CN101336978B - A kind of extraction method of Hovenia dulcis total flavonoids - Google Patents

Abstract

The invention discloses a method for extracting total flavonoids of hovenia dulcis. The method adopts the solvent extraction method and one optional method of the solvent extraction method, the macro-porous resin adsorption method, the supercritical fluid extraction method, the column chromatography or liquid-liquid countercurrent distribution chromatography, or the free combination of the methods. The total flavonoids of hovenia dulcis is extracted from any one part of hovenia acerba, such as fruit stems, fruits, seeds, seed-cases, leaves, flowers, roots, stems, branches, skins and fruit residues, etc., or the combination thereof and is further separated and purified. The prepared total flavonoids is a combination containing two or more flavonoid active components, wherein the total flavonoids comprises the main active components as follows: quercetin, kaempferol, myricetin, (+)-dihydromyricetin derivates, etc.; the total percentage composition of various flavonoid components by weight is 12% to 100%; and the preparation method is applicable to pharmaceutical industry, food industry, cosmetic industry and health-care product industry.

Description

A kind of extraction method of Hovenia dulcis total flavonoids

technical field

The invention relates to an extraction method and a quality control method of total flavonoids of Hovenia dulcis and its application in the fields of medicine, food, cosmetics and health care products, belonging to the field of natural products.

Background technique

Hovenia dulcis Thunb (Hovenia dulcis Thunb) is a plant of the genus Hovenia dulcis Thunb, which is extremely rich in resources in my country. Its fruit or seeds with fleshy stalks are used as medicine, known as "Hovenia dulcis Thunb". Annoyance, anti-vomiting, diuresis. Indications of drunkenness, polydipsia, vomiting, dyspepsia, rheumatic numbness, spasm of hands and feet. Hovenia dulcis leaves, flowers, roots, stems, branches, bark, and sap can sober up the nerves, promote urine excretion, accelerate intestinal peristalsis, dispel wind, dredge collaterals, relieve spasm, quench thirst and annoyance, supplement nutrition, lower blood pressure, and inhibit tumors , sedative, anticonvulsant, liver protection, nourishing liver, hypoglycemic and other effects, folk are also used as medicine.

Flavone is the active ingredient of Hovenia dulcis, and its main structure is flavonol (glycoside), flavanol and dihydroflavonol and other types of compounds, which have the functions of protecting the liver, detoxifying alcohol, antibacterial, and antioxidant.

At present, the solvent extraction method of the total flavonoids of Hovenia dulcis Fructus has been reported in the literature, but it only simply extracts the total flavonoids, and does not combine with other methods to separate and purify the crude extract of total flavonoids. Therefore, the total flavonoids content in the extract are all below 11%, and the purity is very low; there are also reports on the preparation method of monomeric flavonoids in Hovenia dulcis fruit, but it only extracts an active ingredient of flavonoids, and does not involve the preparation method of total flavonoids; and other active parts of Hovenia dulcis There are no literature reports on the preparation methods of flavonoids such as leaves, flowers, roots, stems, branches, skins and pomace, which severely limit the utilization of total flavonoids of Hovenia dulcis and the exertion of medicinal effects.

Contents of the invention

The object of the present invention is to provide a method for extracting total flavonoids from Hovenia dulcis, fruit stalk, fruit, seed, seed shell, leaf, flower, root, stem, branch, skin and pomace.

Another object of the present invention is to provide an application of total flavonoids of Hovenia dulcis as a pharmaceutical preparation, pharmaceutical adjuvant and functional factor in the fields of medicine, food, cosmetics and health care products.

Another object of the present invention is to provide a kind of total flavonoids of Hovenia dulcis, which can be used as a preparation alone or in combination with certain liver protection, hangover, anti-oxidation and antibacterial drugs to make dosage forms, which can be used in the fields of medicine, food, cosmetics and health products .

The total flavonoids of Hovenia dulcis proposed by the present invention are two or two kinds of flavonoids extracted from Hovenia dulcis seeds, fruit stalks, fruits, seeds, seed shells, leaves, flowers, roots, stems, branches, skins or pomace. Combination of the above flavonoid active ingredients, the structures of these compounds are as follows:

The material of the present invention is derived from Hovenia dulcis Thunb, a plant of the genus Hovenia dulcis in the family Rhamnaceae. As the raw material for extracting total flavonoids of Hovenia dulcis, it can be the fruit of Hovenia dulcis on the market, or the fruit stalk, fruit, seed, seed shell, leaf, flower, root, stem, branch, skin and pomace of Hovenia dulcis, etc. Any part or the whole plant, wherein the preferred parts of the medicinal material are the fruit stalks, seeds, leaves and pomace of Hovenia dulcis. The above-mentioned Hovenia dulcis includes raw medicinal materials and decoction pieces without any processing, and also includes various processed products.

The total flavonoids of Hovenia dulcis in the present invention refers to a composition extracted from any part of the above-mentioned plants and containing the following two or more flavonoid active ingredients. These flavonoids include quercetin, quercetin-3-O-α-L-rhamnoside, 4',5,7-trihydroxy-3',5'-dimethoxyflavone, myricetin, Apigenin, taxifolin, 3,4',5,5',7-pentahydroxy-3'-methoxyflavone, Hovenia dulciferol, (+)-3,3',5',5, 7-pentahydroxydihydroflavone, kaempferol, kaempferol-3-O-α-L-rhamnoside, kaempferol-3,7-O-α-L-dirhamnoside, kaempferol -3-O-a-L-rhamnose-(1→6)-O-β-D-glucose (1→2)-O-β-D-glucose, kaempferol-3-o-α-L-rhamnose Sugar-7-O-[β-D-glucose-(1→3)-α-L-rhamnose], dihydrokaempferol, (+)-dihydromyricetin, (+)-pyrocatechu Hovenin, Hovenin I, Hovenin II, Hovenin III, (-)-Catechin, Orangenin and Afotoin etc.

Among the various flavonoid components of Hovenia dulcis according to the present invention, the most important active ingredients are quercetin, kaempferol, myricetin, (+)-dihydromyricetin, hovenia dulcis alcohol, (-) -Catechin, Hovenin I, Hovenin II, Hovenin III, Cucitin, etc. and their derivatives, etc.

As the total flavonoids of Hovenia dulcis, the sum of the percentages of each flavonoid component is 12-100% (W/W) by weight, and preferably 50-100% (W/W).

The prepared total flavonoids of Hovenia dulcis according to the invention can be used in food industry, cosmetic industry and health product industry in addition to the pharmaceutical industry.

The prepared total flavonoids of Hovenia dulcis according to the present invention can be made into preparations alone or combined with other drugs such as detoxification, liver protection, anti-oxidation, antibacterial, etc., for the treatment of alcoholism, liver disease and the treatment of alcoholism caused by free radicals. These preparations have the same or similar pharmacological activities and uses as the total flavonoids of Hovenia dulcis.

The present invention also proposes a method for extracting total flavonoids of Hovenia dulcis, any part of Hovenia dulcis fruit stalk, fruit, seed, seed shell, leaf, flower, root, stem, branch, skin and pomace pomace or a combination thereof, It adopts solvent extraction method and any one of the following methods, or any combination of these methods to extract total flavonoids of Hovenia dulcis: ① solvent extraction method; ② macroporous adsorption resin method; ③ supercritical fluid extraction method; ④ column chromatography; ⑤ liquid-liquid countercurrent distribution chromatography; wherein the preferred macroporous adsorption resin method.

When using these methods to extract total flavonoids from Hovenia dulcis, generally one or more of the following steps are included:

(1) Extraction: the solvent used can be water or any alcohol, ketone or ester solvent, or a mixed solvent composed of these solvents in a certain proportion, or an acidic or Alkaline solvent. The extraction method can be decoction, heating under reflux, Soxhlet extraction, ultrasonic extraction, cold soaking, percolation, microwave extraction, high pressure extraction, etc.

The preferred Soxhlet extraction process is: use 0-95% ethanol solution (V/V), with a solid-liquid ratio of 1:10-50 (g/mL), and continuously heat and reflux at 60-100°C for 5-12 hours.

The preferred ultrasonic extraction process is as follows: the Hovenia dulcis extract material is packed into an ultrasonic extractor, and the ratio of material to liquid is 1: 6 to 15 (g/mL) to add 0 to 95% ethanol solution (V/V). Ultrasonic extraction at a temperature of 30-50°C, a frequency of 40 kHz, and a power of 0.3-0.5 W/cm ² for 15-30 minutes, 2-3 times of extraction, and the combined extracts.

The preferred microwave extraction process is: add 0-95% ethanol solution (V/V) to soak for 20-60 minutes with a solid-liquid ratio of 1:20-40 (g/mL), control the microwave power to 900W, and irradiate for 30-50 Seconds, take it out and cool it down to room temperature quickly, then irradiate for 20-40 seconds, repeat the same operation 3-5 times, filter, and combine the extracts.

(2) Filtration: including centrifugation, suction filtration, ultrafiltration, pressure filtration, etc., with or without the use of any of the following clarifiers or their combination: alcohol precipitation agent, gelatin, kaolin, various resins, polyethylene glycol, Polyethylene glycol, chitosan and natural clarifier products such as 101 juice clarifier, ZTC+1 natural clarifier, etc.

The preferred clarifying agent is diatomaceous earth. According to the weight ratio of diatomite to extract is 1:2~5, diatomite is added to the extract, stirred at 10~50°C for 20~60min, and then filtered to obtain the filtrate, which can be reused Methanol, ethanol, chloroform, ether, or a combination of two or more of them washes the clarifier to obtain an eluate, and the filtrate and eluate are combined.

(3) Concentration: including thin film evaporation, rotary evaporation and decoction concentration under normal pressure or reduced pressure.

(4) Drying: including vacuum drying, spray drying, freeze drying, etc.

When the extraction method is used for preparation, the extract is generally suspended in water first, and then extracted with alkanes (such as petroleum ether, hexane, gasoline, etc.) to remove fat-soluble impurities, and then with a suitable solvent, such as ether, chloroform , ethyl acetate, acetone, n-butanol, etc., or a mixture of these solvents, and extract the total flavonoid components therein to obtain the total flavonoid extract.

When the macroporous adsorption resin method is used for preparation, the macroporous resin used can be any type of non-polar, weak polar, medium polar, weakly alkaline or weakly acidic, such as D101, D4020, D3500, D941, D21, HP-20, HP-20, XDA-1, AB-8, HPD400, S-8, HZ-806, H-50, H-30, LX-38, LX-28, LS-300B, LS- 306, LSD-958, PA, etc. Among them, weak or medium polar resins are preferred, such as LX-38, D101, AB-8, etc. The eluents used are water and ethanol containing water, methanol, acetone, etc., among which 50-95% ethanol solution (V/V) is preferred.

The preferred resin purification process of Hovenia dulcis total flavonoids extract resin is: select LX-38, D101, AB-8 resin as the resin for purification, and the crude total flavonoids of Hovenia dulcis dulcis is based on the crude drug amount (g): the upper part of dispersed milliliters The dilution ratio of the sample solution is 1:4~1:18, the adsorption flow rate is 2~9BV/h, the diameter-to-height ratio of the resin column is 1:3~1:10, the sample loading is 100~500mg/mL (based on crude drug amount), 0 ~20% ethanol solution (V/V) elutes 2~5 times of resin volume for impurity removal, the flow rate of impurity removal is 2~7BV/h, elutes 3~10 times with 50~95% ethanol solution (V/V) Resin volume, elution flow rate is 2~9BV/h.

When supercritical fluid extraction is used for preparation, the raw material of Hovenia dulcis can be directly extracted, and the product obtained by any one of the above methods and steps can also be extracted. Extraction can be performed with or without any of the following solvents and solvent mixtures: water, alcohols, ketones, esters and ether solvents.

When column chromatography is used for preparation, the object of treatment can be the product obtained in the above-mentioned extraction step, or it can be the product obtained by the above-mentioned solvent extraction method, solvent extraction method, macroporous adsorption resin method or supercritical fluid extraction method. product of. The stationary phase used can be silica gel, polyamide, aluminum oxide, dextran (Sephadex series or Sephadex-20 series), C-8, C-18, activated carbon, cellulose, etc., and the eluent used varies depending on the stationary phase. It varies with different solvents, and it is generally a mixed solvent composed of water, methanol, ethanol, acetone, chloroform, ethyl acetate, petroleum ether, etc. Among them, polyamide column chromatography is preferred.

The purification process of polyamide column chromatography is as follows: polyamide is selected, the concentration of the sample solution of the crude extract of Hovenia dulcis is 1:4~1:16 (the amount of crude drug (g): the number of milliliters dispersed), and the adsorption flow rate is 3~8BV/h , the diameter-to-height ratio of the resin column is 1:6～1:20, the loading amount is 100～500mg/mL (based on the amount of crude drug), and the 0～10% ethanol solution (V/V) is eluted with 1～5 times the volume of the resin. Impurity removal, the flow rate of impurity removal is 3-7BV/h, 4-7 times of resin volume is eluted with 50-95% ethanol solution (V/V), the elution flow rate is 2-9BV/h, the eluate is collected, and the The solvent was recovered under pressure, and the residue was dried under reduced pressure to obtain total flavonoids of Hovenia dulcis.

When the liquid-liquid countercurrent extraction method is used for preparation, the object of its treatment can be the product of the above-mentioned extraction step, or it can be the product of the above-mentioned solvent extraction method, solvent extraction method, macroporous adsorption resin method, supercritical fluid extraction method and column. Product after preliminary purification by chromatography. Generally, the extract is suspended in water, and then extracted with a low-polar alkane or ether solvent (such as petroleum ether, hexane, gasoline, etc.) to remove fat-soluble impurities, and then with a suitable polar solvent, such as chloroform, Ethyl acetate, acetone, n-butanol, etc., or a mixture of these solvents, extract the total flavonoids therein to obtain the total flavonoids of Hovenia dulcis.

The total flavonoids of Hovenia dulcis can be used alone or in combination with any other Chinese and Western medicines, foods or auxiliary materials in any proportion to prepare medicines, foods, cosmetics and health products. The obtained medicines, foods, cosmetics and health products can be capsules, Tablets, pills, granules, oral liquids, syrups, granules, liquors, injections, ointments, powders, beverages, etc.

The quality control method of the present invention can comprise one or more in the following assay methods:

1. Total flavonoids

Accurately weigh 250 mg of rutin reference substance dried at 120°C to constant weight, dissolve in anhydrous methanol, transfer to a 250mL volumetric flask, dilute to volume with anhydrous methanol, and shake well. Accurately pipette 5.00mL into a 50mL volumetric flask, dilute to volume with anhydrous methanol, and shake well. The concentration of this rutin reference substance solution is 0.1mg/mL. Accurately draw 0.00mL, 1.00mL, 2.00mL, 3.00mL, 4.00mL, 5.00mL, 6.00mL of 0.1mg/mL rutin reference solution, put them into 25mL volumetric flasks respectively, add 50% ethanol solution (V/V ) to 6mL, add 1.00mL of 5% sodium nitrite solution (W/V) respectively, shake well, stand for 6min, then add 1.00mL of 10% aluminum nitrate solution (W/V), shake well, stand for 6min, then add 10.00mL of 1% sodium hydroxide solution, then dilute to volume with 50% ethanol solution (V/V), shake well, let it stand for 15min, measure the absorbance at a wavelength of 510nm with the blank solution as a reference, and obtain the regression equation.

Accurately draw 3 samples of Hovenia dulcis flavonoids extract, 20 mg each, put in a 50 mL volumetric flask, add 50% ethanol solution (V/V) to ultrasonically dissolve and dilute to the mark, and shake well. Develop color as described above and measure its absorbance A at a wavelength of 510 nm, and calculate the total flavonoid content in the sample by the regression equation.

2. Contents of Hovenia dulcis alcohol, quercetin, coumarin and kaempferol

Chromatographic conditions: chromatographic column: reverse phase C18 (4.6×250mm, 5μm); mobile phase: 30-85% methanol solution (V/V) (adjust pH to 3.0 with phosphoric acid) elution (0-20min); flow rate: 1.0 mL/Min; detection wavelength: 360nm; column temperature: room temperature.

Standard curve drawing: Precisely weigh Hovenia dulciferol, quercetin, coumarin and kaempferol reference substances, and dissolve them in methanol to prepare reference substance solutions with concentrations of 100, 288, and 190 μg/mL respectively, 0, 2 , 5, 10, 15, and 20 μL were injected into the liquid chromatograph, and the peak areas of each chromatographic peak were measured. With the control injection volume (μg) as the abscissa and the chromatographic peak area as the ordinate, a standard curve was drawn.

Content Determination: Accurately weigh 3 batches of total flavonoid extract samples, each about 20mg, put them in a 50mL measuring bottle, add methanol to ultrasonically dissolve, and dilute to the scale, shake well, as Hovenia dulcis, Quercetin For the test sample solution for the determination of the content of acetin, coumarin and kaempferol, 20 μL of the above-mentioned test sample solution is precisely drawn and injected into the liquid chromatograph, and the area of each chromatographic peak is measured, and the content is calculated.

3. Dihydromyricetin

Chromatographic conditions: chromatographic column: reverse phase C18 (4.6×250mm, 5μm); mobile phase: 25-40% methanol solution (V/V) (adjust pH to 3.0 with phosphoric acid) elution (0-50min); flow rate: 1.0 mL/Min; detection wavelength: 294nm; column temperature: room temperature.

Draw the standard curve: draw 0, 2, 5, 10, 15, and 20 μL of (+)-dihydromyricetin reference solution (2.0 μg/μL) precisely and inject them into the liquid chromatograph, measure the peak area of each chromatographic peak, and compare The injection volume (μg) is the abscissa, the chromatographic peak area is the ordinate, and the standard curve is drawn.

Content determination: Accurately weigh 3 parts of 3 batches of total flavonoid extract samples, each about 20 mg, place in a 50 mL measuring bottle, add 20% acetonitrile solution (V/V) for ultrasonic dissolution, and dilute to the mark, shake well, As the test sample solution for the determination of (+)-dihydromyricetin content, 20 μL of the above-mentioned test sample solution was accurately drawn and injected into a liquid chromatograph, and the peak areas of each chromatographic peak were measured to calculate the content.

4. Myricetin

Chromatographic conditions: chromatographic column: reverse phase C18 (4.6×250mm, 5μm); mobile phase: 40-60% methanol solution (V/V) (adjust pH to 3.0 with phosphoric acid) elution (0-50min); flow rate: 1.0 mL/Min; detection wavelength: 370nm; column temperature: room temperature.

Standard curve drawing: Accurately weigh 9.80mg of myricetin reference substance, put it in a 100mL volumetric flask, dissolve it with methanol and set the volume to the mark, shake well, and use it as the reference substance solution. Dilute with methanol and set the volume to the mark, shake well, make myricetin solutions with serial concentrations of 9.8, 19.6, 29.4, 39.2, and 49.0 μg/mL and inject them into the liquid chromatograph, measure the peak area of each chromatographic peak, and compare the injection volume (μg) is the abscissa, the chromatographic peak area is the ordinate, and the standard curve is drawn.

Content determination: Accurately weigh 3 samples of 3 batches of total flavonoid extracts, about 20mg each, put them in a 50mL measuring bottle, add methanol for ultrasonic dissolution, dilute to the scale, shake well, and use it as a test sample for the determination of myricetin content. For the sample solution, 20 μL of the above-mentioned test sample solution was accurately drawn and injected into the liquid chromatograph, and the peak area of each chromatographic peak was measured, and the content was calculated.

4. Catechin, hovenin I, hovenin II, hovenin III

Chromatographic conditions: Chromatographic column: reversed-phase C18 (4.6×250mm, 5μm); mobile phase: 15-60% (0-40min) methanol solution (V/V) elution; flow rate: 1.0mL/Min; detection wavelength: 280nm . Standard curve drawing: 0, 2, 5, 10, 15, 20 μL of reference substance solutions (0.027 μg/μL) of catechin, Hovenia I, Hoveniae II, and Hoveniae III were drawn precisely and injected into the liquid chromatograph , Measure the peak area of each chromatographic peak, take the control injection volume (μg) as the abscissa, and the chromatographic peak area as the ordinate, draw a standard curve.

Content determination: Accurately weigh 3 parts of each of the 3 batches of total flavonoid extract samples, each about 20 mg, place them in a 50 mL measuring bottle, dissolve them ultrasonically with 50% aqueous methanol (V/V), dilute to the mark, shake well, As the test sample solution for the determination of catechin, hovenin I, hovenin II, and hovenin III, accurately draw 20 μL of the above-mentioned test sample solution and inject it into a liquid chromatograph, measure the peak area of each chromatographic peak, and calculate the content .

Detailed ways

Example 1: Extraction of Hovenia dulcis total flavonoids

Take Hovenia dulcis leaf 1Kg and pulverize to 50 meshes, add 75% ethanol solution (V/V) according to the solid-liquid ratio of 1:50 (g/mL), continuously heat and reflux for extraction for 9 hours, recover the solvent to remove ethanol, and adjust the concentration of the extract. pH=6, the processed macroporous resin LSD-958 on the filtrate was washed with 4 times of resin bed volume aqueous solution and 3 times of resin bed volume 20% ethanol solution (V/V) to remove impurities, and the washing solution was discarded and used The 95% ethanol solution (V/V) of 7 times the volume of the resin bed was eluted, the eluate was collected, concentrated to a thick paste and dried to obtain the total flavonoids of Hovenia dulcis. The total flavonoid content of Hovenia dulcis was determined to be 66%, of which quercetin, kaempferol, myricetin, (+)-dihydromyricetin, hovenia dulcis alcohol, (-)-catechin, hovenia dulcisin I, The sum of the contents of Hovenin II, Hovenin III, Coutinin and their derivatives is 51%.

Example 2: Extraction of Hovenia dulcis total flavonoids

Take 1Kg of mature and dried Hovenia dulcis and crush it to 60 mesh, add distilled water and soak for 40 minutes at a solid-liquid ratio of 1:40 (g/mL), control the microwave power to 900W, irradiate for 50 seconds, take it out, cool it to room temperature quickly, and irradiate again For 20 seconds, repeat the same operation 3 times, filter, and combine the extracts. The extract was concentrated to 20% (V/V) under reduced pressure, suspended in water, then extracted with n-hexane and ethyl acetate in sequence, the solvent was recovered and the organic solvent extract was retained. Take the ethyl acetate extract and dissolve it with ethanol, add diatomaceous earth according to 30% by weight, stir at room temperature for 30 minutes, filter and wash with ethanol, combine the filtrates, concentrate and dry in vacuo to obtain total flavonoids of Hovenia dulcis. The content of the total flavonoids was determined to be 87%, of which quercetin, kaempferol, myricetin, (+)-dihydromyricetin, hovendarinol, (-)-catechin, hovenin I, Hovenia dulcis The sum of the content of element II, citronine III, and orangenin and their derivatives is 63%.

Example 3: Extraction of Hovenia dulcis total flavonoids

Get 1Kg of dried pomace after removing the juice from Hovenia dulcis fruit stalk, pulverize, pass through a 100-mesh sieve, filter out impurities such as cellulose, and dry the powder, 8 times the amount (g/mL) of 70% ethanol solution (V/V) 80 Heat reflux extraction at ℃ for 3 times, 2 hours each time, combine the filtrates, concentrate in vacuo to remove ethanol, dissolve the extract in 3 times distilled water by weight, put it on a polyamide column, the adsorption flow rate is 5BV/h, and the polyamide column diameter to height ratio is 1 : 8, the loading amount is 400mg/mL (based on the amount of crude drug), first eluted with water and the outflow is nearly colorless, then rinsed with 4 times the resin bed volume 30% ethanol solution (V/V) to remove impurities, and the rinse solution was discarded , eluted with 95% ethanol solution (V/V) of 5 times the polyamide bed volume, the elution flow rate was 4BV/h, collected the eluate, concentrated under reduced pressure to a thick paste and dried to obtain total flavonoids of Hovenia dulcis. Determination of the content of total flavonoids is 75%, wherein quercetin, kaempferol, myricetin, (+)-dihydromyricetin, Hovenia dulcis alcohol, (-)-catechin, Hovenia dulcis I, Hovenia citrus The sum of the content of element II, citronine III, and orangenin and their derivatives is 52%.

Example 4: Extraction of Hovenia dulcis total flavonoids

Get 1Kg of ripe and dry Hovenia dulcis seeds and pulverize them, pass through a 100-mesh sieve, add 7 times the amount (g/mL) of 70% ethanol solution (V/V), reflux extraction 3 times, each time for 2 hours, the extract is concentrated to remove ethanol, and It is suspended in water, extracted with petroleum ether to remove impurities such as pigments, and the aqueous phase extract is put on a LX-38 macroporous resin column, and the sample is slowly loaded, and the 4 column volumes are first eluted with water, and then 10% ethanol solution (V/ V) Eluting 5 times of column volume, finally eluting with 95% ethanol solution (V/V), when 10% AlCl _solution (W/V) detects negative, stop collecting, combine 95% ethanol solution (V/V) /v) eluting the solvent, concentrating and drying in vacuo to obtain the total flavonoids of Hovenia dulcis. The content of the total flavonoids was determined to be 81%, among which quercetin, kaempferol, myricetin, (+)-dihydromyricetin, hovendarinol, (-)-catechin, hovenin I, Hovenia dulcis The sum of the content of element II, citrus aurantin III and orangenin and their derivatives is 67%.

Example 5: Preparation of Hovenia dulcis Total Flavonoids Tablets

Hovenia dulcis total flavonoids 10g

Starch 10g

The above components are mixed evenly, and an appropriate amount of talcum powder is added, and pressed into 100 tablets.

Example 6: Preparation of Hovenia dulcis total flavonoids compound preparation

Hovenia dulcis total flavonoids 20g

Pueraria flavonoids 15g

The above components are mixed evenly, and packed into hard gelatin capsules, 100 capsules in total.

Example 7: Preparation of Hovenia dulcis total flavonoids compound preparation

Hovenia dulcis total flavonoids 10g

Silverwort Extract 10g

Snakeberry Extract 10g

The above components were mixed uniformly to make 100 granules.

Claims (1)

1. an extraction method of Hovenia dulcis total flavonoids, is characterized in that, comprises the following steps: get Hovenia dulcis leaf 1kg and pulverize to 50 orders, add 75% ethanol solution according to the solid-liquid ratio of 1: 50g/mL (V/ V), continuous heating and reflux extraction for 9h, reclaiming the solvent to remove ethanol, adjusting the pH of the extract=6, the treated macroporous resin LSD-958 on the filtrate, using 4 times the volume of the resin bed solution and 3 times the volume of the resin bed for 20 Rinse with % ethanol solution (V/V) to remove impurities, discard the rinse solution, elute with 95% ethanol solution (V/V) of 7 times the volume of the resin bed, collect the eluate, concentrate to a thick paste and dry to obtain Hovenia dulcis total flavonoids.