Quantitative analysis of free sphingoid bases in the brain and spinal cordtissues by high-performance liquid chromatography with a fluorescence detection

The o-phthaldialdehyde precolumn derivatives of psychosine, sphinganine and sphingosine extracted from brain and spinal cord tissues were determined b y high-performance liquid chromatography-fluorescence detection. This metho d was developed with the purpose of detecting an endogenous amount of psych osine, sphingosine and sphinganine using small aliquots of brain tissues an d spinal cord in rats. These sphingolipid bases were extracted in various r atios of chloroform-methanol and several pH values. Recovery of the method is about 81% in 12 ng/tube (final volume, 320 mu l), 90-95% in 45 ng/tube o f sphingosine and sphinganine within 2-12% relative standard deviation. Det ection limits of these sphingoid bases were about 0.05 pmol/mg brain tissue . In the forebrain, brainstem and spinal cord of rats at three different ag es of postnatal days (PND) 1, PND 13 and 6 months old, the endogenous conce ntrations of psychosine, sphingosine and sphinganine were determined. From these results, this method is suitable for the determination of sphingoid b ases in small aliquot of brain and spinal cord tissues. (C) 1998 Elsevier S cience B.V. All rights reserved.