Higgins T J
J Lipid Res. 1984 Sep;25(9):1007-9.
A simplified procedure for fluorometric determination of sphingosine bases is presented. Nitrogenous bases are reacted with fluorescamine and then extracted into a lower phase solvent, obviating the need to transfer the reaction products prior to quantitation. Consequently, the entire procedure may be carried out in a single set of screw-capped reaction tubes. Chloroform was found to be the best solvent of those tested for the maximal extraction of derivatized sphingosine bases with negligible extraction of nitrogenous contaminants derived from phospholipids and amino sugars. The simplified method retains excellent sensitivity and selectivity and allows for simultaneous processing of large numbers of samples. Furthermore, quantitation of glycolipid-derived hexosamine can be obtained by measuring the fluorescence of the aqueous phase after partitioning.
本文介绍了一种用于荧光测定鞘氨醇碱基的简化方法。含氮碱基与荧光胺反应，然后萃取到下层溶剂中，无需在定量前转移反应产物。因此，整个过程可在一组带螺旋盖的反应管中进行。在测试的溶剂中，发现氯仿是用于最大程度萃取衍生化鞘氨醇碱基且从磷脂和氨基糖衍生的含氮污染物萃取量可忽略不计的最佳溶剂。该简化方法保留了出色的灵敏度和选择性，并且允许同时处理大量样品。此外，通过测量分配后水相的荧光可对糖脂衍生的己糖胺进行定量。
