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Extraction method and application of ergosterol monomer compounds in Armillaria japonica

A technology of Armillaria chrysanthemum and ergosterol, applied in the direction of steroids, organic chemistry, drug combination, etc., can solve the problems of limiting the research and application of ergosterol, less ergosterol content, cumbersome steps, etc., and achieve the benefit of separation The operation and reagent consumption are small and the process is simple

Active Publication Date: 2018-04-17
TIANJIN YAOYU BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But using this method steps are loaded down with trivial details, and cost is high, and the ergosterol content that obtains is few, and yield is relatively low, and 3.5kg dry armillaria fermented culture only obtains 140mg ergosterol, and this limits the further research and application of ergosterol greatly

Method used

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  • Extraction method and application of ergosterol monomer compounds in Armillaria japonica
  • Extraction method and application of ergosterol monomer compounds in Armillaria japonica
  • Extraction method and application of ergosterol monomer compounds in Armillaria japonica

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Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: Extraction of ergosterol monomer compound in Armillaria chrysanthemum

[0052] 1. Raw materials and materials:

[0053] The fruiting bodies of Armillaria chrysogenum were picked from Qilian, Qinghai and identified.

[0054] 2. Reagents:

[0055] Absolute ethanol was purchased from Tianjin North Tianyi Chemical Reagent Factory; chromatographic grade ethyl acetate and n-hexane were purchased from Honeywell Company.

[0056] 3. Instruments and equipment:

[0057] 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 chromatog...

Embodiment 2

[0065] Embodiment 2: Confirmation of ergosterol monomer compound

[0066] 1. Purity analysis by HPLC:

[0067] The object component that embodiment 1 obtains ( figure 2 No. 4-3 peaks) were analyzed by HPLC for purity. After HPLC detection, the purity reaches 95.23%. It is a single compound, and the HPLC spectrum is shown in image 3 .

[0068] 2. Mass spectrometry analysis:

[0069] For the purpose component ( figure 2 4-3 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) + .

[0070] 3. NMR analysis:

[0071] For the purpose component ( figure 2 No. 4-3 peak) in NMR analysis: by carbon spectrum, hydrogen spectrum, COZY correlation spectrum (such as Figure 5-8 ) evidence analysis obtained the following results:

[0072] 1D 1 In the HNMR spectrum (CDCl 3 ,400MHz) data results are as follows: δ: 5.66 (1H, d, J = 5.5Hz, H-6), 5.41 (1H, d, J = 5.5Hz, H-7), 5.22 (2H,...

Embodiment 3

[0076] Embodiment 3: Extraction of ergosterol monomer compound in Armillaria chrysanthemum

[0077] The raw materials, materials, reagents, instruments and equipment used in this embodiment are the same as those in Embodiment 1.

[0078] The extraction of the ergosterol monomer compound in Armillaria chrysanthemum in this embodiment comprises the following steps:

[0079] (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.

[0080] (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 sup...

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Abstract

The present invention provides a method for extracting ergosterol monomer compound in Armillaria chrysanthemum: take Armillaria chrysoderma sporocarp as raw material, extract it with ethyl acetate after being ground into powder, and concentrate to paste; use n-hexane-ethanol solvent soluble; one-dimensional liquid chromatography separation: using Silica chromatographic column, A phase n-hexane, B phase ethanol binary mobile phase, B phase concentration from 0% to 75% gradient elution, collect 8-11min eluent; use ethanol ‑N-hexane solvent dissolution; two-dimensional liquid chromatography separation: use Silica chromatographic column, A phase n-hexane, B phase ethanol binary mobile phase, B phase concentration 2‑6% isocratic elution, collect 12‑15min eluate , Evaporate and concentrate to obtain ergosterol. The separation method is stable, high in repeatability, large in preparation amount, simple and time-saving in operation, low in cost, and high in ergosterol extraction rate; the target substance has inhibitory effect on lung cancer A549 cells and liver cancer Hep‑G2 cells, and has the potential for developing anticancer drugs.

Description

technical field [0001] The invention relates to the technical fields of medicinal chemistry and biomedicine, in particular to a method for extracting ergosterol monomeric compounds from Armillaria chrysanthemum and the application of the ergosterol monomeric compounds in preparing anti-liver cancer drugs and anti-lung cancer drugs. 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 diameter, white or yello...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07J9/00A61P35/00
Inventor 张耀洲王欢欢杜思邈盖其静庞莉韩朝李上文马红杨珊珊陆洪
Owner TIANJIN YAOYU BIOLOGICAL TECH