The Analysis Of Active Chemicals In Yinchenhao Decoction With Hepatocyte Membrane Immobilized Chromatography

Yinchenhao Decoction, a famous recipe from ShangHanLun consisting of three Chinese herbs: Herba Artemisiae Scopariae, Fructus Gardeniae and Radix et Rhizoma Rhei., has an effect on treating jaundice and other hepatitis in clinic. Although Gardenoside and 6, 7-dimethylesculetin of this recipe have been thought to be two active chemicals for its effect, which chemical or chemicals are responsible for its effectiveness is still unknown. To find out these active components, we try to develop a hepatocyte membrane immobilized chromatography, in which the chemicals of Yinchenhao Decoction that would specifically bind to the membrane of hepatocyte HL-7702 were studied.Firstly, The effect of the water extract of Yinchenhao Decoction, which was precipitated with alcohol, on the hepatocyte HL-7702 was studied. It has been testified that 5~0.001mg crude-herb/ml of this extract was no damage for HL-7702, but the hepatocyte has been killed after 24 hours' incubation with the concentration more than 10mg crude-herb/ml; the extract of 5 mg crude-herb/ml had no inhibition to the growth of HL-7702; and the extract of 0.1~1mg crude-herb/ml could improve the growth of HL-7702 significantly (P<0.05) , so the working concentration is between 0.1-1 mg crude-herb/ml.Secondly, the effect of this extract on the damaged HL-7702 induced by D-GalN was observed. The results showed that both ALT, AST in the supernate of HL-7702 increased by treated with D-GalN, and decreased by added this extract. It means that the extract had the property of protecting HL-7702 from being hurt by D-GalN.Thirdly, RP-HPLC was performed to analyze the fingerprints of Yinchenhao Decoction. The results indicated that the optimal wavelengths of detection should be respectively at 254nm, 280nm, 3 10nm, and the best recording time should be retained for 55 minutes, the temperature of column is 25 C.The Hepatocyte membrane immobilized chromatography was used to analyzed the chemical or chemicals in the water extract of Yinchenhao Decoction specifically binded to targets on hepatocyte membrane. The results showed there were fourcharacteristic compounds specifically conjugated to HL-7702, which originated from the herb of Radix et Rhizoma Rhei.