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Agilent Technologies 7100 Technical Note

Capillary electrophoresis system in empower
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Agilent 7100 Capillary
Electrophoresis System
in Empower
Technical Note
This technical guide describes the configuration and implementation of the G7100A
Capillary Electrophoresis in Empower 3.

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  • Page 1 Agilent 7100 Capillary Electrophoresis System in Empower Technical Note This technical guide describes the configuration and implementation of the G7100A Capillary Electrophoresis in Empower 3.

  • Page 2: Table Of Contents

    Extended CE driver features available in Agilent OpenLAB CDS ChemStation Edition Additional Information Waters APPENDIX - Examples Example: Single test run Example: Multiple injections using the IQ Kit for CE Example: Indirect detection using the Plating Bath Buffer Agilent 7100 CE in Empower Environment...

  • Page 3: Introduction

    Waters Instrument Control Software (ICS) is Waters Corporation’s adoption of the Agilent Instrument Control Framework (ICF) for their data systems. This guide describes how to configure and use the 7100 Capillary Electrophoresis (CE) in Waters Empower environment. Empower 3 offers all key functions to control the G7100A Agilent Capillary Electrophoresis system.
  • Page 4 Ensure that the Agilent LC/CE modules in the system meet or exceed the minimum NOTE firmware requirements specified by the 3rd-party CDS software vendor and Agilent's firmware set/firmware interoperability requirements. Agilent recommends using the latest available firmware set. http://www.agilent.com/en-us/firmwareDownload?whid=69761 Agilent 7100 CE in Empower Environment...

  • Page 5: Set Up And Configuration Of Ce In Empower 3

    Select File > New > Project to open the New Project Wizard. Follow the instructions in the wizard screen and proceed with default settings if no other values are given via Standard Operating Procedure (SOP). Agilent 7100 CE in Empower Environment...
  • Page 6 33) Selecting PSI as pressure unit, the CE status dashboard shows the pressure in PSI while NOTE the online plot and the resulting Aux Traces on the Chromatogram offer the values in mbar/bar. Agilent 7100 CE in Empower Environment...
  • Page 7 Enter the IP address or host name of the LAC/E box that your instrument is connected to into the pop-up screen Configuration Directory and click Connect. Do not enter the IP address of the instrument here. Note Figure 4 IP address to connect Agilent 7100 CE in Empower Environment...
  • Page 8 PreConfiguration Utility. Figure 5 Configuration of the CE In the Configuration Editor select the Agilent 7100 CE and click Auto Configure. Enter the IP address of the instrument and click OK. The instrument is detected and the CE and DAD appear on the right-hand side of the configuration window.
  • Page 9 PreConfiguration Utility) is required. Enable Acquire for the Analog In Signal to collect data. Click OK to close the screen. Click OK to close the configuration editor. Click OK to close the PreConfiguration Utility. Agilent 7100 CE in Empower Environment...
  • Page 10 Select Instrument Type AgilentLC and click OK to leave the screen. Figure 7 Entering communication details for CE Access the Nodes Properties in the Empower Configuration Manager and verify that the Instrument is shown as OK. Agilent 7100 CE in Empower Environment...
  • Page 11 Set up and Configuration of CE in Empower 3 Generate a new chromatographic system with the newly configured instrument using File > New > Chromatographic System. Follow the instructions on the screen. Figure 8 Generating a new chromatographic system Agilent 7100 CE in Empower Environment...

  • Page 12: Using The Ce In Empower

    The given space does not allow enlarging the CE status window to full size, therefore a vertical scrollbar is present to adjust the required section of the status dashboard. Figure 10 CE Status dashboard – upper part Agilent 7100 CE in Empower Environment...
  • Page 13 Right-click an icon to access direct control. A context menu appears and an action can be performed. Some contextual actions require further user interaction. Click OK to trigger the action. Figure 12 Example of direct control - actions available for bottle icon Agilent 7100 CE in Empower Environment...
  • Page 14 It is not possible to see the detector signal while flushing the capillary. A feasible NOTE workaround is to check the numerical value of the detection wavelength, which is displayed in the DAD part of the GUI (left side). Agilent 7100 CE in Empower Environment...

  • Page 15: Creating An Instrument Method

    CE or DAD user manual. Pretreatment Method tab: There are no pretreatment steps available for CE. Auxiliary Channels tab: Offers selection of auxiliary traces during a run, e.g. the CE leak current. Agilent 7100 CE in Empower Environment...
  • Page 16 PreConfiguration Utility to apply the new configuration to the physical instrument. In addition, the LAC/E box has to be rebooted and a new autoconfiguration is required in order to adjust the new configuration with the LC Driver and Empower. Agilent 7100 CE in Empower Environment...
  • Page 17 Enter a Stoptime for the CE and/or the DAD. For run execution a Run Time is specified in NOTE Empower later, ensure that both values are synchronized. For more details, see section Executing Runs on page 21. Agilent 7100 CE in Empower Environment...
  • Page 18 Figure 15 Out of range handling The right-hand side of the method screen offers additional features. The online help explains each of the features: Replenishment  Preconditioning  Injection  Timetable  Postconditioning  Agilent 7100 CE in Empower Environment...
  • Page 19 The right side offers additional method parameters, e.g. for spectra collection and a timetable, in order to change parameters during a run. Unlike with other Agilent DAD, there is no VIS-lamp available. Figure 16 DAD method screen Agilent 7100 CE in Empower Environment...
  • Page 20 DAD. To execute the shutdown method, either select the shutdown method in the Run Sample set window or load the shutdown method as the last line of the sequence. Figure 17 Shutdown Method screen Agilent 7100 CE in Empower Environment...
  • Page 21 The method´s Post Time is independently added after the Stop Time, and no data is recorded during the post time. The next sequence run starts after the Post Time of the previous run is finished. Agilent 7100 CE in Empower Environment...
  • Page 22 Click the Inject icon to start the single run. During the run execution, you can stop the run at any time by clicking the red Stop button. Figure 18 Single Run tab Agilent 7100 CE in Empower Environment...
  • Page 23 Entering an injection volume of 0 µl does not result in a blank run. The value is ignored for the run execution, but it is reported. Start the sequence. Reminder: During run execution, the red Stop button allows you to stop the run at any time. Agilent 7100 CE in Empower Environment...
  • Page 24 It is possible to shut down the CE/DAD at the end of the sample set. Load the shutdown method (for details see section General Method Tab on page 20) as the last line of the sequence. Agilent 7100 CE in Empower Environment...

  • Page 25: Replenishment And Conditioning

    The replenishment system provides a quick way to change the buffer automatically. The system removes the used buffer from the vials and transfers it into the waste bottle. Then the vials are filled with fresh buffer from the electrolyte reservoir. Agilent 7100 CE in Empower Environment...
  • Page 26 Clean tubes: flushes the replenishment system to clean the tubes  Replenish vial: empties a vial into the waste and fills it up from the  electrolyte reservoir Wait: waits for a specified time  Figure 19 Replenishment Table Agilent 7100 CE in Empower Environment...
  • Page 27 To efficiently clean the replenishment system after use, the special functions Flush Tubes NOTE and Clean Level Sensor should be performed. This advanced cleaning is only available in the Agilent LabAdvisor software. Agilent 7100 CE in Empower Environment...

  • Page 28: Indirect Detection

    High pressure flush in Preconditioning, Injection and Postconditioning.  External pressure as method setpoint and timetable entry.  CE/p: additional external pressure connected  Agilent 7100 CE in Empower Environment...
  • Page 29 When the External Pressure Installed check box is selected, the gas cylinder is shown in the CE status dashboard accessible via the Instrument Status tab. High pressure can now be applied by right-clicking on the gas cylinder. Figure 21 Access to external pressure Agilent 7100 CE in Empower Environment...
  • Page 30 Verify that other instruments attached to the Workstation or LAC/E are not  collecting data. Rebooting the LAC/E will interrupt data collection on all instruments attached and the lost data will not be able to be recovered. Agilent 7100 CE in Empower Environment...
  • Page 31 Only in CEC mode, the instrument offers an additional initial value Pressure on the left-hand CE method screen. Selecting External, the pressure is applied to both home vials during run execution (see Figure 23). Agilent 7100 CE in Empower Environment...
  • Page 32 Using the CE in Empower new menu Figure 22 Applying external pressure – new menu Figure 23 Applying external pressure during a run Agilent 7100 CE in Empower Environment...

  • Page 33: Empower Features For Ce Instrument Control

    Chromatogram offer the values in mbar/bar. The correct pressure is applied. Analog In  The feature Analog In has not been explicitly tested, but the functionality is present. MS installed  The feature MS installed has not been tested. Figure 24 MS installed Agilent 7100 CE in Empower Environment...

  • Page 34: Extended Ce Driver Features Available In Agilent Openlab Cds Chemstation Edition

    OpenLAB CDS ChemStation Edition offers a sample diagram informing about each vial's purpose. This table is not available in Non-Agilent CDS environment. Vial Table OpenLAB CDS ChemStation Edition offers a vial table, giving an overview of vial position, name and comment. Agilent 7100 CE in Empower Environment...
  • Page 35 Extended CE driver features available in Agilent OpenLAB CDS ChemStation Edition Calibration Curve options The following Calibration Curves are unavailable in Empower: Mobility correction  Calibration Type cIEF  Calibration for determination of isoelectric points or molecular weights  Agilent 7100 CE in Empower Environment...

  • Page 36: Additional Information

    Waters Additional information on the PreConfiguration Utility is provided on the Waters Support Webpage for registered users: http://www.waters.com/waters/support.htm?lid=134936402&cid=511442&t ype=TECN Document reference: TECN134936402 Title: Using the Agilent PreConfiguration Utility with Agilent ICF Support Version 2.2 Agilent 7100 CE in Empower Environment...

  • Page 37: Appendix - Examples

    APPENDIX - Examples APPENDIX - Examples Example: Single test run For testing the functions of the 7100 CE and Empower 3 a test method can be created. The equipment and method parameters can be summarized as follows: Capillary: Fused silica capillary 50 µm ID, 48.5 cm total (40 cm effective), ext.
  • Page 38 APPENDIX - Examples Figure 25 Online plot during the run Figure 26 Resulting electropherogram Agilent 7100 CE in Empower Environment...

  • Page 39: Example: Multiple Injections Using The Iq Kit For Ce

    162.36 3.02950E+02 49187 Injection 6 2.702 162.12 3.05817E+02 49579 Average 49886 2.715 162.89 3.06254E+02 Deviation STDEV: 0.96 % 0.39 % 0.39 % 0.72 % Sixfold sample injection Figure 27 Sequence table for multiple injections Agilent 7100 CE in Empower Environment...
  • Page 40 Figure 28 Overlay of six consecutive injections Using Empower 3 in combination with the Agilent 7100 CE, a good relative standard deviation around 1 % for the peak areas and below 0.5 % for the migration times were obtained. These satisfactory values are achieved by a special self-regulating injection procedure, which significantly improves the injection precision.

  • Page 41: Example: Indirect Detection Using The Plating Bath Buffer

    Comparing the reversed wavelength the peaks can be shown as positive or negative peaks. Signal A: 350 nm/BW 20 nm, Ref 275 nm/BW 10 nm Signal B: 275 nm/BW 10 nm, Ref 350 nm/BW 20 nm Figure 29 Indirect detection Agilent 7100 CE in Empower Environment...
  • Page 42 The figure also shows that the integration and evaluation of the peak areas are possible for positive and negative peaks in the same signal. Figure 30 Integration and reporting of positive and negative peaks *G7102-90200* © Agilent Technologies, Inc 2018 *G7102-90200* G7100-90200 Edition: 01/2018 Edited in Germany Agilent Technologies, Inc...

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