preparation | 1 material type X-4 binocular micro melting point measuring instrument (thermometer uncorrected, Beijing Taike Instrument Co., Ltd.); inovo400 MHz type (Varian Company, USA) nuclear magnetic resonance spectrometer, with TMS as internal standard; HPMS5973 mass spectrometer (HP Company, USA);TENSOR27 infrared spectrometer (Bruker, Germany). Column chromatography silica gel (200~300 mesh), silica gel GF254 for Thin Layer and silica gel H for Thin Layer (Qingdao marine chemical plant). The original plant was identified as Malan by Professor Chen Deyuan of Guiyang College of Traditional Chinese Medicine, and the specimen was deposited in the natural medicine Quality Control Research Center of Guizhou Normal University. 2 extraction and separation of Malan 10kg,80% ethanol reflux extraction 3 times, combined extract, under reduced pressure recovery solvent to no smell, and appropriate amount of water dispersion dissolved, respectively, with petroleum ether, ethyl acetate and n-butanol extraction. The ethyl acetate extraction fraction was subjected to column chromatography eluting with a petroleum ether: ethyl acetate solvent system gradient to give 6 fractions. The first fraction was eluted with petroleum ether-ethyl acetate (30: 1) system on silica gel column; The second fraction was eluted with petroleum ether-ethyl acetate gradient (20: 1~5:1) on silica gel column; the third stream was put on silica gel column and eluted with petroleum ether-acetone (20: 1) system to obtain compound a-povisterol; The fourth stream was put on silica gel column, elution with petroleum ether-ethyl acetate (10: 1~2:1) system; The 5th flow column, with petroleum ether-ethyl acetate (10: 1~2:1) system elution; the 6th fraction was subjected to a silica gel column and eluted with a chloroform-methanol (20: 1) system. The structure of the obtained a-pokesterol was identified as the target compound. |