Preparation method and application of composition with effects of dispelling wind and promoting diuresis

Abstract

The invention provides a preparation method and application of a composition with the efficacy of dispelling wind and promoting diuresis, red beans, coix seeds, ginseng, sea cucumber body walls, poria cocos and fresh lalang grass rhizome are used as main raw materials, and extraction is performed by using an enzymolysis and ultrasonic composite extraction process, so that efficient extraction of active ingredients in the raw materials can be realized; the prepared composition has the effects of resisting oxidation, dispelling wind and promoting diuresis, and an oral preparation prepared from the composition as a raw material is convenient to carry and suitable for daily use.

Description

Technical Field

[0001] The present invention relates to the field of composition preparation, and in particular to a preparation method and application of a composition with wind-dispelling and dampness-eliminating effects. Background Art

[0002] Traditional Chinese medicine believes that rheumatism invades the human body, causing obstruction of qi and blood meridians, which in turn causes a series of symptoms such as joint pain, swelling, and difficulty in flexion and extension. Red beans have the effects of diuresis, swelling, detoxification and pus removal; coix seed can promote diuresis and eliminate dampness, strengthen the spleen and stop diarrhea; ginseng has the pharmacological effects of greatly replenishing vital energy, restoring pulse and strengthening deficiency; sea cucumber peptides, sea cucumber polysaccharides and other ingredients contained in the body wall of sea cucumbers have the effects of anti-inflammatory, enhancing immunity and improving blood circulation; Poria can strengthen the spleen and calm qi, and fresh Imperata root can cool blood and stop bleeding, clear heat and promote diuresis; but there is no existing technology that combines the above components according to the formula principle of monarch, minister, assistant and envoy to form corresponding products. In addition, although a variety of dehumidifying products are disclosed in the prior art, most of them are taken by taking with boiled water or boiling. For example, the dehumidifying and spleen-strengthening meal replacement powder disclosed in patent CN108634201A uses red bean, lentil, Poria cocos, coix seed, Euryale ferox, lotus seed, red bean, red date, wolfberry, tangerine peel, barley, bracken, yam, cassia seed, and sesame seed as raw materials, and is directly taken with water; the Chinese medicine composition with dehumidifying, phlegm-resolving and laxative effects disclosed in patent CN114931614A and its preparation method are dried and crushed Poria cocos, Euryale ferox, coix seed rice, barley tea, red bean, tangerine peel, Sophora japonica and hawthorn, and taken with boiled water; although these methods are convenient to eat, they may cause inconvenience in actual application because a large amount of liquid needs to be taken, especially when the amount of water intake needs to be controlled or when going out, working, etc., the compliance may be reduced. Summary of the invention

[0003] In order to solve the above problems, the present invention provides a preparation method and application of a composition with the effect of dispelling wind and dampness, which takes red bean, coix seed, ginseng, poria, fresh white grass root and sea cucumber body wall as main raw materials, and uses enzymatic hydrolysis and ultrasonic composite extraction process for extraction, which can achieve efficient extraction of active ingredients in the raw materials. The prepared composition has good antioxidant and wind-dispelling and dampness-dispelling effects, and the oral preparation made with the raw materials is easy to carry and suitable for daily use.

[0004] To achieve the above object, the present invention adopts the following technical solutions:

[0005] A method for preparing a composition having the effects of dispelling wind and removing dampness, comprising the following steps:

[0006] S1: crushing and drying red bean, coix seed, fresh Imperata root, ginseng and Poria to obtain plant raw material powder, and grinding the sea cucumber body wall with water to obtain sea cucumber homogenate;

[0007] S2: dissolving cellulase, papain and neutral protease in citric acid buffer to obtain a composite enzyme solution;

[0008] S3: mixing the plant raw material powder obtained in step S1 and the sea cucumber homogenate, adding them to the composite enzyme solution in step S2 for enzymolysis, the enzymolysis temperature is 40-60° C., and the enzymolysis time is 2-4 hours; after the enzymolysis is completed, ultrasonic extraction is performed, the ultrasonic temperature is 85° C., the ultrasonic power is 500 W, and the ultrasonic time is 15-45 minutes to obtain an extract;

[0009] S4: Concentrating the extract obtained in step S3 to obtain a composition having the effect of dispelling wind and removing dampness.

[0010] Red bean has the effects of promoting diuresis and reducing swelling, detoxifying and discharging pus, plays a positive role in regulating the body's water metabolism, can promote urine excretion, reduce the accumulation of moisture in the body, and thus relieve symptoms such as body heaviness and edema caused by dampness; coix seed promotes diuresis and eliminates dampness, strengthens the spleen and stops diarrhea, can inhibit the production and release of inflammatory factors, and reduce joint inflammatory reactions; ginseng has the pharmacological effects of greatly replenishing vital energy, restoring pulse and strengthening deficiency, tonifying the spleen and lungs, promoting fluid and nourishing blood, calming the nerves and improving intelligence, and helps to improve the body's anti-rheumatic ability; the body wall of sea cucumber contains sea cucumber peptides, sea cucumber polysaccharides and other ingredients, which can reduce inflammatory reactions, enhance the body's immunity, promote the removal of dampness and the recovery of damaged tissues; the ingredients such as Poria polysaccharide in Poria can regulate water and salt balance, dilate renal blood vessels, promote urine excretion and promote diuresis; fresh Imperata root has the effects of cooling blood and stopping bleeding, clearing heat and promoting diuresis, and has a good therapeutic effect on urinary problems and edema caused by damp-heat accumulation in the bladder.

[0011] As a further improvement of the above scheme, the weight proportions of the components in step S1 are: 20 parts of red beans, 30 parts of coix seeds, 10 parts of fresh Imperata roots, 5 parts of ginseng, 10 parts of Poria cocos, and 25 parts of sea cucumber body wall.

[0012] As a further improvement of the above scheme, the concentration of the citric acid buffer in step S2 is 2-8 mg / mL, preferably 4 mg / mL; the concentrations of various enzymes in the complex enzyme solution are: cellulase 2 mg / mL, papain 1 mg / mL, neutral protease 1 mg / mL.

[0013] As a further improvement of the above scheme, the enzymatic hydrolysis temperature in step S3 is 50° C. and the enzymatic hydrolysis time is 4 h.

[0014] As a further improvement of the above scheme, the ultrasonic time in step S3 is 30 minutes.

[0015] As a further improvement of the above scheme, the preparation method comprises the following steps:

[0016] S1: According to weight, 20 parts of red beans, 30 parts of coix seeds, 10 parts of fresh Imperata roots, 5 parts of ginseng and 10 parts of Poria cocos are crushed and dried to obtain plant raw material powder, and 25 parts of sea cucumber body wall are added with water and ground to obtain sea cucumber homogenate;

[0017] S2: dissolving cellulase, papain and neutral protease in 4 mg / mL citric acid buffer to obtain a composite enzyme solution, wherein the concentrations of various enzymes in the composite enzyme solution are: 2 mg / mL cellulase, 1 mg / mL papain and 1 mg / mL neutral protease;

[0018] S3: Mix the plant raw material powder obtained in step S1 and the sea cucumber homogenate, add 30 times the weight of the composite enzyme solution in step S2 for enzymolysis, the enzymolysis temperature is 50° C., and the enzymolysis time is 4 hours; after the enzymolysis is completed, ultrasonic extraction is performed, the ultrasonic temperature is 85° C., the ultrasonic power is 500 W, and the ultrasonic time is 30 minutes to obtain an extract;

[0019] S4: Concentrating the extract obtained in step S3 to obtain a composition having the effect of dispelling wind and removing dampness.

[0020] As a further improvement of the above scheme, in step S1, the plant raw materials including red beans, coix seeds, fresh white imperata root, ginseng and Poria cocos are crushed and passed through a 30-80 mesh sieve, and dried in an oven at a drying temperature of 40-60°C and a drying time of 5-20 hours to obtain the plant raw material powder; 0.5-5 times the mass of water is added when the sea cucumber body wall is ground. Preferably, the plant raw materials are crushed and passed through a 60 mesh sieve, and dried in an oven at a drying temperature of 50°C and a drying time of 15 hours, and 2 times the mass of water is added when the sea cucumber body wall is ground.

[0021] As a further improvement of the above scheme, the plant raw material powder and the sea cucumber body wall homogenate in step S3 are mixed and added to the complex enzyme solution at a mass ratio of 1:10-60, preferably 1:30.

[0022] As a further improvement of the above scheme, the extract in step S4 is concentrated to 10%-60% of the original volume at a temperature of 30-60°C and a pressure of -0.04MPa--0.10MPa. Preferably, the extract in step S4 is concentrated to 30% of the original volume at a temperature of 55°C and a pressure of -0.09MPa.

[0023] An application of a composition with the effect of dispelling wind and removing dampness prepared by any of the preparation methods described above, comprising using the composition with the effect of dispelling wind and removing dampness as a raw material to prepare an oral preparation for dispelling wind and removing dampness.

[0024] As a further improvement of the above scheme, the oral preparation comprises, by weight: 60-80 parts of the composition having the effect of dispelling wind and removing dampness, 10-20 parts of excipients, 2-6 parts of sweeteners, 1-4 parts of flavoring agents, 1-3 parts of preservatives and 2-5 parts of thickeners.

[0025] As a further improvement of the above scheme, the excipient is at least one of lactose, maltodextrin, and microcrystalline cellulose; the sweetener is at least one of xylitol, mannitol, maltitol, oligomaltose, and sorbitol; the flavoring agent is at least one of citric acid, malic acid, tartaric acid, and taurine; the preservative is at least one of benzoic acid, sodium benzoate, sorbic acid, and potassium sorbate; and the thickener is at least one of cyclodextrin, pectin, carrageenan, sodium carboxymethyl cellulose, and hydroxypropyl methylcellulose.

[0026] As a further improvement of the above scheme, the oral preparation comprises, by weight: 70 parts of the composition having the effect of dispelling wind and removing dampness, 10 parts of maltodextrin, 6 parts of microcrystalline cellulose, 3 parts of isomaltooligosaccharide, 2 parts of sorbitol, 2 parts of taurine, 2 parts of potassium sorbate, 2 parts of cyclodextrin and 3 parts of pectin.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] (1) Red bean, coix seed, ginseng, Poria cocos, fresh Imperata cylindrica, and sea cucumber body wall are combined according to the principle of monarch, minister, assistant, and envoy, and are compatible and synergistic with each other: coix seed and red bean are monarch drugs, which play the main role in dispelling wind and dampness; fresh Imperata cylindrica root is the assistant drug, which cooperates with the monarch drug to enhance the effect of dispelling dampness and clearing heat; ginseng and sea cucumber body wall are assistant drugs, which are rich in nutrients such as protein and polysaccharides, can replenish the body's positive energy, strengthen the body, and help the body recover better; Poria cocos is the assistant drug, which has a mild medicinal property and plays a role in harmonizing the various drugs;

[0029] (2) The raw material components are processed by using an enzymatic hydrolysis combined with ultrasonic extraction process. Enzymatic hydrolysis can degrade macromolecular substances such as proteins and polysaccharides in the raw materials into small molecules, increasing their solubility and extractability. The cavitation effect and mechanical vibration generated by ultrasonic waves can further destroy the cell structure and strengthen the mass transfer process. The complex enzyme can also be quickly inactivated at high temperatures. The two work together to make the target components in the raw materials easier to dissolve from the raw materials. Moreover, by reasonably controlling various conditions, not only can the extraction efficiency of the active ingredients in the raw materials be improved, but also the extraction time can be shortened, cost can be saved, and energy consumption can be reduced.

[0030] (3) Experiments have shown that the oral preparation prepared by the composition obtained by extracting and concentrating the raw materials of the present invention has a good antioxidant effect, can effectively slow down the weight gain of obese rats with phlegm-damp constitution, and has a lipid-lowering effect. It can also promote urination and sodium and potassium ion excretion in water-loaded mice, has the effect of dispelling wind and dampness, and is easy to carry and suitable for daily consumption. DETAILED DESCRIPTION

[0031] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0032] The weight of the relevant components mentioned in the embodiments of the present invention description can not only refer to the specific content of each component, but also represent the proportional relationship of the weights of the components. Therefore, as long as the content of the relevant components in the embodiments of the present invention description is proportionally enlarged or reduced, it is within the scope disclosed in the embodiments of the present invention description. Specifically, the mass described in the embodiments of the present invention description can be a mass unit known in the chemical industry such as μg, mg, g, kg, etc.

[0033] In order to facilitate the understanding of the present invention, the present invention will be described more comprehensively and carefully below in combination with the preferred embodiments of the specification, but the protection scope of the present invention is not limited to the following specific embodiments.

[0034] Unless otherwise defined, all professional terms used below have the same meanings as those generally understood by those skilled in the art. The professional terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention.

[0035] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0036] Example 1

[0037] S1: According to weight, 20 parts of red beans, 30 parts of coix seeds, 10 parts of fresh Imperata root, 5 parts of ginseng and 10 parts of Poria were washed with water, crushed, passed through a 60-mesh sieve, and dried at 50°C for 15h to obtain plant raw material powder; 25 parts of sea cucumber body wall were washed with water, and then 2 times the weight of water was added to fully grind to obtain sea cucumber homogenate;

[0038] S2: dissolving cellulase, papain and neutral protease in 4 mg / mL citric acid buffer to obtain a composite enzyme solution, wherein the concentrations of various enzymes in the composite enzyme solution are: 2 mg / mL cellulase, 1 mg / mL papain and 1 mg / mL neutral protease;

[0039] S3: Mix the plant raw material powder obtained in step S1 and the sea cucumber homogenate, add 30 times the mass of the composite enzyme solution obtained in step S2 for enzymolysis, the enzymolysis temperature is 50°C, and the enzymolysis time is 2h; after the enzymolysis is completed, ultrasonic extraction is performed, the ultrasonic temperature is 85°C, the ultrasonic power is 500W, and the ultrasonic time is 30min, followed by centrifugal filtration, the centrifugal treatment speed is 12000r / min, the treatment time is 10min, and a ceramic membrane with a pore size of 500nm is selected for filtration and separation to obtain an extract L1;

[0040] S4: The extract L1 obtained in step S3 was concentrated to 30% of the original volume by rotary evaporation under reduced pressure at a temperature of 55° C. and a pressure of minus 0.09 MPa to obtain a coix ginseng and poria composition L1;

[0041] S5: According to weight proportions, 70 parts of Coix ginseng and Poria composition L1 are stirred with 10 parts of maltodextrin, 6 parts of microcrystalline cellulose, 3 parts of isomaltooligosaccharides, 2 parts of sorbitol, 2 parts of taurine, 2 parts of potassium sorbate, 2 parts of epoxidized dextrin and 3 parts of pectin for 30 minutes to obtain Coix ginseng and Poria oral liquid L1, which is then filled into bottles sterilized at high temperature and sterilized again after filling to obtain the final product.

[0042] Example 2

[0043] The enzymatic hydrolysis temperature in step S3 of Example 1 was adjusted to 30° C., and the remaining step parameters were the same as in Example 1, to obtain extract L2 and Coix ginseng and Poria oral liquid L2.

[0044] Example 3

[0045] The enzymatic hydrolysis temperature in step S3 of Example 1 was adjusted to 40° C., and the remaining step parameters were the same as in Example 1, to obtain extract L3 and Coix ginseng and Poria oral liquid L3.

[0046] Example 4

[0047] The enzymatic hydrolysis temperature in step S3 of Example 1 was adjusted to 60° C., and the remaining step parameters were the same as in Example 1, to obtain extract L4 and Coix ginseng and Poria oral liquid L4.

[0048] Example 5

[0049] The enzymatic hydrolysis time in step S3 of Example 1 was adjusted to 1 h, and the parameters of the other steps were the same as those of Example 1, to obtain extract L5 and Coix ginseng and Poria oral liquid L5.

[0050] Example 6

[0051] The enzymatic hydrolysis time in step S3 of Example 1 was adjusted to 3 h, and the parameters of the other steps were the same as those of Example 1, to obtain extract L6 and Coix ginseng and Poria oral liquid L6.

[0052] Example 7

[0053] The enzymatic hydrolysis time in step S3 of Example 1 was adjusted to 4 h, and the parameters of the other steps were the same as those of Example 1, to obtain extract L7 and Coix ginseng and Poria oral liquid L7.

[0054] Example 8

[0055] The ultrasonic time in step S3 of Example 1 was adjusted to 15 min, and the parameters of other steps were the same as those of Example 1, to obtain extract L8 and Coix ginseng and Poria oral liquid L8.

[0056] Example 9

[0057] The ultrasonic time in step S3 of Example 1 was adjusted to 45 min, and the parameters of other steps were the same as those of Example 1, to obtain extract L8 and Coix ginseng and Poria oral liquid L8.

[0058] Comparative Example 1

[0059] The ultrasonic extraction in step S3 of Example 1 was replaced by constant temperature water bath extraction, the water bath temperature was 85° C., the water bath time was 60 min, and the remaining step parameters were the same as in Example 1, to obtain extract D1 and Coix ginseng and Poria oral liquid D1.

[0060] Comparative Example 2

[0061] The enzymatic hydrolysis operation in step S3 of Example 1 was removed, and the plant raw material powder obtained in step S1 and the sea cucumber homogenate were mixed, and then 30 times the mass of ultrapure water were added for ultrasonic extraction. The ultrasonic temperature was 85°C, the ultrasonic power was 500 W, and the ultrasonic time was 30 min. The remaining step parameters were the same as those in Example 1, and the extract D2 and Coix ginseng and Poria oral liquid D2 were obtained.

[0062] Comparative Example 3

[0063] The ultrasonic extraction time in Comparative Example 2 was replaced with 2.5 h, and the remaining step parameters were the same as Comparative Example 2, to prepare extract D3 and Coix ginseng and Poria oral liquid D3.

[0064] Test Example 1: Determination of the yield of polysaccharides and small molecule peptides

[0065] 1. Polysaccharide yield test

[0066] The extracts obtained in Examples 1-9 and Comparative Examples 1-3 were used as samples to test the polysaccharide yields according to the following methods.

[0067] Prepare a standard curve: Accurately weigh 50 mg of standard glucose into a 500 mL volumetric flask, add water to make up to the volume, and obtain a standard solution. Pipette different volumes of standard solution separately and add distilled water to 2.0 mL. Then add 1.0 mL of 6 vol% phenol aqueous solution and 5.0 mL of 98% concentrated sulfuric acid to each sample tube, shake well and let stand for 30 minutes. Use an ultraviolet spectrophotometer to measure the absorbance at a wavelength of 490 nm. Use distilled water as a blank control, and draw a standard curve based on the concentration of standard glucose and the corresponding absorbance.

[0068] Sample determination: Take 1.0 mL of each extract sample, dilute it 100 times with water, take 1 mL of each diluted sample, add 1.0 mL of distilled water, then add 1.0 mL of 6 vol% phenol aqueous solution, and quickly add 5.0 mL of 98% concentrated sulfuric acid. Oscillate on a vortex to mix thoroughly, let it stand for 30 minutes, and measure the absorbance at a wavelength of 490 nm.

[0069] Calculation of results: Substitute the measured sample absorbance into the standard curve equation: y = mx + b, where: y is the sample absorbance (A value), x is the polysaccharide concentration, m is the slope of linear regression, and b is the intercept. Calculate the concentration of polysaccharides in the diluted sample, and convert the actual concentration C after the extraction according to the dilution multiple. Calculate the polysaccharide yield according to the volume V of the solvent and the total mass M of the raw materials: Y1 = C*V / M.

[0070] 2. Small molecule peptide yield test

[0071] The extracts obtained in Examples 1-9 and Comparative Examples 1-3 were ultrafiltered through a hollow cellulose membrane with a molecular weight cutoff of 10,000 and then freeze-dried to obtain a small molecule polypeptide powder, the mass of which was recorded as M'; the total mass of all the raw materials input was recorded as M 总 , Y2=M' / M 总 The freeze-drying conditions were set as follows: temperature -70°C, pressure 50Pa, and processing time 12h.

[0072] The test results of polysaccharide yield and small molecule polypeptide are shown in Table 1. The results show that for the raw material components of the present invention, suitable enzymatic hydrolysis conditions and ultrasonic process play a vital role in the extraction of polysaccharides and small molecule polypeptides. If the conditions are not properly controlled, the extraction rate of polysaccharides and small molecule polypeptides in the raw material will be significantly reduced.

[0073] Table 1 Yield of polysaccharides and small molecule peptides in each group of extracts

[0074] Group Polysaccharide yield Y1 / (mg / g) Small molecule peptide yield Y2 / (mg / g) Example 1 108.5 57.4 Example 2 96.9 48.5 Example 3 103.3 53.6 Example 4 107.4 57.8 Example 5 83.6 43.4 Example 6 111.7 59.2 Example 7 115.5 60.3 Example 8 101.8 52.4 Example 9 106.4 57.5 Comparative Example 1 93.6 47.2 Comparative Example 2 63.4 32.3 Comparative Example 3 90.5 45.7

[0075] Test Example 2: DPPH free radical scavenging test

[0076] The DPPH free radical scavenging test was performed on the Yishenling oral liquid L1, D1 and D2 prepared in Example 1, Comparative Example 1 and Comparative Example 2 respectively according to the following method:

[0077] Set up sample tubes (T), sample background (T0), DPPH tubes (C) and solvent background (C0). For each sample tube (T) of Coix Seed and Poria Oral Liquid, three parallel tubes are required. Use water as the diluent to prepare Coix Seed and Poria Oral Liquid solutions of different concentrations (0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL). For example, take 1 mg / mL Coix Seed and Poria Oral Liquid solution as an example to set up the following sample tubes (T), sample background (T0), DPPH tubes (C) and solvent background (C0):

[0078] Sample tube (T): 1 mL of Yishenling oral solution (concentration of 1 mg / mL) + 2 mL of water + 1 mL of DPPH ethanol solution (concentration of 0.04 mg / mL);

[0079] Sample background tube (T0): 1 mL of Yishenling oral solution (concentration of 1 mg / mL) + 2 mL of water + 1 mL of 95 vol% ethanol solution;

[0080] DPPH tube (C): 3 mL of water + 1 mL of DPPH ethanol solution (concentration 0.04 mg / mL);

[0081] Solvent background tube (C0): 3 mL water + 1 mL 95% vol ethanol solution;

[0082] The solutions in the above reaction tubes were transferred into cuvettes, the absorbance was measured at 517 nm, and the DPPH radical scavenging rate of Coix Seed and Poria Oral Liquid was calculated by the following formula (I). The results are shown in Table 2.

[0083] Formula I:

[0084] The results show that the coix ginseng and poria oral liquid prepared in the present invention has a good scavenging effect on DPPH free radicals, and the coix ginseng and poria oral liquid L1 prepared in Example 1 has better scavenging ability on DPPH free radicals than the coix ginseng and poria oral liquids D1 and D2 prepared in Comparative Examples 1 and 2.

[0085] Table 2 DPPH free radical scavenging rate of different concentrations of Yishenling oral liquid

[0086] Concentration of Yishenling oral solution 0.125mg / mL 0.25mg / mL 0.5mg / mL 1mg / mL 2mg / mL Coix ginseng and ling oral liquid L1 14.56％ 27.35％ 48.57％ 65.53％ 87.57％ Coix ginseng and ling oral liquid D1 8.31％ 19.57％ 36.74％ 54.20％ 72.38％ Coix ginseng and ling oral liquid D2 5.75％ 11.64％ 21.80％ 38.62％ 59.57％

[0087] Test Example 3: Hydroxyl radical scavenging test

[0088] The Coix ginseng and Poria oral liquids L1, D1, and D2 prepared in Example 1, Comparative Example 1, and Comparative Example 2 were respectively mixed with water to prepare Coix ginseng and Poria oral liquid solutions of different concentrations (0.0625 mg / mL, 0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, and 1 mg / mL), and hydroxyl radical scavenging tests were performed according to the following method.

[0089] Set up the sample tube (A 样品 )、Loss pipe (A 损 )、Undamaged(A 未损 ), set up 3 parallel tubes for each sample tube (T) of Yishenling oral liquid, taking 1mg / mL Yishenling oral liquid solution as an example:

[0090] A 样品 : 1mL of o-phenanthroline ethanol solution (concentration of 5mmol / L) + 2mL of phosphate buffer (concentration of 0.2mol / L) + 1mL of Coix Seed and Poria oral solution + 1mL of ferrous sulfate solution (5mmol / L) + 1mL of H2O2 solution (concentration of 0.1% v / v);

[0091] A 损 : 1mL of o-phenanthroline ethanol solution (concentration of 5mmol / L) + 2mL of phosphate buffer (concentration of 0.2mol / L) + 1mL of ultrapure water + 1mL of ferrous sulfate solution (5mmol / L) + 1mL of H2O2 solution (concentration of 0.1% v / v);

[0092] A 未损 : 1 mL of o-phenanthroline ethanol solution (concentration of 5 mmol / L) + 2 mL of phosphate buffer (concentration of 0.2 mol / L) + 1 mL of ferrous sulfate solution (5 mmol / L) + 2 mL of ultrapure water.

[0093] The solutions in the above reaction tubes were transferred into cuvettes, the absorbance was measured at 536 nm, and the hydroxyl radical scavenging rate of Coix Seed and Poria Oral Liquid was calculated by the following formula II. The results are shown in Table 3.

[0094] Formula II:

[0095] The results show that the coix ginseng and poria oral liquid prepared in the present invention has a good scavenging effect on hydroxyl free radicals, and the coix ginseng and poria oral liquid L1 prepared in Example 1 has a much better scavenging ability on hydroxyl free radicals than the coix ginseng and poria oral liquids D1 and D2 prepared in Comparative Examples 1 and 2.

[0096] Table 3 Hydroxyl radical scavenging rate of different concentrations of Yishenling oral liquid in each group

[0097] Concentration of Yishenling oral solution 0.0625mg / mL 0.125mg / mL 0.25mg / mL 0.5mg / mL 1mg / mL Coix ginseng and ling oral liquid L1 23.55％ 36.72％ 49.57％ 68.73％ 85.37％ Coix ginseng and ling oral liquid D1 15.64％ 28.72％ 38.35％ 55.14％ 74.35％ Coix ginseng and ling oral liquid D2 6.55％ 18.84％ 27.36％ 40.71％ 59.32％

[0098] Test Example 4: Weight-loss effect of Yishenling oral liquid on obese rats with phlegm-damp constitution

[0099] 1. Experimental methods

[0100] (1) Fifty male SD rats, 8 weeks old, weighing 150-180 g each, were selected. They were fed with adaptive basal feed for one week before the experiment. Ten rats were randomly selected as the normal diet group and fed with basal feed. The breeding environment was 2-3 rats per cage, the breeding temperature was 22±2℃, the humidity was 60%-70%, and the fluorescent lighting and dark environment were alternated every 12 hours. The rest were used to prepare the rat model of phlegm-damp constitution. The basal feed formula was 55% nitrogen-free extract, 18% crude protein, 10% water, 8% crude ash, 5% crude fiber, and 4% crude fat; the calcium addition was 1.5% and the phosphorus addition was 1%.

[0101] (2) Preparation of obese rat model of phlegm-damp constitution: 40 rats were randomly selected from the above 50 rats and placed in an environment with a temperature of 18±2℃ and a relative humidity of 85%-95% and fed with a high-fat diet. The modeling time was about one month. After the modeling was completed, they were placed in a natural environment and randomly divided into 4 groups according to the random number method, namely, a high-fat diet group, a Coix ginseng and Poria oral liquid L1 group, a Coix ginseng and Poria oral liquid D1 group, and a Coix ginseng and Poria oral liquid D2 group. The high-fat diet formula was basal feed + 10% lard + 10% egg yolk powder. The animals and materials required for the experiment were all from the College of Animal Science and Technology of Hunan Agricultural University.

[0102] The normal diet group was fed with a basic feed, and the other groups were fed with a high-fat diet. The experimental groups were gavaged with 5 g / kg / d of each of the Yishenling oral liquid L1, Yishenling oral liquid D1, and Yishenling oral liquid D2 prepared by the present invention, and the normal diet group and the high-fat diet group were gavaged with an equal amount of distilled water. Each group was given once a day for 28 consecutive days, and the specific grouping is shown in Table 4.

[0103] Table 4 Specific groups of rat diet

[0104]

[0105]

[0106] (3) Collection of serum and sample tissue: After the experiment, the rats were fasted for 12 hours and anesthetized with 3.5% chloral hydrate at a dose of 0.1 ml / 10 g of the rat's body weight. Blood samples were collected from the abdominal aorta. The serum was collected by centrifugation under low temperature conditions and stored at -80°C. After the blood collection operation was completed, the rats were quickly dissected, the portal duct system was ligated and cut, the complete liver tissue was taken out, the blood attached to the liver tissue was rinsed off with physiological saline, placed in a sterile culture dish, weighed and stored at -80°C, and the liver tissue was fixed in 4% paraformaldehyde.

[0107] (4) Body weight measurement: During the experiment, the animal morphology was observed, the rats were weighed weekly, and the initial body weight, weekly body weight, and body weight before sacrifice were recorded.

[0108] (5) Determination of serum biochemical indicators: The levels of triglyceride (TG) and total cholesterol (TC) in serum were determined using a fully automatic biochemical analyzer.

[0109] (6) Determination of body fat: After the experiment, the rats were dissected, and perirenal fat and peritesticular fat were collected, weighed, and recorded. Peritoneal adipose tissue: The rat testicles were found in the lower abdomen, and the white fat attached to them was found at a suitable location. The white fat was peeled off along the vas deferens to the end of the testicles with surgical scissors. Perirenal adipose tissue: The kidneys were found on both sides of the lumbar spine outside the peritoneum, the retroperitoneum was cut open, the kidneys on both sides were removed, and the white fat attached to them was cut off.

[0110] 2. Test results

[0111] (1) Effect of Yishenling Oral Liquid on Body Weight of Obese Rats with Phlegm-Dampness Constitution

[0112] The changes in the body weight of rats in each group are shown in Table 5. The results showed that at the beginning of the experiment, the initial body weights of rats in the high-fat diet group and the Yishenling oral liquid L1, D1, and D2 groups were similar, and the body weights of rats in the normal diet group were lower. During the entire feeding process, the body weights of rats in each group showed an overall upward trend. After 4 weeks of the experiment, the body weights of rats in the Yishenling oral liquid L1, D1, and D2 groups decreased significantly compared with those in the high-fat diet group, among which the body weights of rats in the Yishenling oral liquid L1 group decreased the most.

[0113] Table 5 Body weight changes of rats in each group

[0114] Group Week 0 Week 2 Week 4 Normal diet group 485.7g 521.5g 533.8g High-fat diet group 598.3g 683.4g 736.5g Yishenling oral liquid L1 group 600.6g 625.5g 638.4g Yishenling oral liquid D1 group 599.5g 636.8g 657.2g Yishenling oral liquid D2 group 598.8g 653.1g 678.5g

[0115] (2) Effects of Yishenling Oral Liquid on Serum Indexes of Obese Rats with Phlegm-Dampness Constitution

[0116] After the experiment, the serum triglyceride (TG) and total cholesterol (TC) contents of rats in each group were tested, and the average values ​​were reported, as shown in Table 6. The results showed that the triglyceride content of the normal diet group was lower than that of all other groups; compared with the high-fat diet group, the triglyceride content of rats in the Yishenling Oral Liquid L1, D1, and D2 groups was significantly lower, indicating that Yishenling Oral Liquid can reduce the content of triglycerides in the serum of obese rats with phlegm-damp constitution, among which the effect of Yishenling Oral Liquid L1 group was the most obvious. The total cholesterol content of rats in the high-fat diet group was significantly higher than that of all other groups, and the total cholesterol content of rats in the Yishenling Oral Liquid L1, D1, and D2 groups was not much different from that of the normal diet group, indicating that Yishenling Oral Liquid can restore the total cholesterol of obese rats with phlegm-damp constitution to normal levels, among which the improvement effect of Yishenling Oral Liquid L1 group was the most obvious.

[0117] Table 6 Serum indexes of rats in each group

[0118] Group TG content (mmol / L) TC content (mmol / L) Normal diet group 1.12 1.55 High-fat diet group 1.96 2.64 Yishenling oral liquid L1 group 1.21 1.77 Yishenling oral liquid D1 group 1.25 1.82 Yishenling oral liquid D2 group 1.33 1.90

[0119] (3) Effect of Yishenling Oral Liquid on Body Fat in Obese Rats with Phlegm-Dampness Constitution

[0120] The rats in each group were dissected, and the perirenal fat and peritesticular fat were taken out and weighed. The ratio of fat to body weight was calculated and the average value was reported. The results are shown in Table 7. The results showed that the fat body ratio of rats in the high-fat diet group was the highest among all groups, and the fat body ratio of rats in the Yishenling oral liquid L1, D1, and D2 groups was close to that of the normal diet group, among which the Yishenling oral liquid L1 group was the closest.

[0121] Table 7 Body fat of rats in each group

[0122] Group Lipid body ratio (%) Normal diet group 0.051 High-fat diet group 0.097 Yishenling oral liquid L1 group 0.058 Yishenling oral liquid D1 group 0.063 Yishenling oral liquid D2 group 0.071

[0123] Test Example 5: Diuretic effect of Yishenling oral liquid on water-loaded mice

[0124] 1. Experimental methods

[0125] 60 male mice (SPF) with a body weight of 18-22g were selected and fed for 7 days in an environment with a temperature of 25±2℃, a humidity of 60%-70%, normal light day and night alternation, and free diet. The mice were screened before the experiment. After adaptive feeding, 60 mice were fasted but not watered for 12h and gavaged with 0.02mL / g of normal saline. Mice with a total urine volume of more than 40% of the gavage dose in 2h met the requirements. The 40 mice that met the requirements were randomly divided into the Coixen and Poria oral liquid L1 group, the Coixen and Poria oral liquid D1 group, the Coixen and Poria oral liquid D2 group, the positive drug (hydrochlorothiazide) group, and the blank group, with 8 mice in each group. The animals and materials required for the experiment were all from the College of Animal Science and Technology, Hunan Agricultural University.

[0126] (1) Water load experiment: 0.1 mL of drug was continuously administered by gavage for 5 days per 10 g of mouse body weight. The body weight was measured after fasting for 18 hours in the afternoon of the 5th day, and the water load model experiment was started as the normal control value. Normal saline was administered by gavage at a dose of 0.02 mL / g to cause water and sodium retention. After 0.5 hours, the mouse abdomen was gently pressed to empty the remaining urine. The dose of hydrochlorothiazide was converted to 0.8 mg / mL. The corresponding experimental drugs were gavaged to mice in each experimental group (Coix Seed and Poria Oral Liquid L1, D1, D2 groups and positive drug groups) at a dose of 0.01 mL / g, and normal saline was given to the blank group.

[0127] (2) Urine collection: The urine of mice was collected at 0h, 2h, 4h, and 6h after gavage, and the changes in the body weight of mice at each time period were recorded. During the experiment, mice in each group were fasted and deprived of water.

[0128] (3) ICP-OES determination of Na + , K + Concentration: 20 μL of urine supernatant after centrifugation (3000 r / min, 10 min) was aspirated and the volume was fixed to 6 mL with nitric acid solution (1%). The sodium and potassium ions in the sample were determined according to the ICP-OES method, where the baseline of the sodium element was selected as 589.592 nm and the baseline of the potassium element was selected as 766.490 nm.

[0129] 2. Test results

[0130] (1) Effect of Yishenling oral solution on body weight of water-loaded mice

[0131] The body weight loss of mice in each group is shown in Table 8. The results showed that compared with the body weight at 0h, the body weight of mice decreased to varying degrees; compared with the blank group, the body weight of mice in the Yishenling oral liquid L1, D1, D2 groups and the positive drug group decreased significantly.

[0132] Table 8 Body weight loss of mice in each group at different time

[0133]

[0134] (2) Effect of Yishenling oral solution on urine output in water-loaded mice

[0135] As shown in Table 9, compared with the blank group, the mice in the Coix Seed and Poria Oral Liquid L1, D1, D2 groups and the positive drug group had higher urine output within 6 hours. Among the three Coix Seed and Poria Oral Liquid experimental groups, the urine output of mice in the Coix Seed and Poria Oral Liquid L1 group was the highest, indicating that Coix Seed and Poria Oral Liquid L1 had the best diuretic effect.

[0136] Table 9 Urine output of mice in each group at different times

[0137]

[0138]

[0139] (3) Effect of Yishenling oral liquid on Na in mouse urine + , K + Impact

[0140] As shown in Table 10, the Yishenling oral liquid L1, D1, and D2 groups had significant sodium and potassium benefits compared with the blank group, indicating that the Yishenling oral liquid prepared by the present invention can promote the sodium + , K + The excretion of dandruff can achieve the effect of diuresis and dehumidification, among which the coix ginseng and poria oral liquid prepared in Example 1 has the best effect.

[0141] Table 10 Na in urine of mice in each group + , K + content

[0142] Group <![CDATA[C Na+ (μg / mL)]]> <![CDATA[C K+ (μg / mL)]]> Blank Group 5.21 5.37 Positive drug group 12.25 11.38 Yishenling oral liquid L1 group 7.68 9.52 Yishenling oral liquid D1 group 7.15 9.21 Yishenling oral liquid D2 group 6.73 8.66

[0143] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A method for preparing a composition having the effect of dispelling wind and removing dampness, characterized in that: The following steps are involved: S1: crush and dry red bean, coix seed, fresh Imperata root, ginseng and Poria cocos to obtain plant raw material powder, and grind the sea cucumber body wall with water to obtain sea cucumber homogenate; S2: dissolving cellulase, papain and neutral protease in citric acid buffer to obtain a composite enzyme solution; S3: mixing the plant raw material powder obtained in step S1 and the sea cucumber homogenate, adding them to the composite enzyme solution in step S2 for enzymolysis, the enzymolysis temperature is 40-60° C., and the enzymolysis time is 2-4 hours; after the enzymolysis is completed, ultrasonic extraction is performed, the ultrasonic temperature is 85° C., the ultrasonic power is 500 W, and the ultrasonic time is 15-45 minutes to obtain an extract; S4: Concentrating the extract obtained in step S3 to obtain a composition having the effect of dispelling wind and removing dampness.

2. The preparation method according to claim 1, characterized in that The weight proportions of the components in step S1 are: 20 parts of red beans, 30 parts of coix seeds, 10 parts of fresh Imperata roots, 5 parts of ginseng, 10 parts of Poria cocos, and 25 parts of sea cucumber body wall.

3. The preparation method according to claim 1, characterized in that: The concentrations of various enzymes in the complex enzyme solution in step S2 are: 2 mg / mL cellulase, 1 mg / mL papain, and 1 mg / mL neutral protease.

4. The preparation method according to claim 1, characterized in that: The enzymolysis temperature in step S3 is 50° C. and the enzymolysis time is 4 h.

5. The preparation method according to claim 1, characterized in that: The ultrasonic time in step S3 is 30 minutes.

6. The preparation method according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1: According to weight, 20 parts of red beans, 30 parts of coix seeds, 10 parts of fresh Imperata roots, 5 parts of ginseng and 10 parts of Poria cocos are crushed and dried to obtain plant raw material powder, and 25 parts of sea cucumber body wall are added with water and ground to obtain sea cucumber homogenate; S2: dissolving cellulase, papain and neutral protease in 4 mg / mL citric acid buffer to obtain a composite enzyme solution, wherein the concentrations of various enzymes in the composite enzyme solution are: 2 mg / mL cellulase, 1 mg / mL papain and 1 mg / mL neutral protease; S3: Mix the plant raw material powder obtained in step S1 and the sea cucumber homogenate, add 30 times the weight of the composite enzyme solution in step S2 for enzymolysis, the enzymolysis temperature is 50° C., and the enzymolysis time is 4 hours; after the enzymolysis is completed, ultrasonic extraction is performed, the ultrasonic temperature is 85° C., the ultrasonic power is 500 W, and the ultrasonic time is 30 minutes to obtain an extract; S4: Concentrating the extract obtained in step S3 to obtain a composition having the effect of dispelling wind and removing dampness.

7. An application of a composition having the effect of dispelling wind and removing dampness obtained by the preparation method according to any one of claims 1 to 6, characterized in that: The invention comprises using the composition with the function of dispelling wind and removing dampness as a raw material to prepare an oral preparation for dispelling wind and removing dampness.

8. The use according to claim 7, characterized in that The oral preparation comprises, by weight: 60-80 parts of the composition having the effect of dispelling wind and removing dampness, 10-20 parts of excipients, 2-6 parts of sweeteners, 1-4 parts of flavoring agents, 1-3 parts of preservatives and 2-5 parts of thickeners.

9. The use according to claim 8, characterized in that The excipient is at least one of lactose, maltodextrin, and microcrystalline cellulose; the sweetener is at least one of xylitol, mannitol, maltitol, oligomaltose, and sorbitol; the flavoring agent is at least one of citric acid, malic acid, tartaric acid, and taurine; the preservative is at least one of benzoic acid, sodium benzoate, sorbic acid, and potassium sorbate; and the thickener is at least one of epoxidized dextrin, pectin, carrageenan, sodium carboxymethyl cellulose, and hydroxypropyl methylcellulose.

10. The use according to claim 9, characterized in that The oral preparation comprises, by weight: 70 parts of the composition having the effect of dispelling wind and removing dampness, 10 parts of maltodextrin, 6 parts of microcrystalline cellulose, 3 parts of isomaltooligosaccharide, 2 parts of sorbitol, 2 parts of taurine, 2 parts of potassium sorbate, 2 parts of epoxidized dextrin and 3 parts of pectin.

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