Method for extracting tectorigenin from flos puerariae and application of tectorigenin

Abstract

The invention discloses a method for preparing tectorigenin by using pueraria flower as a raw material and converting tectorigenin by using a tremella sanguinea edible fungus solid-state fermentation method, and the process conditions for preparing tectorigenin by solid-state fermentation of pueraria flower by using tremella sanguinea edible fungus are determined. And analyzing the inhibiting effect concentration of the tectorigenin on the activity of xanthine oxidase.

Description

Technical Field

[0001] The invention relates to the technical field of plant medicinal component extraction, in particular to a method for directly extracting tectorigenin from kudzu flowers. Background Art

[0002] Pueraria lobata is widely distributed in my country, with abundant resources and high medicinal value. Pueraria lobata flower is the dried buds of Pueraria lobata, which is rich in various isoflavone compounds represented by irisin, and has many pharmacological effects, such as antioxidant, estrogen-like and liver protection, and has broad development and application prospects.

[0003] Iridogenin, also known as irisin, is one of the main components of kudzu flower isoflavone compounds. Its chemical name is 5,7,4'-trihydroxy-6-methoxyisoflavone. The isoflavone compounds in kudzu flower are usually in the form of isoflavone glycosides, and only a very small number of isoflavones exist in the form of aglycones. Isoflavone glycosides cannot be directly absorbed and utilized by the human body. They need to be metabolized into isoflavone aglycones by microorganisms in the intestines when passing through the intestines to exert their effects.

[0004] At present, the preparation of irisin is mainly carried out by acid hydrolysis and enzyme hydrolysis. Acid hydrolysis is to break the glucosidic bond in irisin under the action of acid and convert it into irisin. This method is simple to operate and low in cost, but mostly uses strong acid and the reaction temperature is high, which has hidden dangers in terms of safety and is easy to pollute the environment. Enzyme hydrolysis is to break the glucosidic bond in irisin and then form irisin by adding beta-glucosidase. This method is relatively gentle but has high cost. In the research before this application, mainly using irisin extract as raw material, using lactic acid bacteria fermentation, a remarkable effect is obtained. If Pueraria lobata flower can be used as raw material, directly carrying out microbial fermentation conversion, the preparation process of irisin will be further simplified, and the foundation for its industrial production will be laid.

[0005] The present application uses kudzu flower as raw material, screens out edible fungi species that can be used to transform irisin, and after solid-state fermentation, uses ethanol for extraction to directly obtain irisin extract. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a method for extracting tectorigenin from kudzu flowers and application thereof.

[0007] The present invention adopts the following technical scheme: comprising the following steps:

[0008] A method for extracting tectorigenin from kudzu flower, characterized in that the method comprises the following steps: S1: putting 10 g of kudzu flower powder into a 150 mL glass bottle, adding water and stirring evenly until it becomes granular, covering the bottle cap and sterilizing in an autoclave at 121° C. for 20 min, and the kudzu flower powder after sterilization is naturally cooled to room temperature;

[0009] S2: homogenize the activated mycelial pellets of Haemophilus auriculariae and the culture medium in the conical flask, add the homogenized bacterial suspension to each bottle of kudzu flower powder, and add water to a solid-liquid ratio of 1:1-5;

[0010] S3: Place in a 25°C constant temperature incubator for fermentation;

[0011] S4: After the fermentation is completed, the fermented kudzu flower powder is collected, distilled water is added at a solid-liquid ratio of 1:10, stirred on a magnetic stirrer for 5 minutes, centrifuged at 4000 r / min for 10 minutes, and the precipitate is collected;

[0012] S5: Add 120 mL of 70% ethanol to the precipitate, heat and reflux at 80° C. for extraction for 2 h, filter, concentrate the filtrate under reduced pressure, and dry naturally to obtain the fermented tectorius aglycone extract.

[0013] In some embodiments, the amount of the homogenized bacterial suspension added is 10 ml.

[0014] In some embodiments, water is added to S2 to a solid-liquid ratio of 1:3.

[0015] In some embodiments, the S3 fermentation period is 7 days.

[0016] The present application provides a drug for hyperuricemia, characterized in that it contains 0.36 mg / mL of the irisin prepared above.

[0017] The present application uses kudzu flower as raw material, extracts irisin through solid-state fermentation of the edible fungus Sanguisorba officinalis, optimizes the solid-state fermentation process, and then studies the inhibitory effect of irisin on xanthine oxidase.

[0018] This application optimizes the conditions for solid-state fermentation of kudzu flower. Taking the conversion rate of irisin as an indicator, the effects of inoculation amount, liquid-to-liquid ratio, activation time and fermentation time on the conversion rate of irisin were investigated, and the optimal fermentation conditions were determined to be: inoculation amount 10mL, liquid-to-liquid ratio 1:3, activation time 8d, solid-state fermentation time 7d. Under this condition, the conversion rate of irisin can reach 82.3%.

[0019] The extraction method used in this application is 70% ethanol heating reflux extraction, and the water-soluble substances in the kudzu flower and the water-soluble substances in the mycelium of Sanguinea odora will be extracted together. As an important type of water-soluble substance, sugars will not only affect the purity of the extract but also make the extract viscosity high and difficult to dry. Therefore, this application adds a water washing step in advance to the alcohol. Based on the low water solubility of irisin, distilled water is used to remove some water-soluble impurities in kudzu flower and reduce the viscosity of the ethanol extract.

[0020] This application studies the inhibitory kinetics of tectorius aglycone on xanthine oxidase, and its IC50 is 0.36 mg / mL, which has important guiding significance for the drug development of tectorius aglycone. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0022] Figure 1 It is the technical roadmap for the extraction of irisin;

[0023] Figure 2 These are pictures of kudzu flower fermented with different edible fungi in solid state;

[0024] Figure 3 It is the HPLC profile of the extract before and after fermentation;

[0025] Figure 4 This is the result of the effect of inoculum size on the conversion rate of irisin;

[0026] Figure 5 This is the result of the effect of the solid-liquid ratio on the conversion rate of irisin;

[0027] Figure 6 This is the result of the effect of activation time on the conversion rate of irisin;

[0028] Figure 7 It is the result of the effect of fermentation time on the conversion rate of irisin;

[0029] Figure 8 This is the result of the inhibitory effect of irisin and allopurinol on XO. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical scheme and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with the examples. The equipment and reagents used in each embodiment and test example can be obtained from commercial sources unless otherwise specified. The specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0031] Based on the information contained in this application, it is easy for those skilled in the art to make various changes to the precise description of the present invention without departing from the spirit and scope of the appended claims. It should be understood that the scope of the present invention is not limited to the defined process, property or component, because these embodiments and other descriptions are only for illustrating specific aspects of the present invention. In fact, various changes that the technicians in this field or related fields can obviously make to the embodiments of the present invention are all covered within the scope of the appended claims.

[0032] In order to better understand the present invention but not to limit the scope of the present invention, all the numbers used in this application to express the amount, percentage, and other numerical values ​​should be understood as modified by the word "approximately" in all cases. Therefore, unless otherwise specified, the numerical parameters listed in the specification and the appended claims are approximate values, which may be changed according to the different ideal properties attempted to be obtained. Each numerical parameter should at least be regarded as obtained based on the reported significant figures and by conventional rounding methods. In the present invention, "about" means within 10% of a given value or range, preferably within 5%.

[0033] In the following embodiments of the present invention, when the temperature is not particularly limited, it is room temperature. Room temperature refers to the natural room temperature in four seasons without additional cooling or heating treatment, and is generally controlled at 10-30°C, preferably 15-25°C.

[0034] Unless otherwise specified, the reagents and materials used in the embodiments of the present invention can be purchased from the market.

[0035] Pueraria flowers: produced in Anhui, dried, crushed, sealed and stored in a cool place for experimental use. Fungal species: Schizophyllum communieh (CICC 5046), Volvariella volvacea (CCTCC CF20081322), Stropharia rugosoannulata (CCTCC CF 20081452), Ganoderma lucidum (GDMCC NO: 5.535), Hericium erinaceus (CCTCC KF 2008394), Lentinula edodes (CCTCC KF 2008396), Tremella sanguinea Peng (CCTCC HF20081009) were stored in the Institute of Applied Fungi of Huazhong Agricultural University.

[0036] Preparation of CYM solid culture medium: 2g peptone, 2g yeast extract powder, 0.5g MgSO4·7H2O, 0.6g KH2PO4, 1g K2HPO4·3H2O, 22g glucose, 20g agar, add distilled water to 1L, stir evenly, sterilize in an autoclave at 121℃ for 20min. Pour into sterilized culture dishes to make plate culture medium.

[0037] Preparation of CYM liquid culture medium: 2 g of peptone, 2 g of yeast extract powder, 0.5 g of MgSO4·7H2O, 0.6 g of KH2PO4, 1 g of K2HPO4·3H2O, 22 g of glucose, add distilled water to 1 L, fill each conical flask with 100 mL, and sterilize in an autoclave at 121°C for 20 min.

[0038] Activation of strains: Take the fungal slant preserved in the laboratory, under sterile conditions, use an inoculation loop to deduct a small piece and inoculate it into CYM solid culture medium, place it in a constant temperature incubator at 25°C for constant temperature cultivation, stop culturing when the fungus fills the entire culture dish, under sterile conditions, use a hole puncher (inner diameter 7mm) on the plate, use an inoculation loop to pick up a circular fungal block and place it in another CYM solid culture medium for continued cultivation, when the fungus fills the entire culture dish, use a hole puncher (inner diameter 7mm) on the plate, use an inoculation loop to pick up 8 circular fungal blocks and place them in CYM liquid culture medium, and culture at 25°C, 120r / min until a large number of mycelial balls are produced in the conical flask.

[0039] Example 1 Preliminary screening of edible fungi strains

[0040] 10g of kudzu flower powder was placed in a 150mL glass bottle, 25mL of water was added, and the mixture was stirred evenly until it was granular. After the bottle cap was covered, the mixture was sterilized in an autoclave at 121℃ for 20min. After sterilization, the kudzu flower powder was naturally cooled to room temperature. In the clean bench, the activated mycelial balls and culture medium in the conical flask were homogenized, and 5mL of the homogenized bacterial suspension was added to each bottle of kudzu flower powder (i.e., the material-liquid ratio of the entire culture medium was 1:3). It was placed in a 25℃ constant temperature incubator for fermentation. The fermented kudzu flower powder was collected, 120mL of 70% ethanol was added, and the mixture was heated and refluxed at 80℃ for 2h, centrifuged at 4000r / min for 10min, and the supernatant was concentrated under reduced pressure. The fermented irisin conversion ability of the strain was used as an indicator for preliminary screening. Three parallel samples were set for each group of fermentation, and three independent parallel operations were performed.

[0041] Table 1

[0042]

[0043]

[0044] The fermentation strains were preliminarily screened based on the ability of edible fungi strains to transform irisin. Pueraria flowers inoculated with different edible fungi strains were placed in a 25°C constant temperature incubator for culture, and the growth of mycelium of 7 edible fungi was recorded. The results are shown in Table 1. Figure 2 .

[0045] Figure 2 (A) and Figure 2 (B) The pictures are of Stropharia officinalis and Ganoderma lucidum inoculated into kudzu flower 11 days later, and no mycelium was found on the surface of kudzu flower; Figure 2 (C) and Figure 2 (D) These are pictures of the straw mushroom and shiitake mushroom inoculated in kudzu flower for 11 days. It was found that a small amount of hyphae grew on the surface of kudzu flower for both straw mushroom and shiitake mushroom in the early stage of cultivation, but as the cultivation time prolonged, the hyphae stopped growing and began to dry and turn yellow. Figure 2 (E) Figure 2 (F) and Figure 2 (G) are pictures of Hericium erinaceus, white ginseng and blood ear after fermentation. The mycelium of the above three strains grew well and covered the surface of the kudzu flower powder. Among them, Hericium erinaceus can cover the surface of kudzu flower in 11 days, and white ginseng and blood ear can cover the surface of kudzu flower in 7 days. The normal growth of mycelium on kudzu flower is a prerequisite for its conversion to irisin, so Hericium erinaceus, white ginseng and blood ear with good growth were selected for the identification of irisin conversion ability.

[0046] Analysis of the ability to transform irisin from kudzu flowers:

[0047] UPLC-Q-TOF-MS was used for detection. The UPLC conditions were as follows: mobile phase: methanol-water (60:40); flow rate: 0.2 mL / min; detection wavelength: 265 nm; column temperature: 30°C; sample volume: 5 μL. Ion source: electrospray ionization (ESI); positive ion detection mode; drying gas flow rate: 8.0 L / min; dryer temperature: 300°C; spray pressure: 30.0 psi; capillary voltage: 3500 V; molecular weight scan range: 100 amu-800 amu. UNIFI software (Waters, USA) was used for data processing and substance identification.

[0048] (1) Determination of irisin conversion rate

[0049] The extract was dissolved in methanol solution, filtered through a microporous filter membrane (0.45 μm), and the conversion rate was determined by HPLC.

[0050] HPLC analysis conditions: Inert SustainTM AQ-C18 column (4.6 mm × 250 mm, 5 μm); mobile phase: methanol-water (60:40); flow rate: 1.0 mL / min; detection wavelength: 265 nm; column temperature: 30 °C; sample loading: 5 μ L.

[0051] Irisin conversion rate = (1-A1 / A2) × 100% Formula 1

[0052] Wherein: A1: peak area ratio of irisin after hydrolysis, %; A2: peak area ratio of irisin before hydrolysis, %.

[0053] (2) Determination of irisin yield

[0054] Accurately weigh 20 mg of tectorius aglycone standard dried to constant weight at 60°C in vacuum, place in a 10 mL volumetric flask, dissolve with methanol and dilute to 10 mL, shake well, obtain tectorius aglycone standard stock solution (2 mg / mL), and set aside. Dilute the tectorius aglycone standard stock solution to 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.4 mg / mL, 0.6 mg / mL, 0.8 mg / mL, 1.0 mg / mL, respectively. Filter with a microporous filter membrane (0.45 μm), perform HPLC analysis, inject samples in sequence, measure peak area, and plot a standard curve with peak area (mv*min) as ordinate and sample concentration (mg / mL) as abscissa.

[0055] Accurately weigh 20 mg of the extract dried to constant weight and dissolve it in methanol to prepare a 5 mg / mL sample solution. μ m) filtration, HPLC detection, and calculation of the yield of irisin.

[0056]

[0057] Wherein: M1: mass of extract, mg; C: content of irisin in the extract, %; M2: mass of kudzu flower, mg.

[0058] After solid-state fermentation of Hericium erinaceus, white ginseng and Trichosanthes chinensis, the kudzu flower powder was collected for isoflavone extraction, and the irisin conversion capacity of the strain was determined by HPLC. Figure 3 shown.

[0059] Figure 3 (A), (B), (C), (D) and (E) are the HPLC spectra of tectorius aglycone standard, unfermented extract, Hericium erinaceus fermented extract, white ginseng fermented extract and blood fungus fermented extract, respectively. Figure 3 (A) It can be seen that the retention time of irisin is about 15 minutes. Figure 3 In (B), there is a peak with a large area at around 6 min, while there is only a peak with a very small area at around 15 min. Figure 3 (C) The peak area of ​​the extract of Hericium erinaceus after fermentation at around 15 min increased slightly. Figure 3 (D) and Figure 3 In (E), the peak area of ​​the peak with a retention time of about 6 min decreased, and the peak area of ​​about 15 min increased significantly. This indicates that the substances in the kudzu flower were transformed by fermentation, and it can be preliminarily determined that tectorigenin was transformed after fermentation. The tectorigenin conversion rates of the three strains were calculated, among which the tectorigenin conversion rates of white ginseng and blood ear were higher, reaching 73.8% and 80.1%. The conversion ability of Hericium erinaceus was weak, only 21.9%. Therefore, blood ear with stronger conversion ability was selected for further screening.

[0060] Example 2 Optimization of solid-state fermentation conditions

[0061] A single factor experiment was conducted on the solid fermentation conditions with irisin conversion rate as an indicator. 10g of kudzu flower powder was placed in a 150mL glass bottle, stirred evenly after adding water until granular, and sterilized in an autoclave at 121℃ for 20min after the bottle cap was closed. After sterilization, the kudzu flower powder was naturally cooled to room temperature. In the clean bench, the activated blood ear mycelium balls and culture medium in the conical flask were homogenized, and the homogenized bacterial suspension was added to each bottle of kudzu flower powder. It was placed in a 25℃ constant temperature incubator for fermentation. After the fermentation was completed, the fermented kudzu flower powder was collected, distilled water was added at a solid-liquid ratio of 1:10, stirred on a magnetic stirrer for 5min, centrifuged at 4000r / min for 10min, and the precipitate was collected. 120mL of 70% ethanol was added to the precipitate, heated and refluxed at 80℃ for 2h, filtered, the filtrate was concentrated under reduced pressure, and dried naturally to obtain the crude extract of irisin after fermentation.

[0062] Optimize fermentation conditions from the following conditions:

[0063] 1) Effect of the amount of Haemophilus influenzae inoculation on the conversion rate of irisin

[0064] The 7-day-activated radix auriculae was inoculated into 10 g of sterilized kudzu flower solid powder with 5 mL, 10 mL, 15 mL, 20 mL, and 25 mL radix auriculae inoculation amounts, water was added to a solid-liquid ratio of 1:3, and cultured at 25°C for 7 days. Pueraria flower isoflavones were extracted and the irisin conversion rate was determined.

[0065] Depend on Figure 4 It can be seen that with the increase of inoculation size, the conversion rate of irisin shows a slow upward trend. When the inoculation size is low, the mycelium grows slowly and is unevenly distributed, and the conversion rate of irisin is low; when the inoculation size is increased to 10mL, the conversion rate increases significantly and then tends to stabilize. When the inoculation size is too large, the blood ear grows rapidly at first, but the nutrient supply in the kudzu flower is insufficient, which affects the subsequent growth of the mycelium. Therefore, in order to ensure the rapid growth of mycelium and reduce the fermentation cost, an inoculation size of 10mL was selected for solid-state fermentation of kudzu flower.

[0066] 2) Effect of material-liquid ratio on irisin conversion rate

[0067] With an inoculum of 10 mL, the activated 7-day-old tricholoma was inoculated into 10 g of sterilized kudzu flower solid powder, and water was added to a solid-liquid ratio of 1:1, 1:2, 1:3, 1:4, and 1:5, and cultured at 25°C for 7 days. Pueraria flower isoflavones were extracted and the irisin conversion rate was determined.

[0068] In the solid-state fermentation system, the moisture content will affect the growth and metabolism of microorganisms. If the moisture content is too low, the kudzu flower powder will absorb water and swell, making it difficult for the nutrients in it to dissolve and transfer; if the moisture content is too high, the kudzu flower powder will easily clump, the air permeability will be reduced, the microorganisms will not get enough oxygen, and the growth will be slow. Therefore, the appropriate moisture content is the key to improving the conversion rate of irisin. Figure 5 It can be seen that with the change of the material-liquid ratio, the amount of water added increases, and the conversion rate of irisin shows a significant upward trend. The highest conversion rate is 81.8% when the material-liquid ratio is 1:3. Continue to increase the moisture content. When the material-liquid ratio is 1:4, the moisture absorbed by the kudzu flower reaches saturation, the kudzu flower powder is relatively close, the oxygen content available for mycelium growth is low, the mycelium grows poorly, and the irisin conversion rate decreases. Continue to increase the water content. At this time, the fermentation system begins to have freely flowing water, and the irisin conversion rate increases again. When the material-liquid ratio is 1:3, the mycelium growth state is the best and the irisin conversion rate is the highest. Therefore, the material-liquid ratio is selected to be 1:3.

[0069] 3) Effect of activation time of blood ear on the conversion rate of irisin

[0070] With an inoculum of 10 mL, the activated 5d, 6d, 7d, 8d, and 9d scutellariae were inoculated into 10g of sterilized kudzu flower solid powder, water was added to a solid-liquid ratio of 1:3, and cultured at 25°C for 7d. Pueraria flower isoflavones were extracted and the irisin conversion rate was determined.

[0071] Liquid activation of the strain is a very important step before inoculation for solid fermentation. If the activation days are too short, the amount of mycelium is small, and the mycelium grows slowly after inoculation for solid fermentation; and if the activation days are too long, the strain ages, and the growth and enzyme production capacity decreases, which also affects the solid fermentation. Therefore, choosing the appropriate activation days is particularly critical. Figure 6 It can be seen that with the extension of activation days, the conversion rate of irisin gradually increased. When the activation days were 8 days, the solid-state fermentation irisin conversion rate was the highest, and then it decreased. Therefore, 8 days was selected as the activation days of the strain.

[0072] 4) Effect of fermentation time on irisin conversion rate

[0073] With an inoculum of 10 mL, the 8-day-activated radix auriculae was inoculated into 10 g of sterilized kudzu flower solid powder, water was added to a solid-liquid ratio of 1:3, and cultured at 25°C for 5, 6, 7, 8, and 9 days. Pueraria flower isoflavones were extracted and the irisin conversion rate was determined.

[0074] Depend on Figure 7 It can be obtained that the fermentation time will significantly affect the conversion rate of irisin in the extract. In the early stage of fermentation, the mycelium biomass is less, and the microorganisms can continue to grow using the nutrients in the kudzu flower, and the fermentation is not thorough, so the irisin conversion rate is low. As the fermentation time increases, the blood ear grows rapidly, the mycelium extends to the inside of the kudzu flower matrix, and the irisin conversion rate rises rapidly. At 7d, the conversion rate reached 82.3%. Afterwards, as the fermentation days increase, the remaining amount of irisin is already less, and the conversion rate does not increase significantly. Therefore, 7d is selected as the kudzu flower solid-state fermentation days.

[0075] Example 2 Inhibitory effect of tectoriusin on xanthine oxidase

[0076] Determination of half inhibitory concentration:

[0077] Since the product generated by xanthine oxidase (XO) catalyzing the substrate xanthine has a characteristic absorption peak at 290nm, this experiment measures the change of XO activity by measuring the change of ultraviolet absorption value here within a certain period of time, that is, the generation rate of the product. In the PBS (pH7.5, 0.08mol / L) buffer system, the concentration of XO (0.03U / mL) was fixed, and different concentrations of inhibitor irisin solution were added to keep incubation at 37℃ for 10min, and the substrate xanthine (0.1mg / mL) was quickly added to start the reaction. The kinetic section of the microplate reader was used, the wavelength was 290nm, and the detection was performed every 20sec, and the changes in the photometric value within 5min were recorded. The experiment set up 3 groups: sample group (adding sample, substrate and enzyme), blank group (adding only substrate without sample and enzyme) and enzyme group (adding only substrate and enzyme without sample), and each sample was set up 3 parallels.

[0078] Calculation of inhibition rate (%): The enzyme activity was calculated by calculating the change rate of the OD290 nm value of the sample group, blank group and enzyme group, that is, the size of the slope K, and then the inhibition rate was calculated. The inhibition rate formula is:

[0079]

[0080] Among them, K is the slope of OD290 nm. The detection time is 0-5min. From the actual fitting effect, 20sec-200sec is selected, at which time the slope correlation of the enzyme activity curve is better.

[0081] The effects of different concentrations of irisin extract and allopurinol on XO activity were determined. Figure 8 (A) and (B). Figure 8 (A) It can be seen that as the concentration of tectorius aglycone increases, the activity of XO decreases. Tectorius aglycone has a dose-effect relationship in inhibiting XO, and the inhibition rate increases with increasing concentration. The IC50 value of tectorius aglycone for XO is 0.36 mg / mL. This is higher than the IC50 value of the positive control drug allopurinol for XO, which is 3.78 μg / mL. This shows that tectorius aglycone is an effective XO inhibitor.

[0082] At present, allopurinol and febuxostat are commonly used drugs for the treatment of hyperuricemia, but they have many side effects, so it is very important to find effective and less toxic xanthine oxidase inhibitors. Iridogenin is a natural isoflavone compound found in kudzu flowers. Some studies have reported that isoflavone compounds have a good inhibitory effect on xanthine oxidase. Therefore, this experiment explored the inhibitory effect of irisogenin on xanthine oxidase. The IC50 of irisogenin extract was measured to be 0.36 mg / mL.

[0083] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for extracting tectorigenin from kudzu flower, characterized in that: The method comprises the following steps: S1: Put 10g of kudzu flower powder into a 150mL glass bottle, add water and stir evenly until it becomes granular, cover the bottle cap and sterilize in an autoclave at 121℃ for 20min. After sterilization, the kudzu flower powder is cooled naturally to room temperature; S2: homogenize the activated mycelial pellets of Haemophilus auriculariae and the culture medium in the conical flask, add the homogenized bacterial suspension to each bottle of kudzu flower powder, and add water to a solid-liquid ratio of 1:1-5; S3: Place in a 25°C constant temperature incubator for fermentation; S4: After the fermentation is completed, the fermented kudzu flower powder is collected, distilled water is added at a solid-liquid ratio of 1:10, stirred on a magnetic stirrer for 5 minutes, centrifuged at 4000 r / min for 10 minutes, and the precipitate is collected; S5: Add 120 mL of 70% ethanol to the precipitate, heat and reflux at 80° C. for extraction for 2 h, filter, concentrate the filtrate under reduced pressure, and dry naturally to obtain the fermented tectorius aglycone extract.

2. The method according to claim 1, characterized in that The amount of the homogenized bacterial suspension added is 10 ml.

3. The method according to claim 1, characterized in that: Water is added to the S2 to a solid-liquid ratio of 1:

3.

4. The method according to claim 1, characterized in that: The S3 fermentation period is 7 days.

5. A drug for hyperuricemia, characterized in that: Contains 0.36 mg / mL of tectorius aglycone prepared according to claims 1-4.

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