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Rapid, Simple Quantitation in Thin-Layer Chromatography Using a Flatbed Scanner

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Mitchell E. Johnson

Department of Chemistry & Biochemistry, Duquesne University, Pittsburgh, PA 15282-1530

J. Chem. Educ., 2000, 77 (3), p 368

DOI: 10.1021/ed077p368

Publication Date (Web): March 1, 2000

Section:

History, Education, and Documentation

Abstract

A standard flatbed scanner is shown to be a viable tool for quantitative thin-layer chromatography (TLC) plate analysis. Simply scanning a visibly stained TLC plate into a computer substitutes for much more expensive plate readers. With common image analysis software, "elution" profiles can be obtained. The resulting "chromatograms" can be analyzed in the same manner as other chromatograms. Iodine-stained cholesterol and cholesteryl esters are shown to yield nonlinear calibration curves, but the overall sensitivity is excellent for such a simple method. Detection limits are submicrogram for heavily stained spots. Spot intensity, and therefore detection limit, depends strongly on the amount of time the spot is exposed to iodine. Reproducibility is excellent for spots deposited by aspiration from a glass micropipet. Peak area and peak height relative standard deviations (RSDs) were generally below 5%, and retention factor precision was as low as 0.8% RSD.

Keywords (Audience):

Second-Year Undergraduate

Keywords (Domain):

Analytical Chemistry

Keywords (Pedagogy):

Hands-On Learning / Manipulatives

Keywords (Subject):

Chromatography

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