High performance liquid chromatographic analysis of long chain bases in intestinal glycolipids of adult and embryonic Japanese quails.

A new sensitive assay method for sphingoids, 4-D-hydroxysphinganine, sphingosine, and sphinganine, involving reversed phase HPLC was described. Chromatography of p-N-nitrophenylacetyl derivatives of the sphingoids showed sufficient resolution, and each peak showed a linear relationship between molar quantity and area response, from 10 pmol to 10 nmol. The method was applied to the analysis of long chain bases in intestinal glycolipids from embryonic and adult Japanese quails. At the embryonic stage of 12 days' incubation, 4-D-hydroxysphinganine accounted for about 40% of the long chain bases of glycolipids, a concentration comparable to that in adult tissue. Egg yolk, which is an exclusive exogenous nutrient mixture supplied by the mother, contained only a few percent of 4-D-hydroxysphinganine as a glycolipid component. While, [3H]palmitic acid administered to embryos was incorporated into 4-D-hydroxysphinganine as well as sphingosine and sphinganine. These results suggest that sphingoids of intestinal glycolipids including 4-D-hydroxysphinganine are synthesized in the tissue, excluding the possibility of their exogenous origin in the embryonic tissue.