Okumura K, Hayashi K, Morishima I, Murase K, Matsui H, Toki Y, Ito T
Second Department of Internal Medicine, Nagoya University School of Medicine, Japan.
Lipids. 1998 May;33(5):529-32. doi: 10.1007/s11745-998-0237-3.
Ceramides and 1,2-diacylglycerol have been demonstrated in intracellular signaling pathways. A method of simultaneous mass determination of ceramides and 1,2-diacylglycerol in tissues was developed using the latroscan which combines thin layer chromatography and flame ionization detection (TLC/FID) techniques. Because of relatively low amounts of these components in tissues, the fraction of nonpolar lipids, which included ceramides and glycerides, was eluted with chloroform/acetone mixture (3:1, vol/vol) through a silicic acid column to eliminate the polar phospholipids. Development of Chromarods was carried out using three solvent systems in a four-step development technique. The relationship of the peak area ratio to weight ratio compared with cholesteryl acetate added as an internal standard was linear. The amount of ceramides increased with incubation of rat heart homogenate and human erythrocyte membranes in the presence of sphingomyelinase (E.C. 3.1.4.12). The latroscan TLC/FID system provided a quick and reliable assessment of ceramides and 1,2-diacylglycerol.
神经酰胺和1,2 - 二酰基甘油已在细胞内信号通路中得到证实。利用结合了薄层色谱和火焰离子化检测（TLC/FID）技术的latroscan开发了一种同时测定组织中神经酰胺和1,2 - 二酰基甘油的质量方法。由于组织中这些成分的含量相对较低，将包括神经酰胺和甘油酯在内的非极性脂质部分用氯仿/丙酮混合物（3:1，体积/体积）通过硅酸柱洗脱，以去除极性磷脂。使用三步展开技术中的三种溶剂系统对Chromarods进行展开。与作为内标添加的乙酸胆固醇酯相比，峰面积比与重量比的关系呈线性。在鞘磷脂酶（E.C. 3.1.4.12）存在的情况下，大鼠心脏匀浆和人红细胞膜孵育后神经酰胺的量增加。latroscan TLC/FID系统为神经酰胺和1,2 - 二酰基甘油提供了快速可靠的评估。
