Instrument Configuration Editor for the Alliance GPCV 2000 System software by the flow rate (mL/min). Enter the value in the Interdetector Time field.

If you are processing data from the Waters 150CV+ GPCV System, follow the procedure in the “Determining Interdetector Offset” topic in the Empower Help to calculate the interdetector offset value.


2

• Enter 0.000 in the Zero Offset field if you are processing data from the Waters Alliance GPCV 2000 System. If you are processing data from the Waters 150CV+ GPCV System, follow the procedure in the “Determining the Viscometer Transducer Zero Offset (GPCV)” topic in the Empower Help to calculate the viscometer transducer zero offset.

8. Click (LS) to set the light scattering parameters (Figure 2-16).

Figure 2-16 Instrument Method Editor General Tab for the LS Detector

9. In the Concentration Detector area, select Ch1 from the Channel list, then enter 1.000 (or the flow rate for your system) in the Flow Rate field.


11. In the Light Scattering Detector area:

• Enter the manufacturer of your light scattering detector in the Name field.

• Select PD as the detector type from the Detector Type list.

• Enter 0.197 (or the interdetector offset for your system) in the Interdetector Time field.

• Enter 680.0 (or the wavelength of your laser) in the Laser Wavelength field.


2

12. Click the Channels tab (Figure 2-17).

Figure 2-17 Instrument Method Editor Channels Tab for the LS Detector

13. In the first line of the table, select Ch2 from the Channel column, then enter 15.0 in the Geometric Angle column.

14. In the second line of the table, select Ch3 from the Channel column, then enter 90.0 in the Geometric Angle column.

15. Select File > Save to save the instrument method as Import, then click OK.

Note: Any imported file is assigned an Instrument method ID of 1, so selecting File > Save As saves the method with an ID of 1 overwriting the existing method with an ID of 1.

16. Click (Close) or select File > Exit to close the Instrument Method Editor.


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2.6 Performing Light Scattering Calibration and Normalization

The Calibrate Light Scattering System wizard ensures that a GPC processing method and an instrument method are opened before calibrating the LS detector. If the wizard detects that these criteria are not met, it opens other wizards. You must edit the processing method for the light scattering channels, then calibrate the LS detector for each detector-solvent combination used to produce data in the project.

Note: Calibrating your LS system does not create or add points to a calibration curve.

To calibrate and normalize your light scattering system:


2. Select GPCVLS_Default, then click OK. The Project window appears (Figure 2-13 on page 33).

3. Click the Injections tab, select PS 6.2K and PS 43.9K, then click (Review). The Review Main window appears (Figure 2-14 on page 34).

Note: Ensure that the concentration, dn/dc, and molecular weight for each narrow standard used to calibrate the LS detector are entered in the Component Editor.

4. Select Process > Calibrate Light Scattering System. The New Processing Method dialog box appears (Figure 2-18).

Figure 2-18 Selecting a Processing Method Type

5. Ensure that GPC appears in the Processing Type field and the Use Processing Method Wizard check box is selected, then click OK. The Processing Method Wizard opens and the Integration – Peak Detection 1 – Ch1 page appears (Figure 2-19).

Performing Light Scattering Calibration and Normalization 37

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Figure 2-19 Setting the Peak Width

6. Check the peak width. If you are not satisfied, click-drag the cursor to draw a box around the narrowest peak to set the peak width. The view zooms into the selected region. The software determines the peak width setting using the data in the selected region. The value is displayed in the Peak Width field. A peak width value of 120 works well for this RI data. To redraw the box, right-click the chromatogram and select Full View. You can also enter the peak width instead of graphically selecting the integration region.

7. When satisfied with the peak width value, click Next to go to the Integration – Peak Detection 2 – Ch1 page (Figure 2-20) to set the threshold for peak detection.

Figure 2-20 Setting the Peak Threshold


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8. Check your threshold value. If you are not satisfied, click-drag the cursor to draw a box around a section of baseline containing noise that you want to reject. The view zooms into the selected region. The software determines the peak threshold setting using the data contained in the selected region. The value is displayed in the Threshold field. A peak threshold value of 9 works well for this RI data. To redraw the box, right-click the chromatogram and select Full View. You can also enter the threshold instead of graphically selecting the integration region.

9. Click Next to go to the Integration – Integration Region – Ch1 page (Figure 2-21) to define the area where you want to perform integration.

Figure 2-21 Setting the Integration Region

10. Check the integration region. If you are not satisfied, click-drag the cursor to draw a box around the region where you want to perform integration. The view zooms into the selected region. The software determines and displays the start and end settings. Make sure that you include the region up to approximately Vt. A start time of 10 and an end time of 20 work well for this RI data. To redraw the box, right-click the chromatogram and select Full View. You can also enter the start and end times instead of graphically selecting the region.

11. Click Next to go to the Integration – Peak Rejection – Ch1 page (Figure 2-22).


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Figure 2-22 Setting Minimum Area and Height

12. Select Minimum Area, enter 506713, select Minimum Height, then enter 16217. Click Next to go to the GPC – Calibration – Ch1 page (Figure 2-23).

Figure 2-23 Setting GPC Calibration Parameters

13. Enter the following GPC calibration parameters:

• Type of calibration – Universal

• Calibration fit type – 3rd Order

• Calibrate against time or volume – Volume

• Flow rate – 1.000 (or the flow rate of your system)


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14. Click Next to go to the GPC – Column Set – Ch1 page (Figure 2-24). Leave the fields empty.

Figure 2-24 Setting V0 and Vt Values

15. Click Next to go to the Processing Method Name – Ch1 page (Figure 2-25).

Figure 2-25 Naming the Processing Method

16. Enter GPCVLS PM in the Method Name field, then click Finish to complete the wizard and save the processing method. The Integration – Integration Region – Derived Viscometer page appears for the derived viscometer channel (Figure 2-21 on page 40).


2

17. Click Back twice to open the Integration – Peak Detection 1– Derived Viscometer page (Figure 2-19 on page 39).

18. Repeat steps 6 to 12, using appropriate values for the derived viscometer and the LS channels. The following values work well for this data:

Derived Viscometer channel:

Peak width – 90.00

Threshold – 7.000

Minimum Area – 61845

Minimum Height – 1960

LS channels:


Threshold – 5.000



Note: The integration parameters you select for the LS channel apply to all LS channels.

19. Click Finish to close the Processing Method Wizard. The Save current Method Set dialog box appears (Figure 2-26).

Figure 2-26 Saving the Method Set

20. Enter GPCVLS MS in the Name field, then click Save. The Initial LS Calibration page appears (Figure 2-27).


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Note: Clicking the Update existing calibration option button averages the result produced by this wizard session and the result in the existing calibration selected from the table. This choice also enables you to view an existing calibration without modifying it. To view the selected calibration, click Keep Existing normalization in the Normalization Standard page and Do not calibrate now in the Calibration Standard page (Figure 2-32), then click Cancel in the Calibration Constants page (Figure 2-33).

Figure 2-27 Initial LS Calibration Page

21. Ensure that the Create a new calibration option button is selected, then click Next. The LS Calibration Procedures page appears (Figure 2-28).


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Figure 2-28 Selecting LS Calibration Procedures

22. Ensure that the Perform Light Scattering Normalization, Perform Light Scattering Calibration, and Perform RI Calibration check boxes are selected. Click Next. The Detector and Solvent page appears (Figure 2-29).

Figure 2-29 Detector and Solvent Page


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23. The Light Scattering Detector fields are disabled, but match the values in the instrument method. Enter 1.405 or the refractive index of your solvent in the Solvent Refractive Index field, then click Next. The Normalization Standard page appears (Figure 2-30).

Figure 2-30 Normalization Standard Page

24. Click Normalize with isotropic standard, select PS 6.2K or a small, isotropic, narrow standard with a molecular weight less than 50,000, then click Next. The wizard performs the calibration and the Normalization Constants page appears (Figure 2-31).


2

Figure 2-31 Normalization Constants Page

25. Click Calculate to have the wizard calculate the scattering angles and fill in the value for the Scattering Angle for Ch2 on line 1. If the Calculate button is disabled, enter the scattering angle in the table. Click Next. The Calibration Standard page appears (Figure 2-32).

Figure 2-32 Calibration Standard Page


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Tutorial Summary 47

26. Click the Calibrate with mono-dispersed standard option button.

27. Select PS 43.9K from the list. Choose a monodispersed standard with a molecular weight in the mid-range of the molecular weight distribution of your samples. You can use the same standard for normalization and calibration. Ensure that you entered the molecular weight, concentration, and dn/dc of the standard you selected for calibration in the Alter Sample window.

28. Click Next. The Calibration Constants page appears (Figure 2-33).

Figure 2-33 Calibration Constants Page

29. Click Finish to close the wizard. The Review dialog box appears asking if you want to copy the RI Calibration Constant to the instrument. Click Yes. Empower saves this as the current version of the calibration for this detector and solvent combination and uses it in subsequent processing.

2.7 Tutorial Summary

In this chapter, you have learned to:

• Enable the Empower Light Scattering option

• Create an instrument method for acquiring light scattering data

• Create an instrument method for importing light scattering data

• Create a method set using the wizard

• Edit the processing method for LS channels

• Calibrate and normalize your LS system

3

Chapter 3Processing GPCV-LS Data

This chapter steps you through procedures for processing your GPCV-LS data. To process GPC-LS data, follow the same steps, however, you do not need to add the viscometer channel to the method set, integrate it, or check its integration.


This tutorial familiarizes you with the Empower software tools and procedures used to process unknown samples, and shows you how to:

• Prepare standards and unknowns for processing

• Process GPCV-LS triple detection data

• Check the integration for all channels

• Quantitate unknowns

• Check the Distribution, LS Data, Conformation, and, if you process standards, Calibration plots

• Customize plot views

• Save your results

Figure 3-1 outlines the steps to process unknowns.


3

Figure 3-1 Steps in Processing Broad Unknowns

Start

Prepare to Process Data (Section 3.2)

Process GPC/V-LS Detection Data (Section 3.3)

Check Integration for All Channels (Section 3.4)

Integration OK?

Adjust the Processing Method

Check the Molecular Weight Distribution (Section 3.5.1)

Yes

No

Check the LS Data Plot (Section 3.5.2)

Check the Conformation Plot (Section 3.5.3)

Save Your Results (Section 3.6)

End

A

A

Quantitate Unknowns (Section 3.4)

Create Universal

Calibration?

Yes

No

See the Empower Software GPCV Getting Started Guide

Customize Plot Views (Section 3.5.5)

Check the Calibration Plot (Section 3.5.4)

Processing GPCV-LS Data 49

3

3.2 Preparing to Process Data

Processing standards and broad unknown samples involves bringing the broad unknown injections into Review.

1. In the Empower Login dialog box, enter your user name and password, then click OK. The Empower Pro window appears.

2. Click Review Data. The Review Data dialog box appears.

3. Select the GPCVLS_Default project and Injections, then click OK. The Project window appears.

4. Select the Broad PS broad unknown injection (Figure 3-2).

Figure 3-2 Selecting the Broad Unknown Injection

5. Click (Review) or select Tools > Review. The Review Main window appears with the refractometer channel signal of the broad unknown in the Chromatogram plot and the broad unknown injections in the data tree (Figure 3-3).

Review

Preparing to Process Data 50

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Figure 3-3 Broad Unknown Displayed in Review
3.3 Processing GPCV-LS Data

Processing a broad unknown sample involves adjusting the processing method to properly integrate all channels. To process the data from the Review Main window before applying either a processing method or method set:

1. Select Process > GPCV-LS Data for triple detection data processing or GPC-LS Data for dual detection data processing.

Note: If you open a method set or processing method before selecting Process > GPCV-LS Data and the method set contains a GPCV-LS channel set with a universal GPC processing method, the data is processed using the opened methods.

The New Processing Method dialog box appears (Figure 2-18 on page 38).

2. Ensure that GPC appears in the Processing Type field and the Use Processing Method Wizard check box is selected, then click OK.

3. Follow steps 6 to 14 in Section 2.6 on page 38.


3

The following values work well for this data:

RI channel:


Threshold – 5.000



4. Click Next to go to the GPC – Broad Unknowns and Standards page (Figure 3-4).

Figure 3-4 Selecting the Broad Unknown Name

5. Ensure that Broad PS appears in the Name column or click the Name column and select Broad PS from the list.

Note: The name you select must match the component name entered during sample loading or the Alter Sample window (Figure 2-6 on page 27).

6. Click Next to go to the Processing Method Name – Ch1 page (Figure 2-25 on page 42).

7. Enter GPCVLS PM in the Method Name field, then click Finish. The Save current Processing Method dialog box appears (Figure 3-5).


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Figure 3-5 Saving the Processing Method

8. Click Save to save the method and close the wizard. When a message box asks if you want to overwrite the existing processing method with this name, click Yes. The Integration – Integration Region – Derived Viscometer page appears for the derived viscometer channel (Figure 2-21 on page 40).

9. Click Back twice to open the Integration – Peak Detection 1 – Derived Viscometer page (Figure 2-19 on page 39).

10. Repeat step 6. on page 39 through step 12, using appropriate values for the derived viscometer and the LS channels. The following values work well for this data:

Derived Viscometer channel:


Threshold – 3.000



LS channels:


Threshold – 5.000



Note: The integration parameters you select for the LS channel apply to all LS channels.


3

11. Click Finish to close the Processing Method Wizard. The Save current Method Set dialog box appears (Figure 2-26 on page 43).

12. Enter GPCVLS MS in the Name field, then click Save. When a message box asks if you want to overwrite the existing method set, click Yes.

3.4 Checking the Integration for All Channels and Quantitating Unknowns

To verify that the integration is correct for all channels of data and quantitate unknowns:

1. In the Review Main window, click (Apply Method Set) or select Process > Apply Method Set to apply the method set to the data (Figure 3-6).

Figure 3-6 Checking the Integration of Each Channel

2. Click (Next 2D Channel), then evaluate the integration parameters on the viscometer channel. Click (Next 2D Channel), evaluate the integration parameters on the first light scattering channel, then repeat to evaluate integration

Checking the Integration for All Channels and Quantitating Unknowns 54

3

parameters on the second light scattering channel. If you are unsatisfied with the integration parameters:

a. Click (Processing Method Wizard). The GPC Processing Method Wizard dialog box appears (Figure 3-7).

Figure 3-7 GPC Processing Method Wizard Dialog Box

b. Select Edit Existing GPC Processing Method and Keep the Calibration, then click OK. The Review dialog box appears asking if you want to edit the parameters for all channels in the result. Click Yes. The Integration – Integration Region – Ch1 page appears (Figure 2-21 on page 40).

c. A message box asks if you want to edit parameters for all the channels or only the current channel. Click Yes to edit parameters for only the current channel.

d. Click Back twice to open the Integration – Peak Detection 1 – Ch1 page (Figure 2-19 on page 39). Adjust the integration parameters using the wizard, if necessary.

3. Click (Quantitate) and check the values (Figure 3-8).


3

Reviewing Results 56

Figure 3-8 Quantitated Broad PS Sample

4. If you altered the processing method, select File > Save Processing Method, then click (Apply Method Set) to apply the method.

3.5 Reviewing Results

You can review the Distribution, LS Data, Conformation, and Calibration plots for your broad unknown samples. If you processed GPCV-LS data, you can review the Viscosity Law plot. See the Empower GPCV Software Getting Started Guide for details.

3.5.1 Checking the Molecular Weight Distribution

To check the molecular weight distribution curve and molecular weight averages:

1. In the Review Main window, click (Results) or select Window > Results to access the Results window.

2. Click the Peaks tab in the top pane if it does not appear as the default tab, and make sure the peak with the distribution you want to view is selected in the Peaks table.

3. Click the Distribution Plot tab in the bottom pane. The Distribution plot appears (Figure 3-9).

3
Figure 3-9 Molecular Weight Distribution Plot
4. Check the shape of the normalized distribution plot and placement of the molecular weight averages. For details on changing the plot choices, see Section 3.5.5, Customizing Plot Views.

3.5.2 Checking the LS Data Plot

To check the molecular weight values calculated using the light scattering data and the observed Rg values on the LS Data plot:


2. Click the Peaks tab in the top pane if it does not appear as the default tab and make sure the peak with the distribution you want to view is selected in the Peaks table.

3. Click the LS Data Plot tab in the bottom pane. The LS Data plot appears (Figure 3-10).


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Figure 3-10 LS Data Plot
4. Check that the Fitted Rg and Fitted MW values match the Observed Rg and Observed MW values. First, check the Rg fit because the MW fit depends on the results of the Rg fit. The Rg is always fit using a Calibration Fitting Model, so check it when the X-Axis is Slice EV, Slice Adj RT, or Slice RT. The MW can be fit using either a Calibration or Structural Fitting Model. If you used Structural, change the X-Axis to Slice MW before checking the MW fit. For details on changing the plot choices, see Section 3.5.5, Customizing Plot Views.

3.5.3 Checking the Conformation Plot

The Conformation plot is analogous to the Viscosity Law plot in GPCV except that log Fitted Rg and g are plotted instead of log Fitted [η] (intrinsic viscosity) and g′. To check the Conformation plot and molecular weight averages calculated from light scattering data:

1. From Review Main window, click (Results) or select Window > Results to access the Results window.



3

3. Click the Conformation Plot tab in the bottom pane. The Conformation plot appears (Figure 3-11).

Figure 3-11 Conformation Plot

4. Check that the Fitted Rg value matches the Observed Rg. When the Fitting Model is set to Calibration, you can evaluate how well the Fitted Rg data fits the Observed Rg data by changing the X-Axis choice to Slice EV, Slice Adj RT, or Slice RT. For details on changing the plot choices, see Section 3.5.5, Customizing Plot Views.

3.5.4 Checking the Calibration Plot

If there is a universal calibration curve associated with the processing method used to process the GPCV-LS data, the Calibration plot displays Slice Fitted HV and Slice P versus Slice EV. To check the Calibration plot for light scattering data:



3. Click the Calibration Plot tab in the bottom pane. The Calibration plot appears (Figure 3-12).


3
Figure 3-12 Calibration Plot for Processed Standards
4. Check that the Fitted HV matches the Observed HV. For details on changing the plot choices, see Section 3.5.5, Customizing Plot Views.

3.5.5 Customizing Plot Views

To customize your viewing preferences of the plots available in the Results window:

1. Right-click in the plot, then select Properties. The Plot Properties dialog box appears (Figure 3-13).


3

Figure 3-13 Editing Plot Properties

2. Edit the fields as appropriate, then click OK to return to the Results window of Review.

3.6 Saving Your Results

To save the GPCV-LS results, select File > Save > Result.

Note: If you are saving an unknown, the File > Save > Result selection will not be enabled. Select File > Save > All.


In this chapter, you have learned to:

• Prepare standards and unknowns for processing

• Process GPCV-LS triple detection data

• Check the integration for all channels

• Quantitate unknowns

• Check the Distribution, LS Data, Conformation, and Calibration plots

• Customize plot views

• Save your results


4

Chapter 4Automating Your Processing Procedures

This chapter shows you how to automatically process your GPC/V-LS data


This tutorial familiarizes you with Empower software tools and procedures that you can use to automate calibrating your system and processing your unknowns after you have optimized your processing method to process both standards and unknowns. The tutorial shows you how to:

• Use the Background Processing and Reporting function to process GPC/V-LS injections in the background and streamline the calibration of your system and the processing of broad unknowns

• Use the Preview function to examine and print results in selected report formats

4.2 Using Background Processing

To use the Background Processing and Reporting function to process injections:

1. Open the Empower Login dialog box (Figure 1-1 on page 20), then repeat step 2 to step 5. on page 20.

2. Click Process Data. The Process Data dialog box appears.

3. Select the GPCVLS_Default project and Injections, then click OK. The Project window appears (Figure 4-1).


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Figure 4-1 Injections View in the Project Window

4. Select the Broad PS broad unknown sample, then click Process (Figure 4-1). The Background Processing and Reporting dialog box appears (Figure 4-2).

Automating Your Processing Procedures 63

4

Figure 4-2 Background Processing and Reporting Dialog Box

5. Click Use specified method set in the Processing area, then select GPCVLS MS from the list.

6. If you are processing standards to produce a calibration curve, clear the Clear Calibration check box (Figure 4-2).

7. Click OK.

4.3 Viewing and Checking Results

You use Review to view the results of background processing. To view the processed sample sets:

1. In the Project window, click the Results tab. The Result view table appears (Figure 4-3).

Viewing and Checking Results 64

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Figure 4-3 Results View of the Project Window

2. Highlight the result for the current date.

3. Click (Review) or select Tools > Review (Figure 4-3). The refractometer channel of the first processed injection appears in the Chromatogram plot of Review (Figure 4-4). Review allows you to check the:

• Integration of standard and unknown peaks in all the channels of the result

• Molecular Weight Distribution, Conformation, LS Data, and Viscosity Law plots

4. Use the tree view to sequentially look at the results, injection by injection.


4

Figure 4-4 Refractometer Channel of the First Processed Injection

Checking the Broad Unknown Viscosity Law, Molecular Weight Distribution, Conformation, and LS Data Plots

Examine and, if necessary, adjust the Viscosity Law, Molecular Weight Distribution, Conformation, and LS Data plots for the broad unknown injections. In Review, click (Results), then select the plot from the bottom pane of the Results window.

4.4 Examining and Printing Reports Using Preview

To create reports using Preview:

1. In the Project window, click the Results tab.

2. Select the Broad PS result, then click (Preview/Publisher) or select Tools > Preview/Publisher. The Open Report Method dialog box appears prompting you to specify a report method (Figure 4-5).

Examining and Printing Reports Using Preview 66

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Figure 4-5 Open Report Method Dialog Box

3. Click the Use the following Report Method option button, select GPCVLS Summary Report from the list, then click OK. Report Publisher appears with a report for the broad unknown result based on the GPCVLS Results Report method (Figure 4-6).

Figure 4-6 Result Shown in GPCVLS Results Format

4. Click (Print) to print the report displayed in Report Publisher.


4

5. Use Preview to examine and print reports for one or more results in the result set. For details about previewing a report method, refer to the “Previewing a Report from Report Publisher” topic in the Empower Help.

6. Click Close to close Preview, then select File > Exit to close Report Publisher and return to the Project window.


In this chapter, you have learned how to:

• Use the Background Processing and Reporting function to process GPC/V-LS injections in the background and streamline the calibration of your system and the processing of broad unknowns

• Use the Preview function to examine and print results in selected report formats

Tutorial Summary 68

5

Chapter 5Reporting and Backing Up Project Data

This chapter provides procedures for reporting and backing up your projects.


This tutorial familiarizes you with Empower software tools and procedures used to generate reports and back up your GPC/V-LS data, methods, and results. The tutorial shows you how to:

• Generate printed reports of GPC/V-LS results

• Back up your project to protect your data, methods, and results

5.2 Generating Reports

To generate printed reports of your GPC/V-LS results for one or more samples:

1. In the Browse Project window, click the Results tab. A table of processed results appears (Figure 4-3 on page 66).

2. Select the results for which you want to generate reports.

3. Select Tools > Print. The Background Processing and Reporting dialog box appears (Figure 5-1).


5

Figure 5-1 Background Processing and Reporting Dialog Box

4. Select Print in the Reporting area.

5. Click Use specified report method, then select GPCVLS Summary Report from the list.

6. Click OK. The software generates a report for each selected result based on the selected report method.

5.3 Backing Up Data

To protect your data, method, and results, always back up your GPC/V-LS project whenever you change it.

To back up a project:

1. In the Empower Pro window, click Configure System. Configuration Manager appears (Figure 1-3 on page 21).

2. Select Projects from the tree pane.

3. Select GPCVLS_Default as the project to back up.

4. Click (Backup Project) or select File > Backup Project to open the Backup Project Wizard. Follow the instructions on each wizard page. For details, refer to the “Backing Up a Project Using the Wizard” topic in the Empower Help.

Reporting and Backing Up Project Data 70

5


In this chapter, you have learned how to:

• Generate a report of GPC/V-LS results

• Back up your project

Tutorial Summary 71

INDEX

Index

AAcquiring

GPCV data 19light scattering data 28

Adjusting calibration curves 17Alliance GPCV 2000 System 35, 36ASCII file 24, 26Automating processing procedures 63Axial dispersion correction 18

BBackground processing and reporting 64, 70Backing up GPC/V-LS project 71Backup Project Wizard 71Branching index 18Broad unknowns

checking the Calibration plot 60checking the Conformation plot 59checking the LS Data plot 58processing 51reviewing results 57

CCalibrate Light Scattering System wizard 38Calibration curves

adjusting 17fit types 17

Calibration plot, checking 60Calibration standards 16Calibration technique 17

IChannelchecking the integration 55edit processing method for 38–43,

52–55importing ASCII data 26set concentration 30, 35, 36set LS 31, 37set viscometer 35

CheckingCalibration plot 60Conformation plot 59, 67LS Data plot 58, 67Molecular Weight Distribution plot 57, 67Viscosity Law plots 67

Component loading 16Computer configuration 15Conformation plot 18

checking 59, 67Conventions, documentation 12Creating

GPC processing method 38, 52instrument method

acquiring data 28–32importing data 32–37

DData

acquiring GPCV 19acquiring LS 28backing up 71importing light scattering 25–26light scattering 14reporting 70

Default project 19Detection

dual 14triple 14

Detector type 30, 36

Index 72

INDEX

Documentationconventions 12related 9

Dual detection 14

EEditing processing methods 38–43, 52–55Empower Pro interface 19Empower software 15Enabling light scattering 23

FFile extension 26Fit types, calibration curves 17Fitting models 17Flow rate field 30, 35, 36

GGel permeation chromatography

GPC 14GPC/V 14GPC/V/LS 14GPC/V-LS 14GPC-LS 14, 15GPCV 14GPCV-LS 14, 15

Generating reports 70Geometric angle 31, 37GPC/V-LS project, backing up 71GPC-LS dual detection 14, 15GPCV 2000 System 35, 36GPCV channel set 17GPCV-LS triple detection 14, 15GPCVLS_Default project 18, 19

backing up 71restoring 21

I

IImporting light scattering data 25–26Injection number 26Instrument method

creating for acquiring data 28–32creating for importing data 32–37ID 32, 37

Interdetector offset 30, 35, 36

LLaser wavelength 30, 36Light scattering

detector type 30, 36enabling 23GPC/V/LS 14GPC/V-LS 14GPC-LS 14, 15GPCV-LS 14, 15importing data 25–26software option 14

Loadingcomponents 16samples 16

Login defaults 19LS Data plot 18

checking 58, 67LS software option 14, 15, 16, 19LS system

calibrating 38normalizing 38

MMolecular Weight Distribution plot, checking

57, 67

Index 73

INDEX

NNarrow standards

molecular weight 26, 28, 38setting up 26–28

Normalizing the LS system 38–48

OOffset

interdetector 30, 35, 36zero 36

OptionsLS software 14, 16, 19processing 17

Overviewof light scattering 14–18of tutorial 18

PParameters, integration 38Password, default 19Plots

Calibration 60Conformation 18, 59, 67customizing views 61LS Data 18, 58, 67Molecular Weight Distribution 57, 67Viscosity Law 18, 67

Prerequisites 19Preview 67Previewing reports 68Printing reports 70Processing

background 63broad unknowns 51GPC/V-LS data 52

Processing Method Wizard 38, 52, 56Processing methods, editing 38–43, 52–55Processing options 17

I

Processing procedures, automating 63Project data. See Data

RReferences, Empower software 16Related documentation 9Reports

generating 70previewing 67printing 68, 71report groups 17reviewing 69

Restore Projects Wizard 21Restoring GPCVLS default project 19Reviewing reports 69

SSample loading 16SAT/IN modules 15, 24Saving GPC/V-LS results 62Setting up narrow standards 26–28Software, Empower 15System

calibrating 38normalizing 38password 19

TTriple detection 14Tutorial overview 18

VViewing background processing results 65Viscosity Law plots 18, 67

Index 74

INDEX

WWavelength, laser 30, 36Wizards

Backup Project 71Calibrate Light Scattering System 38Processing Method 38, 52, 56Restore Projects 21

ZZero offset 36

I

Index 75

empower light scattering software€¦ · boxes for the base software and software options. also...

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