Watanabe K, Nishiyama M
Division of Biochemistry, Shigei Medical Research Institute, Okayama, Japan.
Anal Biochem. 1995 May 1;227(1):195-200. doi: 10.1006/abio.1995.1270.
A new approach to the analysis of acidic glycosphingolipid molecular species has been developed by using two-dimensional thin-layer chromatography: TLC mapping. TLC mapping was accomplished by use of two types of high-performance thin-layer chromatographic (HPTLC) plates. A NH2-silica gel HPTLC plate developed with chloroform:methanol:1% diethylamine (50:47:15) was used for the first-dimension chromatography. Acidic glycosphingolipids on the plate were transferred to a normal silica gel HPTLC plate by developing the plate with chloroform:methanol:ammonia (4:10:6) after the NH2 plate was placed face to face on top of the silica gel HPTLC plate. Acidic glycophingolipids on the HPTLC plate were then developed with chloroform: methanol:0.2% CaCl2 (60:40:10) for the second-dimension chromatography. The use of the TLC-mapping method allowed us to compare detailed membrane constituents, including minor molecular species with regard to acidic glycophingolipids. The method is rapid and simple and is feasible for determining acidic glycosphingolipid from biological samples.
TLC图谱分析，已开发出一种分析酸性糖鞘脂分子种类的新方法。TLC图谱分析是通过使用两种类型的高效薄层色谱（HPTLC）板完成的。用氯仿：甲醇：1%二乙胺（50:47:15）展开的NH₂-硅胶HPTLC板用于一维色谱分析。在将NH₂板面对面放置在硅胶HPTLC板上方后，用氯仿：甲醇：氨水（4:10:6）展开该板，将板上的酸性糖鞘脂转移到普通硅胶HPTLC板上。然后用氯仿：甲醇：0.2%氯化钙（60:40:10）展开HPTLC板上的酸性糖鞘脂，进行二维色谱分析。TLC图谱分析方法使我们能够比较详细的膜成分，包括酸性糖鞘脂的微量分子种类。该方法快速、简单，对于从生物样品中测定酸性糖鞘脂是可行的。
