High Performance Liquid Chromatography of Monosialogangliosides in Human Plasma with Fluorescence Detection

Abstract

A highly sensitive and simple high performance liquid chromatographic method with fluorescence detection for the determination of monosialogangliosides is described. The gangliosides are converted into the corresponding fluorescent derivatives by reaction with 3-bromomethyl-6, 7-methylenedioxy-1-methyl-2(1H)-quinoxalinone in dimethylformamide. The derivatives are separated isocratically on a normal-phase column (Radial PAK Silica cartridge) and detected fluorimetrically. The detection limits for gangliosides GM₁, GM₂ and GM₃ are 4.0, 1.1 and 0.4ng, respectively, in a 10-μl injection volume at a signal-to-noise ratio of 3. The method was applied to the determination of ganglioside GM₃ in plasma from normal subjects and from gastric cancer patients.