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HPLC separations with micro-bore columns using high-temperature water and flame ionization detection.

Yu Yang1, Toru Kondo, Tina J Kennedy

  • 1Department of Chemistry, East Carolina University, Science & Technology Building, Suite 300, Greenville, NC 27858, USA. yangy@mail.ecu.edu

Journal of Chromatographic Science
|January 28, 2006
PubMed
Summary

High-temperature water chromatography with a flame ionization detector (FID) is stabilized using micro-bore columns. This method enables the separation of carbohydrates and amino acids, overcoming previous flow rate limitations.

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Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Flame ionization detector (FID) can detect analytes in high-temperature water separation.
  • Standard high-performance liquid chromatography (HPLC) columns require high eluent flow rates, causing FID instability.
  • Micro-bore columns offer a solution by requiring lower flow rates.

Purpose of the Study:

  • To investigate the use of micro-bore columns for stable high-temperature water chromatography with FID detection.
  • To separate and analyze carbohydrates, amino acids, and other organic compounds.
  • To determine the performance characteristics (limit of detection, linear range) for amino acid analysis.

Main Methods:

  • Micro-bore columns packed with poly(styrene-divinylbenzene) (PRP-1) or octadecylsilane-bonded phase particles were utilized.

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  • A homemade high-temperature water chromatograph equipped with FID was employed.
  • Both isothermal and programmed temperature methods were applied for separation.

    • Main Results:

    • Micro-bore columns enabled stable FID operation in high-temperature water chromatography due to low eluent flow rates.
    • Successful separation of carbohydrates, amino acids, and other organic acids and bases was achieved.
    • The limit of detection and linear range were determined for tested amino acids.

    Conclusions:

    • Micro-bore columns are effective for achieving stable FID detection in high-temperature water chromatography.
    • This approach facilitates the analysis of various organic compounds, including carbohydrates and amino acids.
    • The method provides quantifiable results for amino acid analysis.