Extraction method and application of genistein monomer compound in armillaria chrysogreen

A technology of Armillaria chrysanthemum and genistein, which is applied in the fields of medicinal chemistry and biomedicine, can solve the problems of limiting genistein research and application, low genistein content, cumbersome steps, etc., and achieves the advantages of separation operation, The effect of large amount of preparation and simple process

Active Publication Date: 2017-12-19
TIANJIN YAOYU BIOLOGICAL TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the steps of this method are cumbersome, the cost is high, the content of genistein obtained is low, and the yield is relatively low. Only 42 mg of genistein can be obtained from 3.5 kg of dry Armillaria fermented culture, which greatly limits the further research on genistein and application

Method used

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  • Extraction method and application of genistein monomer compound in armillaria chrysogreen
  • Extraction method and application of genistein monomer compound in armillaria chrysogreen
  • Extraction method and application of genistein monomer compound in armillaria chrysogreen

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Experimental program
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Effect test

Embodiment 1

[0039] Embodiment 1: Extraction of genistein monomer compound in Armillaria chrysogreen

[0040] 1. Raw materials and materials:

[0041] The fruiting bodies of Armillaria chrysogenum were picked from Qilian, Qinghai and identified.

[0042] 2. Reagents:

[0043] Absolute ethanol was purchased from Tianjin North Tianyi Chemical Reagent Factory; chromatographic grade ethyl acetate and n-hexane were purchased from Honeywell Company.

[0044] 3. Instruments and equipment:

[0045] The ultrafine pulverizer was purchased from Shandong Sanqing Stainless Steel Equipment Co., Ltd.; the desktop refrigerated centrifuge was purchased from Thermo Company; the 30-liter Chinese medicine extraction tank was purchased from Shanghai Shunyi Experimental Equipment Co., Ltd.; the constant temperature blast drying oven was purchased from Shanghai Yiheng Scientific Instruments Co., Ltd.; rotary evaporator Shanghai Yarong Biochemical Instrument Factory; silica normal-phase silica gel chromatograp...

Embodiment 2

[0053] Example 2: Confirmation of Genistein Monomer Compounds

[0054] 1. Purity analysis by HPLC:

[0055] The object component that embodiment 1 obtains ( figure 2 The middle HMG-7.5 peak) was subjected to HPLC for purity analysis. After HPLC detection, the purity reaches more than 95%, and it is a single compound. See the HPLC spectrogram image 3 .

[0056] 2. Mass spectrometry analysis:

[0057] For the purpose component ( figure 2 HMG-7.5 peak) for mass spectrometry analysis: Hanmeng Biotechnology (Tianjin) Co., Ltd., mass spectrogram see Figure 4 . EI-MS m / z: 271(M+H) + , 269 (M-H) + .

[0058] 3. NMR analysis:

[0059] Carry out nuclear magnetic resonance analysis to target component daidinin: by carbon spectrum, hydrogen spectrum, COZY correlation spectrum (such as Figure 5-9 ) evidence analysis obtained the following results:

[0060] 1D 1 The data results in the HNMR spectrum (DMSO-d6, 400MHz) are as follows: δ: 8.28 (1H, s, H-2), 6.23 (1H, d, J=2.1...

Embodiment 3

[0064] Embodiment 3: Extraction of genistein monomer compound in Armillaria chrysogreen

[0065] The raw materials, materials, reagents, instruments and equipment used in this embodiment are the same as those in Embodiment 1.

[0066] The extraction of the genistein monomer compound in Armillaria Chrysanthemum in this embodiment comprises the following steps:

[0067] (1) Armillaria chrysanthemum fruiting body is used as raw material, dried in the shade at room temperature, dried and dehydrated at 70°C, and then ultrafinely pulverized at -20°C for 1 min to obtain Armillaria chrysanthemum ultrafine powder.

[0068] (2) Take 3kg of the above-mentioned Armillaria chrysanthemum superfine powder, soak it in ethyl acetate three times, soak for the first time: add 21L ethyl acetate to 3kg Armillaria chrysanthemum superfine powder, soak for 48h at 25°C, after soaking, 20 Centrifuge at 4,000 rpm for 30 min at ℃, collect the supernatant and residue respectively, concentrate the superna...

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Abstract

The invention provides an extraction method of a genistein monomer compound in Armillarialuteo-virens. The method includes: taking an Armillarialuteo-virens fruit body as the raw material, grinding the raw material into powder, then conducting extraction with ethyl acetate, and carrying out concentration into paste; conducting dissolving with a n-hexane-ethanol solvent; conducting one-dimensional liquid chromatography separation: employing a Silica chromatographic column, adopting a binary mobile phase with n-hexane as the phase A and ethanol as the phase B, performing gradient elution with phase B at a concentration ranging from 0% to 75%, and collecting the 26-29min eluent; performing dissolving with an ethanol-n-hexane solvent; carrying out two-dimensional liquid chromatography separation: adopting a Silica chromatographic column, employing a binary mobile phase with n-hexane as the phase A and ethanol as the phase B, performing isocratic elution with phase B at a concentration of 2%, collecting the 8-11min eluent, and performing evaporation concentration to obtain genistein. The separation method has the advantages of stability, high repeatability, large preparation capacity, simple operation and time saving, low cost, and high genistein extraction rate. The target object has an inhibiting effect on lung cancer A549 cells and liver cancer Hep-G2 cells, and has anticancer drug development potential.

Description

technical field [0001] The present invention relates to the technical fields of medicinal chemistry and biomedicine, in particular to a method for extracting genistein monomeric compounds from Armillaria chrysanthemum, and the use of the genistein monomeric compounds in the preparation of anti-liver cancer drugs and anti-lung cancer drugs application. Background technique [0002] Armillaria luteo-rivens, also known as chanterelles, golden mushrooms, and yellow rings, belongs to Basidiomycotina, Phytomycetes, Agaricaceae, White Mushrooms, and Armillaria. The fruiting body of Armillaria chrysogenum is medium and large, the cap is flat hemispherical to flat, with a diameter of 5-12cm, the edge of the cap is involuted, and the epidermis is cracked to form ciliated posterior phosphorus sheets arranged in a near ring. The gills are similar to the cap color, slightly dense, curved, and unequal in length. The stipe is cylindrical, the stem is enlarged, 2-10cm long, 2-2.5cm in dia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D311/36A61P35/00
CPCC07D311/36
Inventor 张耀洲王欢欢庞莉韩朝李上文马红杨珊珊陆洪杜思邈盖其静
Owner TIANJIN YAOYU BIOLOGICAL TECH
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