APPLICATION OF THIN-LAYER CHROMATOGRAPHY IMAGE ANALYSIS TECHNIQUE IN QUANTITATIVE DETERMINATION OF SPHINGOMYELIN

摘要

A sensitive and convenient method for sphingomyelin determination was developed based on thin-layer chromatography (TLC) and image-processing analysis. The mobile phase composition, detection and quantification conditions were systematically investigated through several trials. The molybdenum blue reaction allowed specific detection of the phospholipid with a high sensitivity and a wide linear range. Digital images of TLC plate chromatograms were captured with flatbed scanner and converted into peakchromatograms using TLSee~R software and quantitative analysis was conducted. The linearity, sensitivity, accuracy and precision of the system were evaluated. The limits of detection and quantification were 0.5 and 1.7 jig/spot, respectively. Separationof the mixture consisting of sphingomyelin, phosphatidylcholine, and phosphatidylethanolamine was also carried out.