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Quality Control of Natural Products by Fingerprinting of Eastern Blotting

Shunsuke Fujii, Nguyen Huu Tung, Takuhiro Uto, Hiroyuki Tanaka, Xiao-Wei Li, Shao-Qing Cai, Waraporn Putalum and Yukihiro Shoyama

Eastern blotting was developed for small molecule compounds which have no affinity of conjugation to membrane instead of western blotting. Crude extracts of medicinal plants were developed by TLC plate followed to be blotted to polyvinylidene fluoride or polyether sulfone membrane by heating. Components on membrane were conjugated with carrier protein resulting in fixing to membrane. Staining was followed to western blotting system using monoclonal antibody (MAb). Four types of natural products, solasodine glycosides, aristrochic acid, glycyrrhizin-liquiritin and ginsenosides were discussed in this review. 1) Solasodine glycosides are selected as an example of staining for almost all solasodine glycosides using the anti-solamargine MAb having wide cross-reactivity. 2) Aristrochic acids possess no capacity of conjugation with membrane and their aqueous solubility is quite poor. To improving them, aristrochic acids should be synthesized for conjugation with carrier protein in order to combine to membrane. 3) Two different types of marker components in licorice, triterpene glycoside; glycyrrhizin and flavonoid glycoside; liquiritin can be detected by double eastern blotting. 4) Ginsenosides are also detected by double eastern blotting and this system might be possible to clear the distribution of ginsenosides in tissue for understanding of pharmacological activity.