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CN-122010973-A - Flavonoid compound and extraction method thereof

CN122010973ACN 122010973 ACN122010973 ACN 122010973ACN-122010973-A

Abstract

The invention belongs to the technical field of natural product chemistry and medicine extraction, and provides a flavonoid compound and an extraction method thereof. The flavonoid compound provided by the invention has a structure shown in the following formula. The flavonoid compound provided by the invention has novel structure, and widens the variety of natural bioactive substances. Meanwhile, the flavonoid compound provided by the invention has a rapid antidepressant effect. The invention also provides an extraction method of the flavonoid compound, which has the advantages of high extraction efficiency, reasonable purification route, high product purity and good reproducibility.

Inventors

  • LAI REN
  • ZHOU SHENGWEN
  • YANG JINSONG
  • LV QIUMIN
  • CHEN YIFAN

Assignees

  • 中国科学院昆明动物研究所

Dates

Publication Date
20260512
Application Date
20260204

Claims (10)

  1. 1. A flavonoid compound, which is characterized by having a structure shown in formula 1: Formula 1.
  2. 2. The method for extracting flavonoid compounds according to claim 1, comprising the steps of: Mixing the fruiting body of Phlebopus portentosus with ethanol water solution, sequentially performing ultrasonic extraction, solid-liquid separation and liquid phase concentration to obtain crude extract; Re-dissolving the crude extract to obtain a crude extract dispersion liquid, sequentially adopting petroleum ether and chloroform for extraction to remove impurities in the crude extract dispersion liquid to obtain a purified water phase, extracting the purified water phase by using ethyl acetate, and concentrating the obtained ethyl acetate phase to obtain an ethyl acetate part extract; Performing silica gel column chromatography on the ethyl acetate part extract, wherein an eluent of the silica gel column chromatography is a chloroform-methanol system, the volume ratio of chloroform to methanol in the chloroform-methanol system is sequentially 1:0, 2:0.8-1.2, 1:0.8-1.2 and 0:1, and eluting fractions with the volume ratio of chloroform to methanol of 1:0.8-1.2 are concentrated to obtain a crude product; performing high performance liquid chromatography on the crude product to obtain the flavonoid compound; The high performance liquid chromatography separation comprises sequentially performing a first high performance liquid chromatography separation and a second high performance liquid chromatography separation; the conditions for the first high performance liquid chromatography separation include: the chromatographic column is Waters XBridge BEH OBD C preparation columns; The mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is water, and the mobile phase B is acetonitrile; The elution mode is gradient elution; the gradient elution procedure was: 0-5min, wherein the volume fraction of the mobile phase A is 95%; 5-30min, wherein the volume fraction of the mobile phase A is changed from 95% to 5%; the conditions for the second high performance liquid chromatography separation include: The chromatographic column is Waters XSelect CSH C chromatographic column; The mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is water, and the mobile phase B is acetonitrile; The elution mode is gradient elution; the gradient elution procedure was: 0-5min, wherein the volume fraction of the mobile phase A is 90%; 5-15min, wherein the volume fraction of the mobile phase A is changed from 90% to 40%.
  3. 3. The extraction method according to claim 2, wherein the volume concentration of the aqueous ethanol solution is 80-95%; the dosage ratio of the boletus ranunculatus fruiting bodies to the ethanol aqueous solution is 1 g:3-10 mL.
  4. 4. The extraction method according to claim 2 or 3, wherein the number of times of ultrasonic extraction is 1 to 3; The power of ultrasonic extraction is 80-120W, and the frequency is 28-40 kHz; The ultrasonic extraction working mode is an intermittent mode, the intermittent mode is ultrasonic for 4-6 min and stop for 4-6 min, and single ultrasonic extraction comprises 2-4 intervals.
  5. 5. The extraction method according to claim 2, wherein the number of times of extraction of petroleum ether, chloroform and ethyl acetate is independently 2 to 4 times.
  6. 6. The extraction method of claim 2, wherein the sample loading mode of the silica gel column chromatography is dry sample loading, and the dry sample loading comprises the steps of mixing the ethyl acetate part extract with silica gel, loading after a solvent is volatilized, wherein the mass ratio of the ethyl acetate part extract to the silica gel is 1:8-12, and the particle size of the silica gel is 100-200 meshes; The particle size of silica gel filled in a chromatographic column used for silica gel column chromatography is 80-100 meshes.
  7. 7. The extraction method according to claim 2 or 6, wherein the flow rate of the eluent is 0.8-1.2 ml/min in the silica gel column chromatography process.
  8. 8. The extraction method according to claim 2, wherein the conditions for the first hplc separation include: the flow rate is 1-2 mL/min; The column temperature is 25-35 ℃; ultraviolet detection wavelengths are 215nm and 254nm; and collecting the components with the retention time of 15-18 min.
  9. 9. The method of claim 8, wherein the conditions for the second hplc separation include: the flow rate is 0.4-0.6 mL/min; The column temperature is 25-35 ℃; ultraviolet detection wavelengths are 215nm and 254nm; The main peak fractions were collected.
  10. 10. The method according to claim 2, 8 or 9, wherein the high performance liquid chromatography further comprises a post-treatment after the target fraction is obtained, wherein the post-treatment comprises sequentially concentrating and freeze-drying the target fraction to obtain the flavonoid compound.

Description

Flavonoid compound and extraction method thereof Technical Field The invention relates to the technical field of natural product chemistry and medicine extraction, in particular to a flavonoid compound and an extraction method thereof. Background The boletus ranunculaceae (Lanmaoa asiatica) is edible wild fungi with characteristics of Yunnan province, has unique flavor and is rich in various natural products with biological activity. Studies show that some secondary metabolites such as strophasterol E contained in the bacterium have an inhibitory effect on staphylococcus aureus (Staphylococcus aureus) and bacillus subtilis (Bacillus subtilis), and the other compound 6 beta-methoxy ergosterol-7, 9 (11), 22E-triene-3 beta, 5 alpha-diol shows anti-tumor activity. The method is significant in developing natural bioactive substances by excavating active ingredients in boletus ranunculaceae. Disclosure of Invention Accordingly, the present invention is directed to a flavonoid compound and an extraction method thereof. The flavonoid compound provided by the invention has novel structure, and expands natural bioactive substances. In order to achieve the above object, the present invention provides the following technical solutions: the invention provides a flavonoid compound, which has a structure shown in a formula 1: Formula 1. The invention also provides an extraction method of the flavonoid compound, which comprises the following steps: Mixing the fruiting body of Phlebopus portentosus with ethanol water solution, sequentially performing ultrasonic extraction, solid-liquid separation and liquid phase concentration to obtain crude extract; Re-dissolving the crude extract to obtain a crude extract dispersion liquid, sequentially adopting petroleum ether and chloroform for extraction to remove impurities in the crude extract dispersion liquid to obtain a purified water phase, extracting the purified water phase by using ethyl acetate, and concentrating the obtained ethyl acetate phase to obtain an ethyl acetate part extract; Performing silica gel column chromatography on the ethyl acetate part extract, wherein an eluent of the silica gel column chromatography is a chloroform-methanol system, the volume ratio of chloroform to methanol in the chloroform-methanol system is sequentially 1:0, 2:0.8-1.2, 1:0.8-1.2 and 0:1, and eluting fractions with the volume ratio of chloroform to methanol of 1:0.8-1.2 are concentrated to obtain a crude product; performing high performance liquid chromatography on the crude product to obtain the flavonoid compound; The high performance liquid chromatography separation comprises sequentially performing a first high performance liquid chromatography separation and a second high performance liquid chromatography separation; the conditions for the first high performance liquid chromatography separation include: the chromatographic column is Waters XBridge BEH OBD C preparation columns; The mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is water, and the mobile phase B is acetonitrile; The elution mode is gradient elution; the gradient elution procedure was: 0-5min, wherein the volume fraction of the mobile phase A is 95%; 5-30min, wherein the volume fraction of the mobile phase A is changed from 95% to 5%; the conditions for the second high performance liquid chromatography separation include: The chromatographic column is Waters XSelect CSH C chromatographic column; The mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is water, and the mobile phase B is acetonitrile; The elution mode is gradient elution; the gradient elution procedure was: 0-5min, wherein the volume fraction of the mobile phase A is 90%; 5-15min, wherein the volume fraction of the mobile phase A is changed from 90% to 40%. Preferably, the volume concentration of the ethanol water solution is 80-95%; the dosage ratio of the boletus ranunculatus fruiting bodies to the ethanol aqueous solution is 1 g:3-10 mL. Preferably, the ultrasonic extraction times are 1-3 times; The power of ultrasonic extraction is 80-120W, and the frequency is 28-40 kHz; The ultrasonic extraction working mode is an intermittent mode, the intermittent mode is ultrasonic for 4-6 min and stop for 4-6 min, and single ultrasonic extraction comprises 2-4 intervals. Preferably, the extraction times of the petroleum ether, the chloroform and the ethyl acetate are independently 2-4 times. The dry sample loading method comprises the steps of mixing the ethyl acetate part extract with silica gel, loading the mixture after a solvent is volatilized, wherein the mass ratio of the ethyl acetate part extract to the silica gel is 1:8-12, and the particle size of the silica gel is 100-200 meshes; The particle size of silica gel filled in a chromatographic column used for silica gel column chromatography is 80-100 meshes. Preferably, in the silica gel column chromatography process, the fl