Method for separating and purifying natural product using three-phase counter current chromatograph
A technology of countercurrent chromatography and natural products, which is applied in the field of separation and purification of natural products using three-phase countercurrent chromatography, which can solve the problems of low overall resolution and inability to realize compounds, and achieve the effect of solving the problem of low overall resolution
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Embodiment 1
[0037] Embodiment 1, separation and purification of Scutellaria baicalensis water extract
[0038] Use agarose matrix gel Superose 12 and n-hexane-ethyl acetate-methanol-acetic acid-water (volume ratio is 1-6-1.5-1.5-6) system to constitute the three phases of the three-phase countercurrent chromatography system of the present invention, to Separating and purifying the water extract of Scutellaria baicalensis, the specific steps are:
[0039] 1) According to the volume ratio of 1:6:1.5:1.5:6, mix n-hexane, ethyl acetate, methanol, acetic acid and water, and balance in a separatory funnel for 2 hours, the mixed solution is divided into upper and lower phases, and n-hexane Alkane-ethyl acetate-methanol-acetic acid-water solvent system; the upper and lower phases are separated;
[0040] 2) Use Superose 12 gel (GE Healthcare, average particle diameter 10 μm) as the solid stationary phase; the lower phase in the solvent system in step 1) whose polarity is similar to the solid st...
Embodiment 2
[0045] Embodiment 2, the separation and purification of Danshen fat-soluble compound
[0046] Use reverse-phase bonded phase silica gel C18 and n-hexane-ethanol-water (volume ratio is 10-5.5-4.5) system to constitute the three phases of the three-phase countercurrent chromatography system of the present invention to separate and purify the fat-soluble compounds of Salvia miltiorrhiza, and the specific steps are as follows: :
[0047] 1) According to the volume ratio of 10-5.5-4.5, mix n-hexane, ethanol and water, and balance in a separatory funnel for 2 hours, the mixed solution is divided into upper and lower phases, and obtain n-hexane-ethanol-water solvent system; phase separation;
[0048] 2) Use reverse-phase bonded phase silica gel C18 (carbon 18) (GE Healthcare, particle average diameter 5 μm) as the solid stationary phase; use the upper phase in the solvent system in step 1) whose polarity is similar to that of the solid stationary phase as the liquid stationary phase...
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