Preparation method of ganoderic acid B

Preparation method of ganoderic acid B

  • CN 106,928,305 B
  • Filed: 03/13/2017
  • Issued: 11/20/2020
  • Est. Priority Date: 03/13/2017
  • Status: Active Grant
First Claim
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1. A method for preparing ganoderic acid B from Ganoderma fruiting body extract comprises:

  • (1) taking 50kg of ganoderma lucidum fruit bodies, slicing, putting into pilot plant extraction equipment, adding 87% absolute ethyl alcohol with 8 times of volume, and extracting for 2 times, 2 hours for the first time and 1 hour for the second time;

    the extraction temperature is 65 ℃


    mixing the two extractive solutions, concentrating in a cooking pot, and concentrating with 20L rotary evaporator to obtain Ganoderma encarpium extract;

    eluting Ganoderma fruiting body extract with D101 macroporous resin, eluting with 60% ethanol, eluting with 3-5 times of solution volume, and concentrating the eluate to obtain component rich in ganoderic acid B;

    (2) preparing a V/V two-phase solvent system of n-hexane, ethyl acetate, methanol and water in a separating funnel, wherein the ratio of the n-hexane to the ethyl acetate to the methanol to the water is 5;




    pumping the upper phase and the stationary phase into a high-speed counter-current chromatograph, and rotating the main machine at the rotating speed of 900r/min after the pipeline is filled with the upper phase;

    pumping the lower phase and the mobile phase at the flow rate of 2ml/min, wherein the detection wavelength is 248 nm;

    after the two phases reach balance, namely the base line is stable, 20mL of sample with the concentration of 20mg/mL is prepared by using a flowing phase, the sample is injected by a sample injection valve, an ultraviolet detector is used for on-line monitoring, and the detection wavelength is 248 nm;

    collecting each fraction, and analyzing and detecting separation condition by high performance liquid chromatography;

    comparing with ganoderic acid B standard product, collecting and mixing fractions containing ganoderic acid B, and measuring with high performance liquid area normalization method to obtain ganoderic acid B29.6mg with purity of 88.3%;

    in the high performance liquid chromatography detection of the ganoderic acid B, a diode array detector is utilized to perform full-wave-band scanning of a chromatographic peak, so that the purity of the peak is determined;

    the solvent elution conditions were determined as follows;

    high performance liquid chromatography of ganoderic acid BThe chromatographic column Agilent ZORBAX SB-Aq4.6mm X250 mm, 5 μ

    m,mobile phase a was 0.01% glacial acetic acid, B was acetonitrile gradient elution;













    the sample volume is 10 mu L;

    the column temperature is 30 ℃


    the detection wavelength is 248 nm;

    flow rate;

    1 ml/min;

    under the condition, two chromatographic peaks are displayed in the chromatogram;

    the chromatographic peak with the peak-emergence time of 31min, namely the target compound ganoderic acid B, has a relative peak area of 88.3-89.1%;

    the chromatographic peak with the peak-off time of 37min is an impurity peak, and the relative peak area is 10.9-11.7%;

    the high performance liquid phase diagram of ganoderic acid B is shown in figure 4 of the specification.

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