Waters Alliance GPC 2000 Series System

Transcription

1 Waters Alliance GPC 2000 Series System Quick Start Guide 34 Maple Street Milford, MA , Revision A

2 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 manual 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 manual WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA. ALL RIGHTS RESERVED. THIS BOOK OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER. Millennium, Alliance, Stragel, and Waters are registered trademarks and PowerStation and Ultrahydrogel are trademarks of Waters Corporation. Ethernet is a registered trademark of Compaq Corporation. Microsoft and Windows NT are registered trademarks of Microsoft Corporation. Zip is a registered trademark of Iomega Corporation. All other trademarks are the sole property of their respective owners.

3 Quick Summary The following figure summarizes the information in this guide. Start Power on the Alliance GPC 2000 Series System (Section 2.1) Label the Columns (Section 2.4.4) Create a Sample Set Method (Section 2.6) Prime the System (Section 2.2) Create a New Instrument Method ( Section ) Prepare and Cap Sample Vials (Section 2.7) Equilibrate the System (Section 2.3) Specify Solvent Manager Parameter Values ( Section ) Load the Carousel (Section 2.7.2) Configure the Mobile Phase (Section 2.4.1) Specify Vapor Sensor Parameter Values (Section 2.5.3) Run the Sample Set (Section 3) Configure the Temperature Ramp Rate (Section 2.4.2) Specify Sample Manager Parameter Values (Section 2.5.4) Shut down the Alliance GPC 2000 Series System (Section 4.1) Configure the Injector (Section 2.4.3) Specify Detector Parameter Values (Section 2.5.5) End

4 Related Adobe Acrobat Reader Documentation For detailed information about using the Adobe Acrobat Reader, refer to the Adobe Acrobat Reader Online Guide. This Online Guide covers procedures such as viewing, navigating and printing electronic documentation from Adobe Acrobat Reader. Printing From This Electronic Document Adobe Acrobat Reader lets you easily print pages, pages ranges, or the entire electronic document by selecting Print from the File menu. 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. Conventions Used in This Guide This guide uses the following conventions to make text easier to understand. Purple Text indicates user action. For example: Press 0, then press Enter for the remaining fields. When you are instructed to press the X key, press the indicated keypad key. When you are instructed to press the X screen key, press the keypad key directly below the key name displayed on the screen. Italic text denotes new or important words, and is also used for emphasis. For example: An instrument method tells the software how to acquire data. Underlined, Blue Color text indicates hypertext cross-references to a specific chapter, section, subsection, or sidehead. Clicking this topic using the hand symbol automatically brings you to this topic within the electronic document. Right-clicking and selecting Go Back from the popup context menu brings you back to the originating topic. For example: To adjust temperature settings in both sample compartment zones and at the preheater, see Section 2.4, Configuring the System, and Section 2.5, Creating an Instrument Method.

5 Notes and Warnings This guide uses the following note and warning conventions: Notes call out information that is important to the operator. For example: Note: Record your result before you proceed to the next step. Attentions provide information about preventing possible damage to the system or equipment. For example: STOP Attention: To avoid damaging the detector flow cell, do not touch the flow cell window. Cautions provide information essential to the safety of the operator. For example: Caution: To avoid chemical or electrical hazards, always observe safe laboratory practices when you operate your system. Caution: To avoid electrical shock and possible injury, remove the power cord from the rear panel of the instrument before you perform the procedures in this section.

6 Table of Contents Chapter 1 Introduction System Description System Components System Compartments System Hardware and Software Chapter 2 Preparing the System for Operation Starting the System Priming the System Equilibrating the System Configuring the System Configuring the Mobile Phase Configuring the Temperature Ramp Rate Configuring the Injector Labeling the Columns Creating an Instrument Method Specifying Storage Server Parameter Values Specifying Solvent Manager Parameter Values Specifying Vapor Sensor Parameter Values Specifying Sample Manager Parameter Values Specifying Detector Parameter Values Creating a Sample Set Method Table of Contents 6

7 2.7 Preparing Samples Preparing and Capping Sample Vials Loading the Carousel Chapter 3 Running a Sample Set Checking the Noise And Drift Starting the Run Chapter 4 Powering Off the System Shutting Down the System Powering Off Table of Contents

8 List of Figures 1-1 Waters Alliance GPC 2000 Series System (Front View) Steps for Preparing the System Alliance GPC 2000 Series Login Window Alliance GPC 2000 Series Interactive Mode Window Using the Priming Syringe Auto Prime Dialog Box Mobile Phase A Property Tab in Instrument Configuration Editor Temperature Control Property Tab in Instrument Configuration Editor Injector Property Tab in Instrument Configuration Editor Columns Property Tab in Instrument Configuration Editor Storage Server Property Tab in Instrument Method Editor Solvent Manager Property Tab in Instrument Method Editor Vapor Sensor Property Tab in Instrument Method Editor Sample Manager Property Tab in Instrument Method Editor Detector Property Tab in Instrument Method Editor Sample Set Method Editor Completed Sample Set Method Table Save Sample Set Method Dialog Box Opening the Sample Compartment Door Accessing the Carousel Loading or Unloading a Vial Attaching the Carousel Handle to the Carousel Removing the Carousel Sample Set Method Table Start Sample Set Dialog Box Table of Contents 8

9 List of Tables 1-1 Major System Components Shutdown Tasks Table of Contents 9

10 1 Introduction The Waters Alliance GPC 2000 Series System Quick Start Guide introduces you to the basic features of the Waters Alliance GPC 2000 Series system and describes how to make a run. Audience This guide is intended for both novice and experienced chromatographers who need to operate the Waters Alliance GPC 2000 Series system. What Is In This Guide? The Waters Alliance GPC 2000 Series System Quick Start Guide contains basic procedural information to help you set up the Waters Alliance GPC 2000 Series system and make a run. Note: This guide is not designed to teach you chemistry and does not contain background or reference information. For additional information, or if you want to learn how to modify any of the procedures in this guide, refer to the Waters Alliance GPC 2000 Series System Online Help or the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide. Before You Begin Before you perform the procedures in this guide, ensure that: Your system is properly installed (refer to the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Chapter 2). Your system software and software options are properly installed. 10 Introduction

11 1.1 System Description The Waters Alliance GPC 2000 Series system (Figure 1-1) is a fully integrated liquid chromatography system designed for use in a wide variety of gel permeation chromatography (GPC) applications. The Alliance GPC 2000 Series system is available in two configurations: Alliance GPC 2000 System, which includes a differential refractive index (RI) detector Alliance GPCV 2000 System, which includes a differential RI detector and a multicapillary viscometry detector These systems perform polymer separations and use Millennium 32 GPC and GPCV software to characterize the molecular weight distribution of the polymers. The GPCV 2000 system also uses universal calibration techniques to determine molecular weights and branching index and frequency information. The solvent management and sample management systems of the Alliance GPC 2000 Series system are integrated within an insulated environment. Controlled temperatures are user-programmable. Three levels of independent thermal monitoring ensure maintenance of the selected temperatures and initiation of the corresponding shutdown routines in the event of a malfunction. The system is compatible with typical GPC solvents such as tetrahydrofuran (THF), toluene, 1,2,4-trichlorobenzene (TCB), and dimethylformamide (DMF), as well as aggressive solvents such as hexafluoroisopropanol (HFIP). Programmable vapor sensors alert the operator to rising levels of vapors in the analysis and sample compartments. 11 System Description

12 1.2 System Components Table 1-1 describes the major components of the Alliance GPC 2000 Series system. Table 1-1 Major System Components Component Solvent management system Sample management system Columns Description Provides degassing, selection, prewarming (up to 50 C), and inline filtering of solvent. The system manages the solvents isocratically. Provides preheating, a mix/spin cycle, and a filter/inject cycle for samples. An internal wash station washes the needle to minimize sample carryover. A wide selection of Styragel (polystyrene divinylbenzene) columns for organic-soluble polymers and Ultrahydrogel (hydroxylated methacrylate) columns for aqueous-soluble polymers can be installed. Column lengths range from 250 mm to 500 mm; widths from 4.6 mm ID (narrow bore) to 7.8 mm ID (standard bore). Detectors A differential refractometer is present in the Alliance GPC 2000 and Alliance GPCV 2000 systems. The Alliance GPCV 2000 system also has the patented multicapillary viscometer. System software The Windows NT operating system provides networking capabilities. The Alliance GPC 2000 Series system software controls all system hardware components, runs sample sets, and acquires data. The Millennium 32 software with GPC and/or GPCV software provides data processing and report generation. 12 Introduction

13 1.3 System Compartments There are four compartments accessed from the front of the system: Syringe compartment Analysis compartment Solvent Management/Communications compartment Sample Management compartment Figure 1-1 shows a front view of the Alliance GPC 2000 Series system. Status LEDs Analysis Compartment Syringe Compartment Sample Compartment Door Button Sample Management Compartment Power Connector Solvent Management/ Communications Compartment Monitor Figure 1-1 Waters Alliance GPC 2000 Series System (Front View) Syringe Compartment The syringe compartment contains: 5-ml syringe used to draw sample into the sample loop Solvent dispenser, which can be used to add solvent to a sample vial Waste manifold, which is used to minimize noise from the detector waste as it exits the system 13 System Compartments

14 Analysis Compartment The analysis compartment contains: Inject valve and sample loop Column bypass valve Refractometer Optional viscometer Column rack, which can hold a maximum of six columns up to 50 centimeters in length Solvent Management/Communications Compartment The Solvent Management portion of the compartment contains: The solvent delivery module, which is similar to the assembly used in the Waters 2690 Separations Module. If the system has the seal wash option, it is built into the solvent delivery module. The inline vacuum degasser, which removes dissolved gasses from the solvents and exhausts the gasses through a vent line. The solvent select valves, which control which solvent (A or B) goes on to the solvent management system. The low temperature stabilization module (LTSM). The module has a preheater loop which can be used to preheat the degassed solvent as it travels to the solvent management system. The Communications portion of the compartment contains: The communications panel for the system control module. The connectors used on this panel are the mouse, the keyboard, a network connector, a serial port, the monitor connector, and the printer port. The ON/OFF/Reset switch for the system control module. A CD-ROM drive. A 3 1/2-inch disk drive. A Zip drive. Sample Management Compartment The Sample Management Compartment contains the sample carousel. Three types of vials can be used in the carousel: 10-mL glass vial 4-mL glass vial with insert 7-mL stainless steel filter vial 14 Introduction

15 1.4 System Hardware and Software The Alliance GPC 2000 Series system software controls all hardware components of the system, runs sample sets, and acquires data. The processing of data and reporting of results are performed by the Millennium 32 software with the GPC and/or GPCV software options. Hardware Features System hardware features include: PC-based central processing unit (CPU) 10.2 gigabyte hard drive Disk drive for 3.5-inch diskettes Zip drive CD-ROM drive I/O card for configuring electrical inputs and outputs Ethernet connection Video, audio, and USB ports The system also includes a separate color monitor, keyboard, and mouse and can accommodate an optional printer. Software Features System software features include: A graphical user interface with self-prompting menus A comprehensive online Help system to assist in operating the system, checking status, and diagnosing problems Capability to remotely network multiple Alliance GPC 2000 Series systems by Ethernet to a Millennium 32 remote server and/or workstation A real-time status display on the system front panel and in the software status bar Real-time monitoring of multiple parameters, of which any two may be viewed at one time Data processing and reporting options using either Millennium 32 software or third-party software (using the appropriate analog-to-digital [A/D] interface and the I/O card) For details about the system software, refer to the Alliance GPC 2000 Series System Overview topic in the Alliance GPC 2000 Series System Online Help Find tab. 15 System Hardware and Software

16 Data Management Options The Alliance GPC control software collects the data from the detector(s). While the Alliance GPC 2000 Series system software acquires data, you can send the data to: Millennium 32 GPC or GPCV software for processing and reporting. For details, refer to the Millennium 32 Chromatography Manager Overview and Millennium 32 GPC and GPCV Overview topics in the Alliance GPC 2000 Series System Online Help Find tab. A third-party data system for processing and reporting (using the I/O card). Refer to the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Section 2.5, Making I/O Card Connections, to connect other devices and Section 3.3, Creating the Current Instrument Method, to configure the I/O card. For more details, refer to the I/O Card Property Tab topic in the Alliance GPC 2000 Series System Online Help Find tab. You can use the Viewer button to display the resulting chromatogram after a sample set is complete. 16 Introduction

17 2 Preparing the System for Operation This section contains step-by-step procedures to prepare the system for operation. Figure 2-1 summarizes the phases of this system preparation. Start Start the System (Section 2.1) Create an Instrument Method (Section 2.5) Prime the System (Section 2.2) Create a Sample Set Method (Section 2.6) Equilibrate the System (Section 2.3) Prepare Samples and Load the Carousel (Section 2.7) Configure the System (Section 2.4) Preparation Complete Figure 2-1 Steps for Preparing the System 17 System Hardware and Software

18 2.1 Starting the System To start up the Alliance GPC 2000 Series system: 1. If the peripheral devices are powered off, power on the color monitor and any other peripheral devices. 2. Move the power switch on the right side panel of the Alliance GPC 2000 Series system to the On (I) position. 3. Open the solvent management compartment door, press and release the inner power button on the communication panel, then close the solvent management compartment door. When the status LEDs on the front of the Alliance GPC 2000 Series system stop flashing, the system is initialized. This takes approximately 5 to 10 minutes. 4. When the Windows NT Begin Logon dialog box appears, simultaneously press Control+Alt+Delete to access the Windows NT operating system. 5. Enter your user name and password. The default Windows NT user name is Administrator (no password). Note: Names and passwords are case-sensitive; be sure to use correct uppercase and lowercase letters. 18 Preparing the System for Operation

19 6. Click OK. The Alliance GPC 2000 Series software starts and the Alliance GPC 2000 Series Login window appears (Figure 2-2). Figure 2-2 Alliance GPC 2000 Series Login Window 7. If you are connecting to a remote Alliance GPC 2000 Series system, click Browse and select the GPC 2000 Node to which you want to connect from the Available Nodes dialog box. If you want to connect to the local Alliance GPC 2000 Series system, proceed to step 8. Note: If the Alliance GPC 2000 Series Login window does not appear after a few minutes, select Programs, GPC2000 Chromatography, GPC2000 Console from the Windows NT Start menu. 8. Click OK. The Alliance GPC 2000 Message Board displays the progress of system startup. 9. When startup is complete, the interactive mode window appears (Figure 2-3). The status indicator at the bottom-right corner of the window shows the current system status and matches the status LED on the front of the system. If a problem occurs, or if the status indicator is red, see the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Chapter 5, Troubleshooting. 19 Starting the System

20 Sample Set Mode Button Carousel Mode Button Diagnostic Mode Button Interactive Mode Button Interactive Mode Control Pane Status Indicator Figure 2-3 Alliance GPC 2000 Series Interactive Mode Window 2.2 Priming the System Prime the solvent management system with a low-viscosity solvent when the flow path is dry or when the system tubing has air bubbles that purging does not remove. Priming the system involves performing a manual prime, performing an Auto Pump Prime, then purging the injector, refractometer, and viscometer (GPCV 2000 system only). Caution: To avoid chemical hazards, always observe safe laboratory practices when you operate the Alliance GPC 2000 Series system and when you handle solvents. Refer to the Material Safety Data Sheets for the solvents you use. 20 Preparing the System for Operation

21 Caution: To prevent overheating a solvent, ensure that you select the correct solvent when you configure a solvent in the Mobile Phase A property tab in the Instrument Configuration Editor. Caution: If you use a solvent that is not in the drop-down list of the Mobile Phase A or B property tab and you select Other in the Instrument Configuration Editor, keep a record identifying the solvent along with its associated operating range and boiling point. STOP Attention: To avoid damaging the columns, ensure that the prime/vent valve is open before you prime the solvent management system. Opening the prime/vent valve prevents the high flow rate during Auto Pump Prime from exerting pressure on the columns. Auto Pump Prime does not ramp up or down when changing the flow rate. Note: To maintain the efficiency of the solvent management system, and to obtain accurate, reproducible chromatograms, use only filtered, HPLC-grade solvents. The following materials are required to prime the system: Priming syringe (Startup Kit) Suitable solvent (THF for the example in this guide) Wide-mouth container that is resistant to your solvent To prime the system: 1. Set up the solvent reservoirs as described in the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Section 2.3.1, Connecting the Solvent Reservoirs. Gently shake the filters in the reservoirs to remove any trapped bubbles. Ensure that all waste tubing drains into the waste container. 2. Click (Interactive Mode) in the Alliance GPC 2000 Series system window. 3. Select Manual Pump Prime from the Operations menu. The Manual Prime wizard appears for either solvent A or B. Note: To prime using the alternate solvent, click Exit, select Current Instrument Method from the Setup menu, then select the solvent you want to prime (A or B) in the Instrument Method Editor. 4. Follow the instructions in the wizard, then click Next. 5. Connect the priming syringe to the prime/vent valve (Figure 2-4), then open the valve by turning it counterclockwise one-half to one turn. 21 Priming the System

22 Note: Because the priming syringe does not lock onto the prime/vent valve, hold the syringe body in place while you pull the syringe plunger. Syringe Plunger Syringe Body Prime/Vent Valve Figure 2-4 Using the Priming Syringe 6. Pull the priming syringe plunger to draw solvent from the reservoir and through the tubing. You may need to use some force to pull the solvent through the system. Repeat several times until you pull all the air into the priming syringe (up to 60 ml or more) and solvent flows out of the prime/vent valve. 7. Click Exit to close the Manual Prime wizard. 8. Select Auto Pump Prime from the Operations menu. The Auto Prime dialog box appears (Figure 2-5). The Flow Rate parameter controls the solvent flow rate during Auto Pump Prime. You can set a flow rate of 0 to 10 ml/min. The default value is 1 ml/min. The Time parameter specifies the duration of the Auto Pump Prime. The default value is 60 seconds, and the suggested value is 300 seconds. 22 Preparing the System for Operation

23 Figure 2-5 Auto Prime Dialog Box 9. Review the Flow Rate and Time parameter values and change them if necessary. 10. Click Start, then continue to pull solvent into the syringe until no air bubbles flow into the syringe. 11. Place a container under the prime/vent valve to catch drips, then remove the syringe. 12. After the solvent management system enters Idle mode, close the prime/vent valve hand-tight by turning the valve fully clockwise. 2.3 Equilibrating the System Before you run samples and acquire data, equilibrate the Alliance GPC 2000 Series system at initial run conditions until the baseline is stable. In this section, we will equilibrate using interactive mode, but you can also equilibrate as part of a sample set using an Equilibrate function. For details, refer to the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Section 3.7, Running a Sample Set. Note: Small variations in parameters such as flow rate, temperature, and solvent viscosity can adversely affect chromatographic results. Allow at least four hours to equilibrate the solvents in the system. Bringing the system from ambient to high temperatures may require up to 24 hours for equilibration and a stable baseline. Note: Before you equilibrate your system, select the appropriate temperature settings (set points, limits, and ramp rates) for the solvent in the current instrument method. To adjust temperature settings in both sample compartment zones and at the preheater, see Section 2.4, Configuring the System, and Section 2.5, Creating an Instrument Method. For details, refer to the Temperature Control Property Tab topic in the Alliance GPC 2000 Series System Online Help Find tab. 23 Equilibrating the System

24 STOP To equilibrate the system in interactive mode: 1. Click (Interactive Mode) in the Alliance GPC 2000 Series system window. 2. Specify an appropriate set of instrument operating parameters for the current instrument method. For details, refer to the Creating/Modifying an Instrument Method for Interactive Mode Operation and the Temperature Control Property Tab topics in the Alliance GPC 2000 Series System Online Help Find tab. 3. Enter the solvent flow rate in the Flow text box of the Interactive Mode control pane (Figure 2-3). Refer to the column care and use manuals for recommended ranges of flow rates. Attention: To prevent damage to your columns from sudden changes in solvent flow rate, use the Limit Ramp Rate function in the Instrument Configuration Editor (for solvent reservoir A). Increase the flow rate in increments of no more than 0.1 ml/min. 4. Click Set Flow in the Interactive Mode control pane to download the current instrument method and to activate baseline monitoring. Set Flow uses the initial conditions in the instrument method to perform equilibration. Plots of the refractometer baseline and viscometer baseline (Alliance GPCV 2000 system only) appear in the Channel Plot panes when equilibration begins. 5. Equilibrate until the baseline is stable. If the baseline does not stabilize, see the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Chapter 5, Troubleshooting. Note: You can also equilibrate your system by inserting an Equilibrate row at the start of a sample set method. Select Equil from the Function drop-down list in the Sample Set Method table (Figure 3-1), then select an appropriate instrument method from the Method drop-down list. When you run the sample set method, equilibration automatically occurs. Using an Equil function in your sample set method allows you to equilibrate for a specified run time. 24 Preparing the System for Operation

25 2.4 Configuring the System Configuring the Alliance GPC 2000 Series system involves specifying parameters that are required for basic operation of the system. See the Windows NT documentation for instructions on making the initial setup and configuration settings in the Windows NT software. To specify generic settings that affect how you use the system and how the system operates: 1. Configure the system parameters in the Instrument Configuration Editor during initial system installation and after making changes to the system components and solvents. Specify parameters in the Instrument Configuration Editor for: Mobile Phase A (see Section 2.4.1) Temperature Ramp Rate (see Section 2.4.2) Injector (see Section 2.4.3) Columns (see Section 2.4.4) 2. After you configure the components, click OK to save your settings and close the Instrument Configuration Editor Configuring the Mobile Phase For this example, THF is used as the mobile phase. See Section 2.2, Priming the System, for details about preparing the flow path for use with THF. For information on commonly used solvents, see the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Appendix D, Solvent Considerations. To configure the mobile phase: 1. Click (Interactive Mode) in the Alliance GPC 2000 Series system window. 2. Select Instrument Configuration from the Setup menu. The Instrument Configuration Editor appears with the Mobile Phase A property tab active (Figure 2-6). 25 Configuring the System

26 Figure 2-6 Mobile Phase A Property Tab in Instrument Configuration Editor 3. From the Solvent drop-down list, select THF. Review the suggested operating range, boiling point, and recommended temperature shutdown limits of the solvent. Caution: If you use a solvent that is not listed and you select Other from the Solvent drop-down list, keep a record identifying the solvent along with its associated operating range and boiling point. STOP 4. Specify the flow ramp rate and maximum operating flow rate: a. Select Limit Ramp Rate to control the rate at which the flow rate increases. b. Select 0.50 from the Ramp Rate (ml/min/min) drop-down list. c. Specify the maximum flow rate by entering 2.0 in the Operating Max Flow (ml/min) field. Attention: A flow rate that is too high can damage columns and cause solvent to overflow from the solvent dispenser. The appropriate flow rate depends on the solvent viscosity and temperature. d. Select Immediately in the Stop Flow section to stop solvent flow immediately after a Stop Flow command. 5. Specify the pressure shutdown limits: a. Enter 2000 in the High Pressure (psi) field. b. Enter 0 in the Low Pressure (psi) field. 26 Preparing the System for Operation

27 6. Select the options for the solvent. See Table D-2 in the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide to determine which options to select for your solvent. a. Select Degasser to vacuum degas the solvent. b. If the seal-wash option is installed, select Seal Wash to wash the piston seals. Note: The seal-wash option must be installed and connected to a seal-wash reservoir. See the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Section 2.3.4, Connecting the Piston Seal Wash. c. Select Bubble Detection to detect bubbles in the flow path of the solvent management system. 7. Continue with Section Configuring the Temperature Ramp Rate The temperature ramp rate controls the rate at which the analysis compartment temperature increases during startup and decreases when cooling down. This parameter in the Instrument Configuration Editor affects changes to the temperature set point in the instrument method (see Section 2.5.5, Specifying Detector Parameter Values). To set the temperature ramp rate: 1. Click the Temperature Control property tab in the Instrument Configuration Editor (Figure 2-7). Figure 2-7 Temperature Control Property Tab in Instrument Configuration Editor 2. Select Enable Temperature Ramp Rate to control the rate of temperature increase when the analysis compartment is warming up and cooling down. 3. Select 0.50 in the Temperature Ramp Rate (C/min) drop-down list. 4. Continue with Section Configuring the System

28 2.4.3 Configuring the Injector Configuring the injector involves specifying the exact sample loop volume, setting the syringe size, selecting the draw rate, and selecting the dispense rate. Note: To configure the injector, you must know the exact injector loop volume. The exact volume is stamped on a tag attached to the injector sample loop in the analysis compartment. To configure the injector: 1. Click the Injector property tab in the Instrument Configuration Editor (Figure 2-8). Figure 2-8 Injector Property Tab in Instrument Configuration Editor 2. In the Injector Loop Volume (ul) field, enter the exact injector loop volume. 3. Select 5 ml from the Syringe Size drop-down list. 4. Select 5.0 from the Syringe Dispense Rate (ml/min) drop-down list. The dispense rate is used when dispensing solvent to wash the needle or to provide solvent to the solvent dispenser (sample make-up station). 5. Select 5.0 from the Syringe Solvent Draw Rate (ml/min) drop-down list. The draw rate is used when drawing fresh solvent from the solvent reservoir. The system displays the aspirate volumes of the samples and the maximum number of injections from the vials. 6. Continue with Section Preparing the System for Operation

29 2.4.4 Labeling the Columns The Columns property tab of the Instrument Configuration Editor allows you to identify the columns in the analysis compartment (for reference only). To enter column identification information in the software: 1. Click the Columns property tab in the Instrument Configuration Editor (Figure 2-9). Figure 2-9 Columns Property Tab in Instrument Configuration Editor 2. Select the Column 1 check box. 3. Select Styragel HT 6E Linear from the Column Style drop-down list. 4. Enter the column serial number and description (up to 250 alphanumeric characters) in the Column Serial Number field. 5. Repeat steps 2 through 4 for each column. 6. Click OK to save the instrument configuration and close the dialog box. 29 Configuring the System

30 2.5 Creating an Instrument Method The instrument method specifies the operating parameters for the solvent manager, sample manager, vapor sensors, and detectors. Creating an instrument method involves setting parameter values for these components in the property tabs of the Instrument Method Editor dialog box: Storage Server property tab (Section 2.5.1). For details, refer to the Storage Server Property Tab topic in the Alliance GPC 2000 Series System Online Help Find tab. Solvent Manager property tab (Section 2.5.2). For details, refer to the Solvent Manager Property Tab topic in the Alliance GPC 2000 Series System Online Help Find tab. Vapor Sensors property tab (Section 2.5.3). For details, refer to the Vapor Sensors Property Tab topic in the Alliance GPC 2000 Series System Online Help Find tab. Sample Manager property tab (Section 2.5.4). For details, refer to the Sample Manager Property Tab topic in the Alliance GPC 2000 Series System Online Help Find tab. Detector property tab (Section 2.5.5). For details, refer to the Detector Property Tab topic in the Alliance GPC 2000 Series System Online Help Find tab Specifying Storage Server Parameter Values Use the Storage Server tab to start creating the new instrument method and to save the completed instrument method. Creating a New Instrument Method To create a new instrument method: 1. Select Instrument Method Editor from the Tools menu in the Alliance GPC 2000 Series system window. The Instrument Method Editor appears with the Storage Server property tab active (Figure 2-10). 30 Preparing the System for Operation

31 Figure 2-10 Storage Server Property Tab in Instrument Method Editor 2. Click New to create a new instrument method. The new method is based on the default instrument method. You can modify the instrument method by selecting or entering parameter values in each property tab of the current instrument method, as described in Section 2.5.2, Specifying Solvent Manager Parameter Values, through Section 2.5.5, Specifying Detector Parameter Values. Saving an Instrument Method When you have specified the operating parameters for the solvent manager, sample manager, vapor sensors, and detectors, return to the Storage Server tab to save the completed instrument method. To save a new instrument method: 1. With the Storage Server tab active, click Save As. The Save As dialog box appears. 2. Enter a name for the newly created instrument method, then click OK. 3. Click OK to close the Instrument Method Editor and to activate the new values in the current instrument method. 31 Creating an Instrument Method

32 2.5.2 Specifying Solvent Manager Parameter Values To specify the solvent manager parameter values: 1. Click the Solvent Manager property tab in the Instrument Method Editor dialog box (Figure 2-11). Figure 2-11 Solvent Manager Property Tab in Instrument Method Editor 2. Click A-THF in the Solvent Source section to select the solvent. 3. Enter a flow rate of 1 ml/min in the Flow (ml/min) field. 4. Select Enable in the Solvent Temperature section, to activate the solvent temperature settings of the solvent preheater, then: Select 40 in the Set Point (C) field. Select 45 in the High Limit (C) field. Select 30 in the Low Limit (C) field. 5. Continue with Section 2.5.3, Specifying Vapor Sensor Parameter Values. 32 Preparing the System for Operation

33 2.5.3 Specifying Vapor Sensor Parameter Values Use the Vapor Sensors property tab to enable or disable the vapor sensors or to change the vapor shutdown limit or the vapor warning limit. To specify the vapor sensor parameter values: 1. Click the Vapor Sensors property tab in the Instrument Method Editor dialog box (Figure 2-12). Figure 2-12 Vapor Sensor Property Tab in Instrument Method Editor 2. Select Enable to activate monitoring by each vapor sensor: Ambient Vapor (near the right side vents) Carousel Vapor (in the sample compartment near the carousel) Detector Vapor (on the back wall of the analysis compartment) 3. Leave the shutdown and warning limits at the default values. 4. Continue with Section 2.5.4, Specifying Sample Manager Parameter Values Specifying Sample Manager Parameter Values Use the Sample Manager property tab to specify the settings for mixing, preheating, and filtering samples. The sample compartment has temperature zones for the carousel and the injector (needle). You can change the default temperature set point and temperature limits for each zone. For details on specifying temperatures in an instrument method, refer to the Setting Up a New Instrument Method topic in the Alliance GPC 2000 Series System Online Help Find tab. To specify the sample manager parameter values: 1. Click the Sample Manager property tab in the Instrument Method Editor dialog box (Figure 2-13). 33 Creating an Instrument Method

34 Figure 2-13 Sample Manager Property Tab in Instrument Method Editor 2. To specify the method for mixing samples, select Enable in the Mixer section, then: a. Select Medium in the relative speed drop down list. b. Select 5.0 in the mix duration drop down list. c. Select Agitate (alternating between clockwise and counterclockwise) in the mix direction drop down list. d. Select 5 seconds in the mix spin time and mix stop time drop down lists. 3. To specify the rate at which the needle travels, select 50 mm/min for the travel rate in the Filter section. 4. To control the carousel temperature, select Enable in the Carousel Temperature section, then: a. Enter 40 in the Set Point (C) field. b. Enter 45 in the High Limit (C) field. c. Enter 35 in the Low Limit (C) field. 5. To specify the rate at which the syringe moves when aspirating a sample, select 0.8 ml/min for the draw rate in the Syringe section. 6. To control the injector (needle) temperature, select Enable in the Injector Temperature section, then: a. Enter 40 in the Set Point (C) field. b. Enter 45 in the High Limit (C) field. c. Enter 35 in the Low Limit (C) field. 7. Continue with Section 2.5.5, Specifying Detector Parameter Values. 34 Preparing the System for Operation

35 2.5.5 Specifying Detector Parameter Values To specify the parameters for the refractometer and the viscometer (GPCV 2000 system only): 1. Click the Detector property tab in the Instrument Method Editor dialog box (Figure 2-14). Figure 2-14 Detector Property Tab in Instrument Method Editor 2. In the RI section, select Autozero to automatically zero the refractometer baseline. 3. If you have an Alliance GPCV 2000 system, select Visc Autozero in the Viscometer section to automatically zero the viscometer baseline. 4. If you want to control the analysis compartment temperature, select Enable in the Column Temperature section, then: a. Enter 40 in the Set Point (C) field. b. Enter 45 in the High Limit (C) field. c. Enter 35 in the Low Limit (C) field. 5. Select 1 samples/sec for the sample rate. 6. Follow the instructions in Saving an Instrument Method on page 31 to save the changes and close the Instrument Method Editor. 35 Creating an Instrument Method

36 2.6 Creating a Sample Set Method You create a sample set method to automatically perform system operations and make the sample injections. The sample set method you create in this section will: Purge the injector Purge the refractometer Purge the viscometer Calibrate the viscometer Inject two samples from each vial Note: You can also perform the purge and calibrate functions in interactive mode. For details, see the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide, Section 3.4, Preparing the Flow Path. 1. Click (Sample Set Mode), then click the Editor button. The Sample Set Method Editor appears (Figure 2-15). Figure 2-15 Sample Set Method Editor 2. Click New to create a new sample set method. 36 Preparing the System for Operation

37 3. Before running samples, you may want to repurge the system. To automate this process, you can add the purge functions to your sample set. To do this, click the Sample field in the first line of the method table, then type Inject Purge for the sample name. 4. Click the Method field and select the instrument method you created in Section 2.5, Creating an Instrument Method. 5. Click the Function field and select Inj Purge from the drop-down list. 6. Click the Time field and enter 5 minutes. 7. Move to the second line and: a. Enter RI Purge for the sample name. b. Select the instrument method you created in Section 2.5, Creating an Instrument Method. c. Select RI Purge in the Function field. d. Enter 5 minutes in the Time field. 8. If you have a GPCV 2000 system, move to the third line and: a. Enter Visc Purge for the sample name. b. Select the instrument method you created in Section 2.5, Creating an Instrument Method. c. Select Visc Purge in the Function field. d. Enter 40 minutes in the Time field. 9. If you have a GPCV 2000 system, move to the fourth line and: a. Enter Equilibrate for the sample name. b. Select the instrument method you created in Section 2.5, Creating an Instrument Method. c. Select Equil in the Function field. d. Enter 60 minutes in the Time field. 10. If you have a GPCV 2000 system, move to the fifth line and: a. Enter Visc Calibrate for the sample name. b. Select the instrument method you created in Section 2.5, Creating an Instrument Method. c. Select Visc Calibrate in the Function field. 11. Enter the following sample parameters on the next available line: a. Enter 1 in the Vial # field. b. Enter PSSTD190K in the sample field. 37 Creating a Sample Set Method

38 c. Select the instrument method you created in Section 2.5, Creating an Instrument Method. d. Select Narrow Std in the Function field. e. Select 2 in the Inject (number of injections) field. f. Enter 30 minutes in the Time (run time) field. g. Select 10 in the vial size field. 12. Fill in the rest of the sample set table for the remaining vials (Figure 2-16). Follow these steps to quickly complete the table: a. Click the row number to select the first vial row. b. Right-click the highlighted row and select Copy from the context menu. c. Right-click again and select Fill Down, 11 to copy the first vial row into the next 11 rows. d. Enter the correct vial number in each new row. Figure 2-16 Completed Sample Set Method Table 38 Preparing the System for Operation

39 13. Click Save As. The Save Sample Set Method dialog box appears (Figure 2-17). Figure 2-17 Save Sample Set Method Dialog Box 14. In the File Name area, enter a name for the sample set method, then click OK. A dialog box prompting you to load the sample set appears. 15. Click Yes, then click OK to close the Sample Set Method Editor window. 2.7 Preparing Samples The startup procedures in this guide are based on injections of a narrow polystyrene standard, which has a molecular weight of 190K plus or minus 10%. A bottle with a premeasured amount of this standard is provided with each system Preparing and Capping Sample Vials STOP Attention: To prevent damaging the sample management system, use only Alliance GPC 2000 sample filter vials to filter samples in the Alliance GPC 2000 Series system. The following materials are required to prepare and cap the sample vials: Twelve 10-mL glass vials (standard) 20-mm crimp/seal caps Crimper (Startup Kit) Large vial holders 50 mg of the narrow standard (Startup Kit or ordered in bulk) 100mL of THF 39 Preparing Samples

40 STOP To prepare samples: 1. Follow the appropriate procedure to dissolve the narrow standard in THF: If you have the bottle with a premeasured amount of the narrow standard provided with the system, add 100 ml of THF to the bottle to obtain the correct concentration. If you do not have the premeasured standard, order the standard in bulk, put 50 mg of the standard in a suitable container, then add 100 ml of THF. The mixture generally takes three to four hours for the standard to completely dissolve into the solvent. To check the mixture, gently swirl the bottle. If you see particles in the mixture, it is not ready. When the mixture is clear, it is ready to use. 2. When the mixture is ready, add approximately 8 ml to each vial. Note: To allow for solvent expansion, fill a vial only to the bottom of its shoulder. Alternatively, calculate the expected solvent volume at high temperature using the expansion coefficient of the solvent and reduce the volume of solvent in a vial accordingly. 3. Use a crimper to seal a 20-mm crimp cap onto each vial. Attention: Ensure that the fluoropolymer (shiny) side of the seal faces down toward the sample and that the cap is crimped tightly enough to prevent it from turning. 4. Place each vial in a large vial holder, then continue with Section 2.7.2, Loading the Carousel Loading the Carousel The sample compartment door button on the front panel indicates the status of the sample compartment. When the button is: Lit You can press the button to open the sample compartment door. Flashing The sample compartment door is open. You can pull the door completely open, or you can push the button to close and lock the door. Off The system is either powered off or running a sample set, and the sample compartment door cannot be opened. Note: If the sample compartment door button flashes intermittently, power off the system and the Windows NT software, then restart. Caution: To avoid possible injury from burns, wear protective clothing whenever you open the sample compartment door. 40 Preparing the System for Operation

41 To load vials in the carousel: 1. Open the sample compartment door by pressing the sample compartment door button (Figure 2-18). The door moves outward 0.25 in. (0.6 cm) and the button flashes. Sample Compartment Door Button Handle Unlocked Door Position TP01646 Figure 2-18 Opening the Sample Compartment Door 41 Preparing Samples

42 2. Pull the handle outward 8 in. (20 cm) from the system cabinet, then downward to an angle of 30 degrees from vertical (Figure 2-19). As the sample compartment door opens, the tray rotates toward the door for access to the sample vials. Handle Sample Compartment Door Carousel Tray Sliding Door Mechanism TP01648 Figure 2-19 Accessing the Carousel 42 Preparing the System for Operation

43 3. If a vial is accessible, you can use gloves and/or tongs to load or remove the vial without removing the carousel (Figure 2-20). Carousel 10-mL Vial with Cap Rumble Strips Large Vial Holder Tray TP01649 Figure 2-20 Loading or Unloading a Vial 4. If the vial positions are not accessible, remove the carousel using the carousel handle provided with the system: a. Slide the notch at the end of the carousel handle over the bottom peg of the carousel center post. b. While pushing the handle down on the washer at the bottom of the center post, move the handle upright so that the top notch on the handle engages the top peg on the center post (Figure 2-21). c. With the handle in the vertical position, pull up to lift the carousel off the tray (Figure 2-22). 43 Preparing Samples

44 2 Carousel Handle Carousel Center Post Bottom Peg 1 Notch Washer Figure 2-21 Attaching the Carousel Handle to the Carousel Top Notch Top Peg Figure 2-22 Removing the Carousel 44 Preparing the System for Operation

45 5. Load the vials in the desired positions. 6. When you finish loading and/or removing vials, install the carousel if you removed it, then remove the carousel handle by pushing it down, tilting it away from the center post, then lifting it out. 7. Close the sample compartment door until it reaches a stop (within 0.5 in. (1.3 cm) of the cabinet) and ensure the door is vertical. 8. Press the sample door button to completely close and lock the door. The carousel rotates to its home position. 45 Preparing Samples

46 3 Running a Sample Set This section describes the procedures for performing a run. You can run a sample set with or without Millennium 32 software. For details, refer to the Interactively Creating/Modifying a Sample Set Method topic in the Alliance GPC 2000 Series System Online Help Find tab. 3.1 Checking the Noise And Drift Before you run the sample set, you should check the noise and drift for the refractometer. To check the noise and drift: 1. In interactive mode, right-click the top plot and select RI from the cascade menu. 2. Zoom in on a 6 minute time window of the flattest portion of the baseline, then select Noise and Drift from the cascade menu. 3. In the Time Interval section of the Noise and Drift dialog box, select Screen. 4. Click Calculate. The system calculates the noise and drift. 5. Check the noise and drift to make sure they are within the specifications listed in Appendix A of the Waters Alliance GPC 2000 Series System Installation and Maintenance Guide. 6. If your system has the optional viscometer, repeat this process on the bottom plot to check the noise and drift for the viscometer. If the noise and drift meet specifications, the system is ready to run. If the noise and drift exceed the specifications, right-click the channel pane and examine the other data channels to troubleshoot the problem. 46 Running a Sample Set

47 3.2 Starting the Run To run a sample set: 1. Click (Sample Set) in the Alliance GPC 2000 Series system window. The Sample Set Mode window appears with the Sample Set Method table (Figure 3-1). Figure 3-1 Sample Set Method Table 2. To open the sample set: a. Click Open. b. Select the sample set method you created in Section 2.6, Creating a Sample Set Method. c. Click OK. 3. Click Run in the Sample Set Operations section. The Start Sample Set dialog box appears (Figure 3-2). Figure 3-2 Start Sample Set Dialog Box 4. Ensure the Start at line number field is set to In the Millennium Login section, ensure the Enable check box is selected. 47 Starting the Run