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Complexities in lipid quantitation using thin layer chromatography for separation and flame ionization for detection.

R T Crane, S C Goheen, E C Larkin

    Lipids
    |January 1, 1983
    PubMed
    Summary
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    Quantifying lipids using thin-layer chromatography with flame ionization detection (TLC-FID) is challenging. Different lipids exhibit varying response factors, and detector response depends on sample quantity, volatility, and scan speed, complicating accurate lipid mixture analysis.

    Area of Science:

    • Analytical Chemistry
    • Chromatography
    • Lipidomics

    Background:

    • Thin-layer chromatography (TLC) is a common separation technique.
    • Flame ionization detection (FID) is used for detecting organic compounds.

    Purpose of the Study:

    • To evaluate the suitability of TLC coupled with FID for quantitative lipid analysis.
    • To investigate the factors affecting detector response in TLC-FID.

    Main Methods:

    • Separation of lipids using silica gel coated quartz rods in TLC.
    • Detection of lipids using flame ionization detection (FID).
    • Analysis of response factors (RF) and detector response under varying conditions.

    Main Results:

    • Response factors for lipids varied significantly and were dependent on the amount of material analyzed.

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  • Detector response was influenced by scan speed and sample volatility.
  • Complex relationships were observed between detector response, sample quantity, and heat available per microgram of sample.

    • Conclusions:

    • TLC-FID is not a reliable method for accurate quantitation of lipid mixtures due to inconsistent response factors and complex dependencies.
    • The relationship between detector response and sample quantity is not directly correlated, hindering quantitative applications.