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Evaluating MISER chromatography as a tool for characterizing HILIC column equilibration.

Kenneth D Berthelette1, Thomas H Walter1, Martin Gilar1

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Hydrophilic Interaction Liquid Chromatography (HILIC) column equilibration is faster with higher water content in mobile phases. A new method, MISER, improves characterization of this process for polar analyte analysis.

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Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Hydrophilic Interaction Liquid Chromatography (HILIC) retains polar analytes using polar stationary phases and acetonitrile-rich mobile phases.
  • A key challenge in HILIC is the long column equilibration time, which is not fully understood.
  • Current methods for characterizing HILIC column equilibration offer limited data and time resolution.

Purpose of the Study:

  • To evaluate the Multiple Injections in a Single Experimental Run (MISER) technique for characterizing HILIC column equilibration.
  • To investigate the effect of mobile phase composition on HILIC column equilibration time.
  • To identify methods for reducing HILIC column equilibration duration.

Main Methods:

  • Utilized the MISER technique for high-resolution HILIC column equilibration analysis.
  • Varied mobile phase water content to assess its impact on equilibration.
  • Employed a BEH Amide column for experimental validation.

Main Results:

  • MISER provides improved time resolution for column equilibration studies.
  • HILIC column equilibration is significantly faster when transitioning to mobile phases with higher aqueous content.
  • Equilibration to 10% aqueous mobile phase was approximately 5-fold faster than to 3% aqueous mobile phase.

Conclusions:

  • MISER is a valuable tool for detailed characterization of HILIC column equilibration.
  • Reducing HILIC column equilibration times is achievable by optimizing mobile phase composition.
  • Understanding and optimizing HILIC equilibration is crucial for efficient polar analyte analysis.