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HPLC determination of sphinganine and sphingosine in serum and urine of healthy individuals

Sphingolipids are defined as compounds having a long-chain sphingoid base as the backbone. The prevalent long-chain bases of most mammals are D-erythro-sphinganine and sphingosine. They are found in all eukaryotic cells, primarily on the external leaflet of the plasma membrane and have a numerous functions. The aim of our research was to determine the concentrations and ratio of two sphingoid bases, sphinganine and sphingosine in serum and urine of healthy individuals in order to establish the referent values. The second aspect of the research was to ascertain whether there was any significant difference in the concentrations of sphinganine and sphingosine between men and women. The third goal was to establish if there was any alteration in the sphingoid bases levels induced by hormonal changes in women during pregnancy and after menopause. Sphingolipids were extracted from serum and urine. Free and total sphingoid bases were obtained by base and acid hydrolysis respectively. Sphinganine and sphingosine were derivatized with the OPA (O-Phtaldialdehyde) reagent and analysed by high performance liquid chromatography. In the urine of healthy individuals, the concentrations of free sphinganine and sphingosine, total sphinganine and total sphinganine/sphingosine ratio were significantly different between men and women. Such results have not been obtained for the serum samples. The concentrations of sphingoid bases in the urine of the healthy women after menopause and pregnant women were lower than the referent values, which could be consequence of the hormonal changes.

sphingoid bases; serum; urine

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