Thermo Scientific CarboPac PA200 Product Manual
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Thermo Scientific CarboPac PA200 Product Manual

Thermo Scientific CarboPac PA200 Product Manual

Columns for dual egc ics-6000
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6
CarboPac PA200 Columns
For Dual EGC ICS-6000
065734 Revision 01
•
January 2018
For Research Use Only. Not for use in diagnostic procedures.

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Summary of Contents for Thermo Scientific CarboPac PA200

  • Page 1 CarboPac PA200 Columns For Dual EGC ICS-6000 065734 Revision 01 • January 2018 For Research Use Only. Not for use in diagnostic procedures.
  • Page 2 CarboPac PA 200, 0.4 x 250 mm - Item # 302863 CarboPac PA 200, 1 x 250 mm - Item # 302861 Thermo Scientific Product Manual for CarboPac PA200 Columns Page 2 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.
  • Page 3 For Research Use Only. Not for Use in Diagnostic Procedures. Revision History: Revision 01, January, 2018, Original Publication; Thermo Scientific Product Manual for CarboPac PA200 Columns Page 3 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.
  • Page 4 Highlights helpful information that can make a task easier. Thermo Scientific Product Manual for CarboPac PA200 Columns Page 4 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 5: Table Of Contents

    The Dionex CarboPac PA200 Guard Column ..................... 11 System Start-up ............................11 2.6.1 ICS-6000 Dual EG Start-Up Procedure for CarboPac PA200, 1 mm Column ........12 2.6.2 ICS-6000 Dual EG Start-Up Procedure for CarboPac PA200, 0.4 mm Column ........ 13 2.6.3...
  • Page 6 Excessive Gradient Rise ........................33 Reconditioning or Replacement of the Gold (disposable) Electrodes or Replacement of the Reference Electrode ......................33 Appendix A – Dionex CarboPac PA200 Column Care ........34 Recommended Operation Pressures ......................34 Column Start-Up ............................34 Column Storage ............................34 Dionex CarboPac PA200 Column Cleanup ....................

  • Page 7: Introduction

    1. Introduction 1.1 CarboPac PA200 The CarboPac PA200 column is the latest addition to the CarboPac family of columns for carbohydrate separations. This column has been specially developed to provide high resolution separations of charged and neutral oligosaccharides and is the recommended column for these applications.

  • Page 8: Installation

    – Installation Installation Read the instrument manuals. This manual assumes that you are using Thermo Scientific Dionex instrumentation and are familiar with the installation and operation of the Thermo Scientific Dionex Ion Chromatograph (IC). If you do not understand the operation of the...

  • Page 9: System Configuration

    Cap EGC Injector Degasser Degasser Guard Column Pump Vent Vent Vacuum Analytical Column Pump Amperometric Waste Detector Thermo Scientific Product Manual for CarboPac PA200 Columns Page 9 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 10: System Requirements

    >5000 psi. The use of precut tubing, complete with high pressure fitting and ferrules, is recommended for easier and more secure installation. For 1-mm as well as for 0.4mm Dionex CarboPac PA200 Columns, 0.0025” i.d. PEEK (Item # 071870) tubing is required for all the connections after the EGC 400 MSA cartridge. Make sure that extra care is taken to minimize dead volume.

  • Page 11: The Injection Volume

    2.5 The Dionex CarboPac PA200 Guard Column A Dionex CarboPac PA200 Guard Column is normally used with the Dionex CarboPac PA200 Analytical Column. Retention times will increase by approximately 20% when a guard column is placed in-line before the analytical column under isocratic conditions. A guard column is utilized to prevent sample contaminants from eluting onto the analytical column.

  • Page 12: Ics-6000 Dual Eg Start-Up Procedure For Carbopac Pa200, 1 Mm Column

    NOTE 2.6.1 ICS-6000 Dual EG Start-Up Procedure for CarboPac PA200, 1 mm Column A. Fill a 2 L eluent reservoir bottle with ASTM filtered, Type I (18-megohm) degassed deionized water. Connect the reservoir to the eluent inlet line of the pump.

  • Page 13: Ics-6000 Dual Eg Start-Up Procedure For Carbopac Pa200, 0.4 Mm Column

    Let the system equilibrate for 30 min. 2.6.2 ICS-6000 Dual EG Start-Up Procedure for CarboPac PA200, 0.4 mm Column A. Fill a 2 L eluent reservoir bottle with ASTM filtered, Type I (18-megohm) degassed deionized water. Connect the reservoir to the eluent inlet line of the pump.
  • Page 14 GG. Connect the OUTLET of the column to the CELL IN of the ED cell, and direct the CELL OUT to waste. Thermo Scientific Product Manual for CarboPac PA200 Columns Page 14 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 15: Verification Of Column Cleanliness

    Once you obtain the expected chromatographic performance, you are ready to proceed to running your application. Thermo Scientific recommends that the system suitability standard be run whenever you reinstall a column after long-term storage.

  • Page 16: Operation

    – Operation 3. Operation 3.1 Dionex CarboPac PA200 column Operational Parameters  0-14  Temperature limit: 4-60 °C  Pressure limit: 4500 psi  Organic Solvent Limit: 100% compatible  Typical Eluents: potassium hydroxide, potassium methanesulfonate  Standard Flow Rate: 1 mm: 0.063 mL/min...

  • Page 17: Purity Requirements And Preparation Of Eluents And Standards

    0.2 μm Nylon filters (Fisher Cat. # 0974046) is a good alternative to vacuum degassing under sonication and is sufficient for the majority of cases. On-line eluent and EG- produced eluents degassing is supported through the use of Thermo Scientific pumping systems. 3.2.2...

  • Page 18: Method Development

    ELUENT OUT of the DIonex RFIC EGC 400 KOH Degasser to P port of the injection valve. K. Install the CarboPac PA200 (1 x 50 mm) guard column and the CarboPac PA200 (1 x 250 mm) separator column to C port of the injection valve. Direct the OUTLET of the column to waste.

  • Page 19: System Re-Start Procedure For Dionex Carbopac Pa200 0.4-Mm Column

    60202) to 2 port of the injection valve. K. Install the CarboPac PA200 (0.4 x 50 mm) guard column and the CarboPac PA200 (0.4 x 250 mm) separator column to 3 port of the injection valve. Direct the OUTLET of the column to waste.

  • Page 20: Applications

    The CarboPac PA200, 3 mm column (Document No. 031992) is NOT compatible with the ICS- 6000 Dual EG mode but it is recommended to be used with manually-prepared sodium acetate/sodium hydroxide eluents.
  • Page 21 CarboPac PA200, 0.4-mm: 0.010 mL/min Detection: CarboPac PA200, 3-mm: PAD, Au on PTFE, 2 mil gasket, Ag/AgCl ref. CarboPac PA200, 1-mm: PAD, Au on PTFE, 1 mil gasket, Ag/AgCl ref. CarboPac PA200, 0.4-mm: PAD, Au on PTFE, 1 mil gasket, Ag/AgCl ref.

  • Page 22: Profiling Of Inulin

    DP80 are possible. The CarboPac PA200, 3 mm column (Document No. 031992) is NOT compatible with the ICS- 6000 Dual EG mode but it is recommended to be used with manually-prepared sodium acetate/sodium hydroxide eluents.
  • Page 23 – Applications Figure 4 Inulin Profiles: CarboPac PA200, 3-mm vs. 1-mm vs. 0.4-mm Thermo Scientific Product Manual for CarboPac PA200 Columns Page 23 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 24: Profiling Of Galacto-Oligosaccharides

    The CarboPac PA200, 3 mm column (Document No. 031992) is NOT compatible with the ICS- 6000 Dual EG mode but it is recommended to be used with manually-prepared sodium acetate/sodium hydroxide eluents.
  • Page 25 – Applications Figure 5 GOS Profiles: CarboPac PA200 3-mm vs. 1-mm vs. 0.4-mm DP1- DP3 1,400 37.0 1,300 36.0 3-mm 3-mm 1,200 35.0 Zoomed in 34.0 1,100 33.0 1,000 32.0 31.0 30.0 29.0 28.0 Wash step 27.0 26.0 25.0 DP4- DP13 24.0...

  • Page 26: Troubleshooting Guide

    5.1.1 Finding the Source of High System Pressure Column pressure, (after subtracting the system pressure) for the Dionex CarboPac PA200 Analytical Column should be close to the pressure listed in the QAR when using the test chromatogram conditions.` If a Dionex CarboPac guard and analytical column are both installed, column pressure will increase by approximately 20% over the pressure listed in the QAR for the column.

  • Page 27: Replacing Column Bed Support Assemblies For 1 Mm Column

    Samples containing particulate material may clog the column inlet bed support. Filter samples through a 0.45 µm filter prior to injection. Thermo Scientific Product Manual for CarboPac PA200 Columns Page 27 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 28: High Background

    5.2.1 Preparation of Eluents A. The Dionex CarboPac PA200 1 mm and 0.4 mm columns are not compatible with manually prepared eluents. The use of a Dionex EGC 400 MSA cartridge and a Dionex EGC 400 KOH cartridge is required.

  • Page 29: A Contaminated Guard Or Analytical Column

    This will avoid any slippage of the ferrule under high pressure conditions which can cause low peak efficiencies. Thermo Scientific Product Manual for CarboPac PA200 Columns Page 29 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 30: Shortened Retention Times

    If a 10% dilution of the eluent is not sufficient to obtain the desired peak resolution, or if the resulting increase in retention times is unacceptable, clean the column (see Appendix A, Dionex CarboPac PA200 Column Care). After cleaning the column, follow the system re-start procedure (see Section 3.4 System Re-start) BEFORE reinstall the column in the system and let it equilibrate with eluent for about 30 minutes directing the column effluent to waste.

  • Page 31: Loss Of Resolution For Early Eluting Peaks

    E. Service the injection valve (check for leaks, Tefzel fragments, or sediments inside the valve). Thermo Scientific Product Manual for CarboPac PA200 Columns Page 31 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 32: Large Baseline Dip In The Chromatogram

    1mM and at 12 mM, peak heights will increase at 12mM. Hence you will see a decrease in peak heights whenever going to lower eluent concentrations as well. Thermo Scientific Product Manual for CarboPac PA200 Columns Page 32 of 37 065734-01...

  • Page 33: Excessive Gradient Rise

    Chromatography System Manual Doc. No. 065342 and User’s Compendium for Electrochemical Detection Doc. No. 065340 for any help necessary with electrochemical detection, working and reference electrodes. Thermo Scientific Product Manual for CarboPac PA200 Columns Page 33 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

  • Page 34: Appendix A - Dionex Carbopac Pa200 Column Care

    100 mM KOH. More stubborn contamination problems may necessitate thoroughly cleaning the column. Use the following steps to thoroughly clean the Dionex CarboPac PA200 at a flow rate similar to application flow rate or half the application flow rate to avoid the over-...
  • Page 35 C. Wash the column with deionized water for about 30 minutes. D. Then clean the Dionex CarboPac PA200 with 100 mM KOH for at least one hour. E. Reconnect column to the cell and equilibrate the Dionex CarboPac PA200 to the desired initial conditions and test the performance using QAR standard and eluent.

  • Page 36: Appendix B - Quality Assurance Report (Qar)

    Appendix B – Quality Assurance Report (QAR) Appendix B – Quality Assurance Report (QAR) Thermo Scientific Product Manual for CarboPac PA200 Columns Page 36 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.
  • Page 37 Appendix B – Quality Assurance Report (QAR) Thermo Scientific Product Manual for CarboPac PA200 Columns Page 37 of 37 065734-01 For Research Use Only. Not for use in diagnostic procedures.

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