论文部分内容阅读
组织神经节苷脂(Gls)传统的分析方法,是用薄层层析,在分析之前要先进行操作步骤繁杂的分离纯化。Harth等于1978年报告用组织粗提液直接点样于层析薄板,再以三种不同的溶剂系统展开的方法,得清晰的薄板层析图谱。它们的展开剂为:1.氯仿;2.氯仿-甲醇-水(70:30:4,V/V/V);3.氯仿-甲醇-0.25%KCl(60:35:8,V/V/V)。此法简便,但未能推广采用。根据我们的经验它未能推广的原因是粗提液颜色较深,对唾液酸测定干扰甚大,薄板上的点样量较难掌握。我们用硅胶G层析薄板(5×20厘米),将组织的氯仿-甲醇粗提液直接点样于其上,以Harth的前两种展开剂展开后,用问苯二酚显色,参考定位,刮下含Gls部分硅胶,经洗脱、离心、氮气吹干后,溶于C:M(1:1),以唾液酸含量为参数,点样于高效层析(HPTLC)薄板上,在氯仿-甲醇-0.15%CaCl_2(60:40:9,V/V/V)展开,可得良好的图谱。这一改良可去掉呈色物质,大大减少HPTLC板的用量,降低了成本,同时本法未经Folch分配,有可能获得比传统方法更为真实的结果。
Tissue gangliosides (Gls) traditional analytical methods, is the use of thin-layer chromatography, the analysis before the first step for the complicated steps of purification. Harth et al., 1978, report directly on the chromatographic plate using the tissue crude extract, followed by three different solvent systems to obtain a clear plate chromatogram. Their development agents are: 1. chloroform; 2. chloroform-methanol-water (70: 30: 4, V / V / / V). This method is simple, but failed to promote the use. According to our experience, it failed to promote the reason is the dark color of the crude extract, the determination of sialic acid interference, the spot on the sheet is difficult to grasp the amount of sample. We use silica gel G chromatography plate (5 × 20 cm), the tissue chloroform-methanol crude extract directly spotted on it, with Harth’s first two developing agent after development, with quinone phenol color, reference Positioning and scraping off the Gls part of the silica gel, after elution, centrifugation, nitrogen drying, dissolved in C: M (1: 1), sialic acid content as a parameter, spotting in high performance chromatography (HPTLC) In chloroform - methanol - 0.15% CaCl 2 (60: 40: 9, V / V / V) expand, get a good map. This improvement removes the color material, drastically reduces the amount of HPTLC used, reduces costs, and this method is not partitioned by Folch, making it possible to obtain more realistic results than conventional methods.