Kwon O S, Newport G D, Slikker W
Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock 72211, USA.
J Chromatogr B Biomed Sci Appl. 1998 Dec 11;720(1-2):9-14. doi: 10.1016/s0378-4347(98)00449-6.
The o-phthaldialdehyde precolumn derivatives of psychosine, sphinganine and sphingosine extracted from brain and spinal cord tissues were determined by high-performance liquid chromatography-fluorescence detection. This method was developed with the purpose of detecting an endogenous amount of psychosine, sphingosine and sphinganine using small aliquots of brain tissues and spinal cord in rats. These sphingolipid bases were extracted in various ratios of chloroform-methanol and several pH values. Recovery of the method is about 81% in 12 ng/tube (final volume, 320 microl), 90-95% in 45 ng/tube of sphingosine and sphinganine within 2-12% relative standard deviation. Detection 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 ages of postnatal days (PND) 1, PND 13 and 6 months old, the endogenous concentrations of psychosine, sphingosine and sphinganine were determined. From these results, this method is suitable for the determination of sphingoid bases in small aliquot of brain and spinal cord tissues.
采用高效液相色谱 - 荧光检测法测定从脑和脊髓组织中提取的鞘氨醇、二氢鞘氨醇和鞘氨醇的邻苯二甲醛柱前衍生物。开发该方法的目的是使用大鼠脑组织和脊髓的小份样本检测内源性的鞘氨醇、鞘氨醇和二氢鞘氨醇。这些鞘脂碱以不同比例的氯仿 - 甲醇和几个pH值进行提取。该方法在12 ng/管（最终体积320微升）中的回收率约为81%，在鞘氨醇和二氢鞘氨醇45 ng/管中的回收率为90 - 95%，相对标准偏差在2 - 12%以内。这些鞘氨醇碱基的检测限约为0.05 pmol/mg脑组织。在出生后第1天、第13天和6个月大的三个不同年龄大鼠的前脑、脑干和脊髓中，测定了鞘氨醇、鞘氨醇和二氢鞘氨醇的内源性浓度。根据这些结果，该方法适用于测定脑和脊髓组织小份样本中的鞘氨醇碱基。
