CN111096983A - Phellinus igniarius phenolic extract with hypoglycemic activity and preparation and application thereof - Google Patents
Phellinus igniarius phenolic extract with hypoglycemic activity and preparation and application thereof Download PDFInfo
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- CN111096983A CN111096983A CN202010052874.7A CN202010052874A CN111096983A CN 111096983 A CN111096983 A CN 111096983A CN 202010052874 A CN202010052874 A CN 202010052874A CN 111096983 A CN111096983 A CN 111096983A
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- extract
- phellinus igniarius
- ethyl acetate
- phellinus
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Abstract
The invention relates to a phellinus igniarius phenolic extract with hypoglycemic activity, a preparation method and application thereof.A ethyl acetate extract with higher phenolic purity (more than or equal to 70 percent) is obtained by ultrasonic ethanol aqueous solution auxiliary extraction and solvent fractional extraction, and mainly comprises substances such as osmoracetone, hispidin, davallialactone, 2,5-bis (4,7-dihydroxy-8-methyl-2-oxo-2H-chromen-3-yl) cyclohexa-2,5-diene-1,4-dione, phytomine B and exocavirn A2The cell glucose consumption has obvious promotion effect, and has good hypoglycemic activity and potential diabetes prevention and treatment efficacy; the invention has simple process and no fine separation stepAnd the preparation efficiency is high, the cost is low, and the method is suitable for subsequent pilot plant test and industrial production.
Description
(I) technical field
The invention relates to a phellinus igniarius phenolic extract with hypoglycemic activity, and a preparation method and application thereof.
(II) background of the invention
Diabetes Mellitus (DM) is a chronic endocrine metabolic disease in which carbohydrate, fat, protein, water, and electrolytes are disturbed due to absolute or relative hyposecretion of insulin in the body, and is associated with genetic factors as well as various environmental factors. Its typical manifestations are "more than three and one less" -polydipsia, polyphagia, polyuria, emaciation, and elevated blood sugar.
Diabetes Mellitus (DM) is a chronic endocrine metabolic disease in which carbohydrate, fat, protein, water, and electrolytes are disturbed due to absolute or relative hyposecretion of insulin in the body, and is associated with genetic factors as well as various environmental factors. Its typical manifestations are "more than three and one less" -polydipsia, polyphagia, polyuria, emaciation, and elevated blood sugar.
With the development of modern society, bad life and working modes are increased, the aging of population is intensified continuously, and the incidence of diabetes mellitus in the world is on a rapid rising trend for decades. According to the statistics of the international diabetes union (IDF), about 4.63 hundred million 20-79 years old adults suffer from diabetes in 2019 globally; it is predicted that by 2030, diabetics will reach 5.784 billion; it is expected that by 2045 years, diabetics will reach 7.002 billion. The number of diabetic patients is the first in China, the total number is about 1.164 hundred million, hyperglycemia can cause heart disease, hypertension, renal failure, blindness, ulcer or necrosis of lower limbs and other diabetic complications, so that the patients are disabled or dead, the number of diabetic patients is the third killer second to tumor and cardiovascular diseases, the number of diabetic patients who die from diabetes and the diabetic complications in 2019 is up to 83 thousands, and the related medical expenses in China are measured in billions every year.
For the treatment of type I diabetes, the research is directed to the development of insulin preparations and substitutes that are convenient and effective to administer, whereas for the treatment of type II diabetes, there is no need to rely solely on insulin supplementation, and oral hypoglycemic agents such as insulin secretagogues (e.g., sulfonylureas), insulin sensitizers (e.g., thiazolidinediones), glucose metabolism enhancers (e.g., biguanides), and α -glucosidase inhibitors (e.g., Acarbose, Voglibose, and Miglitol, which are clinically used at present, have a major hypoglycemic mechanism that acts as a competitive, dose-dependent inhibition on small intestine α -glucosidase, thereby preventing poorly absorbable polysaccharide compounds from being converted into readily absorbable monosaccharides).
Phellinus igniarius (also called Phellinus igniarius), Phellinus igniarius, Hoffia sciaefolia and Morus alba (Linn.) Kuntze, is a medicinal fungus of the genus Phellinus igniarius, the order of compendia, the order of Polyporales, the family of Polyporaceae, and the genus Phellinus. Wild phellinus igniarius, which is called forest gold, is widely distributed in China, especially in north China, northeast China, northwest China, Yunnan China, Sichuan China and the like. Over 1000 years ago, the first herbal of our country has a record of phellinus igniarius, and the Li Shizhen' compendium of herbal has a clear record of the origin, property and function of phellinus igniarius.
Phellinus linteus facultative parasitic, mainly saprophytic, is a high-temperature medicinal fungus, and is parasitic on the withered branches of broadleaf trees such as white birch, gutter, and the like of poplar and willow, so as to cause white rot of heartwood. Phellinus igniarius is prepared by spore germination to generate hypha, the hypha is gathered to form the mycelium, the mycelium forms sporocarp, the sporocarp can produce brood seed, and the Phellinus igniarius is continuously reproduced to have slightly bitter taste, but has the effects of benefiting five internal organs, expelling toxin, stopping bleeding, promoting blood circulation and the like, wherein the sporocarp is used as the medicine in the 'medicinal herbs': the record of "treating metrorrhagia and leukorrhagia, amenorrhea, postpartum hemagglutination, and male scrofula" is described. Modern researches show that phellinus igniarius has the effects of improving immunity, resisting tumors and oxidation, protecting the liver, resisting inflammation, bacteria and mutation, reducing blood sugar, resisting diabetes and the like. The main chemical components of the composition are polysaccharides, and the composition also contains substances such as fatty acid, sterol, triterpene, flavone, aromatic acid, melanin, phenols and the like.
In recent years, large-scale artificial cultivation of phellinus igniarius sporocarp is successful, and the problem of scarcity of wild phellinus igniarius resources is solved.
The existing literature and patents of phellinus linteus are mainly focused on the components of phellinus linteus polysaccharide in the aspects of reducing blood sugar and resisting diabetes.
As demonstrated by the Zhangioli research, Phellinus linteus mycelium polysaccharide can reduce blood sugar concentration of diabetic mice caused by streptozotocin. The sugar-reducing rates of the high, medium and low dose groups of phellinus igniarius mycelium polysaccharide to the diabetic mice are 0.92%, 0.90% and 1.99% respectively, and the sugar-reducing rate of the positive control group is 3.31% (Zhangbao, research on the biological activities of the phellinus igniarius mycelium polysaccharide for reducing blood sugar and protecting liver function, Master thesis of northeast Master university, 2007).
Liaozunsheng research shows that the phellinus igniarius hypha polysaccharide has a good blood sugar reducing effect on a tetraoxypyrimidine mouse hyperglycemic model (Liaozunsheng, research on separation and purification and blood sugar reducing effects of phellinus igniarius hypha polysaccharide, university of Fujian agriculture and forestry, Master academic paper, 2013).
Zhang Xuedan research shows that the Phellinus linteus mycelium obtained by solid state fermentation method is subjected to hot water extraction and deproteinization, and the extract is subjected to intragastric administration, so that the blood sugar level of type I diabetic mice can be effectively regulated, and the hyperglycemia symptoms of the mice can be improved (Zhang Xuedan, research on the hypoglycemic activity of Phellinus linteus mycelium extract and chitosan oligosaccharide, Master academic thesis of Huazhong agriculture university, 2015).
The study of dynasty emperor cloud crane shows that the polysaccharide of the mycelium of the phellinus igniarius through gastric perfusion can reduce the blood sugar of a diabetic mouse and improve the sugar tolerance (the study of the antioxidation and the blood sugar reducing functions of the polysaccharide of the phellinus igniarius, a university Master academic paper, 2017).
Studies of Shidejun et al in Jilin found that a high dose of phellinus linteus polysaccharide extract is helpful for improving diabetes mellitus type I (DM) mice (Shidejun et al, the influence of phellinus linteus polysaccharide on a diabetes mellitus type I mouse model, university of Yanbian, 2017) in Streptozotocin (STZ) modeling, studies of Huangqian et al in Fujian found that phellinus linteus polysaccharide can regulate MMP-2/TIMP-2 balance, reduce renal interstitial fibrosis (Huangqian et al, the study of phellinus linteus polysaccharide through P311/TGF- β 1/Snail1 pathway in diabetes mellitus mouse renal interstitial fibrosis, pharmacology and clinic of traditional Chinese medicines, 2019), while in terms of phenols, results of Shanghai Zhangiesu et al in Shanghai showed that phellinus linteus rice from Phellinus baumii3 compounds of protocatechualdehyde, naringenin and baicalein separated from fermented mycelium are used for HepG2The cell glucose consumption has obvious promotion effect (Zhang super, Phellinus baumii compound on HepG)2Investigation of the effect of cellular glucose consumption and its mechanism of action, bacteriological report, 2016).
Liu bin et al invented "a compound hypoglycemic oral liquid of phellinus igniarius polysaccharides and traditional Chinese medicines and a preparation method thereof", showed that the compound of phellinus igniarius solid fermentation mycelium polysaccharides and traditional Chinese medicines with hypoglycemic effect is very strong (application publication No. CN 103405639A).
The invention discloses a medicinal composition for preparing a medicament for treating and preventing hyperglycemia, and provides application of the medicinal composition in preparing the medicament for treating and preventing hyperglycemia, wherein the medicinal composition is prepared by mixing or extracting schisandra chinensis, phellinus igniarius sporocarp and radix puerariae (application publication number, CN 104383084A).
Pengliang 'a medicinal fungus prescription for preventing and treating diabetes and its preparation method', states that a medicinal fungus prescription can prevent and treat type II diabetes, the prescription includes: inonotus obliquus, Phellinus linteus, Coriolus versicolor, Ganoderma sinense, Ganoderma applanatum, Phellinus linteus and Ganoderma capense (application publication No. CN 106389483A).
The above patent documents of the phellinus igniarius for reducing blood sugar and resisting diabetes are combined, the phellinus igniarius polysaccharides are mainly concentrated on the aspect of phellinus igniarius polysaccharides, the extracted phellinus igniarius polysaccharides are crude polysaccharide mixtures, specific components and structures are not clear, and the characterization of the fine structures of macromolecular polysaccharides is still difficult at present. Only one phenol, 3 compounds of protocatechuic aldehyde, naringenin and baicalein were shown to be present in HepG2The cell glucose consumption has obvious promotion effect, but the cost is high for obtaining the three pure compounds.
Disclosure of the invention
The invention aims to provide a phellinus igniarius phenolic high-content extract with an in-vitro blood sugar reducing effect, and a preparation method and application thereof, aiming at overcoming the defects of the prior art.
The technical scheme adopted by the invention is as follows:
phellinus igniarius phenolic extract with hypoglycemic activity, wherein the purity of phenols is more than or equal to 70%, and is obtained by the following method:
(1) vacuum drying Phellinus igniarius solid cultured fruiting bodies at 40-60 ℃, crushing and sieving to 40-80 meshes, adding 60-80% ethanol aqueous solution according to a solid-to-liquid ratio of 1: 10-1: 30, carrying out ultrasonic-assisted extraction for 1-3 times, each time for 20-40 min, centrifuging, and taking supernate;
(2) performing rotary evaporation on the supernatant obtained in the step (1), concentrating, freeze-drying to obtain a phellinus igniarius ethanol extract, adding purified water, performing ultrasonic suspension mixing, adding petroleum ether with the same volume, extracting at 30-60 ℃ for 1-3 times, collecting the petroleum ether extract, adding ethyl acetate with the same volume into the residual phase, extracting for 1-3 times, and combining and collecting ethyl acetate extract phases;
(3) and (3) collecting the ethyl acetate extract phase in the step (2), performing vacuum rotary evaporation and concentration until no organic solvent smell exists, and performing freeze drying to obtain the phellinus igniarius phenolic extract with the hypoglycemic activity.
The invention adopts ultrasonic ethanol water solution to assist extraction and solvent fractional extraction to obtain an ethyl acetate extract with higher phenol purity (more than or equal to 70 percent), the ethyl acetate extract has obvious inhibition effect on α -glycosidase in vitro, and the main substances in the extract are phenol components through LC-MS analysis, wherein the phenol components mainly comprise substances such as osmoracene, hispidin, davallialactone, 2,5-bis (4,7-dihydroxy-8-methyl-2-oxo-2H-chromen-3-yl) cyclohexoxa-2, 5-diene-1, 4-diene, hydrochromine B, inoscaivin A and the like.
The present invention also relates to a method for preparing the phellinus linteus phenolic extract, which comprises:
(1) vacuum drying Phellinus igniarius solid cultured fruiting bodies at 40-60 ℃, crushing and sieving to 40-80 meshes, adding 60-80% ethanol aqueous solution according to a solid-to-liquid ratio of 1: 10-1: 30, carrying out ultrasonic-assisted extraction for 1-3 times, each time for 20-40 min, centrifuging, and taking supernate;
(2) performing rotary evaporation on the supernatant obtained in the step (1), concentrating, freeze-drying to obtain a phellinus igniarius ethanol extract, adding purified water, performing ultrasonic suspension mixing, adding petroleum ether with the same volume, extracting at 30-60 ℃ for 1-3 times, collecting the petroleum ether extract, adding ethyl acetate with the same volume into the residual phase, extracting for 1-3 times, and combining and collecting ethyl acetate extract phases;
(3) collecting the ethyl acetate extract phase in the step (2), performing rotary evaporation and concentration until no organic solvent smell exists, and performing freeze drying to obtain the phellinus igniarius phenolic extract with hypoglycemic activity.
The rotary evaporation in the steps (2) and (3) is carried out at 40-60 ℃ and under the vacuum degree of 0.05-0.1 Mpa.
In the step (1), the ultrasonic extraction power is 80-100W.
The extract has obvious inhibition effect on α -glycosidase in vitro, which shows that the extract has good blood sugar reducing and potential diabetes preventing and treating effects.
Therefore, the phellinus igniarius phenolic extract can be used for preparing health care products for reducing blood sugar or medicines for preventing and treating diabetes.
The method has the advantages that the phellinus igniarius phenolic extract has high phenolic content, has a remarkable inhibiting effect on α -glycosidase, has good hypoglycemic activity and a potential effect of preventing and treating diabetes, is simple in process, free of fine separation steps, high in preparation efficiency and low in cost, and is suitable for subsequent pilot plant test and industrial production.
(IV) description of the drawings
FIG. 1 is a diagram of Phellinus linteus ethyl acetate components LC-MS (A) and structure (B);
FIG. 2 is a flow chart of the preparation process of Phellinus linteus phenolic extract.
(V) detailed description of the preferred embodiments
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:
the preparation process comprises the following steps:
the phellinus igniarius solid cultivation sporocarp (provided by sericulture institute of academy of agricultural sciences in Zhejiang province) → 40-60 ℃ vacuum drying (Dejangde Binder VD23 vacuum drying oven) → crushing and sieving to 40-80 mesh → 50g of the fruit is weighed and added with 1: 10-1: 30 (solid-to-liquid ratio, W/W), 60-80% (W/W) of ethanol aqueous solution → ultrasonic assisted extraction (FS-1200 type ultrasonic cell disruptor, Shanghai Biotechnology corporation) with extraction power of 80-100W for 20-40 min, extraction times of 1-3 → 5000R/min centrifugal residue removal TD5A-WS, No.4 rotor, 4 × 100mL → combined supernatant liquid → rotary evaporation (rotary evaporator, R2002B, combined with circulating Water pump, Shanghai Shensheng limited company), 40-60 ℃ vacuum degree of 0.05-0.1 MPa, concentration to original volume 1/5 of dry phellinus igniarius ethanol extract → EE) and adding Bu Water phase extraction solvent → 100mL of Bu 2, adding ethyl acetate to obtain residual ethyl acetate phase extraction solvent (PET-3), and concentrating, and collecting residual ethyl acetate phase, respectively, adding ethyl acetate phase extraction solvent before vacuum phase extraction, concentrating to obtain the residual ethyl acetate phase extraction solvent (PET-3 → PET-3), and concentrating to obtain the residual ethyl acetate phase.
Description of the main operating steps:
(1) phellinus Linteus fruiting body is obtained by culturing ramulus Mori solid, and is provided by Bombycis Mori of Nostoc of Zhejiang province.
(2) Oven drying Phellinus igniarius sporophore, pulverizing, sieving to obtain Phellinus igniarius powder, and refrigerating at 4 deg.C in double-layer sealed bag.
(3) Vacuum rotary evaporation and concentration: the components are concentrated before final freeze-drying, the organic solvent is completely evaporated, part of water is reserved for subsequent freeze-drying, and if the organic solvent is not completely evaporated, the subsequent in vitro activity determination and the application of medical food are seriously influenced.
(4) α -glucosidase inhibition in vitro was evaluated by adding 100. mu.l α -glucosidase (0.5U/mL) dissolved in 100mM phosphate buffer pH 6.9 and 50. mu.l of a sample dissolved in 10% methanol solution to a 96-well plate, incubating at room temperature for 10min, thereafter, adding 50. mu.l p-NPG dissolved in 5mM phosphate buffer, reacting at each well, maintaining at room temperature for 10min, and then measuring OD at 405. mu.nm with a microplate reader.
Inhibition (%) - (control-a blank) -sample-a blank) ]/(control-a blank) × 100%
Wherein "a control" represents the absorbance of a 10% methanol solution and the enzyme system; "blank" represents the absorbance of 10% methanol; "sample" represents the absorbance of the extract, phosphate buffer and enzyme system; "blank sample" represents the extract and phosphate buffer, the inhibitory activity being at half the Inhibitory Concentration (IC)50) The values are represented.
(5) Analytical testing
① extract yield (dry extract/dry Phellinus linteus fruiting body raw material) x 100%
② Total phenol content is determined by Folin-phenol method, taking appropriate amount of extract, dissolving with 80% methanol, adding 200 μ l of 0.1N Folin phenol reagent and 2000 μ l of ultrapure water into the extract, shaking, standing for 3min, adding 1000 μ l of sodium carbonate (20%, w/v) solution, shaking, standing at room temperature in dark for 1.0h, determining absorbance at 765nm, setting distilled water as blank control, expressing result as equivalent amount of gallic acid contained in per gram dry weight (mg GAE/g dry weight).
③ LC-MS analysis was performed with an LC-MS system (Waters UPLC Synapt G2-Si HDMS, Milford, MA, USA), Waters C18 column (5 μm, 4.6X 150mm) analysis, mobile phase 0.1% formic acid in pure water (A) and acetonitrile (B), gradient elution procedure 0-20min, 5-20% B, 20-60min, 20-50% B, 60-95min, 50-100% B, 5min equilibration with initial mobile phase between each injection, 1.0mL min flow rate-1The amount of the sample was 10.0. mu.L. ESI-MS anion mode. The capillary voltage is 2500v, the taper hole voltage is 40v, the sample extraction voltage is 5.0v, the ion source temperature is 120 ℃, and the temperature of the solvent gas is 350 ℃. The spraying gas is high-purity nitrogen (N)2) The collision gas is high purity argon (Ar). The reverse gas flow rate was 80L/h, and the solvent-removing gas flow rate was 800L/h. The mass spectrum scanning range is 100-1200amu, and the scanning time is 0.3 s. Compounds were identified by comparison with literature data and quantified as relative peak areas.
The yield and in vitro inhibition α -glycosidase activity of each collected phellinus igniarius component are determined, and the results are as follows:
TABLE 1 Phellinus linteus fruiting body component yield and α -glycosidase inhibitory activity
As can be seen from Table 1, α -glucosidase inhibitory activity was the highest among the various fractions of Phellinus linteus fruiting body, and the ethyl acetate fraction (EtOAc), IC thereof was the highest50The value is 45.1-53.2 mu g/mL, is far higher than other components, and is especially higher than the activity of positive control acarbose by more than one order of magnitude. The total phenol content is 75.2-82.3% by determination, and the total phenol content mainly comprises the compounds of osmocene (1), hispidin (2), davallialactone (3), 2,5-bis (4,7-dihydroxy-8-methyl-2-oxo-2H-chromen-3-yl) cyclohexa-2,5-diene-1,4-dione (4), chlorylene B (5) and inosavin A (6) (see figure 1).
Table 2: phellinus igniarius sporophore ethyl acetate component (EtOAc) phenol composition
Example 1:
referring to fig. 2, a phellinus igniarius cultivated sporocarp → 45 ℃ vacuum drying → crushing and sieving to 50 meshes → weighing 50g, adding 1:10 (solid-to-liquid ratio, W/W), 60% (W/W) ethanol aqueous solution → ultrasonic auxiliary extraction, the extraction power is 80W, the extraction time is 20in, the extraction time is 3 times → 5000r/min centrifugal deslagging → combining supernatant liquid → rotary evaporation, 50 ℃ vacuum degree is 0.1Mpa, concentrating to the original volume 1/5 → freeze drying → phellinus igniarius Ethanol Extract (EE) → adding 200mL of purified water, ultrasonic suspension mixing, adding petroleum ether of the same volume for 2 times, collecting petroleum ether extract (PET) → residual phase, adding ethyl acetate of the same volume for 2 times, combining and collecting ethyl acetate extract phase (EtOAc) → residual phase adding n-equal volume for 2 times of extraction, combining and collecting n-BuOH extract phase → EtOAc → vacuum rotary evaporation concentrating (same conditions before) until no organic solvent is removed, and carrying out in vitro flavor inhibition α → MS analysis → LC analysis.
Inhibition of the EtOAc fraction by analysis α -glycosidase Activity IC50The yield was 2.5% and the purity of the phenols was 72.3%, 52.5 μ g/mL.
Example 2:
phellinus igniarius cultivated sporocarp → 45 ℃ vacuum drying → crushing and sieving to 50 meshes → weighing 50g, adding 1:20 (solid-to-liquid ratio, W/W), 70% (W/W) ethanol aqueous solution → ultrasonic assisted extraction, the extraction power is 90W, the time is 30min, the extraction frequency is 3 times → 5000r/min centrifugal deslagging → merging supernatant liquid → rotary evaporation, 50 ℃, the vacuum degree is 0.1Mpa, concentrating to the original volume 1/5 → freeze drying → Phellinus igniarius Ethanol Extract (EE) → adding 300mL purified water for ultrasonic suspension, adding petroleum ether with the same volume for 2 times, collecting petroleum ether extract (PET) → residual phase, adding ethyl acetate with the same volume for 2 times, collecting ethyl acetate extract phase (EtOAc) → residual phase, adding n-BuOH), collecting the above phases → vacuum rotary evaporation concentrating (before) until no organic solvent is smelled → performing in vitro inhibition of enzyme activity of α -MS → performing LC analysis.
Inhibition of the EtOAc fraction by analysis α -glycosidase Activity IC50Yield was 3.2% and purity of phenols was 74.6% at 49.3 μ g/mL.
Example 3:
phellinus igniarius cultivated sporophore → 45 ℃ vacuum drying → crushing and sieving to 60 meshes → weighing 50g, adding 1:30 (solid-to-liquid ratio, W/W), 80% (W/W) ethanol water solution → ultrasonic assisted extraction, with the extraction power of 100W, time of 30min, the extraction frequency of 3 times → 5000r/min centrifugal deslagging → combining supernatant liquid → rotary evaporation, 50 ℃, the vacuum degree of 0.05Mpa, concentrating to the original volume 1/5 → freeze drying → Phellinus igniarius Ethanol Extract (EE) → adding 300mL of purified water for ultrasonic suspension, adding petroleum ether of the same volume for 3 times, collecting petroleum ether extract (PET) → residual phase, adding ethyl acetate of the same volume for 3 times, collecting ethyl acetate extract phase (EtOAc) → residual phase, adding n-BuOH), collecting the above phases → vacuum rotary evaporation concentrating (before) to no organic solvent flavor → performing in vitro inhibition of α -MS → analyzing the glycosidase activity → LC analysis.
Inhibition of the EtOAc fraction by analysis α -glycosidase Activity IC50When the yield was 45.3 μ g/mL, the phenol purity was 79.6% and the yield was 3.2%.
Claims (6)
1. Phellinus igniarius phenolic extract with hypoglycemic activity, wherein the purity of phenols is more than or equal to 70%, and is obtained by the following method:
(1) vacuum drying Phellinus igniarius solid cultured fruiting bodies at 40-60 ℃, crushing and sieving to 40-80 meshes, adding 60-80% ethanol aqueous solution according to a solid-to-liquid ratio of 1: 10-1: 30, carrying out ultrasonic-assisted extraction for 1-3 times, each time for 20-40 min, centrifuging, and taking supernate;
(2) performing rotary evaporation on the supernatant obtained in the step (1), concentrating, freeze-drying to obtain a phellinus igniarius ethanol extract, adding purified water, performing ultrasonic suspension mixing, adding petroleum ether with the same volume, extracting at 30-60 ℃ for 1-3 times, collecting the petroleum ether extract, adding ethyl acetate with the same volume into the residual phase, extracting for 1-3 times, and combining and collecting ethyl acetate extract phases;
(3) and (3) collecting the ethyl acetate extract phase in the step (2), performing vacuum rotary evaporation and concentration until no organic solvent smell exists, and performing freeze drying to obtain the phellinus igniarius phenolic extract with the hypoglycemic activity.
2. A method of preparing a phellinus linteus phenolic extract of claim 1, the method comprising:
(1) vacuum drying Phellinus igniarius solid cultured fruiting bodies at 40-60 ℃, crushing and sieving to 40-80 meshes, adding 60-80% ethanol aqueous solution according to a solid-to-liquid ratio of 1: 10-1: 30, carrying out ultrasonic-assisted extraction for 1-3 times, each time for 20-40 min, centrifuging, and taking supernate;
(2) performing rotary evaporation on the supernatant obtained in the step (1), concentrating, freeze-drying to obtain a phellinus igniarius ethanol extract, adding purified water, performing ultrasonic suspension mixing, adding petroleum ether with the same volume, extracting at 30-60 ℃ for 1-3 times, collecting the petroleum ether extract, adding ethyl acetate with the same volume into the residual phase, extracting for 1-3 times, and combining and collecting ethyl acetate extract phases;
(3) collecting the ethyl acetate extract phase in the step (2), performing rotary evaporation and concentration until no organic solvent smell exists, and performing freeze drying to obtain the phellinus igniarius phenolic extract with hypoglycemic activity.
3. The method of claim 2, wherein: the rotary evaporation in the steps (2) and (3) is carried out at 40-60 ℃ and under the vacuum degree of 0.05-0.1 Mpa.
4. The method of claim 2, wherein: in the step (1), the ultrasonic extraction power is 80-100W.
5. The use of Phellinus linteus phenolic extract as claimed in claim 1 in preparing health products for lowering blood sugar.
6. The use of Phellinus linteus phenolic extract as claimed in claim 1 in the preparation of medicine for preventing and treating diabetes.
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