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A fluorescence detector for high-pressure liquid chromatography.

W Slavin, A T Williams, R F Adams

    Journal of Chromatography
    |April 1, 1977
    PubMed
    Summary

    This study introduces a sensitive fluorescence spectrophotometer for liquid chromatography, achieving picogram-level detection of various compounds in natural samples. The enhanced sensitivity and specificity often eliminate the need for extensive sample preparation.

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    Area of Science:

    • Analytical Chemistry
    • Spectroscopy
    • Chromatography

    Background:

    • Modern liquid chromatography (LC) requires sensitive detection methods.
    • Traditional spectrophotometers may lack the sensitivity for trace analysis.
    • Fluorescence detection offers high sensitivity for specific compounds.

    Purpose of the Study:

    • To develop and characterize a fluorescence spectrophotometer for LC.
    • To evaluate its sensitivity and specificity for analyzing various compounds.
    • To demonstrate its utility in analyzing real-world samples.

    Main Methods:

    • Adaptation of a fluorescence spectrophotometer with a micro quartz flow cell.
    • Implementation of a double-beam optical system with a second photomultiplier to cancel light source variations.

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  • Scanning of emission spectra by stopping flow and scanning the fluorescence detector.

    • Main Results:

    • Achieved picogram-level detectability for numerous fluorescing compounds in real samples.
    • Demonstrated high sensitivity and specificity, often obviating the need for sample preparation.
    • Successfully analyzed polycyclic aromatic hydrocarbons, vitamins, porphyrins, and methyl anthranilate.

    Conclusions:

    • The developed fluorescence spectrophotometer offers superior sensitivity compared to variable-wavelength ultraviolet spectrophotometers.
    • Its specificity and sensitivity make it ideal for analyzing complex natural samples.
    • The system enables highly sensitive trace analysis in fields like environmental monitoring and pharmaceutical research.