A rapid, sensitive method for accurate determination of the lecithin/sphingomyelin ratio by thin-layer chromatography and reflectance spectrofluorometry.
A highly reproducible thin-layer chromatographic procedure has been developed for accurate determination of the lecithin/sphingomyelin ratio. Two interfering compounds, phosphatidyl inositol and phosphatidyl serine, have been investigated and eliminated by adsorption onto DEAE-cellulose. A uniform fluorescence staining procedure employing 2',7'-dichlorofluorescein has been developed. Accurate quantitation was performed by direct measurement of the reflected fluorescence intensity of the lecithin and sphingomyelin fluorophore spots with a spectrofluorometer equipped with a thin-layer scanning attachment. Stability and reproducibility studies are reported.
D002855:Chromatography, Thin Layer / Q000379:methods; D010713:Phosphatidylcholines / Q000032:analysis*; D010743:Phospholipids / Q000032:analysis; D013050:Spectrometry, Fluorescence / Q000379:methods; D013109:Sphingomyelins / Q000032:analysis*
D010713:Phosphatidylcholines; D010743:Phospholipids; D013109:Sphingomyelins