A New HPLC-analytical Method for Total Sphingosine Contents as an Indirect Index for the Ganglioside Contents of Deer Antlers

Routinely applicable HPLC assay procedures for the ganglioside content in various deer antler preparations were established through the creation of a UV-absorbing chromophoric substance - trans--unsaturated-hexadecene-aldehyde - from the sphingosine moiety in ganglioside molecules by two step chemical reactions. In order to guarantee the assay's accuracy and sensitivity, the HPLC-assay procedure adopted internal reference procedures by mixing cis--unsaturated-hexadecene aldehyde[V] or cis-3-heptadecene- 1,2-diol[IV] to assay samples. The internal reference compound [IV] or [V] was synthesized in our laboratory starting from mannitol-diacetonide through three or four step organic reactions. This new HPLC-assay procedure was successfully applied to deer antler extracts with good dose-dependent calibration curves at the picomole level of gangliosides.