Walnut peptide compound plant solid beverage capable of reducing uric acid as well as preparation method and application of walnut peptide compound plant solid beverage

Abstract

The invention discloses a walnut peptide compound plant solid beverage capable of reducing uric acid as well as a preparation method and application of the walnut peptide compound plant solid beverage. The walnut enzymolysis mixed peptide is taken as a core, so that the absorption efficiency of active ingredients is remarkably improved, and the overall functional effect can be enhanced. By combining with multiple plant functional components such as sour cherry, chicory, dandelion, celery and nuciferine, a synergistic effect can be formed in multiple links such as inhibiting generation of uric acid, promoting excretion and improving a metabolic environment, so that the uric acid reducing effect better than that of a single plant component is realized. Freeze-drying, low-temperature processing and proper plant extraction processes are adopted, so that plant active substances are reserved to the greatest extent, and the product stability is improved; the preparation process is simple, suitable for large-scale production, stable in product form, good in solubility and convenient to carry and eat for a long time.

Description

Technical Field

[0001] This invention relates to a uric acid-lowering walnut peptide compound plant solid beverage, its preparation method, and its application. Background Technology

[0002] Hyperuricemia is a metabolic disease characterized by elevated blood uric acid levels due to disordered purine metabolism or reduced uric acid excretion. In recent years, its prevalence has been steadily increasing, particularly among younger people, due to changes in dietary structure and irregular lifestyles. Hyperuricemia is not only a direct risk factor for gout but is also closely related to various chronic diseases such as metabolic syndrome, obesity, kidney damage, and cardiovascular disease. Therefore, controlling blood uric acid levels is of significant health importance.

[0003] Currently, the main drugs used to lower blood uric acid include allopurinol, febuxostat, and benzbromarone. However, these drugs generally have adverse effects such as gastrointestinal discomfort, liver and kidney toxicity, and allergic reactions. Furthermore, long-term reliance on drug treatment can easily burden the body's metabolism. Therefore, nutritional intervention programs based on natural plant ingredients are gradually gaining attention due to their advantages such as wide availability, high safety, and low side effects.

[0004] Walnut peptides are low-molecular-weight active peptides obtained from walnut protein through enzymatic hydrolysis. They have good absorption and bioavailability, and related studies have shown that they have bioactivities such as regulating metabolism, anti-oxidation, and improving kidney function. Plant raw materials such as sour cherry, dandelion, chicory, celery, coix seed, and poria cocos contain polyphenols, flavonoids, sesquiterpenes, polysaccharides, and various bioactive substances. They have potential effects such as promoting uric acid metabolism, inhibiting xanthine oxidase activity, and enhancing renal excretion capacity, making them important material sources for constructing plant-based uric acid-lowering products. Lotus leaf alkaloids, as an important alkaloid component in lotus leaves, have a certain research basis in regulating lipid metabolism and improving internal environment homeostasis.

[0005] However, existing uric acid-lowering products generally have the following problems: (1) The raw materials are mostly single plant components, with insufficient synergistic effect and limited overall uric acid lowering effect; (2) Some products have poor stability of active substances, and unreasonable process conditions lead to loss of functional components; (3) There is a lack of research on compound formulations that combine active peptides with multiple plant functional components; (4) In the development of solid beverage products, research on solubility, taste and activity retention is still incomplete.

[0006] Therefore, developing a compound solid beverage based on walnut peptides, combined with multiple plant active components, with synergistic uric acid-lowering effects, scientific formulation, high safety, and easy processing and preservation, has significant research and development value and application prospects. Summary of the Invention

[0007] This invention aims to overcome the problems of existing uric acid-lowering products, such as single raw materials, insufficient synergistic effect of active components, low bioavailability, and unfavorable preparation processes for maintaining active substances. It provides a compound plant-based solid beverage with walnut peptides as the core active ingredient, combined with multiple plant extracts for synergistic uric acid lowering, and its preparation method. By optimizing the formula structure and process flow, the resulting product has good solubility, stability, and superior uric acid-lowering function, making it suitable for industrial production and long-term consumption.

[0008] To achieve the above objectives, the present invention provides a uric acid-lowering walnut peptide compound plant solid beverage, characterized by comprising the following raw materials in parts by weight: (1) 15-30 parts of walnut peptide: The peptide obtained by enzymatic hydrolysis, impurity removal and freeze drying of walnut protein has the activity of inhibiting xanthine oxidase (XOD) and free radical scavenging. (2) 5-15 parts of sour cherry powder: rich in anthocyanins and polyphenols, which can improve purine metabolism disorders and oxidative stress; (3) 4-8 parts of chicory powder: regulates the ABCG2 uric acid excretion pathway and promotes uric acid excretion; (4) Celery powder 4-8 parts: contains flavonoids such as apigenin, which inhibit XOD activity; (5) Dandelion powder 5-10 parts: It helps the kidneys excrete acid through its diuretic effect; (6) Lotus leaf alkaloids 4-8 parts: regulate lipid metabolism and multi-pathway metabolic stress; (7) 4-8 parts of Job's tears powder: synergistically improves kidney function and metabolic balance; (8) 4-8 parts of Poria cocos powder: regulates intestinal flora and metabolic homeostasis; (9) Angelica dahurica 0.5-1 part: to harmonize flavor and provide antioxidant activity.

[0009] According to the present invention, the preparation method of the above-mentioned walnut peptide includes the following steps: dispersing walnut protein in distilled water at a solid-liquid ratio of 1:50 (w / v), heating to 90°C and holding for 15 minutes, cooling to 53°C and adjusting the pH to 7.0-7.2, adding neutral protease and enzymatically hydrolyzing at 53°C for 120 minutes, removing impurities by passing through a 0.2-micron membrane, and freeze-drying to obtain walnut peptide powder.

[0010] According to the present invention, the preparation method of the above-mentioned sour cherry powder is as follows: fresh sour cherries are pitted, crushed and pre-frozen (-18°C, 2 hours), then freeze-dried for 36 hours under the conditions of cold trap temperature -45°C and vacuum pressure 0.01 kPa, and then crushed through an 80-100 mesh sieve.

[0011] According to the present invention, the preparation method of the above-mentioned chicory powder is as follows: chicory root is chopped and added to 30% ethanol solution at a solid-liquid ratio of 1:15, and extracted by hot reflux at 70-80°C for 2 hours. After concentration, it is spray dried (inlet air temperature 160°C, outlet air temperature 80°C).

[0012] According to the present invention, the preparation method of the above-mentioned celery powder is as follows: fresh celery is pulverized and pre-frozen (-18°C, 2 hours), freeze-dried for 36 hours under the conditions of cold trap temperature -45°C and vacuum pressure 0.01 kPa, and then pulverized through an 80-100 mesh sieve.

[0013] According to the present invention, the preparation method of the above-mentioned dandelion powder is as follows: the dried whole dandelion herb is pulverized and added to 50% ethanol solution at a solid-liquid ratio of 1:15. The mixture is then refluxed twice at 60-70℃ for 1.5 hours each time. After concentration, the mixture is spray-dried (inlet air temperature 150℃, outlet air temperature 75℃).

[0014] According to the present invention, the preparation method of the above-mentioned lotus leaf alkaloid is as follows: dried lotus leaves are pulverized and added to 70% ethanol at a solid-liquid ratio of 1:10, and extracted by reflux at 75°C for 1.5 hours. After purification by macroporous adsorption resin column, it is mixed with maltodextrin at a ratio of 1:1 and spray-dried (inlet air temperature 140°C, outlet air temperature 70°C). The lotus leaf alkaloid content is ≥10%.

[0015] According to the present invention, the preparation method of the above-mentioned Angelica dahurica powder is as follows: Angelica dahurica slices are pre-frozen (-18°C, 2 hours), freeze-dried for 48 hours under the conditions of cold trap temperature -45°C and vacuum pressure 0.01 kPa, and then pulverized through an 80-100 mesh sieve.

[0016] On the other hand, the present invention also provides a method for preparing a uric acid-lowering walnut peptide compound plant solid beverage, comprising the following steps: (1) Weigh out the following ingredients according to the above proportions: walnut peptide, sour cherry powder, chicory powder, celery powder, dandelion powder, lotus leaf alkaloid, coix seed powder, poria powder and angelica powder; (2) Add the above raw materials to the mixer and mix evenly for 20–40 min; (3) Dry and sieve under vacuum conditions to obtain solid beverage powder product.

[0017] On the other hand, the present invention also provides the application of uric acid-lowering walnut peptide compound plant solid beverage in the preparation of food or medicine for lowering serum uric acid levels.

[0018] Compared with the prior art, the present invention has the following advantages: By using enzymatically hydrolyzed walnut peptides as the core, the absorption efficiency of active ingredients is significantly improved, enhancing the overall functional effect. Combined with multiple plant functional components such as sour cherry, chicory, dandelion, celery, and lotus leaf alkaloids, it achieves synergistic effects in multiple aspects, including inhibiting uric acid production, promoting excretion, and improving the metabolic environment, thus achieving a better uric acid-lowering effect than single plant ingredients. Utilizing freeze-drying, low-temperature processing, and suitable plant extraction techniques, the product retains the active substances of the plants to the greatest extent, improving product stability. The preparation process is simple, suitable for large-scale production, and the product has a stable form, good solubility, and is easy to carry and consume long-term.

[0019] Animal experiments have shown that the product of this invention can significantly reduce serum uric acid levels in hyperuricemia model mice (comparable to the positive control drug benzbromarone), improve renal function (reducing serum creatinine and blood urea nitrogen), and repair pathological damage to kidney / liver / intestinal tissues (HE staining showed restoration of glomerular structure, relief of hepatic sinusoidal dilation, and improved intestinal villus integrity). The compound formula of this invention is derived from natural plants, has high safety, and is suitable for use as a daily nutritional intervention product for people with hyperuricemia. Attached Figure Description

[0020] The accompanying drawings, which are included to provide a further understanding of this application and form part of this application, illustrate exemplary embodiments and are used to explain this application, but do not constitute an undue limitation of this application. In the drawings: Figure 1 The figure shows the effect of the uric acid-lowering walnut peptide compound plant solid beverage of the present invention on blood urea nitrogen content, kidney coefficient, serum creatinine and serum uric acid in mice. Figure 2 The figure shows the effect of the uric acid-lowering walnut peptide compound plant solid beverage of the present invention on the appearance of mouse kidneys. Figure 3 The figure shows the effect of the uric acid-lowering walnut peptide compound plant solid beverage of the present invention on mouse kidney tissue; Figure 4 The figure shows the effect of the uric acid-lowering walnut peptide compound plant solid beverage of the present invention on mouse liver tissue; Figure 5 This is a diagram showing the effect of the uric acid-lowering walnut peptide compound plant solid beverage of the present invention on the colon tissue of mice. Detailed Implementation

[0021] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0022] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0023] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Example 1: Preparation of a uric acid-lowering walnut peptide compound plant-based solid beverage

[0024] (1) Preparation of walnut peptide: Walnut protein was dispersed in distilled water at a solid-liquid ratio of 1:50 (w / v), heated to 90°C and maintained for 15 min to denature the protein; then the system was cooled to 53°C, the pH was adjusted to 7.2, and neutral protease (9000 U / g protein) was added. Under these conditions, the enzyme was hydrolyzed for 120 min, and the pH was maintained at 7.2 throughout the process; after the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 95°C for 10 min, and the supernatant was collected by centrifugation; after vacuum freeze-drying, the walnut peptide was obtained.

[0025] (2) Preparation of sour cherry powder: After pitting fresh European sour cherries, they are crushed into sour cherry pulp. The sour cherry pulp is then pre-frozen at -18℃ for 2 hours, and then dried in a freeze dryer at -45℃ and 0.01kPa for 36 hours. The pulp is then crushed and passed through a 100-mesh sieve.

[0026] (3) Preparation of chicory powder: The dried chicory root was chopped and 30% ethanol solution was added at a solid-liquid ratio of 1:15. The extract was extracted by hot reflux at 80°C for 2 hours. After centrifugation to remove residue, the extract was concentrated into a paste under reduced pressure at 60°C and then spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain chicory powder.

[0027] (4) Preparation of celery powder: Wash fresh celery, cut it into sections, crush it into celery pulp, then pre-freeze the celery pulp at -18℃ for 2 hours, then dry it in a freeze dryer at -45℃ and 0.01kPa for 36 hours, and then crush it through a 100-mesh sieve.

[0028] (5) Preparation of dandelion powder: The dried whole dandelion herb was pulverized and added to 50% ethanol solution at a solid-liquid ratio of 1:15. The mixture was refluxed twice at 70℃ for 1.5 hours each time. The extracts were combined, filtered, and concentrated under reduced pressure at 60℃ to a relative density of 1.10-1.15 (measured at 60℃). Then, the mixture was spray-dried (inlet air temperature 150℃, outlet air temperature 75℃) to obtain dandelion powder.

[0029] (6) Preparation of lotus leaf alkaloid powder: The dried lotus leaves were crushed and 70% ethanol was added at a solid-liquid ratio of 1:10. The mixture was refluxed at 75°C for 1.5 hours and extracted twice. The extracts were combined, filtered and the ethanol was recovered under reduced pressure to obtain the crude extract of lotus leaves. The crude extract was purified by passing it through a macroporous adsorption resin column. The elution fraction rich in lotus leaf alkaloid was collected, concentrated and mixed with maltodextrin at a ratio of 1:1. The mixture was then spray-dried (inlet air temperature 140°C, outlet air temperature 70°C) to obtain lotus leaf alkaloid powder (lotus leaf alkaloid content ≥10%).

[0030] (7) Preparation of Job's tears powder: After peeling, washing and steaming Job's tears for 40 minutes, they are initially dried, then pulverized and passed through a 100-mesh sieve.

[0031] (8) Preparation of Poria powder: The dried Poria is pulverized by a pulverizer and passed through a 100-mesh sieve to obtain fine Poria powder.

[0032] (9) Preparation of Angelica dahurica powder: wash and slice Angelica dahurica slices, pre-freeze at -18℃ for 2 hours, then dry in a freeze dryer at -45℃ and 0.01kPa for 48 hours, and then pulverize through a 100-mesh sieve.

[0033] (10) Add 30 parts by weight of walnut peptide, 15 parts by weight of sour cherry powder, 8 parts by weight of chicory powder, 8 parts by weight of celery powder, 10 parts by weight of dandelion powder, 8 parts by weight of lotus leaf alkaloid powder, 8 parts by weight of coix seed powder, 8 parts by weight of poria powder and 1 part by weight of angelica powder to a mixer and mix evenly for 30 minutes. After passing through a 60-mesh sieve, package the mixture to obtain the uric acid-lowering walnut peptide compound plant product powder. Example 2: Preparation of Uric Acid-Lowering Walnut Peptide Compound Plant Solid Beverage

[0034] (1) Preparation of walnut peptide: Walnut protein was dispersed in distilled water at a solid-liquid ratio of 1:50 (w / v), heated to 90°C and maintained for 15 min to denature the protein; then the system was cooled to 53°C, the pH was adjusted to 7.0, and neutral protease (9000 U / g protein) was added. Under these conditions, the enzyme was hydrolyzed for 120 min, and the pH was maintained at 7.0 throughout the process; after the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 95°C for 10 min, and the supernatant was collected by centrifugation; after vacuum freeze-drying, the walnut peptide was obtained.

[0035] (2) Preparation of sour cherry powder: After pitting fresh European sour cherries, they are crushed into sour cherry pulp. The sour cherry pulp is then pre-frozen at -18℃ for 2 hours, and then dried in a freeze dryer at -45℃ and 0.01kPa for 36 hours. The pulp is then crushed and passed through an 80-mesh sieve.

[0036] (3) Preparation of chicory powder: The dried chicory root was chopped and 30% ethanol solution was added at a solid-liquid ratio of 1:15. The extract was extracted by hot reflux at 80°C for 2 hours. After centrifugation to remove residue, the extract was concentrated into a paste under reduced pressure at 60°C and then spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain chicory powder.

[0037] (4) Preparation of celery powder: Wash fresh celery, cut it into sections, crush it into celery pulp, then pre-freeze the celery pulp at -18℃ for 2 hours, then dry it in a freeze dryer at -45℃ and 0.01kPa for 36 hours, and then crush it through an 80-mesh sieve.

[0038] (5) Preparation of dandelion powder: The dried whole dandelion herb was pulverized and added to 50% ethanol solution at a solid-liquid ratio of 1:15. The mixture was refluxed twice at 70℃ for 1.5 hours each time. The extracts were combined, filtered, and concentrated under reduced pressure at 60℃ to a relative density of 1.10-1.15 (measured at 60℃). Then, the mixture was spray-dried (inlet air temperature 150℃, outlet air temperature 75℃) to obtain dandelion powder.

[0039] (6) Preparation of lotus leaf alkaloid powder: The dried lotus leaves were crushed and 70% ethanol was added at a solid-liquid ratio of 1:10. The mixture was refluxed at 75°C for 1.5 hours and extracted twice. The extracts were combined, filtered and the ethanol was recovered under reduced pressure to obtain the crude extract of lotus leaves. The crude extract was purified by passing it through a macroporous adsorption resin column. The elution fraction rich in lotus leaf alkaloid was collected, concentrated and mixed with maltodextrin at a ratio of 1:1. The mixture was then spray-dried (inlet air temperature 140°C, outlet air temperature 70°C) to obtain lotus leaf alkaloid powder (lotus leaf alkaloid content ≥10%).

[0040] (7) Preparation of Job's tears powder: After peeling, washing and steaming Job's tears for 40 minutes, they are initially dried, then pulverized and passed through a 100-mesh sieve.

[0041] (8) Preparation of Poria powder: The dried Poria is pulverized by a pulverizer and passed through a 100-mesh sieve to obtain fine Poria powder.

[0042] (9) Preparation of Angelica dahurica powder: Wash and slice Angelica dahurica slices, pre-freeze at -18℃ for 2 hours, then dry in a freeze dryer at -45℃ and 0.01kPa for 48 hours, and then pulverize through an 80-mesh sieve.

[0043] (10) Add 15 parts by weight of walnut peptide, 5 parts by weight of sour cherry powder, 4 parts by weight of chicory powder, 4 parts by weight of celery powder, 5 parts by weight of dandelion powder, 4 parts by weight of lotus leaf alkaloid powder, 4 parts by weight of coix seed powder, 4 parts by weight of poria powder and 0.5 parts by weight of angelica powder to a mixer and mix evenly to obtain the uric acid-lowering walnut peptide compound plant finished powder. Example 3: Preparation of Uric Acid-Lowering Walnut Peptide Compound Plant Solid Beverage

[0044] (1) Preparation of walnut peptide: Walnut protein was dispersed in distilled water at a solid-liquid ratio of 1:50 (w / v), heated to 90°C and maintained for 15 min to denature the protein; then the system was cooled to 53°C, the pH was adjusted to 7.2, and neutral protease (9000 U / g protein) was added. Under these conditions, the enzyme was hydrolyzed for 120 min, and the pH was maintained at 7.2 throughout the process; after the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 95°C for 10 min, the supernatant was collected by centrifugation, and the walnut peptide was obtained by vacuum freeze-drying.

[0045] (2) Preparation of sour cherry powder: After pitting fresh European sour cherries, they are crushed into sour cherry pulp. The sour cherry pulp is then pre-frozen at -18℃ for 2 hours, and then dried in a freeze dryer at -45℃ and 0.01kPa for 36 hours. The pulp is then crushed and passed through a 100-mesh sieve.

[0046] (3) Preparation of chicory powder: The dried chicory root was chopped and 30% ethanol solution was added at a solid-liquid ratio of 1:15. The extract was extracted by hot reflux at 70°C for 2 hours. After centrifugation to remove the residue, the extract was concentrated into a paste under reduced pressure at 60°C and then spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain chicory powder.

[0047] (4) Preparation of celery powder: Wash fresh celery, cut it into sections, crush it into celery pulp, then pre-freeze the celery pulp at -18℃ for 2 hours, then dry it in a freeze dryer at -45℃ and 0.01kPa for 36 hours, and then crush it through a 100-mesh sieve.

[0048] (5) Preparation of dandelion powder: The dried whole dandelion herb was pulverized and added to 50% ethanol solution at a solid-liquid ratio of 1:15. The mixture was refluxed twice at 60℃ for 1.5 hours each time. The extracts were combined, filtered, and concentrated under reduced pressure at 50℃ to a relative density of 1.10-1.15 (measured at 50℃). Then, the mixture was spray-dried (inlet air temperature 150℃, outlet air temperature 75℃) to obtain dandelion powder.

[0049] (6) Preparation of lotus leaf alkaloid powder: The dried lotus leaves were crushed and 70% ethanol was added at a solid-liquid ratio of 1:10. The mixture was refluxed at 75°C for 1.5 hours and extracted twice. The extracts were combined, filtered and the ethanol was recovered under reduced pressure to obtain the crude extract of lotus leaves. The crude extract was purified by passing it through a macroporous adsorption resin column. The elution fraction rich in lotus leaf alkaloid was collected, concentrated and mixed with maltodextrin at a ratio of 1:1. The mixture was then spray-dried (inlet air temperature 140°C, outlet air temperature 70°C) to obtain lotus leaf alkaloid powder (lotus leaf alkaloid content ≥10%).

[0050] (7) Preparation of Job's tears powder: After peeling, washing and steaming Job's tears for 40 minutes, they are initially dried, then pulverized and passed through a 100-mesh sieve.

[0051] (8) Preparation of Poria cocos powder: The dried Poria cocos was pulverized with a pulverizer and passed through a 100-mesh sieve to obtain fine Poria cocos powder.

[0052] (9) Preparation of Angelica dahurica powder: The Angelica dahurica slices were washed, sliced, pre-frozen at -18 °C for 2 h, then dried in a freeze dryer at a cold trap temperature of -45 °C, a pressure of 0.01 kPa for 48 h, and pulverized and passed through a 100-mesh sieve.

[0053] (10) 20 parts by mass of walnut peptide, 10 parts by mass of tart cherry powder, 6 parts by mass of chicory powder, 6 parts by mass of celery powder, 8 parts by mass of dandelion powder, 6 parts by mass of nuciferine powder, 6 parts by mass of coix seed powder, 6 parts by mass of Poria cocos powder and 0.8 parts by mass of Angelica dahurica powder were added to a mixer and mixed evenly to obtain the finished powder of the compound plant of walnut peptide for reducing uric acid.

[0054] Animal experiment To illustrate the uric acid-lowering effect of the compound plant solid beverage of walnut peptide in a hyperuricemia mouse model, the specific experiment is as follows: Example 4: Functional verification of the compound plant solid beverage of walnut peptide for reducing uric acid

[0055] 1 Experimental materials and methods A total of 60 male KunMing mice of Specific Pathogen Free (SPF) grade, with a body weight of (22 ± 2) g, were randomly selected and provided by Hangzhou Qizhen Laboratory Animal Technology Co., Ltd. The production license number of the experimental animals is: SCXK(Zhe)2022-0005, and the animal experiment was approved by the Animal Ethics Committee of Zhejiang A&F University (approval number: ZAFUAC***2541) to ensure that all animal experiments comply with relevant regulations. All mice were placed in a stable environment with experimental feeding room conditions (temperature (25 ± 1) °C, humidity 50 ± 5%, and a light cycle of 12 h (9:00 - 21:00)) for adaptive cultivation for seven days, during which water and food were freely available.

[0056] 1.3 Experimental grouping The mice were divided into a control group, a model group, a positive drug group (benzbromarone), a high-concentration compound group and a low-concentration compound group, with 12 mice in each group.

[0057] 1.4 Modeling and functional verification Mice in the NC group were administered 0.5% CMC-Na solution via gavage and injection. Mice in all other groups were administered 100 mg / kg adenine via gavage and 150 mg / kg potassium oxonate via intraperitoneal injection, once daily for three weeks. During the modeling period, the model group was administered 0.5% CMC-Na solution via gavage, the positive control group was administered 10 mg / kg benzbromarone via gavage, the high-concentration compound group was administered 3 g / kg of urate-lowering walnut peptide compound plant-based solid beverage via gavage, and the low-concentration compound group was administered 1.5 g / kg of urate-lowering walnut peptide compound plant-based solid beverage via gavage. The volume of both gavage and intraperitoneal injection was 10 ml / kg. The health status of the mice was checked daily, and their weight, food intake, and water intake were recorded regularly.

[0058] 1.5 Sample Collection Mouse serum sample collection: Mice were intraperitoneally injected with 50 mg / kg of 1% sodium pentobarbital. After the corneal reflex disappeared and the limbs showed no spontaneous movement, blood samples were collected using the retro-orbital venous plexus method via capillary tube. Approximately 0.8-1.0 mL of blood was collected from each mouse and transferred to centrifuge tubes. The centrifuge tubes containing the blood samples were placed in a 4°C freezer for 30 min, and then centrifuged in a high-speed refrigerated centrifuge at 4°C and 3000×g for 15 min. After centrifugation, the supernatant serum was aspirated and transferred to sterile EP tubes, which were stored at -80°C for subsequent serum marker detection. Tissue sample collection and processing: After serum collection, mice were immediately euthanized by cervical dislocation, and the mice were quickly dissected. The kidneys, liver, and colon tissues were separated and removed, and the surface bloodstains were washed off with PBS. The dissected kidney, liver, and colon tissues were placed in light-proof EP tubes pre-filled with 4% paraformaldehyde fixative (tissue to fixative volume ratio of 1:10) and fixed at room temperature for 24 hours. They were then embedded in paraffin, sectioned, and used for subsequent HE staining and pathological observation.

[0059] 2 Experimental Results like Figure 1 As shown, the serum uric acid level in the model group was significantly elevated during the experiment, indicating uric acid metabolism disorder. Simultaneously, blood urea nitrogen, serum creatinine, and kidney coefficient were all significantly elevated, suggesting abnormal renal function and pathological damage to the kidneys during the modeling process.

[0060] Serum uric acid levels in mice of each group were measured. The results showed that serum uric acid levels in both the high-concentration compound group and the low-concentration compound group were lower than those in the model group. The high-concentration compound group had a similar level to the benzbromarone group, indicating that both the high-concentration and low-concentration compound groups had uric acid-lowering effects, and the effect of the high-concentration compound group was similar to that of the positive control drug.

[0061] In the assessment system of hyperuricemia, serum creatinine and blood urea nitrogen (BUN) are important indicators for evaluating renal function, and they are closely related to serum uric acid levels. Elevated serum creatinine and BUN levels indicate abnormal renal function, suggesting pathological damage to the kidneys. Blood urea nitrogen levels in each group were measured. The results showed that the high-concentration and low-concentration compound groups had significantly lower BUN levels than the model group, indicating that both treatments possess renal protective capabilities and can improve elevated BUN caused by hyperuricemia. Serum creatinine levels were significantly lower in both the high-concentration and low-concentration compound groups, indicating that they can improve renal metabolic capacity under hyperuricemia damage, thereby assisting in lowering uric acid. The renal coefficients in both the high-concentration and low-concentration compound groups were significantly lower than those in the model group, further confirming their ameliorative effect on renal pathological damage.

[0062] In summary, both high-concentration and low-concentration compound preparations not only have good uric acid-lowering effects, but can also help lower uric acid by protecting kidney function and improving kidney metabolism. The effect of the high-concentration compound preparation is closer to that of the positive control drug benzbromarone, while the low-concentration compound preparation also shows certain uric acid-lowering and kidney-protective effects.

[0063] Hyperuricemia causes changes in the appearance and morphology of the kidneys and liver in mice, thus reflecting the damage and recovery caused by hyperuricemia. Figure 2 The results showed that the kidneys of the control group mice exhibited typical normal physiological morphology: regular shape, rosy color, and smooth surface. The kidneys of the model group mice showed significant pathological changes, including significantly reduced volume, pale and dull color, and a rough and uneven surface. The kidneys of the positive drug group mice showed a significant improvement in appearance compared to the model group, with morphological abnormalities effectively alleviated. The kidneys of both the high-concentration and low-concentration compound groups showed varying degrees of repair effect compared to the model group, exhibiting a healthy appearance with a rosy, full, and smooth surface, suggesting that this compound preparation has a clear ameliorative effect on hyperuricemia-induced kidney morphological damage.

[0064] Paraffin sections of the kidneys, livers, and colons of mice in each group were stained with hematoxylin and eosin (HE). Kidney sections were observed under 1x and 20x optical microscopes, and the results are as follows: Figure 3 The analysis revealed that the kidneys in the NC group had intact structures, with normal morphology of glomeruli, tubules, and interstitium, and no inflammation or pathological changes were observed. The kidneys in the model group showed severe structural damage, with significant large-scale inflammatory cell infiltration, glomerular cell proliferation with severe cell damage, tubular cell edema and enlarged lumen, and significant interstitial fibrosis. In the groups treated with benzbromarone, the lesions in the kidney tissue were significantly reduced compared to the model group. Both the high-concentration and low-concentration compound groups showed less damage to kidney sections than the model group, and the kidney edges were smoother, demonstrating a certain degree of recovery. Figure 4Liver sections were taken from mice. The control group showed intact liver structure, orderly cell arrangement, and no obvious pathological changes; the livers were healthy and normal. The model group showed severe liver damage with extensive inflammatory cell infiltration and disordered cell arrangement. The benzbromarone group showed slight structural abnormalities, mild sinusoidal dilation, and inflammatory cell infiltration. Both the low-concentration and high-concentration compound groups showed significant improvement, with orderly cell arrangement and no obvious inflammatory cell infiltration or abnormal sinusoidal dilation. This indicates that the combined use of adenine and potassium oxonate can induce hyperuricemia and liver damage in mice. Oral administration of the walnut peptide compound plant-based solid beverage can alleviate liver damage to some extent, reduce internal liver inflammation, and promote recovery from hyperuricemia-induced damage. Figure 5 The images show colon sections from mice. In the control group, the intestinal structure was normal, with normal villus length and structure, no pathological changes, and normal structure. In the model group, intestinal damage was significant. Villus length was shortened, and a large number of epithelial cells sloughed off and died. The crypt structure was disordered, and plasma cells at the base were increased. In the benzbromarone group, intestinal damage was significantly improved compared to the model group. Villus atrophied slightly, and a small amount of epithelial cells sloughed off. Some structural abnormalities appeared in the crypts. In the high-concentration compound group, intestinal damage was significantly improved compared to the model group. Villus atrophied slightly, and a small amount of epithelial cells sloughed off. There was no significant structural disorder in the crypts.

[0065] All the above results indicate that the uric acid-lowering walnut peptide compound plant solid beverage prepared in this invention exhibits significant uric acid-lowering activity. Serum biological index detection results clearly show that changes in blood urea nitrogen, serum creatinine, and kidney coefficient clearly demonstrate the outstanding effect of this product in lowering uric acid. Kidney morphology and histological observations further confirm that after product intervention, the kidneys of model mice showed a rosy and smooth appearance close to that of healthy mice, and the integrity of the tissue microstructure was significantly improved. The morphological characteristics of the glomeruli and renal tubules approached the normal physiological state, and the extent of inflammatory cell infiltration and the degree of tissue fibrosis showed a trend of alleviation. Liver histopathological analysis showed that the walnut peptide compound plant solid beverage can alleviate hepatocellular damage caused by hyperuricemia and effectively improve the pathological abnormalities of liver tissue. Simultaneously, colon histopathological evaluation results showed that the walnut peptide compound plant solid beverage can specifically reverse the intestinal barrier function damage caused by hyperuricemia: the length of intestinal villi in model mice was significantly restored, structural integrity was remodeled, epithelial cell shedding was significantly reduced, crypt structure approached the normal physiological configuration, and the degree of inflammatory cell infiltration was significantly reduced. This result indicates that the walnut peptide compound plant solid beverage not only has highly efficient uric acid-lowering activity, but also excellent protective and damage-repairing functions.

[0066] In summary, this invention, a compound plant-based solid beverage containing walnut peptides, has multiple effects such as lowering uric acid, protecting the kidneys, liver, intestines, and reducing inflammation. It provides an innovative approach for the daily management and prevention of hyperuricemia and has broad prospects for industrialization.

[0067] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A compound plant-based solid beverage containing walnut peptides to lower uric acid, characterized in that, The solid beverage comprises the following ingredients by weight: 15-30 parts walnut peptide, 5-15 parts sour cherry powder, 4-8 parts chicory powder, 4-8 parts celery powder, 5-10 parts dandelion powder, 4-8 parts lotus leaf alkaloid, 4-8 parts coix seed powder, 4-8 parts poria cocos powder, and 0.5-1 part angelica dahurica.

2. The uric acid-lowering walnut peptide compound plant solid beverage according to claim 1, characterized in that, The preparation method of the walnut peptide includes the following steps: dispersing walnut protein in distilled water at a solid-liquid ratio of 1:50 (w / v), heating to 90°C and holding for 15 minutes, cooling to 53°C and adjusting the pH to 7.0-7.2, adding neutral protease and enzymatically hydrolyzing at 53°C for 120 minutes, removing impurities by passing through a 0.2-micron membrane, and freeze-drying to obtain walnut peptide powder.

3. The uric acid-lowering walnut peptide compound plant solid beverage according to claim 1, characterized in that, The preparation method of the sour cherry powder is as follows: pit and crush fresh sour cherries, pre-freeze (-18℃, 2 hours), then freeze-dry for 36 hours under the conditions of cold trap temperature -45℃ and vacuum pressure 0.01kPa, and then crush through an 80-100 mesh sieve.

4. The uric acid-lowering walnut peptide compound plant solid beverage according to claim 1, characterized in that, The preparation method of the chicory powder is as follows: chicory root is chopped and added to 30% ethanol solution at a solid-liquid ratio of 1:

15. It is then extracted by hot reflux at 70-80℃ for 2 hours, concentrated, and spray-dried (inlet air temperature 160℃, outlet air temperature 80℃).

5. The uric acid-lowering walnut peptide compound plant solid beverage according to claim 1, characterized in that, The method for preparing the celery powder is as follows: fresh celery is pulverized and pre-frozen (-18℃, 2 hours), then freeze-dried for 36 hours under the conditions of cold trap temperature -45℃ and vacuum pressure 0.01kPa, and then pulverized through an 80-100 mesh sieve.

6. The uric acid-lowering walnut peptide compound plant solid beverage according to claim 1, characterized in that, The preparation method of the dandelion powder is as follows: after pulverizing the dried whole dandelion, add 50% ethanol solution at a solid-liquid ratio of 1:15, reflux extract twice at 60-70℃ for 1.5 hours each time, concentrate and spray dry (inlet air temperature 150℃, outlet air temperature 75℃).

7. The uric acid-lowering walnut peptide compound plant solid beverage according to claim 1, characterized in that, The method for preparing lotus leaf alkaloid is as follows: dried lotus leaves are pulverized and then added to 70% ethanol at a solid-liquid ratio of 1:

10. The mixture is refluxed at 75°C for 1.5 hours, purified by macroporous adsorption resin column, and then mixed with maltodextrin at a ratio of 1:

1. The mixture is then spray-dried (inlet air temperature 140°C, outlet air temperature 70°C) to obtain a lotus leaf alkaloid content ≥10%.

8. The uric acid-lowering walnut peptide compound plant solid beverage according to claim 1, characterized in that, The preparation method of Angelica dahurica powder is as follows: Angelica dahurica slices are pre-frozen (-18℃, 2 hours), freeze-dried for 48 hours under cold trap temperature of -45℃ and vacuum pressure of 0.01kPa, and then pulverized through an 80-100 mesh sieve.

9. A method for preparing a uric acid-lowering walnut peptide compound plant solid beverage as described in any one of claims 1-8, characterized in that, Includes the following steps: (1) Weigh walnut peptide, sour cherry powder, chicory powder, celery powder, dandelion powder, lotus leaf alkaloid, coix seed powder, poria powder and angelica powder according to the proportions described in claim 1; (2) Add the above raw materials to the mixer and mix evenly for 20–40 min; (3) Dry and sieve under vacuum conditions to obtain the solid beverage powder product.

10. The use of the uric acid-lowering walnut peptide compound plant solid beverage according to claim 1 in the preparation of food or medicine for lowering serum uric acid levels.

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