Pu Huang 2kg loaded into the extraction kettle of the supercritical extraction device, supercritical CO2 as a solvent, supercritical extraction of Pu Huang, extraction pressure of 28MPa, temperature of 45 , extraction time of 3 hours, CO2 flow rate of 25L / h, to be extracted in the tank environment stabilization, add 65% acetone carrying agent (20% of the amount of herbs) to the carrying agent tank, adjust the temperature of the resolution axe is 55 , the pressure is 8MPa, collect the extract released from the discharge port of the resolution kettle. The extract was extracted with n-butanol and concentrated under reduced pressure to obtain 244g of extract. Take 50g of the extract and silica gel (extract: silica gel = 1:3) stirred uniformly, low temperature drying, and then loaded with new silica gel column, mixed samples of silica gel on the column, chloroform-ethanol mixed solution according to the volume ratio of 9:1, 7:1, 4:1, 2:1, 1:1, 1:2 gradient elution, each gradient of the dosage of the eluent for the 200 ml, the flow rate of elution for the 4 ml/min, the collection of each part of the eluent, TLC Tracking monitoring, collect the flow fraction of chamosin, reduce pressure concentration to get chamosin crude product, take the crude product with methanol to formulate into 10mg/ml concentration of the solution, with preparative high performance liquid chromatography purification, collect the main component peaks, rotary evaporation and concentration, vacuum freeze-drying, to get the chamosin purity of 264mg.
The above conditions of the preparative high performance liquid chromatograph are as follows: the preparative column packing is Octadecylsilane-bonded silica gel, the column temperature was room temperature, the mobile phase was methanol: 0.1% acetic acid aqueous solution = 18:32 (v/v), the flow rate was 5 ml/min, and the detection wavelength was 254 nm.
