[Separation and purification of an endogenous inhibitor of sodium pump from chansu by thin-layer chromatography and reversed-phase high performance liquid chromatography].
An endogenous inhibitor of the sodium pump from the Chinese medication Chansu was purified. The dry substance Chansu was extracted with methanol. The dry residue dissolved in water and filtered subsequently through membrane filters with the exclusion size of 1000 Da, 3000 Da and 10000 Da in a Filtron Pro Vario-3-System and applied to thin-layer chromatographic plate made of Silica gel 60 F254 + 366 developed with a mixture of CHCl3-MeOH-H2O(75:20:5, volume ratio). The fractions with Rf 0.55 inhibiting the sodium pump were purified on an HPLC C18-RP column using a linear H2O-methanol gradient with 220 nm and 300 nm DAD detection. The bioactivity was measured by 86Rb-uptake into human red blood cells. The results showed that a low molecular weight, water soluble compound, which inhibited the sodium pump activity in the red blood cells and had a maximum absorbance at 250 nm was isolated from the Chinese medication Chansu. Several mg of the compound in pure state could be obtained from 1 kg Chansu. It was different from ouabain and proscillaridin A in chemical structure, because ouabain and proscillaridin A show a UV maximum absorption at 220 nm and 300 nm, while the new inhibitor at 250 nm.
Li
S Q
SQ
Department of Biochemistry, Institute of Experimental Medicine, Hebei Academy of Medical Sciences, Shijiazhuang 050021, China. lisuqin@heinfo.net
D000818:Animals; D002018:Bufanolides / Q000737:chemistry*; D002025:Bufonidae; D002298:Cardenolides; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D004077:Digoxin / Q000302:isolation & purification*; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D012503:Saponins / Q000302:isolation & purification*; D026941:Sodium Channel Blockers / Q000302:isolation & purification*; D000254:Sodium-Potassium-Exchanging ATPase / Q000302:isolation & purification
D002018:Bufanolides; D002298:Cardenolides; D004365:Drugs, Chinese Herbal; D012503:Saponins; D026941:Sodium Channel Blockers; C095591:chan su; C034504:digoxin-like factors; D004077:Digoxin; D000254:Sodium-Potassium-Exchanging ATPase