Study On Purification Of Rana Chensinensis Extract From Water Solution And Pharmacological Effect

There are many bioactive substances in the skin of frogs, which have important value in improvement of cell growth and broad-spectrum, as well as analgesic and anti-inflammatory function. In this paper, the crude extraction method is optimized by iron exchange chromatography and gel-filtration chromatography, and its analgesic and anti-inflammatory effect is also examined.The optimization of extraction experiment are studied through single factor and orthogonal test. Meanwhile the study have demonstrated its pharmacological actions. The active compound are purified from the water extraction of frog skin by DEAE-52 ion exchange chromatography which are optimized in terms of 5mmol/PBS (pH4.8, 1mol/L NaCl) in a gradient elution. And then the active integrents are purified by Sephadex G-50 column chromatography before eluting salt bounded with column by Sephadex G-10 column.The various integrents are screened by coronary occlusion, including acute toxicity, analgesic and anti-inflammatory method. Here, the analgesic effect is studied by using hot plate, acetic acid-induced wringing and heat stimulation test in mice. The antipyretic effect evaluated by ear swelling experiment shows a good linear relationship. The first and second part of bioactive integrents are screened by acetic acid-induced wringing and anti-inflammatory method. The two activity results show a liner increase, prior to the water-extracted integrents. The effective integrents purified Sephadex G-50 gel column chromatography are successfully screened by acetic acid-induced wringing and anti-inflammatory experiments by three models in rats ans rabbits. LD50 of water-extracted frog skin is 0.027, and its 95% confidient interval is 473.15~603.809mg/kg by acute toxicity experiment.