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Related Experiment Videos

Cavity ring-down spectroscopy as a detector for liquid chromatography.

Kate L Snyder1, Richard N Zare

  • 1Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.

Analytical Chemistry
|September 11, 2003
PubMed
Summary
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Cavity ring-down spectroscopy (CRDS) is now a viable detector for high-performance liquid chromatography (HPLC). A novel flow cell enables CRDS analysis of microliter liquid volumes, offering sensitive and reproducible HPLC separations.

Area of Science:

  • Analytical Chemistry
  • Spectroscopy
  • Chromatography

Background:

  • High-performance liquid chromatography (HPLC) is a cornerstone of chemical analysis.
  • Sensitive and robust detection methods are crucial for advancing HPLC capabilities.
  • Cavity ring-down spectroscopy (CRDS) offers high sensitivity but its application in liquid chromatography is limited.

Purpose of the Study:

  • To demonstrate the utility of cavity ring-down spectroscopy (CRDS) as a detector for high-performance liquid chromatography (HPLC).
  • To develop and implement a specialized flow cell for CRDS-HPLC analysis of small liquid volumes.
  • To evaluate the performance of the CRDS-HPLC system in terms of dynamic range, noise, reproducibility, and detection limits.

Main Methods:

  • Design and implementation of a Brewster's angle flow cell for CRDS analysis of microliter liquid volumes.

Related Experiment Videos

  • Integration of the flow cell with a pulsed CRDS system for HPLC detection.
  • Characterization of the system's performance using static measurements and HPLC separations with quinalizarin as a test analyte.
  • Determination of linear dynamic range, baseline noise, peak area reproducibility, and mass detection limits.

    • Main Results:

    • The CRDS-HPLC system achieved a linear dynamic range of 3 orders of magnitude for static measurements (30 nM to 30 microM) and 2 orders of magnitude for HPLC (0.5 microM to 50 microM).
    • Baseline noise levels were comparable to commercial UV-vis detectors, with root-mean-square (RMS) noise of 2.8 x 10(-6) AU for static and 3.2 x 10(-6) AU for HPLC measurements after smoothing.
    • HPLC peak areas showed reproducibility within 2-3%, and a mass detection limit of 2.5 x 10(-8) g/mL was achieved for a molecule with a molar absorptivity of 9 x 10(3) M(-1) cm(-1).

    Conclusions:

    • Cavity ring-down spectroscopy (CRDS) is a highly effective and sensitive detection method for high-performance liquid chromatography (HPLC).
    • The developed Brewster's angle flow cell enables CRDS analysis of minute liquid volumes, overcoming previous limitations.
    • The CRDS-HPLC system demonstrates performance comparable or superior to conventional UV-vis detectors, offering a promising alternative for sensitive chemical analysis.