preparation | The dried two-tip herbs were crushed with 80% ethanol (ethanol/water, the same as below) reflux extraction 2 times, each time 3 hours, combined extract, under reduced pressure to recover solvent ethanol extract 580g. It was suspended in 5L water and extracted with equal amount of petroleum ether, ethyl acetate and n-butanol, respectively. After recovering each part of solvent, the petroleum ether extract, ethyl acetate extract and n-butanol extract were obtained. The n-butanol extract was eluted with water, 10% ethanol, 30% ethanol, 50% ethanol, 70% ethanol and 95% ethanol by AB-8 macroporous resin column, 50% and 70% ethanol eluent, after recovering ethanol under reduced pressure and drying to obtain dry paste, was then subjected to silica gel column chromatography, chloroform-methanol-water (8: 2: 1 volume ratio) elution, thin layer chromatography detection combined into 8 parts, the fourth part of the silica gel column chromatography, chloroform-methanol-water (7: 3: 1 volume ratio) elution, thin layer chromatography detection combined into 4 parts, the second was subjected to Sephadex LH20 column chromatography (60% methanol-water) to give compound 1(24mg), and the third was subjected to Sephadex LH20 column chromatography (60% methanol-water) to give compound 2(12mg). Compound 2 that is more than 20 anemarrhena glycosides. |