NANOGRAM DETECTION OF PHOSPHOLIPIDS ON THIN-LAYER CHROMATOGRAMS

An extremely sensitive method for the quantitative determination of ph ospholipids on silica gel-precoated thin-layer chromatography plates i s reported, The procedure is based on the successive application of tw o lipid spray reagents. Firstly, phospholipid chromatograms are spraye d with the lipophilic fluorochrome 1,6-diphenyl-1,3,5-hexatriene, foll owed by application of the phosphorus-specific molybdenum blue (Dittme r-Lester) reagent, Compared to the single use of the respective dyes, the consecutive utilization of both spray reagents resulted in an enor mous enhancement in sensitivity of at least one order of magnitude. Th e procedure was proved with eight phospholipids of animal origin, and visual detection limits of down to 10 ng were achieved. The described technique provides a sensitive and specific means for the analysis of phospholipids on the nanogram scale by augmenting the molybdenum blue staining with the fluorochrome 1,6-diphenyl-1,3,5-hexatriene operating as an enhancer. (C) 1998 Academic Press.