A water-soluble astragalus extract, an oral fast-dissolving film, and a preparation method and application thereof

Abstract

The application discloses a water-soluble astragalus extract, an oral quick-dissolving film and a preparation method and application thereof, and belongs to the technical field of medicines. The technical problem to be solved is to provide a water-soluble astragalus extract which is free of alcohol, has high water solubility, and has clear and controllable effective component content, and an oral quick-dissolving film which has clear and high active component content, rapid disintegration and complete dissolution. The preparation method comprises the following steps: taking dried astragalus slices, crushing and sieving to obtain astragalus coarse powder; adding water to the astragalus coarse powder, adjusting the pH to 4.5-6.0, and adding a composite enzyme composed of cellulase, pectinase and xylanase for enzymolysis; after the enzymolysis, ultrasonic-dynamic countercurrent extraction is carried out, and the extraction liquid is combined; the extraction liquid is cooled to room temperature, centrifuged, and the supernatant is sequentially subjected to 0.45 mu m microfiltration membrane and 10 kDa ultrafiltration membrane, and the permeate is collected; vacuum concentration and freeze-drying are carried out, and the water-soluble astragalus extract is obtained.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a water-soluble Astragalus extract, an oral instant film, its preparation method, and its application. Background Technology

[0002] Astragalus is the dried root of *Astragalus membranaceus* (Fisch.) Bge. var. *mongholicus* (Bge.) Hsiao or *Astragalus membranaceus* (Fisch.) Bge., belonging to the legume family. It has the effects of tonifying qi and strengthening the exterior, promoting diuresis and eliminating toxins. Modern research shows that astragalus contains active substances such as astragalus polysaccharides, flavonoids (such as verbascoside), and amino acids (such as arginine and proline).

[0003] Currently available astragalus products on the market, such as "Astragalus Extract" produced by Yangtze River Pharmaceutical Group, typically use ethanol extraction or alcohol-containing processes, resulting in a certain amount of alcohol in the finished product. These products are unsuitable for people with alcohol allergies, children, pregnant women, drivers, and patients with certain diseases (such as liver disease), thus limiting their application population. Furthermore, traditional astragalus extracts have poor water solubility (often below 80%), which can easily lead to precipitation, turbidity, and unpleasant taste when preparing oral solid or liquid dosage forms, and the content of active ingredients is unstable.

[0004] The existing Chinese invention patent application CN118344501A discloses a method for preparing a high-content Astragalus polysaccharide extract. First, Astragalus membranaceus is enzymatically hydrolyzed. Then, ethanol is added as an organic solvent to extract astragalus saponins. Following adsorption and enzymatic hydrolysis using macroporous adsorption resin, ultrafiltration to remove proteins, concentration, and drying, the yield of Astragalus polysaccharides reaches 15.6%, and the total polysaccharide content in the extract product reaches 76.2%. However, this method suffers from drawbacks such as complex preparation steps and low yield of Astragalus polysaccharides, therefore, the preparation process needs further improvement. Furthermore, testing showed that the water solubility of the Astragalus polysaccharides obtained by the above process is approximately 85%, indicating that water solubility needs further improvement.

[0005] Orally dissolving film (ODF) is a dosage form in oral mucosal drug delivery systems, typically an ultrathin film the size of a postage stamp (50-150 μm). However, existing Astragalus membranaceus raw materials and film formulation technologies suffer from four major pain points, severely restricting product development: Pain Point 1: Extremely high water-insoluble content, severely deteriorating taste. Ordinary astragalus powder has a water-insoluble content as high as 98.8%. After being made into a film, a large number of insoluble fiber particles are suspended in the oral cavity, producing a noticeable gritty feeling. Patient compliance is extremely poor, and it cannot meet the basic requirements of ODF (Original Dietary Formulation) to be "fine and quick-dissolving". Pain Point 2: Low utilization rate of free amino acids and weak efficacy basis. The total amount of free amino acids in traditional extraction processes and ordinary powders is low (measured <5 mg / g) and is not quantitatively controlled, which prevents Astragalus membranaceus from exerting its rapid anti-fatigue and mucosal repair effects, and key amino acids such as arginine and proline are not effectively enriched; Pain Point 3: Astragaloside A and flavonoids dissolve slowly and have low bioavailability. Conventional films use ordinary astragalus powder or crude extract, which hinders the dissolution of hydrophobic saponins. The dissolution rate is often less than 50% within 5 minutes, failing to achieve the rapid effect of oral instant films. Pain Point 4: Traditional processes have complex formulations. Existing technologies often use compound formulas or multiple excipients to mask defects. For example, Chinese invention patent application CN105055659A introduces multiple medicinal materials (in addition to Astragalus membranaceus, it also contains Atractylodes macrocephala, ginseng, wolfberry and licorice) and cross-linked polyvinylpyrrolidone and other excipients, which not only increases the difficulty of quality control, but also does not fundamentally solve the problem of insoluble matter.

[0006] Therefore, there is an urgent need to provide a method for preparing a water-soluble Astragalus extract that is alcohol-free, highly water-soluble, has a clearly defined and controllable content of active ingredients, and has a simple preparation process, and to use it to prepare an oral instant-dissolving film with clearly defined and high content of active ingredients, rapid disintegration, and complete dissolution, which has important clinical and market value. Summary of the Invention

[0007] To address the technical problems in existing technologies, this invention provides a water-soluble Astragalus extract, an oral instant film, its preparation method, and its application. The extract involves enzymatic hydrolysis of crude Astragalus powder using a complex enzyme composed of cellulase, pectinase, and xylanase. The resulting extract contains ≥99% water-soluble components, ≥7.0 mg / g arginine, ≥20.0 mg / g proline, and ≥0.04% verbascoside glucoside. Simultaneously, the film-forming materials and plasticizers for the oral instant film were screened to obtain an optimal formulation. The resulting oral instant film exhibits clearly defined and high content of active ingredients, rapid disintegration, and complete dissolution.

[0008] Terminology Explanation: Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Unless otherwise stated, all patents, patent inventions, and disclosures cited throughout this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms herein, the definitions provided in this chapter shall prevail.

[0009] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0010] Definitions of raw materials and related terms can be found in the reference "Pharmacopoeia of the People's Republic of China (2025 Edition)": China Medical Science and Technology Press: March 2025: 1st Edition".

[0011] Astragalus membranaceus, a legume, is a plant called Astragalus mongholicus. Astragalus membranaceus (fisch.)Bge.var. Mongolian (Bge.) Hsiao or Astragalus membranaceus Astragalus membranaceus The dried root of (Fisch.) Bge. Harvested in spring and autumn, the fibrous roots and root heads are removed, and then dried in the sun. It is sweet in nature and slightly warm in flavor; it enters the lung and spleen meridians; it has the effects of tonifying qi and raising yang, consolidating the exterior and stopping sweating, promoting diuresis and reducing swelling, generating fluids and nourishing blood, promoting circulation and relieving pain, promoting pus drainage and detoxification, and promoting wound healing and tissue regeneration. It is used to treat qi deficiency and fatigue, poor appetite and loose stools, qi sinking, chronic diarrhea and rectal prolapse, hematochezia and metrorrhagia, spontaneous sweating due to exterior deficiency, edema due to qi deficiency, internal heat and thirst, blood deficiency and chlorosis, hemiplegia, numbness and pain, carbuncles that are difficult to heal, and chronic ulcers that do not heal.

[0012] Unless otherwise stated, conventional methods within the scope of the art shall be used.

[0013] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0014] The terms “optional / arbitrary” or “optionally / arbitrarily” mean that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0015] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a method for preparing a water-soluble Astragalus extract, the method comprising the following preparation steps: (1) Raw material pretreatment: Take dried Astragalus membranaceus slices, crush and sieve them to obtain Astragalus membranaceus coarse powder; (2) Enzyme-assisted extraction: Add water to the crude Astragalus powder, adjust the pH to 4.5-6.0, add a complex enzyme composed of cellulase, pectinase and xylanase, and perform enzymatic hydrolysis; (3) Ultrasonic-dynamic countercurrent extraction: After enzymatic hydrolysis, the temperature is raised to 80-95℃, and dynamic countercurrent extraction is performed under ultrasonic assistance. The extracts are then combined. (4) Centrifugation and membrane separation: The extract was cooled to room temperature, centrifuged, and the supernatant was passed through a 0.45 μm microfiltration membrane and a 10 kDa ultrafiltration membrane in sequence. The permeate was collected. (5) Concentration and drying: The permeate is concentrated under reduced pressure at ≤60℃ and then freeze-dried to obtain the final product.

[0016] Preferably, the sieving in step (1) is sieving through a 40-60 mesh sieve.

[0017] Preferably, the amount of water added in step (2) is 8-15 times the mass of the crude Astragalus powder.

[0018] Preferably, the mass ratio of cellulase, pectinase and xylanase in step (2) is 3-5:1-3:1-3, the amount of enzyme added is 0.5%-1.5% of the mass of Astragalus membranaceus crude powder, and the enzyme is stirred and hydrolyzed at 45-55℃ for 1-3 hours.

[0019] Preferably, the frequency of the ultrasound in step (3) is 40-60 kHz and the power is 300-500 W.

[0020] Preferably, the dynamic countercurrent extraction in step (3) is performed 2-3 times, with each extraction lasting 30-60 minutes.

[0021] Preferably, the centrifugation in step (4) is performed at 4000-6000 rpm for 10-20 minutes.

[0022] Preferably, the concentration in step (5) is to concentrate the permeate to a relative density of 1.05-1.15 at 60°C.

[0023] A second aspect of the present invention relates to a water-soluble astragalus extract, which is prepared according to the above-described preparation method.

[0024] The third aspect of the present invention relates to an oral quick-dissolving film, the oral quick-dissolving film comprising the following components in parts by weight: 20-40 parts of water-soluble Astragalus extract, 30-50 parts of film-forming material, 5-15 parts of plasticizer, and 0.5-3 parts of flavoring agent; The water-soluble Astragalus extract was prepared according to the above preparation method.

[0025] Preferably, the film-forming material is selected from one or more of hydroxypropyl methylcellulose, pullulan, polyvinyl alcohol, and polyvinylpyrrolidone.

[0026] More preferably, the film-forming material is composed of hydroxypropyl methylcellulose and polyvinyl alcohol in a mass ratio of 1-3:1.

[0027] Preferably, the plasticizer is selected from at least one of glycerin, polyethylene glycol, and honey.

[0028] More preferably, the plasticizer is composed of glycerol and polyethylene glycol-400 in a mass ratio of 2-4:1.

[0029] Preferably, the flavoring agent is selected from at least one of sucralose, aspartame, xylitol, sucrose, acesulfame potassium, sorbitol, isomaltitol, and fruit flavoring.

[0030] A fourth aspect of the present invention relates to a method for preparing the above-mentioned oral quick-dissolving film, the method comprising the following preparation steps: S1: Dissolve water-soluble Astragalus membranaceus extract and flavoring agent in water to obtain a medicinal solution; S2: Add the film-forming material and plasticizer to water, heat and swell to obtain a gel solution; S3: Mix the drug solution and the adhesive solution, vacuum degas, coat, dry, demold, and cut to obtain the final product.

[0031] Preferably, in step S1, the mass ratio of the water-soluble Astragalus extract to the flavoring agent and the volume ratio to water is 1g:3-4mL.

[0032] Preferably, in step S2, the mass ratio of the film-forming material to the plasticizer and the volume ratio of the plasticizer to water is 1g:3-6mL, and the swelling time is 1-3h.

[0033] Preferably, in step S3, the vacuum degassing time is 10-20 minutes.

[0034] Preferably, in step S3, the coating thickness is 0.4-0.6 mm.

[0035] Preferably, in step S3, the drying is performed at 50-70°C for 1-3 hours.

[0036] The fifth aspect of the present invention relates to the application of the water-soluble Astragalus extract or oral instant film containing the water-soluble Astragalus extract prepared by the above preparation method in the preparation of products for replenishing qi, relieving fatigue, and repairing oral mucosa.

[0037] Preferably, the product is a pharmaceutical, food, functional food, or health product.

[0038] Compared with the prior art, the present invention has at least the following beneficial effects: (1) By enzymatically hydrolyzing Astragalus membranaceus crude powder with a complex enzyme composed of cellulase, pectinase and xylanase, the content of water-soluble components in the obtained extract is ≥98%, arginine content is ≥7.0 mg / g, proline content is ≥20.0 mg / g, and verbascoside isoflavone glucoside content is ≥0.04%; (2) By screening the film-forming materials and plasticizers of the oral quick-dissolving film, the optimal formulation composition was obtained, and the obtained oral quick-dissolving film has the characteristics of clear and high content of active ingredients, rapid disintegration and complete dissolution. Detailed Implementation

[0039] To make the objectives, technical solutions, and beneficial effects of this invention clearer, detailed explanations are provided below through specific embodiments. It should be noted that these embodiments are for illustrative purposes only and do not constitute any limitation on the scope of the invention. Unless otherwise stated, the experimental methods described in the embodiments are conventional techniques in the art, and the materials and reagents used are commercially available.

[0040] Example 1: Take 1 kg of Astragalus membranaceus slices, pulverize and pass through a 50-mesh sieve. Add 12 times the amount of water, adjust the pH to 5.2, add 10 g of a compound enzyme (cellulase:pectinase:xylanase = 2:1:1), and enzymatically hydrolyze at 50℃ for 2 hours. Raise the temperature to 90℃ and extract twice under dynamic countercurrent under ultrasonication at a frequency of 50 kHz and a power of 400 W, 45 minutes each time, and combine the extracts. Centrifuge at 5000 rpm for 15 minutes, and pass the supernatant through a 0.45 μm microfiltration membrane and a 10 kDa ultrafiltration membrane successively. Concentrate the permeate under reduced pressure at 55℃ to a relative density of 1.10 (60℃), and freeze-dry to obtain water-soluble Astragalus membranaceus extract.

[0041] Example 2: Take 1 kg of Astragalus membranaceus slices, pulverize and pass through a 50-mesh sieve. Add 8 times the amount of water, adjust the pH to 4.5, add 15 g of a compound enzyme (cellulase:pectinase:xylanase = 5:3:3), and enzymatically hydrolyze at 45℃ for 3 hours. Raise the temperature to 90℃ and extract twice under dynamic countercurrent under ultrasonication at a frequency of 50 kHz and a power of 400 W, 45 minutes each time, and combine the extracts. Centrifuge at 5000 rpm for 15 minutes, and pass the supernatant through a 0.45 μm microfiltration membrane and a 10 kDa ultrafiltration membrane successively. Concentrate the permeate under reduced pressure at 55℃ to a relative density of 1.15 (60℃), and freeze-dry to obtain water-soluble Astragalus membranaceus extract.

[0042] Example 3: Take 1 kg of Astragalus membranaceus slices, pulverize and pass through a 50-mesh sieve. Add 15 times the amount of water, adjust the pH to 6, add 10 g of a compound enzyme (cellulase:pectinase:xylanase = 3:1:1), and enzymatically hydrolyze at 55℃ for 1 hour. Raise the temperature to 90℃ and extract twice under dynamic countercurrent under ultrasonication at a frequency of 50 kHz and a power of 400 W, 45 minutes each time, and combine the extracts. Centrifuge at 5000 rpm for 15 minutes, and pass the supernatant through a 0.45 μm microfiltration membrane and a 10 kDa ultrafiltration membrane successively. Concentrate the permeate under reduced pressure at 55℃ to a relative density of 1.05 (60℃), and freeze-dry to obtain water-soluble Astragalus membranaceus extract.

[0043] Comparative Example 1: The difference from Example 1 lies in the composition of the complex enzyme. The complex enzyme in Comparative Example 1 contains only cellulase and pectinase in a mass ratio of 2:1. Everything else is the same as in Example 1.

[0044] Comparative Example 2: The difference from Example 1 lies in the composition of the complex enzyme. The complex enzyme in Comparative Example 2 contains only cellulase and xylanase, with a mass ratio of 2:1. Everything else is the same as in Example 1.

[0045] Comparative Example 3: The difference from Example 1 lies in the composition of the complex enzyme. The composition of the complex enzyme in Comparative Example 3 is: cellulase:papain:xylanase = 2:1:1. The rest is the same as in Example 1.

[0046] Comparative Example 4: The difference from Example 1 lies in the composition of the complex enzyme. The composition of the complex enzyme in Comparative Example 4 is: cellulase: pectinase: β-glucanase = 2:1:1. The rest is the same as in Example 1.

[0047] Experimental Example 1: Determination of water solubility and indicative components of the extract (1) Method for determining the water solubility of extracts Weigh 1.0 g of the extract into 20 mL of purified water, stir at 25°C for 5 minutes, filter through a 0.22 μm filter membrane, dry and weigh the residue, and calculate the solubility.

[0048] (2) Method for determining the total amount of free amino acids: The ninhydrin colorimetric method (spectrophotometric method) is used, specifically as follows: Preparation of reference solution: Accurately weigh an appropriate amount of leucine reference standard dried to constant weight at 105℃, and add water to prepare a solution containing 100 μg per 1 mL.

[0049] Preparation of test solution: Take appropriate amounts of Astragalus extract powder from Examples 1-3 and Comparative Examples 1-4 of this invention, accurately weigh them, dissolve and dilute them with water to a suitable concentration, filter, and collect the filtrate.

[0050] Colorimetric reaction: Accurately measure 1 mL of the reference solution and 1 mL of the test solution into separate stoppered test tubes. Add 0.5 mL of pH 6.8 phosphate buffer and 0.5 mL of 2% ninhydrin solution to each tube, shake well, stopper, and heat in a boiling water bath for 15 minutes. Remove and cool rapidly to room temperature.

[0051] Determination: After cooling, add 5.0 mL of water to each sample, shake well, and use the corresponding reagent as a blank. Measure the absorbance at a wavelength of 570 nm. Calculate the total amount of free amino acids in the test solution (calculated as leucine) based on the absorbance using the external standard method.

[0052] (3) Methods for determining the content of arginine and proline Chromatographic conditions: Octadecylsilane-bonded silica gel (C18, 4.6 × 250 mm, 5 μm) was used as the stationary phase; acetonitrile-0.1 mol / L sodium acetate buffer (pH 6.4) was used as the mobile phase with gradient elution; the detection wavelength was 254 nm; the flow rate was 1.0 mL / min; and the column temperature was 40 °C.

[0053] Preparation of reference solution: Take appropriate amounts of arginine and proline reference standards, accurately weigh them, and add 0.1 mol / L hydrochloric acid solution to prepare a mixed solution containing 0.5 mg of each per 1 mL.

[0054] Preparation of test solution: Take appropriate amounts of Astragalus extract from Examples 1-3 and Comparative Examples 1-4 respectively, weigh accurately, and add 0.1 mol / L hydrochloric acid solution to prepare Astragalus extract containing 100 mg per 1 mL.

[0055] Derivatization: Accurately measure 1 mL each of the reference solution and the test solution, place them in a derivatization flask, add 0.5 mL of phenyl isothiocyanate (PITC) acetonitrile solution and 0.5 mL of 1 mol / L triethylamine acetonitrile solution, shake well, let stand at room temperature for 1 hour, add 1 mL of n-hexane, shake, let stand to separate the layers, take the lower layer solution and filter to obtain the final product.

[0056] Determination: Accurately pipette 5 μL each of the derivatized reference solution and the test solution into the liquid chromatograph, measure the peak area, and calculate the result based on the peak area using the external standard method.

[0057] (4) Determination of the content of verbascoside isoflavone glucoside: The HPLC method was used, referring to the method in Part I of the 2020 edition of the Chinese Pharmacopoeia.

[0058] The measurement results are shown in Table 1.

[0059] Table 1: Determination results of water-soluble and index components of Astragalus extract samples in each group

[0060] Water solubility: The water solubility of Examples 1-3 was all above 99%, significantly higher than that of Comparative Examples 1-4. This indicates that the synergistic use of cellulase, pectinase, and xylanase in this invention can effectively disrupt the cell wall structure of Astragalus membranaceus and promote the release of water-soluble components; while the absence of any one of these enzymes (Comparative Examples 1 and 2) or its substitution with other enzymes (Comparative Examples 3 and 4) will lead to a significant decrease in water solubility.

[0061] Total free amino acids: The total free amino acid content in Examples 1-3 was 32.8-34.1 mg / g, which is much higher than that in Comparative Examples 1-4 (16.9-22.4 mg / g). This proves that the process of the present invention can effectively enrich small molecule amino acid components.

[0062] Arginine and proline content: The arginine content of Examples 1-3 was 7.1-7.4 mg / g and the proline content was 20.1-21.2 mg / g, both of which met the quality control lower limit set in the invention (arginine ≥ 7.0 mg / g, proline ≥ 20.0 mg / g); while Comparative Examples 1-4 did not meet this standard.

[0063] Verbena isoflavone glucoside content: The content of Examples 1-3 was 0.041%-0.045%, which met the quality requirement of ≥0.04%; the content of Comparative Examples 1-4 was 0.025%-0.031%, which did not meet the standard.

[0064] The water-soluble Astragalus extract used in the following examples and comparative examples was prepared using the preparation method of Example 1.

[0065] Example 4: An oral instant dissolving film, wherein the oral instant dissolving film, by weight, comprises: 30 parts water-soluble astragalus extract, 30 parts hydroxypropyl methylcellulose E15, 10 parts polyvinyl alcohol, 8 parts glycerin, 2 parts polyethylene glycol-400, 1 part sucralose, and 300 parts purified water.

[0066] Preparation method: Dissolve water-soluble Astragalus membranaceus extract and sucralose in 100 parts of purified water to obtain a drug solution; add hydroxypropyl methylcellulose E15, polyvinyl alcohol, glycerin, and polyethylene glycol-400 to the remaining purified water, stir and swell for 2 hours to obtain a gel solution; mix, degas under vacuum for 15 minutes; coat on PET release film with a coating thickness of 0.5 mm, dry at 60℃ for 2 hours, demold, and cut into 20×30 mm film sheets.

[0067] Comparative Example 5: The difference from Example 4 is that the film-forming material composition is different. In Comparative Example 5, an equal amount of polyvinyl alcohol is used instead of hydroxypropyl methylcellulose E15.

[0068] An oral instant dissolving film, wherein the oral instant dissolving film, by weight, comprises: 30 parts water-soluble astragalus extract, 40 parts polyvinyl alcohol, 8 parts glycerin, 2 parts polyethylene glycol-400, 1 part sucralose, and 300 parts purified water.

[0069] Preparation method: Dissolve water-soluble Astragalus membranaceus extract and sucralose in 100 parts of purified water to obtain a drug solution; add polyvinyl alcohol, glycerin, and polyethylene glycol-400 to the remaining purified water, stir and swell for 2 hours to obtain a gel solution; mix, degas under vacuum for 15 minutes; coat on PET release film with a coating thickness of 0.5 mm, dry at 60℃ for 2 hours, demold and cut into 20×30 mm film sheets.

[0070] Comparative Example 6: The difference from Example 4 is that the film-forming material composition is different. In Comparative Example 6, an equal amount of hydroxypropyl methylcellulose E15 is used instead of polyvinyl alcohol.

[0071] An oral instant dissolving film, wherein the oral instant dissolving film, by weight, comprises: 30 parts of water-soluble astragalus extract, 40 parts of hydroxypropyl methylcellulose E15, 8 parts of glycerin, 2 parts of polyethylene glycol-400, 1 part of sucralose, and 300 parts of purified water.

[0072] Preparation method: Dissolve water-soluble Astragalus membranaceus extract and sucralose in 100 parts of purified water to obtain a drug solution; add hydroxypropyl methylcellulose E15, glycerin, and polyethylene glycol-400 to the remaining purified water, stir and swell for 2 hours to obtain a gel solution; mix, degas under vacuum for 15 minutes; coat on PET release film with a coating thickness of 0.5 mm, dry at 60℃ for 2 hours, demold and cut into 20×30 mm film sheets.

[0073] Comparative Example 7: The difference from Example 4 lies in the composition of the film-forming material. In Comparative Example 7, the film-forming material is composed of polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP), with a mass ratio of PVA:PVP=3:5.

[0074] An oral instant dissolving film, wherein the oral instant dissolving film, by weight, comprises: 30 parts water-soluble Astragalus extract, 15 parts polyvinyl alcohol (PVA), 25 parts polyvinylpyrrolidone (PVP), 8 parts glycerin, 2 parts polyethylene glycol-400, 1 part sucralose, and 300 parts purified water.

[0075] Preparation method: Dissolve water-soluble Astragalus membranaceus extract and sucralose in 100 parts of purified water to obtain a drug solution; add polyvinylpyrrolidone, polyvinyl alcohol, glycerin, and polyethylene glycol-400 to the remaining purified water, stir and swell for 2 hours to obtain a gel solution; mix, degas under vacuum for 15 minutes; coat on PET release film with a coating thickness of 0.5 mm, dry at 60℃ for 2 hours, demold and cut into 20×30 mm film sheets.

[0076] Comparative Example 8: The difference from Example 4 is that the composition of the plasticizer is different; in Comparative Example 8, an equal amount of glycerol is used instead of polyethylene glycol-400.

[0077] An oral instant dissolving film, wherein the oral instant dissolving film, by weight, comprises: 30 parts of water-soluble astragalus extract, 30 parts of hydroxypropyl methylcellulose E15, 10 parts of polyvinyl alcohol, 10 parts of glycerin, 1 part of sucralose, and 300 parts of purified water.

[0078] Preparation method: Dissolve water-soluble Astragalus extract and sucralose in 100 parts of purified water to obtain a drug solution; add hydroxypropyl methylcellulose E15, polyvinyl alcohol and glycerin to the remaining purified water, stir and swell for 2 hours to obtain a gel solution; mix, degas under vacuum for 15 minutes; coat on PET release film with a coating thickness of 0.5 mm, dry at 60℃ for 2 hours, demold and cut into 20×30 mm film sheets.

[0079] Comparative Example 9: The difference from Example 4 is that the composition of the plasticizer is different; in Comparative Example 8, an equal amount of polyethylene glycol-400 was used instead of glycerin.

[0080] An oral instant dissolving film, wherein the oral instant dissolving film, by weight, comprises: 30 parts of water-soluble astragalus extract, 30 parts of hydroxypropyl methylcellulose E15, 10 parts of polyvinyl alcohol, 10 parts of polyethylene glycol-400, 1 part of sucralose, and 300 parts of purified water.

[0081] Preparation method: Dissolve water-soluble Astragalus membranaceus extract and sucralose in 100 parts of purified water to obtain a drug solution; add hydroxypropyl methylcellulose E15, polyvinyl alcohol, and polyethylene glycol-400 to the remaining purified water, stir and swell for 2 hours to obtain a glue solution; mix, degas under vacuum for 15 minutes; coat on PET release film with a coating thickness of 0.5 mm, dry at 60℃ for 2 hours, demold and cut into 20×30 mm film sheets.

[0082] Experimental Example 2: This test case measures the disintegration time, 5-minute free amino acid dissolution rate, and taste score (blind review by 10 people) of the oral instant films obtained in Example 4, Comparative Examples 5-9.

[0083] (1) The method for determining the disintegration time of oral instant dissolving film is as follows: Take one sample of oral instant dissolving film (20×30 mm) to be tested and place it in a petri dish containing 10 mL of purified water at 37℃±0.5℃. Record the time (seconds) required for the film to completely disintegrate from contact with the water surface. Three samples of each type are tested and the average value is taken.

[0084] (2) Method for determining the dissolution rate of free amino acids in 5 minutes Preparation of simulated artificial saliva: Take 2.38g of disodium hydrogen phosphate, 0.19g of potassium dihydrogen phosphate, and 0.8g of sodium chloride, dissolve them in an appropriate amount of purified water, adjust the pH to 6.8 with dilute hydrochloric acid, and then add purified water to make up to 1000 mL. Shake well to obtain the dissolution medium.

[0085] Accurately weigh one sample of oral instant dissolving film (20×30 mm) and place it in a dissolution vessel. Start timing when the film contacts the dissolution medium. At 5 minutes, take 10 mL of sample from a fixed sampling point and immediately filter it through a 0.22 μm microporous membrane. Discard the initial filtrate and collect the subsequent filtrate. Determine the free amino acid content in the filtrate according to the "Determination of Total Free Amino Acids" method (ninhydrin colorimetric method) in Experiment Example 1.

[0086] 5-minute free amino acid dissolution rate (%) = 5-minute free amino acid amount ÷ 33.4 × 100% (3) Method for determining taste score (blind taste test by 10 people) A blinded scoring method was used, selecting 10 healthy volunteers (half male and half female, aged 25-45 years, without taste dysfunction). Each volunteer evaluated the taste of each oral instant film sample. The scoring criteria are shown in Table 2.

[0087] Procedure: Volunteers placed one sample of the oral instant-dissolving film (20×30 mm) on their tongue, closed their mouths naturally, and recorded the sensation of the film dissolving in their mouths. Scores were given based on the following four dimensions, with each item scored from 0-3 points, for a total score of 0-12 points. After each sample was tested, volunteers rinsed their mouths with purified water, and the next sample was tested after a 5-minute interval. The final taste score was the average of the scores from 10 volunteers (out of 12 points).

[0088] Table 2: Taste Scoring Criteria for Oral Instant Films

[0089] The results of the oral instant film disintegration time, free amino acid dissolution rate and taste score are shown in Table 3.

[0090] Table 3: Disintegration time, free amino acid dissolution rate and taste score of oral instant film

[0091] Disintegration time: Example 4 had the shortest disintegration time (28s), which was significantly faster than Comparative Examples 5-9, demonstrating that the film-forming material composed of hydroxypropyl methylcellulose and polyvinyl alcohol had the best synergistic effect with the plasticizer composed of glycerol and PEG-400.

[0092] 5-minute free amino acid dissolution rate: The dissolution rate of Example 4 is as high as 96.2%, which is close to complete release and far exceeds that of Comparative Examples 5-9, demonstrating the rapid effect of the oral dissolution film of the present invention.

[0093] Taste rating: Example 4 had the highest taste rating (9.2 / 12), with volunteers generally reporting that it "melted quickly and had no gritty feeling". In contrast, Comparative Examples 5 to 9 all had varying degrees of slow disintegration and grainy texture.

[0094] In summary, the formulation of Example 4 achieves a comprehensive effect of rapid disintegration, efficient dissolution, and excellent taste.

[0095] Experimental Example 3: Pharmacological Experiment The anti-fatigue effects of the water-soluble extracts of Astragalus membranaceus obtained in Examples 1-3 and Comparative Examples 1-4 were investigated. Meanwhile, the extract prepared in Example 1 of Chinese Invention Patent Application CN105055659A was used as Comparative Example 10. The specific steps for preparing the extract in Comparative Example 10 were as follows: Step 1: Add 3 kg of Atractylodes macrocephala, 0.5 kg of ginseng, 4 kg of wolfberry, and 2 kg of licorice to water and decoct for 1 hour. The amount of water added is 8 times the total weight of the herbs. Concentrate the decoction under reduced pressure to obtain a clear extract with a relative density of 1.14 (50℃). Then add 95 vol% ethanol, with the weight of ethanol being 1.5 times the weight of the clear extract. After cooling and standing, take the supernatant. Heat the supernatant to recover the ethanol and concentrate it to obtain a thick extract with a relative density of 1.37 (50℃). Step 2: Take 10 kg of Astragalus membranaceus and add it to a 70 vol% ethanol aqueous solution for reflux decoction for 1 hour. The amount of ethanol aqueous solution added is 4 times the weight of Astragalus membranaceus. Concentrate under reduced pressure to obtain a clear extract with a relative density of 1.06 (50℃). Add water and stir. The amount of water added is 1.2 times the weight of the clear extract. After cooling and standing, take the supernatant. Heat the supernatant to recover the solvent and concentrate to obtain a thick extract with a relative density of 1.32 (50℃). Step 3: Combine the thick pastes obtained in Step 1 and Step 2, mix them evenly, and spray dry them into powder to obtain the final product.

[0096] The pharmacological experiment was conducted as follows: Eight-week-old SPF-grade male Kunming mice, provided by the Beijing Experimental Animal Center, were randomly divided into four groups: a blank control group, Example 1-Example 3 groups, Comparative Example 1-Comparative Example 4 groups, and Comparative Example 10 group, with 10 mice in each group. During the rearing period, mice were provided with ample feed and unlimited water, and the room temperature was maintained at approximately 25°C. The experiment began after 7 days of acclimatization. Except for the blank control group, each group received the drug via gavage at a dose equivalent to 2 g / kg of raw drug, once daily for 7 consecutive days. After drug administration, mice in each group were given a lead weight (5% of their body weight) to their tails and swam until exhaustion. The time to exhaustion was recorded. Exhaustion was defined as the mouse sinking to the bottom for 8 seconds and being unable to surface for respiration. Simultaneously, before swimming and 60 minutes after exhaustion, the tail tip was disinfected with an alcohol swab, the tail was cut off, blood was collected, the first drop of blood was wiped away, and whole blood was drawn using a test strip. Blood lactate levels were measured using a Kyoto portable blood lactate analyzer before swimming and 60 minutes after exhaustion.

[0097] The time to exhaustion in mice after ingestion of different samples is shown in Table 4. The results of blood lactate content measurements in each group of mice before swimming and at 60 minutes after exhaustion are shown in Table 5.

[0098] Table 4: Time to exhaustion in mice after different sample intakes

[0099] Note: Compared with the blank control group ## This indicates that P < 0.01. ### This indicates that P < 0.001; compared with Example 1 group, the differences between the sample groups are... & This indicates that P < 0.05. && This indicates that P < 0.01.

[0100] The data above show that, compared with the blank control group, the extracts of Examples 1-3, Comparative Examples 1-4, and Comparative Example 10 all significantly prolonged the time to exhaustion in mice. Among them, the effect of Examples 1-3 on prolonging the time to exhaustion in mice was significantly better than that of Comparative Examples 1-4 and Comparative Example 10.

[0101] Table 5: Blood lactate levels at different times after exhaustive exercise in mice of different groups

[0102] Note: In the same column of data, compared with the blank control group, # This indicates that P < 0.05. ## This indicates that P < 0.01; compared with Example 1 group, the differences between the sample groups are small. & This indicates that P < 0.05.

[0103] The data in the table show that, compared with the blank control group, the extracts of Examples 1-3, Comparative Examples 1-4, and Comparative Example 10 all significantly reduced the blood lactate content in mice after 60 minutes of exhaustive exercise. However, the blood lactate content of the mice in Examples 1-3 after 60 minutes of exhaustive exercise was significantly lower than that in the comparative groups. Meanwhile, the blood lactate content of mice in Comparative Examples 1-4 and Comparative Example 10 was still significantly higher than that before swimming 60 minutes after exhaustive exercise, while the blood lactate content of the mice in Examples 1-3 almost returned to the level before swimming 60 minutes after exhaustive exercise. Therefore, Examples 1-3 showed significantly better anti-fatigue and post-fatigue recovery effects on mice than the comparative groups.

[0104] The above detailed description is a specific illustration of one feasible embodiment of the present invention, and this embodiment is not intended to limit the patent scope of the present invention. It should be noted that all equivalent implementations or modifications made without departing from the present invention should be included within the scope of the technical solution of the present invention. Therefore, the protection scope of the present invention should be determined by the appended claims.

Claims

1. A method for preparing a water-soluble Astragalus extract, characterized in that, The preparation method includes the following preparation steps: (1) Raw material pretreatment: Take dried Astragalus membranaceus slices, crush and sieve them to obtain Astragalus membranaceus coarse powder; (2) Enzyme-assisted extraction: Add water to the crude Astragalus powder, adjust the pH to 4.5-6.0, add a complex enzyme composed of cellulase, pectinase and xylanase, and perform enzymatic hydrolysis; (3) Ultrasonic-dynamic countercurrent extraction: After enzymatic hydrolysis, the temperature is raised to 80-95℃, and dynamic countercurrent extraction is performed under ultrasonic assistance. The extracts are then combined. (4) Centrifugation and membrane separation: The extract was cooled to room temperature, centrifuged, and the supernatant was passed through a 0.45 μm microfiltration membrane and a 10 kDa ultrafiltration membrane in sequence. The permeate was collected. (5) Concentration and drying: The permeate is concentrated under reduced pressure at ≤60℃ and then freeze-dried to obtain the final product.

2. The preparation method according to claim 1, characterized in that, In step (2), the amount of water added is 8-15 times the mass of the crude Astragalus powder, the mass ratio of cellulase, pectinase and xylanase is 3-5:1-3:1-3, the amount of enzyme added is 0.5%-1.5% of the mass of the crude Astragalus powder, and the enzyme is stirred and hydrolyzed at 45-55℃ for 1-3 hours.

3. The preparation method according to claim 1, characterized in that, The frequency of the ultrasound in step (3) is 40-60kHz and the power is 300-500 W. The number of times the dynamic countercurrent extraction is performed is 2-3 times, and the extraction time for each time is 30-60 min.

4. The preparation method according to claim 1, characterized in that, The centrifugation in step (4) is 4000-6000 rpm for 10-20 min.

5. The preparation method according to claim 1, characterized in that, The concentration in step (5) is to concentrate the permeate to a relative density of 1.05-1.15 at 60°C.

6. A water-soluble Astragalus extract, characterized in that, The water-soluble Astragalus extract was prepared according to any one of claims 1-5.

7. An oral instant-dissolving film, characterized in that, The oral instant film comprises the following components in parts by weight: 20-40 parts of water-soluble Astragalus extract, 30-50 parts of film-forming material, 5-15 parts of plasticizer, and 0.5-3 parts of flavoring agent; The water-soluble Astragalus extract was prepared according to any one of claims 1-5.

8. The oral instant-dissolving film according to claim 7, characterized in that, The film-forming material is selected from one or more of hydroxypropyl methylcellulose, pullulan, and polyvinyl alcohol; the plasticizer is selected from at least one of glycerin, polyethylene glycol, and honey; and the flavoring agent is selected from at least one of sucralose, aspartame, xylitol, sucrose, acesulfame potassium, sorbitol, isomaltitol, and fruit flavoring.

9. A method for preparing the oral instant-dissolving film according to any one of claims 7-8, characterized in that, The preparation method includes the following preparation steps: S1: Dissolve water-soluble Astragalus membranaceus extract and flavoring agent in water to obtain a medicinal solution; S2: The film-forming material is swollen with a plasticizer to obtain a gel solution; S3: Mix the liquid medicine and the adhesive, remove bubbles, coat, dry, demold, and cut to obtain the final product.

10. The application of a water-soluble Astragalus extract prepared by the preparation method according to any one of claims 1-5 or an oral instant film according to any one of claims 7-8 in the preparation of products for replenishing qi, relieving fatigue, and repairing oral mucosa.

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