Chromatographic resolution of glucosidic compounds, ginsenosides on polyethersulphone membrane, and its application to the quantitative immunoassay for ginseng saponins.
Chromatographic resolution of glucosidic compounds, ginsenosides on polyethersulphone membrane, and its application to the quantitative immunoassay for ginseng saponins.
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DOI：
10.1093/glycob/cwi096
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发表时间：
2005-10
期刊：
影响因子：
4.3
通讯作者：
O. Morinaga;N. Fukuda;Hiroyuki Tanaka;Y. Shoyama
中科院分区：
文献类型：
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作者：
O. Morinaga;N. Fukuda;Hiroyuki Tanaka;Y. Shoyama
建立了一种在聚醚砜（PES）膜上分离人参皂苷类糖苷类化合物的方法。该方法对各种人参粗提物中人参皂苷Rb1 （G-Rb1）、G-Rc和G-Rd具有较好的分辨率和定量免疫分析效果。新建立的方法简单，适用于定量分析。采用乙腈-水-乙酸溶剂体系制备的人参皂苷在PES膜上直接用NaIO4溶液和牛血清白蛋白（BSA）处理，得到了人参皂苷-BSA在PES膜上的偶联物。结合抗g - rb1单克隆抗体（MAb），然后用标记过氧化物酶的第二抗体靶向第一抗体。最后，一种底物与酶反应并染色。对染色膜进行扫描，用NIH Image软件对斑点进行定量分析。G-Rb1、G-Rc和G-Rd分别至少62.5 ng可检出。三种人参皂苷的定量分析范围在0.125 ~ 2.0 μ g之间。
A method has been devised for the chromatographic resolution of glucosidic compounds, ginseng saponins, on polyethersulphone (PES) membrane. The method results in good resolution and quantitative immunoassay for ginsenoside Rb1 (G-Rb1), G-Rc, and G-Rd in crude extracts of various ginsengs. The newly established method is simpler and applies for quantitative analysis. Ginsenosides developed by acetonitrile-water-acetic acid solvent system on a PES membrane were directly treated with a NaIO4 solution followed by bovine serum albumin (BSA), resulting in a ginsenoside-BSA conjugate on a PES membrane. Anti-G-Rb1 monoclonal antibody (MAb) was bound, and then a second antibody labeled with peroxidase directed against the first antibody. Finally a substrate reacted to the enzyme and gave staining. The stained membrane was scanned, and spots were analyzed quantitatively using NIH Image software. At least 62.5 ng of G-Rb1, G-Rc, and G-Rd were clearly detectable individually. Three ginsenosides can be analyzed quantitatively between 0.125 and 2.0 microg.