Quantitative Densitometry in Situ of Lipids Separated by thin Layer Chromatography

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Abstract
Operating parameters are described for a densitometric method to determine in situ 8 lipid classes separated by TCL. The separated lipids, visualized on the TLC plate by a cupric acetate-phosphoric acid charring method, were quantitatively determined by spectrodensitometry using the Shimadzu CS-910 Dual Wavelength TCL Scanner. Plates were scanned in either a linear scanning mode or in a zigzag scanning mode (flying spot). Reproducibility of sample application (spotting) and the lipid separation procedure were determined by scanning. Transmittance measurements yielded response areas that were 2.8 .times. higher than reflectance measurements. Operating parameters such as scanning direction, wavelength, single and dual-wavelength measurement, scanning speed and slit geometry were studied. Optimal conditions were established for quantitative densitometry of lipids on thin layer plates.