High-performance liquid chromatographic analysis of fucoganglioside hydrolysis by alpha-L-fucosidase

A sensitive HPLC method has been developed for monitoring fucoganglioside hydrolysis by purified alpha-L-fucosidase. The high-resolution method employs a Lichrosorb-NH2 column, a 10-min isocratic elution with potassium phosphate/acetonitrile buffer, detection of ganglioside products with a uv monitor at 195 nm, and quantification of low picomolar amounts of these gangliosides with an integrator. The usefulness of the HPLC method has been exemplified by using it to demonstrate the hydrolysis of gangliosides fucosyl-GM1 and fucosyl-GD1b by purified human liver alpha-L-fucosidase in the absence of activator proteins and/or detergents.