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Related Experiment Videos

Microchip-based capillary electrochromatography using packed beds.

Abebaw B Jemere1, Richard D Oleschuk, D Jed Harrison

  • 1Department of Chemistry, University of Alberta, Edmonton, AB, Canada.

Electrophoresis
|September 16, 2003
PubMed
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This study presents a novel microchip packed column for capillary electrochromatography (CEC), achieving high resolution for fluorescent dyes and enabling sensitive detection of amino acids and thiourea.

Area of Science:

  • Analytical Chemistry
  • Separation Science
  • Microfluidics

Background:

  • Capillary electrochromatography (CEC) offers high separation efficiency.
  • Microchip-based separations are desirable for miniaturization and portability.
  • Developing integrated packed columns on microchips is crucial for advancing CEC technology.

Purpose of the Study:

  • To design and evaluate a microchip-integrated packed column for capillary electrochromatography.
  • To assess the separation performance and reproducibility of the microchip CEC system.
  • To demonstrate the capability for indirect fluorescence detection of analytes.

Main Methods:

  • Fabrication of a quartz microchip device with a cross-injector and double weir trapping design.
  • Construction of 1, 2, and 5 mm packed CEC columns on the microchip.

Related Experiment Videos

  • Separation and analysis of three fluorescent dyes (BODIPY 493/503, rhodamine 123, acridine orange).
  • Performance evaluation using plate numbers, height equivalent to a theoretical plate (H), and Van Deemter plots.
  • Assessment of peak height, area, and retention time relative standard deviations (RSD).
  • Demonstration of indirect fluorescence detection for thiourea and amino acids.

    • Main Results:

    • Baseline resolution of three fluorescent dyes was achieved with high plate numbers (up to 360,000 plates/m) and low H values (as low as 2.8 microm).
    • A 2 mm column provided approximately 1.8 times more theoretical plates than a 1 mm column at the same flow rate.
    • Van Deemter analysis indicated increased plate height for the 5 mm column.
    • Excellent reproducibility was observed with RSD values of 2.5% for peak height, 3.3% for peak area, and 1-3% for retention times.
    • Indirect fluorescence detection enabled a detection limit of 10 microM for amino acids.

    Conclusions:

    • The microchip-integrated packed column system is effective for high-performance capillary electrochromatography.
    • The device demonstrates excellent separation efficiency, resolution, and reproducibility.
    • The system shows potential for sensitive analysis of various analytes, including amino acids and thiourea.