CN109467581B

CN109467581B - Method for extracting flavonoid glycoside from lotus leaves and pharmaceutical preparation containing flavonoid glycoside from lotus leaves

Info

Publication number: CN109467581B
Application number: CN201811558120.8A
Authority: CN
Inventors: 吴祖栋; 杨丽; 齐炼文; 邢宸; 邢为藩
Original assignee: Nanjing Chenxiang Medical Research Co ltd
Current assignee: Nanjing Chenxiang Medical Research Co ltd
Priority date: 2018-12-19
Filing date: 2018-12-19
Publication date: 2020-09-25
Anticipated expiration: 2038-12-19
Also published as: CN109467581A

Abstract

The invention provides a method for extracting and purifying flavonoid glycoside from lotus leaves and a preparation containing the flavonoid glycoside from lotus leaves, belonging to the technical field of extraction of traditional Chinese medicines, and the method comprises the following steps: adding water with the temperature of 60-80 ℃ into lotus leaf powder, stirring and extracting, separating an extracting solution and filter residues, extracting for 3 times, throwing the filter liquor, cooling to room temperature, adjusting the pH to 2-3, standing for 2 hours, and performing cross-flow filtration to obtain a fine filter liquor; adjusting the pH of the fine filtration liquid to 5-6.5, adsorbing, washing and eluting by using macroporous resin, collecting eluent with the purity of the lotus leaf flavonoid glycoside being more than or equal to 99%, and completing the extraction and purification of the lotus leaf flavonoid glycoside; the lotus leaf flavonoid glycoside obtained by purification in the invention is quercetin-3-O-beta-D glucuronide, wherein the purity of the quercetin-3-O-beta-D glucuronide is more than 99.22%, the yield is high, and the cost is low. The extraction method is simple, convenient and quick, and the extraction process can realize automation, intellectualization, greening and digitization.

Description

Method for extracting flavonoid glycoside from lotus leaves and pharmaceutical preparation containing flavonoid glycoside from lotus leaves

Technical Field

The invention belongs to the technical field of traditional Chinese medicine extraction and application, and particularly relates to a method for extracting flavonoid glycoside from lotus leaves and a pharmaceutical preparation containing the flavonoid glycoside from lotus leaves.

Background

Nymphaeaceae plant lotus (Nelumbonaccidae Gaertn) is planted in many places in China, is widely planted in Jiangsu, Zhejiang, Anhui and other provinces, can be appreciated and eaten, is also a medicinal plant, and has application in the industries of medicines, foods, health care products and the like. Folium Nelumbinis contains chemical components such as flavone, alkaloid, polyphenol, volatile oil, steroid, terpene, fatty acid, fatty alcohol, protein, tannin, etc., and flavone and alkaloid compounds are main pharmacologically active components. The flavonoids include about 30 kinds of monoglucuronide (quercetin-3-O-beta-D-glucuronide, Q3GA), diglucoside (quercetin-3-O-beta-D-diglucoside), hyperoside (quercetin-3-O-beta-D-galactopyranoside), isoquercitrin (isoquercitrin-3-O-beta-D-glucoside) and astragalin (kaempferol-3-O-glucoside, kaempferol-3-glucoside), etc., and the rest is polyphenol, volatile oil, etc. At present, the supply of lotus leaves is larger than the demand, the use amount is less, the application of the lotus leaves in the industries of medicine, food, health care products and the like is in a relatively initial stage, and the development breadth and the development depth can not meet the requirements of the times. How to develop new application of lotus leaves, especially for preventing and treating frequently encountered diseases and certain serious diseases, is the mission of pharmaceutical workers.

The folium Nelumbinis is dry leaf of Nymphaeaceae plant flos Nelumbinis, and has mild nature, bitter taste, and no toxicity (mouse LD)₅₀More than 5g/kg), has effects of clearing summer-heat, dispersing pathogenic factors, ascending the clear and descending the turbid, activating spleen and promoting appetite, stopping bleeding and removing blood stasis. The lotus flavonoid glycoside is used for reducing fat and losing weight, resisting oxidation, eliminating free radicals, resisting aging, protecting liver, inhibiting hepatic fibrosis, resisting hepatotoxicity, resisting bacteria and diminishing inflammation and the like, and enters heart, liver, gallbladder, spleen and stomach channels. The traditional medical books are recorded, and the secret handed down syndrome treatment key in Ming dynasty with Yuan Li records the weight-reducing effect of lotus leaves: it is thin and inferior when taken with lotus leaves. The book Ben Cao Shi Yi (Chinese materia Medica) is also cloud: "thin after long-term eating". Along with the improvement of the living standard of people, the daily intake of fat and sugar is increased day by day, patients with coronary heart disease, AD (Alzheimer's disease), hyperlipidemia and obesity are also increased day by day, higher requirements are also put forward for pharmaceutical research workers, and the research and development of lipid-lowering and weight-reducing medicines are urgent day by day.

The study on lipid lowering and weight losing of a single lotus leaf water extract for intragastric administration of a rat and a mouse model with hyperlipidemia is carried out, the influence on serum cholesterol and triglyceride is observed, and the result shows that: the lotus leaf water can reduce TC of hyperlipidemic rats by 25.6-39.3 percent and TG by 18.9-39.2 percent, has no obvious influence on HDL-C, but can reduce low-density lipoprotein cholesterol (LDL-C) obviously along with the reduction of TC and TG, and meanwhile, the lotus leaf water decoction can reduce the specific viscosity of whole blood and the hematocrit (%), thereby improving the thick and viscous state of blood. The lotus leaf flavone has obvious inhibition effect on high cholesterol; the experiment for synthesizing TC by in vitro cultured liver cells shows that: the lotus leaf flavone can reduce TC in liver cells and obviously increase TC in a culture solution. Its action is mainly manifested in promoting lipid metabolism, and its synthesis inhibition is weak. The main active site is concentrated on flavone, the animal activity is normal, and the phenomena of obvious diarrhea and appetite suppression do not occur.

In recent years, many pharmaceutical workers pay attention to the extraction and purification of effective components in lotus leaves, and the research on the quality, dosage form, new indications, action mechanism and the like of the effective components. However, the existing lotus leaf flavonoid glycoside extraction method or purification process is complicated and tedious, takes time and labor, has high production cost, low product purity or low yield, is not satisfactory, and has no large-scale production.

Disclosure of Invention

In view of the above, the present invention aims to provide an extraction method of flavonoid glycoside of lotus leaf with simple extraction method, low cost, high extraction yield and high product purity, and a pharmaceutical composition comprising flavonoid glycoside of lotus leaf.

Based on the above purpose, the invention provides the following technical scheme:

a method for extracting flavonoid glycoside from lotus leaf comprises the following steps: 1) adding water with the temperature of 60-80 ℃ into the lotus leaf powder, carrying out primary stirring extraction, and carrying out primary filter-spinning to obtain a filtrate 1 and a filter residue 1; 2) adding water with the temperature of 60-80 ℃ into the filter residue 1, stirring and extracting for the second time, and performing filter throwing for the second time to obtain a filtrate 2 and a filter residue 2; 3) adding water with the temperature of 60-80 ℃ into the filter residue 2, stirring and extracting for the third time, and performing filter spinning for the third time to obtain a filtrate 3 and a filter residue 3; 4) mixing the filtrate 1, the filtrate 2 and the filtrate 3, cooling to 25-30 ℃, adjusting the pH to 2-3, standing for 2 hours, and performing cross-flow filtration to obtain a fine filtrate; 5) adjusting the pH value of the fine filtration liquid to 5-6.5, adsorbing, washing and eluting by using macroporous resin, collecting eluent with the purity of the lotus leaf flavonoid glycoside being more than or equal to 99%, and completing the extraction and purification of the lotus leaf flavonoid glycoside; the lotus leaf flavonoid glycoside is quercetin-3-O-beta-D glucuronide.

Preferably, the mass ratio of the lotus leaf powder to water in the step 1) is 1: (7-10).

Preferably, the mass ratio of the filter residue 1 in the step 2) and the filter residue 2 to water in the step 3) is independently 1: (5-8).

Preferably, the time for the first stirring extraction in the step 1) is 12-18 min.

Preferably, the time for the second stirring extraction in the step 2) or the third stirring extraction in the step 3) is independently 5-10 min.

Preferably, the step 5) of collecting the eluent further comprises adjusting the pH value of the eluent to 1.5-3.5, washing the eluent with purified water and 30% ethanol, and then eluting the eluent with an ethanol solution with the volume concentration of 50-95% to realize concentration.

The invention also provides lotus leaf flavonoid glycoside extracted by the method, wherein the purity of the lotus leaf flavonoid glycoside, namely the quercetin-3-O-beta-D glucuronide, is more than or equal to 99%.

The invention also provides a pharmaceutical preparation containing the lotus leaf flavonoid glycoside.

According to the extraction method of the lotus leaf flavonoid glycoside provided by the invention, lotus leaf powder is stirred, extracted, subjected to spin filtration for three times to obtain a rough filtrate, the rough filtrate is cooled to room temperature, the pH value is adjusted to 2-3, the rough filtrate is placed for 2 hours, cross-flow filtration is carried out to obtain a fine filtrate, the pH value of the fine filtrate is adjusted to 5-6.5, the fine filtrate is adsorbed, washed and eluted by macroporous resin to obtain the lotus leaf flavonoid glycoside, namely quercetin-3-O-beta-D glucuronide, the purity of the lotus leaf flavonoid glycoside is more than or equal to 99 percent; the extraction method is simple, convenient and quick, the extraction process can realize automation, intellectualization, greening and digitization, and compared with other methods for extracting and separating and purifying resin columns or different packed columns for multiple times, the extraction method can obviously save energy, reduce emission, save time and labor, save plants and equipment, improve the yield, reduce the cost, and ensure stable and controllable quality of products, thereby being convenient for industrial production.

Furthermore, a combined method of throw filtration and cross-flow filtration is adopted for filtration, so that the filtration speed is increased, and meanwhile, the precise filtration is realized.

Furthermore, the extraction method realizes an overall process quality on-line control system through parameter setting of each step, solves the problems of large intra-batch uniformity and large inter-batch repeatability difference of the produced products, and fundamentally ensures stable and controllable product quality.

Drawings

FIG. 1 is HPLC fingerprint chart of unseparated and purified folium Nelumbinis powder;

FIG. 2 is an HPLC chart of a flavonoid glycoside of Nelumbo Nucifera (Q3GA) control;

FIG. 3 is an HPLC chart of flavonoid glycoside of Nelumbo nucifera (Q3GA) in example 1;

FIG. 4 is an HPLC chart of a solvent of flavonoid glycoside of Nelumbo Nucifera (Q3 GA);

FIG. 5 is an HPLC chart of flavonoid glycoside of Nelumbo nucifera (Q3GA) in example 4.

Detailed Description

The invention provides a method for extracting and purifying flavonoid glycoside from lotus leaves, which comprises the following steps: 1) adding water with the temperature of 60-80 ℃ into lotus leaf powder, stirring and extracting, and performing first-time filter throwing to obtain coarse filtrate 1 and filter residue 1; 2) adding water with the temperature of 60-80 ℃ into the filter residue 1, stirring and extracting, and performing filter throwing for the second time to obtain a coarse filter solution 2 and a filter residue 2; 3) adding water with the temperature of 60-80 ℃ into the filter residue 2, stirring and extracting, and performing filter throwing for the third time to obtain rough filtrate 3 and filter residue; the filter residue can be used for preparing methane and green manure; 4) mixing the

crude filtrates

1, 2 and 3, cooling to room temperature, adjusting pH to 2-3, standing for 2-3 hr, cross-flow filtering to obtain fine filtrate, and extracting; 5) adjusting the pH value of the fine filtration liquid to 5-6.5, adsorbing, washing and eluting by using macroporous resin, collecting eluent with the purity of the lotus leaf flavonoid glycoside being more than or equal to 99%, and completing the purification of the lotus leaf flavonoid glycoside; the lotus leaf flavonoid glycoside is quercetin-3-O-beta-D glucuronide.

In the invention, lotus leaf powder is mixed with water for the first stirring extraction. In the invention, the mass ratio of the lotus leaf powder to water is preferably 1: (7-10), more preferably 1: (8-9); the time for the first stirring extraction is preferably 12-18 min, more preferably 13-17 min, and most preferably 15 min. The temperature of the water is preferably 60-80 ℃, and specifically can be 65-79 ℃, 68-75 ℃, 70-73 ℃, 69 ℃, 71 ℃ or 78 ℃; the temperature of the material liquid for the first stirring extraction is preferably 55-60 ℃. The source and the granularity of the lotus leaf powder are not specially limited, the lotus leaf powder is usually adopted, the water is not specially limited, and the purified water for conventional extraction in the field is adopted.

After the filter residue 1 is obtained, water with the temperature of 60-80 ℃ is added, and stirring and extraction are carried out for the second time. In the invention, the mass ratio of the filter residue 1 to the water is preferably 1: (5-8), more preferably 1: (6-7). In the invention, the time for the second stirring extraction is preferably 5-10 min, more preferably 6-9 min, and most preferably 8 min. The temperature of the water is preferably 60-80 ℃, and specifically can be 65-79 ℃, 68-75 ℃, 70-73 ℃, 69 ℃, 71 ℃ or 78 ℃; the actual temperature of the second stirring extraction is preferably 55-60 ℃. After the second stirring extraction, the second filtration is carried out to obtain filtrate 2 and filter residue 2.

After the filter residue 2 is obtained, water is added again, and stirring and extraction are carried out for the third time. The main parameters and operations of the slag-water ratio, the extraction time, the water temperature, the actual feed liquid temperature and the like are the same as those of the second extraction. The coarse separation of the slag-liquid in the extraction can be realized by using throwing filtration or filter pressing, preferably throwing filtration, and is simple, convenient and quick.

The stirring speed in the three stirring extraction processes is not particularly limited, and is preferably high according to the possible setting of specific equipment, so that the stirring is favorable for dissolving out the active ingredients. Generally, the laboratory equipment is small, the rotation speed is hundreds of revolutions per minute, and the rotation speed is more between 30 and 60 revolutions per minute for different types of production equipment and stirrers.

The invention combines the filtrate 1-3, then cools the filtrate to room temperature, adjusts the pH value to 2-3, and stands for 2h to obtain the fine filtrate by cross flow filtration and fine filtration, wherein the spin filtration is the pre-filtration of the fine filtration, the fine filtration can not be carried out without spin filtration, and the fine filtration is the target filtration.

After the pH of the fine filtrate is adjusted to 5-6.5, the fine filtrate is subjected to macroporous resin adsorption, gradient washing and elution, and the eluent with the purity of the lotus leaf flavonoid glycoside being more than or equal to 99% is collected, so that the separation and purification of the traditional Chinese medicine extracting solution are completed. The reagent for regulating the fine filtrate is not particularly limited, hydrochloric acid and sodium hydroxide are commonly used, and the concentration is determined according to the requirement. In the invention, the model of the macroporous resin is preferably FL-2, SP-3, D101 or D301; the diameter-height ratio of the macroporous resin column is preferably 1: 8-33; the sampling flow rate of the fine filtration liquid is preferably 1-2 BV/h, and more preferably 1.5 BV/h. After the fine filtrate is subjected to sample loading, gradient washing and elution are carried out. Before elution, the method preferably further comprises a washing step, particularly the raw material for injection is prepared, after the washing step is preferably carried out by gradient washing with ethanol with the volume fraction of 0-12%, then the elution is carried out by using an ethanol water solution with the volume concentration of 15-25%, and the eluent with the purity of the flavonoid glycoside of lotus leaf more than or equal to 99% is collected, so that the extraction, separation and purification of the flavonoid glycoside of lotus leaf are completed.

After the dilute eluent is collected, the pH value of the eluent is preferably adjusted to be 1.5-3.5, preferably 2-3, macroporous resin is used for adsorption and enrichment, purified water and 30% ethanol are used for washing, then the eluent is eluted by an ethanol solution with the volume concentration of 50-95%, the elution speed is consistent with that of the macroporous resin adsorption and elution of the fine filtrate, and concentration is achieved.

After the concentration, the invention preferably further comprises drying the lotus leaf flavonoid glycoside concentrated solution obtained by concentration to obtain lotus leaf flavonoid glycoside solid powder. The drying is preferably reduced pressure drying and/or negative pressure spray drying. In view of the fact that the solubility of the lotus leaf flavonoid glycoside in water is very low and is an insoluble level, the lotus leaf flavonoid glycoside concentrated solution is preferably placed and filtered, the solid phase component is collected to be fresh yellow powdery lotus leaf flavonoid glycoside, the filtrate is subjected to negative pressure spray drying, then fresh yellow powdery lotus leaf flavonoid glycoside powder is obtained, the fresh yellow powdery lotus leaf flavonoid glycoside powder is obtained twice, and the purity of the fresh yellow powdery lotus leaf flavonoid glycoside powder is not obviously different.

The invention also provides a lotus leaf flavonoid glycoside extracted by the method, namely the purity of quercetin-3-O-beta-D glucuronide is more than 99.5%.

The invention also provides a medicinal preparation of the lotus leaf flavonoid glycoside. The content of quercetin-3-O-beta-D glucuronide in the pharmaceutical preparation is preferably 25-45%. The invention has no special requirements on the auxiliary materials and the dosage forms of the medicinal preparation. In the specific implementation process of the invention, the auxiliary materials of the pharmaceutical preparation comprise one or more of a sweetening agent, a filling agent, a slow release agent and a coloring agent. The dosage forms of the pharmaceutical preparation provided by the invention are dripping pills, capsules, micro-pills, tablets, sustained-release micro-pills, sustained-release capsules and sustained-release tablets. The medicinal preparation has the effects of clearing summer-heat evil, dispersing pathogenic factors, ascending the clear and descending the turbid, activating the spleen and stimulating the appetite, stopping bleeding and dissipating blood stasis and the like, and enters heart, liver, gallbladder, spleen and stomach channels. The Chinese medicinal composition is used for reducing fat and losing weight, resisting oxidation, removing free radicals, enhancing immunity, resisting aging, protecting liver, inhibiting hepatic fibrosis, resisting liver toxicity, resisting bacteria and diminishing inflammation and the like, and provides a new choice for clinical treatment of patients with liver diseases and stroke.

The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.

Example 1 preparation of flavonoid glycoside of Lotus leaf/Quercetin-3-O-. beta. -D glucuronide

Extracting 100g of lotus leaf powder, placing the lotus leaf powder in a 2000ml three-necked bottle, adding 800ml of water at 80 ℃, stirring for 20min, filtering while hot, compressing and draining, adding 600ml of water at 80 ℃ to filter residue, stirring for 10min, filtering while hot, compressing and draining. Mixing the three filtrates, cooling to room temperature, adjusting pH to 2.6, standing for 2 hr, fine filtering, and adjusting pH to 5.5.

Separating and purifying to obtain a fine filtrate with the pH value of 5.5, passing through an FL-2 macroporous resin column at the flow rate of 1.5BV/h, wherein the height ratio of the resin column diameter is 1:26, then washing with 0-12% ethanol at the flow rate of 1.5BV/h in a gradient manner, and eluting with 15% -25% ethanol by volume concentration. At this point, purification is completed only by passing through a macroporous resin column once.

Enriching the dilute eluent with the pH of 2-3 by using SP-3 macroporous resin, washing the concentrated eluent by using purified water and 30% ethanol, eluting the concentrated eluent by using an ethanol solution with the volume concentration of 50-95%, and collecting the concentrated solution of the product.

Drying considering that the solubility of lotus leaf flavonoid glycoside in water is small and is insoluble, after the concentrated eluent is placed, about 40% of Q3GA is separated out, and the concentrated eluent can be distilled under reduced pressure to be dry, thus obtaining fresh yellow powdery Q3GA, the HPLC chart is shown in figure 3, the purity is 99.51%, the weight is 1.39g, and the yield is 69.50%.

Example 2 preparation of flavonoid glycoside of Lotus leaf/Quercetin-3-O-. beta. -D glucuronide

Extracting folium Nelumbinis powder 20kg, placing in extraction tank, adding 80 deg.C hot water 160kg, stirring for 20min, hot filtering, adding 80 deg.C water 120kg to the residue, stirring for 10min, and hot filtering. Mixing the three filtrates, cooling to room temperature, adjusting pH to 2.6, standing for 2 hr, cross-flow filtering to obtain fine filtrate, and adjusting pH to 5.5.

Separating and purifying to obtain a fine filtrate with pH of 5.5, passing through FL-2 macroporous resin column at flow rate of 1.5BV/h with height ratio of 1:8, washing with 0-12% ethanol at flow rate of 1.5BV/h, and eluting with 15% -25% ethanol. At this time, the separation and purification can be completed only by passing through the macroporous resin column once.

Enriching the dilute eluent with the pH value of 2-3 by using SP-3 macroporous resin, washing by using purified water and 30% ethanol, eluting by using an ethanol solution with the volume concentration of 50-95%, collecting a concentrated product solution, standing to obtain more bright yellow precipitates, and filtering to obtain a bright yellow powdery product I and a bright yellow filtrate, wherein the purity of the product I is 99.48% (normalization).

Drying to obtain fresh yellow filtrate, drying with negative pressure spray dryer to obtain fresh yellow powdered flavonoid glycoside Q3GA II with purity of 99.46% (normalization), mixing product I and II, drying with purity of 99.48%, weight of 284.73g, and yield of 71.18%.

Example 3 preparation of flavonoid glycoside of Lotus leaf/Quercetin-3-O-. beta. -D glucuronide

Extracting folium Nelumbinis powder 30kg, placing in an extraction tank, adding 80 deg.C hot water 240kg, stirring for 20min, hot filtering, adding 80 deg.C water 180kg to the residue, stirring for 10min, and hot filtering. Mixing the three filtrates, cooling to room temperature, adjusting pH to 2.6, standing for 2 hr, fine filtering, and adjusting pH to 5.5.

Separating and purifying to obtain a fine filtrate with pH of 5.5, passing through FL-2 macroporous resin column at flow rate of 1.5BV/h with height ratio of 1:8, washing with 0-12% ethanol at flow rate of 1.5BV/h, and eluting with 15% -25% ethanol. And (5) completing purification by passing through a macroporous resin column once.

Enriching and taking dilute eluent with the pH of 2-3, passing the dilute eluent through SP-3 macroporous resin for enrichment, washing the dilute eluent with purified water and 30% ethanol, eluting the dilute eluent with ethanol solution with the volume concentration of 50-95%, collecting product concentrated solution, standing the product concentrated solution to obtain a bright yellow precipitate, and filtering the product concentrated solution to obtain a bright yellow powdery product I and a bright yellow filtrate.

Drying the filtrate of the product I by using a negative pressure spray dryer to obtain a fresh yellow powdery product II, combining the product I and the product II, and drying to obtain the product with the purity of 99.68 percent, the weight of 441.25g and the yield of 73.54 percent.

Example 4 preparation of Lotus leaf flavonoid glycoside/Quercetin-3-O-. beta. -D glucuronide

Extracting folium Nelumbinis powder 40kg, placing in extraction tank, adding 80 deg.C hot water 320kg, stirring for 20min, hot filtering, adding 80 deg.C water 240kg to the residue, stirring for 10min, and hot filtering. Mixing the three filtrates, cooling to room temperature, adjusting pH to 2.6, standing for 2 hr, fine filtering, and adjusting pH to 5.5.

Separating and purifying, collecting the fine filtrate with pH of 5.5, passing through FL-2 macroporous resin column at flow rate of 1.5BV/h with height ratio of 1:8, washing with 0-12% ethanol at flow rate of 1.5BV/h, eluting with 15-25% ethanol, and passing through macroporous resin column once to complete purification.

Enriching the dilute eluent with the pH of 2-3 by using SP-3 macroporous resin, washing by using purified water and 30% ethanol, eluting by using an ethanol solution with the volume concentration of 50-95%, collecting the product concentrated solution, standing to obtain more bright yellow precipitates, and filtering to obtain a bright yellow powdery product I and a bright yellow filtrate.

Drying to obtain filtrate of product I, spray drying under negative pressure to obtain fresh yellow powder product II, mixing I and II, drying, and obtaining product with purity of 99.59%, weight of 588.94g and yield of 73.62% as shown in HPLC chart 5.

Example 5 Lotus flavonoid glycoside pellets

Pill prescription

Prescription of coating liquid

Process for the preparation of a catalyst

The lotus leaf flavonoid glycoside, the carboxymethyl starch sodium and the microcrystalline cellulose are respectively taken for pretreatment and sieved by a sieve of 80 meshes.

The preparation method comprises the steps of uniformly mixing the raw materials and the auxiliary materials according to the principle that the quantity of the lotus leaf flavonoid glycoside, the carboxymethyl starch sodium and the microcrystalline cellulose is equivalent and gradually increased, adding purified water, and repeatedly kneading to prepare the soft material.

Extruding and rounding the soft material into strips by an extruder, and cutting and rounding by a cutting and rounding machine to obtain the wet pellets.

Drying, vacuum drying wet pellet at 40 deg.C below in vacuum drying oven, and removing fine powder to obtain pellet.

Coating the coated pellet with gastric film to increase weight by 2.53%.

The sample is inspected according to the quality standard for clinical research, and all the inspection indexes are in accordance with the regulations.

Example 5 Lotus flavonoid glycoside sustained Release pellets

Preparation of lotus leaf flavonoid glycoside sustained and controlled release pellet

Prescription 1 preparation of normally dissolved pellet

Pill prescription 1

Coating liquid formula 1

Process for the preparation of a catalyst

Pretreating to obtain starch, pregelatinized starch, carboxymethyl starch sodium (2.5 g), and microcrystalline cellulose, and mixing to obtain adjuvant mixture; collecting flavonoid glycoside and PVPK of folium Nelumbinis₁₂Sieving with 120 mesh sieve and mixing. Dissolving with 75% ethanol to obtain solution.

Preparing soft material from flavonoid glycoside and PVPK₁₂Adding 75% ethanol solution into the adjuvant mixture, mixing while adding, and repeatedly kneading to obtain soft material.

Coating the pellet with green gastric-soluble film. Thus obtaining the pellet which is normally dissolved out.

Prescription 2 preparation of sustained-release 6h pellet

Pill prescription 2

Coating liquid prescription 2

Process for the preparation of a catalyst

Preprocessing raw and auxiliary materials, respectively sieving lactose, sodium cyclamate, hydroxypropyl methylcellulose and carbomer with a 80-mesh sieve, and uniformly mixing to obtain an auxiliary material mixture; mixing folium Nelumbinis flavonoid glycoside and polyvidone K₁₂Sieving with 100 mesh sieve, mixing, dissolving with appropriate amount of 75% ethanol to obtain solution, and keeping.

Preparing soft material from flavonoid glycoside of folium Nelumbinis and polyvidone K₁₂Adding 75% ethanol solution into the adjuvant mixture, mixing while adding, and repeatedly kneading to obtain soft material.

Extruding and rounding the soft material into strips by an extruder, and cutting and rounding by a cutting and rounding machine. To obtain wet micro-pills

Drying, vacuum drying at 40 deg.C or below for about 40 min, and removing fine powder.

Coating with yellow acrylic resin IV film to obtain sustained release pellet of 6 hr.

Prescription 3 pellet capable of slowly releasing for 12h

Pill prescription 3

Coating liquid prescription 3

Process for the preparation of a catalyst

Pretreating raw materials and adjuvants, sieving hydroxypropyl methylcellulose K100M CR (HPMC K100M CR) with 80 mesh sieve

Using; collecting flavonoid glycoside and polyvidone K from folium Nelumbinis₁₂Sieving with 100 mesh sieve, mixing, dissolving with appropriate amount of 75% ethanol to obtain solution, and keeping.

Preparing soft material from flavonoid glycoside of folium Nelumbinis and polyvidone K₁₂Adding 75% ethanol solution into hydroxypropyl methylcellulose while mixing, repeatedly kneadingForming a soft material.

Extruding and rounding into strips by an extruder, and cutting and rounding by a cutting and rounding machine to obtain the wet pellets.

Drying the wet pellets in a vacuum drying oven, drying at 40 deg.C or below (about 40 min), and removing fine powder.

Coating with red acrylic resin I film to obtain the delayed release pellet of 12 hr.

Lotus leaf flavonoid glycoside sustained and controlled release pellet capsule

The mixed pills are prepared by respectively weighing 160g, 240g and 250g of the pellets in the

formulas

1, 2 and 3 or adjusting the weight of each pellet with a required dissolution-release curve according to the dissolution and release values of the three pellets, and fully and uniformly mixing.

Loading the mixed pellets into capsules of proper type to obtain the sustained-release pellets for 12 h.

Subpackaging, packaging, labeling, packaging and warehousing.

The sample is inspected according to the quality standard for clinical research, and all indexes are in accordance with the regulations.

The clinical application of the pharmaceutical composition is mainly used for preventing and treating liver diseases (fatty liver and liver cancer) and stroke, and provides a new choice for clinical medication.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims

1. A method for extracting flavonoid glycoside from lotus leaf comprises the following steps:

1) mixing lotus leaf powder with water at the temperature of 60-80 ℃, stirring and extracting for the first time, and performing first filtering by throwing to obtain filtrate 1 and filter residue 1;

2) mixing the filter residue 1 with water at the temperature of 60-80 ℃, stirring and extracting for the second time, and performing filter spinning for the second time to obtain a filtrate 2 and a filter residue 2;

3) mixing the filter residue 2 with water at the temperature of 60-80 ℃, stirring and extracting for the third time, and performing filter spinning for the third time to obtain a filtrate 3 and a filter residue 3;

4) mixing the filtrate 1, the filtrate 2 and the filtrate 3, cooling to 25-30 ℃, adjusting the pH to 2-3, standing for 2 hours, and performing cross-flow filtration to obtain a fine filtrate;

5) adjusting the pH value of the fine filtration liquid to 5-6.5, adsorbing, washing and eluting by using macroporous resin, and collecting eluent with the purity of the lotus leaf flavonoid glycoside being more than or equal to 99% to complete the extraction and purification of the lotus leaf flavonoid glycoside; the lotus leaf flavonoid glycoside is quercetin-3-O-beta-D glucuronide.

2. The extraction method according to claim 1, wherein the mass ratio of the lotus leaf powder to water in the step 1) is 1: (7-10).

3. The extraction method according to claim 1 or 2, wherein the mass ratio of the residue 1 in step 2) and the residue 2 to water in step 3) is independently 1: (5-8).

4. The extraction method according to claim 1, wherein the time for the first stirring extraction in the step 1) is 12-18 min.

5. The extraction method according to claim 4, wherein the time for the second stirring extraction in step 2) or the third stirring extraction in step 3) is independently 5-10 min.

6. The extraction method according to claim 1, wherein the step 5) of collecting the eluate further comprises adjusting the pH value of the eluate to 1.5-3.5, enriching by a macroporous resin adsorption method, washing by purified water and 30% ethanol, and eluting by an ethanol solution with a volume concentration of 50-95% to realize concentration.