薄层色谱法分离僵蚕中蛋白质和氨基酸类成分的研究

The Seperation of Proteins and Amino Acids in Bombys Batryticatus (Bai Jiang Can) by Thin Layer Chromatography

目的建立僵蚕蛋白质和氨基酸类成分的薄层色谱分离鉴别方法,为僵蚕的质量控制提供依据。方法用不同的展开剂、硅胶薄层板的干燥温度等对僵蚕化学成分进行分离。结果以羧甲基纤维素钠(CMC—Na)为粘合剂制备的硅胶G薄层板,2 0℃以下自然干燥,用含0 .1 %的茚三酮的正丁醇—冰醋酸—水( 4∶1∶1 )作展开剂,展开90min,60℃左右烘烤2min,分离得到1 0个不同组分的斑点,与氨基酸对照品在相同位置上有相同颜色的斑点。结论薄层色谱法稳定、重现性好、操作方便、快速。

Objective To establish methods for the seperation and identification of proteins and amino acids in Bombys batryticatus by thin layer chromatography (TLC). Methods This study was carried out by investigating the effects of various seperating conditions, such as different developers and temperature levels for drying the plates used in TLC, on seperating effectiveness of chemical constituents in Bombys batryticatus. Rusults The ideal conditions for this goal were determined from this study,including preparation of silicon G plate with CMC Na as cohesive, drying the plate below 20?℃ under natural condition, developing for 90 min with the developer composed of 0.1?% ninhydrin in the solution of n butyl alcohol acetic acid water (4∶1∶1) and heating at about 60?℃ for 2 min. Ten spots were developed from the sample of Bombys batryticatus in this procedure, being at the same bands with similar color as the controlling spots of standard amino acids. Conclusions This combination of conditions can be used to seperate and identify proteins and amino acids in Bombys batryticatus, with such advantages as higher stability, better repeadity, simple in operating and shorter period of developing.