Sugar-free spleen-tonifying and kidney-tonifying granules and preparation method thereof

Abstract

The invention discloses sugar-free spleen-tonifying and kidney-tonifying granules and a preparation method thereof, and relates to the field of traditional Chinese medicine preparations. The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 180-240 parts of codonopsis pilosula, 180-240 parts of fructus lycii, 180-240 parts of glossy privet fruit, 100-150 parts of bighead atractylodes rhizome, 60-80 parts of semen cuscutae, 35-50 parts of salted fructus psoraleae, 60-100 parts of rhizoma polygonati, 80-120 parts of Chinese yam and 30-60 parts of salvia miltiorrhiza. The technology is innovated, the quality and efficiency are improved, according to the polarity difference of active ingredients of medicinal materials, an alcohol-water double extraction method is adopted, so that the extraction of fat-soluble ingredients and water-soluble ingredients is more targeted and more complete, and the technical bottleneck that the fat-soluble ingredients and ingredients wrapped in cell walls are not fully extracted is solved. The water extraction part is subjected to enzymolysis treatment, plant cell walls can be effectively destroyed, dissolution of effective components such as lycium barbarum polysaccharide and glossy privet fruit glycoside in cells is promoted, the extraction rate is remarkably increased, and a core technical foundation is laid for achieving better drug effects of the product.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of traditional Chinese medicine preparation, in particular to a sugar-free spleen and kidney tonifying granule and a preparation method thereof. BACKGROUND

[0002] Spleen and kidney tonifying granule is a classical prescription of traditional Chinese medicine, which is commonly used for related diseases caused by spleen and kidney deficiency. Traditional preparation has high sugar content, which is not suitable for patients who need to limit sugar intake such as diabetes and obesity. Existing sugar-free improvement researches are mostly focused on the replacement of excipients and molding process, which solves the problem of sugar-free, but still has deficiencies in improving the intrinsic efficacy of the product and enhancing the bioavailability. The original standard only controls the quality of a few components, which is difficult to fully reflect the material basis of efficacy. Therefore, it is of great significance to develop a sugar-free spleen and kidney tonifying granule with more comprehensive efficacy, more controllable quality and more suitable for a wider range of people.

[0003] Therefore, we propose a sugar-free spleen and kidney tonifying granule and a preparation method thereof to solve the above-mentioned problems.

[0004] The above information disclosed in this BACKGROUND section is only for increasing the understanding of the background of the application, and therefore, it can include information known by those of ordinary skill in the art. SUMMARY

[0005] One of the purposes of the present application is to provide a sugar-free spleen and kidney tonifying granule, which enhances the comprehensive effects of spleen tonifying, kidney tonifying and blood nourishing by optimizing the classical prescription and introducing specific efficacy components.

[0006] The second purpose of the present application is to provide a preparation method of the sugar-free spleen and kidney tonifying granule, which improves the transfer rate and stability of effective components by introducing modern extraction and purification technology, and ensures the uniformity and controllability of product quality.

[0007] In order to achieve the above-mentioned purposes, the present application provides the following technical scheme: a sugar-free spleen and kidney tonifying granule is prepared from the following raw materials by weight: 180-240 parts of radix codonopsis, 180-240 parts of medlar, 180-240 parts of ligustrum lucidum aiton, 100-150 parts of atractylodes, 60-80 parts of semen cannabis, 35-50 parts of salted psoralea, 60-100 parts of polygonatum, 80-120 parts of dioscorea, and 30-60 parts of salvia miltiorrhiza.

[0008] Preferably, it also includes the following excipients by weight: 200-400 parts of dextrin, 8-12 parts of steviol glycoside, 30-60 parts of microcrystalline cellulose, and 2-5 parts of silicon dioxide.

[0009] Preferably, the preferred raw material ratio is: 210 parts of radix codonopsis, 210 parts of medlar, 210 parts of ligustrum lucidum aiton, 126 parts of atractylodes, 70 parts of semen cannabis, 42 parts of salted psoralea, 80 parts of polygonatum, 100 parts of dioscorea, and 45 parts of salvia miltiorrhiza.

[0010] The application also provides a preparation method of the above-mentioned sugar-free spleen-strengthening and kidney-nourishing granules, comprising the following steps:

[0011] Step 1, raw material pretreatment and group extraction:

[0012] The radix codonopsitis, rhizoma atractylodis macrocephalae, rhizoma polygonati and rhizoma dioscoreae are mixed, 60-70% ethanol solution in an amount of 8-12 times the total weight of the mixture is added, and the mixture is heated to reflux and extracted for 2-3 times, each time for 1-2 hours. The alcohol extract is combined, filtered, and concentrated to an alcohol extract concentrate A with a relative density of 1.10-1.15 at 60°C, which is prepared for use.

[0013] The fructus lycii, ligustrum lucidum fruit, cuscuta, salted psoralea and salvia miltiorrhiza are mixed, water in an amount of 8-10 times the total weight of the mixture is added, and the mixture is boiled for 2-3 times, each time for 1-1.5 hours. The decoction is combined, filtered, and concentrated to a water extract concentrate B with a relative density of 1.10-1.15 at 60°C, which is prepared for use.

[0014] Step 2, complex enzymatic hydrolysis:

[0015] The water extract concentrate B obtained in step 1 is cooled to 40-50°C, the pH is adjusted to 4.5-5.5, and complex enzymes are added for enzymatic hydrolysis. The enzymatic hydrolysis time is 1-2 hours, and then the temperature is raised to 90°C for sterilization for 10 minutes. The mixture is filtered to obtain an enzymatic hydrolysis concentrate B'.

[0016] Step 3, mixing and concentration:

[0017] The alcohol extract concentrate A and the enzymatic hydrolysis concentrate B' are combined and stirred uniformly, and then concentrated to a thick paste C with a relative density of 1.25-1.30 at 60°C.

[0018] Step 4, vacuum belt drying and crushing:

[0019] The thick paste C is dried by a vacuum belt dryer until the water content is less than or equal to 5%. The dried paste is crushed into a fine powder that passes through an 80-100 mesh sieve to obtain a dried paste powder D.

[0020] Step 5, granulation:

[0021] The dried paste powder D is mixed with the prescription amount of dextrin, microcrystalline cellulose, silicon dioxide and stevioside for 20 minutes to obtain a uniform mixture. 85% (v / v) ethanol is used as a wetting agent, and 1-2% povidone K30 by weight of the dried paste powder is added to the wetting agent as a binder. The mixture is made into a soft material, and granulated using a 16 mesh sieve.

[0022] Step 6, drying and granulation:

[0023] The wet granules are placed in a boiling drying bed and dried at 65-75°C until the water content is less than or equal to 6.0%. The granules are sieved using a 16 mesh sieve, and the qualified granules are collected.

[0024] Step 7, total mixing and packaging:

[0025] The whole granules are placed in a three-dimensional mixer for total mixing for 20-30 minutes to ensure uniform content; and then, according to the specifications, the granules are packed to obtain the finished product of the sugar-free kidney-yi granules.

[0026] Preferably, in the step 2, the complex enzyme is a mixture of cellulase and pectinase at a mass ratio of 1:1-1:2.

[0027] Preferably, in the step 4, the drying temperature is 70-85℃, and the vacuum degree is -0.08 to -0.10 MPa.

[0028] Preferably, in the step 5, the ratio of the dry paste powder D to dextrin is 1:2.

[0029] Preferably, in the step 7, the packing specification is 10g per bag.

[0030] Compared with the prior art, the present application has the following beneficial effects:

[0031] 1. The present application innovatively adds huangji (tonifying qi and nourishing yin, invigorating the spleen and nourishing the kidney), shanyao (tonifying the spleen and stomach, generating fluid and benefiting the lung, and tonifying the kidney and astringing essence) and danshen (promoting blood circulation to remove blood stasis) on the basis of the classic prescription, not only strengthens the spleen and kidney, but also improves microcirculation and promotes drug absorption through the blood-activating effect of danshen, forming a synergistic effect of "spleen and kidney tonification and blood-activating", and the efficacy is synergistically enhanced, especially suitable for patients with spleen and kidney deficiency with blood stasis.

[0032] 2. The present application is innovative in process, improves quality and efficiency, and adopts alcohol-water double extraction method according to the polarity difference of the effective components of medicinal materials, so that the extraction of fat-soluble components and water-soluble components is more targeted and complete, solving the technical bottleneck of insufficient extraction of fat-soluble components and cell wall-enclosed components.

[0033] 3. The present application performs enzymatic hydrolysis on the water extraction part, which can effectively destroy the plant cell wall and promote the dissolution of effective components in the cells such as gordonii polysaccharide and ligustrum lucidum glycoside, significantly improving the extraction rate and laying a core technical foundation for the product to achieve better efficacy.

[0034] 4. The present application adopts vacuum belt drying, which has lower drying temperature and shorter drying time compared with traditional oven drying, can maximize the protection of heat-sensitive components from being destroyed, and the product has better solubility.

[0035] 5. The present application adds povidone K30 in the wetting agent, which can significantly improve the formability and hardness of the granules, reduce the amount of fine powder, improve the yield, and improve the solubility of the granules.

[0036] 6. The sugar-free product has wide applicability, completely removes sucrose, uses stevioside for flavoring, and is supplemented with microcrystalline cellulose and silicon dioxide to improve flowability and moisture resistance, and has good product stability, and is suitable for diabetics and the general population pursuing low sugar intake. BRIEF DESCRIPTION OF DRAWINGS

[0037] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description only constitute some embodiments described in the present application, and other drawings can also be obtained by those skilled in the art based on these drawings.

[0038] Figure 1 Thin layer chromatogram for isopsoralen;

[0039] Figure 2 TLC chromatogram of isopsoralen on different thin layer plates;

[0040] Figure 3 TLC chromatogram of isopsoralen at different temperatures;

[0041] Figure 4 TLC chromatogram of isopsoralen at different humidities;

[0042] Figure 5 Thin layer chromatogram for Medlar;

[0043] Figure 6 TLC chromatogram of Medlar on different thin layer plates;

[0044] Figure 7 TLC chromatogram of Medlar at different temperatures;

[0045] Figure 8 TLC chromatogram of Medlar at different humidities. DETAILED DESCRIPTION

[0046] In order to enable those skilled in the art to better understand the technical solutions of the present application, the present application will be further described in detail below in combination with the drawings. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without making creative efforts fall within the protection scope of the present application.

[0047] Example 1:

[0048] A sugar-free spleen-strengthening and kidney-nourishing granule is made from the following raw medicinal materials by weight: Radix Codonopsis 180-240 parts, Medlar 180-240 parts, Ligustrum lucidum 180-240 parts, Atractylodes 100-150 parts, Semen Falcatum 60-80 parts, Salted Psoralen 35-50 parts, Polygonatum 60-100 parts, Dioscorea 80-120 parts, and Salvia miltiorrhiza 30-60 parts.

[0049] Also include the following weight parts of adjuvant: dextrin 200-400 parts, stevioside 8-12 parts, microcrystalline cellulose 30-60 parts, silicon dioxide 2-5 parts.

[0050] The preferred raw material ratio is: Dangshen 210 parts, Medlar 210 parts, Female Privet Fruit 210 parts, Baizhu 126 parts, Gomphrena globosa 70 parts, salted Psoralea 42 parts, Huangjing 80 parts, Shanyao 100 parts, Danshen 45 parts.

[0051] Example 2:

[0052] A preparation method of a sugar-free spleen and kidney granule, characterized in that it comprises the following steps:

[0053] Step 1, raw material pretreatment and grouping extraction:

[0054] Mix Dangshen, Baizhu, Huangjing, and Shanyao, add 8-12 times the total weight of 60-70% ethanol solution, heat reflux extraction 2-3 times, 1-2 hours each time, combine the alcohol extract, filter, recover ethanol, concentrate to 60°C under the relative density of 1.10-1.15 alcohol extract clear paste A, ready for use;

[0055] Mix Medlar, Female Privet Fruit, Gomphrena globosa, salted Psoralea, and Danshen, add 8-10 times the total weight of water, decoct 2-3 times, 1-1.5 hours each time, combine the decoction, filter, concentrate to 60°C under the relative density of 1.10-1.15 water extract clear paste B, ready for use;

[0056] Step 2, compound enzymolysis:

[0057] Cool the water extract clear paste B obtained in step 1 to 40-50°C, adjust the pH to 4.5-5.5, add compound enzyme for enzymolysis, enzymolysis time is 1-2 hours, then heat to 90°C for 10 minutes, filter, get enzymolysis clear paste B';

[0058] The compound enzyme is a mixture of cellulase and pectinase in a mass ratio of 1:1-1:2;

[0059] Step 3, mixing and concentrating:

[0060] Combine alcohol extract clear paste A and enzymolysis clear paste B', stir evenly, continue to concentrate to 60°C under the relative density of 1.25-1.30 thick paste C;

[0061] Step 4, vacuum belt drying and crushing:

[0062] The thick paste C is dried by a vacuum belt dryer, the drying temperature is 70-85℃, the vacuum degree is -0.08 to -0.10 MPa, and the moisture content is less than 5% after drying; the dried thick paste is crushed into a fine powder which can pass through an 80-100 mesh sieve to obtain a dried thick paste powder D;

[0063] Step 5, granulation:

[0064] The dried thick paste powder D is mixed with the prescription amount of dextrin, microcrystalline cellulose, silicon dioxide and stevioside for 20 minutes to obtain a uniform mixture, the ratio of the dried thick paste powder D to dextrin is 1:2, 85% (v / v) ethanol is used as a wetting agent, and 1-2% povidone K30 by weight of the dried thick paste powder is added to the wetting agent as a binder, a soft material is prepared, and granulation is performed using a 16 mesh sieve;

[0065] Step 6, drying and granulation:

[0066] The wet granules are placed in a fluidized bed dryer and dried at 65-75℃ until the moisture content is less than 6.0%, and then the granules are sieved using a 16 mesh sieve to obtain qualified granules;

[0067] Step 7, total mixing and packaging:

[0068] The granules after sieving are placed in a three-dimensional mixer for 20-30 minutes to ensure uniform content, and then each bag contains 10g of the granules according to the specifications, thereby obtaining the finished product of the sugar-free spleen-strengthening and kidney-nourishing granules.

[0069] I. The effect of the formula and preparation process of the application is verified by the following experiments.

[0070] 1. Selection of excipients

[0071] According to the different characteristics of the excipients, different excipient prescriptions are designed using common excipients such as dextrin, lactose, sorbitol, mannitol and microcrystalline cellulose, and the sugar-free preparation excipients are screened. The molding rate, dissolution rate, hygroscopicity and flowability of the preparation are used as indicators for screening the optimal excipients. The prescription design is shown in Table 1.

[0072] Table 1 Screening of different preparation prescription ratios

[0073]

[0074] Index determination:

[0075] Molding property: the prepared granules are first passed through a No. 1 sieve and then a No. 5 sieve, the granules that can pass through the No. 1 sieve but cannot pass through the No. 5 sieve are collected and weighed, and the granule molding rate is calculated. Molding rate = (weight of granules passing through the No. 1 sieve and not passing through the No. 5 sieve / total mass of granules) x 100%.

[0076] Hygroscopicity: Take the prepared granules, place them in a 30℃ oven for 48h, and add NaCl to the bottom of the glass desiccator until a supersaturated solution (RH = 75%) is formed. Place the granules, which have been constant in weight for 48h, on the bottom of a flat weighing bottle. Open the weighing bottle and weigh the mass accurately. Place the weighing bottle in the above-mentioned desiccator and weigh the mass after 48h. Calculate the hygroscopicity. Hygroscopicity = [(mass after hygroscopicity - mass before hygroscopicity) / mass before hygroscopicity] x 100%.

[0077] Solubility: According to the 2020 edition of "Chinese Pharmacopoeia (Volume IV)", the solubility of granules is detected. Solubility = (mass of dissolved granules / mass of granules) x 100%.

[0078] Flowability: Measured by the angle of repose. Using the fixed funnel method, connect 3 funnels in series and fix them on the coordinate paper at a height of 1cm. Carefully pour the granules along the funnel wall into the uppermost funnel until the tip of the granule cone formed on the coordinate paper touches the funnel mouth. Measure the radius R of the cone bottom from the coordinate paper and calculate the angle of repose α, formula: tgα = H / R. Each prescription is repeated 3 times, and the average value is taken.

[0079] Comprehensive score: The comprehensive score is evaluated based on the molding rate, solubility, hygroscopicity and flowability of the preparation. The comprehensive score criteria are as follows:

[0080] Comprehensive score = molding rate x 70% + solubility x 15% + hygroscopicity x 10% + angle of repose x 5%

[0081] Hygroscopicity: 0% for 100 points, ≥10% for 0 points;

[0082] Angle of repose: ≤20° for 100 points, ≥30° for 0 points. The results are shown in Table 2.

[0083] Table 2 Comprehensive score of excipients

[0084]

[0085]

[0086] From Table 2, among the 5 kinds of excipients, the comprehensive score of dextrin is the highest, which is 69.12. Therefore, dextrin is selected as the diluent of sugar-free spleen and kidney granules.

[0087] 2. Selection of flavoring agents

[0088] According to the characteristics of different excipients, with dextrin as the diluent, aspartame and stevioside as the flavoring agents, different formulation prescriptions are designed to optimize the best ratio of excipients. The specific situation of the prescription design is shown in Table 3 and Table 4.

[0089] Table 3 Flavoring agent screening prescription design

[0090]

[0091] Table 4 Prescription research results

[0092]

[0093] From Table 4, by comparing the quality attributes of the granules prepared from each prescription, prescription 5 is selected as the best prescription, i.e. steviol glycoside is selected as the flavoring agent of the preparation, and the dosage is 10 g.

[0094] 3. Selection of flow aid

[0095] On the basis of determining dextrin as the diluent and steviol glycoside as the flavoring agent, in order to further improve the flowability of the granules and reduce caking and adhesion, two flow aids, microcrystalline cellulose (MCC) and pregelatinized starch, are selected for screening. The angle of repose, molding rate and bulkiness of the granules are used as the evaluation indexes, and a comprehensive score is made. The prescription design is shown in Table 5.

[0096] Table 5 Flow aid screening prescription design

[0097]

[0098] Table 6 Prescription research results

[0099]

[0100] *Note: Bulkiness scoring standard: severe caking - 0 points; slight caking - 60 points; basically loose - 80 points; completely loose - 90-100 points.

[0101] From Tables 5 and 6, the granules with microcrystalline cellulose added perform best in terms of flowability, molding rate and bulkiness, and have the highest comprehensive score. Therefore, microcrystalline cellulose is selected as the flow aid of the preparation.

[0102] 4. Screening of anti-hygroscopic agent

[0103] In order to solve the problem that the sugar-free granules are more prone to moisture absorption after the removal of sucrose, two anti-hygroscopic agents, silicon dioxide and magnesium stearate, are investigated. The hygroscopic rate and the flowability (angle of repose) of the granules are used as the main evaluation indexes. The test is carried out on the basis of the best prescription G, and the prescription design is shown in Table 7.

[0104] Table 7 Anti-hygroscopic agent screening prescription design

[0105]

[0106] Table 8 Prescription research results

[0107]

[0108] From table 7, 8, it can be seen that the addition of 0.2% of silicon dioxide (prescription G2) can significantly reduce the moisture absorption rate of the particles, and further improve the flowability, the effect is better than magnesium stearate. Therefore, the silicon dioxide is finally determined as the anti-hygroscopic agent of the preparation.

[0109] In summary, the optimal excipient composition of the product is determined as follows: dextrin as a diluent, stevioside as a flavoring agent, microcrystalline cellulose as a flow aid, and silicon dioxide as an anti-hygroscopic agent.

[0110] 5. Advantages of the grouping extraction process

[0111] To investigate the advantages of grouping extraction compared with traditional mixed water extraction, two groups of processes were set up for comparison:

[0112] Control group (traditional process): combine all medicinal materials, and decoct three times according to the traditional process.

[0113] Test group (process of the application): according to the requirements of the application, the medicinal materials are divided into two groups.

[0114] The contents of different polar active ingredients in the final products of the two groups were determined respectively, the transfer rates from medicinal materials to finished products were calculated, and the dry extract yields were compared. The results are shown in table 9.

[0115] Table 9 Comparison of the effects of grouping extraction and traditional extraction process

[0116]

[0117] The data show that the grouping extraction process adopted in the application can significantly improve the transfer rate of fat-soluble active ingredients, while ensuring that the transfer rate of water-soluble ingredients is not affected or even slightly improved. The slight reduction in dry extract yield actually indicates that the process is more selective, reduces the extraction of invalid impurities, and is beneficial to improve the purity of the preparation and reduce the dosage. This process effectively solves the common technical problem of insufficient extraction of fat-soluble ingredients in traditional water extraction.

[0118] 6. Advantages of complex enzymatic treatment

[0119] To investigate the effect of enzymatic treatment on the extraction of active ingredients from water extraction group medicinal materials, two groups were set up for comparison based on the water extraction process:

[0120] Control group (no enzymolysis): water extraction clear paste B is directly concentrated.

[0121] Test group (enzymolysis treatment): water extraction clear paste B is subjected to complex enzymolysis according to the method of the application, cellulase: pectinase = 1:1, pH 5.0, enzymolysis at 50°C for 1.5 hours, and enzyme-treated clear paste B' is obtained.

[0122] The contents of main water-soluble effective components in two groups of the paste were determined, and the extraction rates were calculated. The results are shown in Table 10.

[0123] Table 10 Influence of enzymatic hydrolysis on water extraction effect comparison

[0124]

[0125] The data fully prove that the composite enzymatic hydrolysis process can significantly destroy the plant cell wall structure, so that the effective components such as polysaccharides, glycosides and phenolic acids in the cells are more fully released into the solvent. Compared with the control group without enzymatic hydrolysis, the extraction rates of key effective components such as wolfberry polysaccharide, specnuezhenide and salvianolic acid B are significantly improved. This shows that the enzymatic hydrolysis process not only improves the utilization rate of raw materials, but also fundamentally ensures the sufficient existence of effective components in the finished product, which provides a solid material basis for the curative effect of the product.

[0126] In summary, the extraction and purification process combined with grouping extraction and composite enzymatic hydrolysis adopted by the present application can more comprehensively and efficiently extract various effective components in the prescription compared with the traditional single water extraction process, especially solving the technical bottleneck of insufficient extraction of fat-soluble components and cell wall-encapsulated components. The process is scientific and reasonable, and has significant advantages, which lays a core technical foundation for the product to achieve better efficacy.

[0127] 7, Selection of granule forming process

[0128] Orthogonal test design: Through pre-test, the ratio of dry paste powder D to dextrin, ethanol concentration and mixing time were selected for 3-factor and 3-level investigation, and L9(3 4 ) orthogonal table was used for test, with granule qualified rate as evaluation index. The factor level table is shown in Table 11, and the test results are shown in Tables 12-13.

[0129] Table 11 Factor level table

[0130]

[0131]

[0132] Table 12 L9(3 4 ) orthogonal test table and test results

[0133]

[0134] Table 13 Variance analysis table

[0135]

[0136] From the above variance analysis, the primary and secondary order of the influence of different factors on the qualified rate of particles is A (dry paste powder D: dextrin) > C (mixing time) > B (ethanol concentration). The optimal granulation process conditions are A2B3C3, that is: dry paste powder D:dextrin ratio 1:2, ethanol concentration 85%, mixing time 20 min.

[0137] 8、Verify the test results of 3 batches of samples

[0138] On the basis of the prescription and the optimal forming process of the application, 3 batches of samples were prepared from dry paste powder D, and the materials were added according to 20 times the prescription amount for process verification. The moisture and solubility of the obtained granules should meet the requirements. The total sum of particles that do not pass through No. 1 sieve and can pass through No. 5 sieve should not exceed 15%; the moisture should not exceed 6.0%. The solubility should be completely dissolved. The content determination of each bag of female privet fruit is calculated by taking speciosin (C 31 H 42 O 17 ) as the standard, which should not be less than 3.4 mg. The verification test results are shown in Table 14.

[0139] Table 14 Verification results of 3 batches of tests

[0140]

[0141] As can be seen from Table 14, the verification results of 3 batches of tests show that each index meets the requirements in the Granules item in the 2020 edition of Chinese Pharmacopoeia, and does not affect the content of speciosin in the finished product. Therefore, the forming process can be used for the actual production of sugar-free spleen and kidney granules.

[0142] II. To ensure the controllability of the quality of the medicinal materials of the application, the following UV identification and thin layer chromatography (TLC) identification methods of each raw material of the product are established.

[0143] 1. UV identification

[0144] Take 17 mg of the powder of the product, add 10 ml of ethanol in a 10 ml stoppered test tube, and immerse in a 60℃ water bath for 3 hours, shaking once every hour, and filter. The filtrate is measured by UV-visible spectrophotometry, and has maximum absorption at a wavelength of 283±3 nm and minimum absorption at a wavelength of 258±3 nm. Different batches of samples are checked, and the results are shown in Table 15.

[0145] Table 15 UV identification results of spleen and kidney granules

[0146]

[0147]

[0148] The UV inspection item is as follows: the maximum absorption is at the wavelength of 283±3 nm, and the minimum absorption is at the wavelength of 258±3 nm. As shown in the table, the six batches of samples all meet the requirements.

[0149] 2. Identification of Psoralea corylifolia by TLC

[0150] 2.1 Specificity test

[0151] Preparation of test solution: take 10 g of the product, add 10 ml of 20% hydrochloric acid ethanol solution and 60 ml of chloroform, reflux for 40 minutes, filter, and concentrate the filtrate to 1 ml as the test solution.

[0152] Preparation of control solution: take the isopsoralen control, add ethyl acetate to prepare a solution containing 1 mg per 1 ml as the control solution.

[0153] Preparation of negative solution of Psoralea corylifolia (salted): take 10 g of negative sample of Psoralea corylifolia (salted), and prepare the negative solution of Psoralea corylifolia (salted) according to the preparation method of test solution.

[0154] According to the TLC test, take 5 μl of each of the above solutions and spot them on the same silica gel GF254 thin layer plate, use n-hexane-ethyl acetate (8:2) as the developing agent, develop, take out, dry, and observe under the ultraviolet light (254 nm). The results are shown in Figure 1 .

[0155] Figure 1 1-8 are: 1. batch number 43220701, 2. batch number 43220702, 3. batch number 43220703, 4. isopsoralen control, 5. negative of Psoralea corylifolia (salted), 6. batch number 43220917, 7. batch number 43221004, and 8. batch number 43221005. As shown in Figure 1 , in the test solution chromatogram, there are spots of the same color at the positions corresponding to the control solution chromatogram. The negative of Psoralea corylifolia (salted) has no interference. There is no obvious difference between the samples of different batches, and they have consistency.

[0156] 2.2 Durability test

[0157] Comparison of different thin layer plates: take self-made silica gel plates and pre-made silica gel plates, and perform experiments respectively. The results show that there is no significant difference in separation effect between the self-made silica gel plates and the pre-made silica gel plates. See Figure 2 .

[0158] Comparison of different temperatures: take the thin layer plates after spotting, and perform experiments under the conditions of temperature 10℃ and temperature 35℃ respectively. The results show that there is no significant difference in separation effect under different temperatures. See Figure 3 .

[0159] Comparison of different humidity: take the sample after the thin layer plate, respectively, in the relative humidity 42%, relative humidity 88% conditions test, the results of different humidity conditions separation effect no significant difference. See Figure 4 .

[0160] 3. Gouqi thin layer identification

[0161] 3.1 specificity test

[0162] Preparation of test solution: take 10g of the product, fine grinding, add water 40ml shaking to dissolve, then add ether 40ml shaking extraction, take the ether liquid, dry, the residue add ethyl acetate 1ml to dissolve as test solution.

[0163] Preparation of control drug solution: take 0.5g of Gouqi control drug, add water 35ml boiling 15 minutes, cool, filter, filter with ether 15ml shaking extraction, take the ether liquid, dry, the residue add ethyl acetate 1ml to dissolve as control drug solution.

[0164] Preparation of Gouqi negative solution: take 10g of Gouqi negative sample, according to the preparation method of test solution, the same method to make Gouqi negative solution.

[0165] According to the test of thin layer chromatography, take 5ul of the above solution respectively, point on the same silica gel G thin layer plate, with petroleum ether (30-60℃)-ethyl acetate-formic acid (20:20:0.1) as developing agent, develop, take out, dry, under the ultraviolet light (365nm) inspection, the results are shown in Figure 5 .

[0166] Figure 5 1-8 are: 1. batch number 43220701, 2. batch number 43220702, 3. batch number 43220703, 4. Gouqi negative, 5. Gouqi control drug 6. batch number 43220917, 7. batch number 43221004, 8. batch number 43221005. The results are known from Figure 5 the test sample chromatogram, in the corresponding position with the control sample chromatogram, the same color fluorescent spots. Gouqi negative without interference. There is no obvious difference between batches of samples, with consistency.

[0167] 3.2 durability test

[0168] Comparison of different thin layer plate: take the self-made silica gel plate and pre-made silica gel plate, respectively, the results of self-made silica gel plate and pre-made silica gel plate separation effect no significant difference. See Figure 6 .

[0169] Comparison of different temperatures: Take the sample after the thin layer plate, respectively, under the condition of temperature 10℃, temperature 35℃ test, the results of different temperature separation effect no significant difference. See Figure 7 .

[0170] Comparison of different humidity: Take the sample after the thin layer plate, respectively, under the condition of relative humidity 42%, relative humidity 88% test, the results of different humidity separation effect no significant difference. See Figure 8 .

[0171] III. Stability analysis

[0172] According to the "Chinese Pharmacopoeia" 2020 edition general 9001 raw materials and preparation stability test guidelines, the stability test of the product.

[0173] 1. Test materials

[0174] Jianpi Yishen Granules (batch number: 43220701, 43220702, 43220703, 43220917, 43221004, 43221005) were provided by Lanzhou Taibao Pharmaceutical Co., Ltd.

[0175] 2. Test method

[0176] 2.1. Investigation items

[0177] Characteristics, identification, particle size, moisture, loading difference, microbial limit, content determination.

[0178] 2.2. Accelerated test

[0179] Six batches of samples were placed under the condition of temperature 40℃±2℃, relative humidity 75%±5% for 6 months, and samples were taken at 0 months, 1 month, 2 months, 3 months, 6 months for detection, to investigate its stability, the results are shown in Tables 16-21.

[0180] 2.3. Long-term test

[0181] Six batches of samples were placed under the condition of temperature 25℃±2℃, relative humidity 60%±5%, and samples were taken at 0 months, 3 months, 6 months, 9 months, 12 months, 18 months, 24 months, 36 months for detection, to investigate its stability, the results are shown in Tables 22-27.

[0182] 3. Test results

[0183] After 6 months of accelerated test, all the investigation indexes were still within the qualified range. After 18 months of long-term test, all the investigation indexes were still within the qualified range. Stability investigation is still under investigation. The test showed that the product had good stability.

[0184] Table 16 Accelerated stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43220701

[0185]

[0186]

[0187] Table 17 Accelerated stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43220702

[0188]

[0189] Table 18 Accelerated stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43220703

[0190]

[0191] Table 19 Accelerated stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43220917

[0192]

[0193]

[0194] Table 20 Accelerated stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43221004

[0195]

[0196] Table 21 Accelerated stability test results of Spleen-Kidney Nourishing Granules (after change) Batch No.: 43221005

[0197]

[0198]

[0199] Table 22 Long-term stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43220701

[0200]

[0201] Table 23 Long-term stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43220702

[0202]

[0203]

[0204] Table 24 Long-term stability test results of Spleen-Kidney Nourishing Granules Batch No.: 43220703

[0205]

[0206] Table 25 Results of long-term stability test of Spleen-strengthening and Kidney-nourishing Granules Batch No.: 43220917

[0207]

[0208] Table 26 Results of long-term stability test of Spleen-strengthening and Kidney-nourishing Granules Batch No.: 43221004

[0209]

[0210] Table 27 Results of long-term stability test of Spleen-strengthening and Kidney-nourishing Granules Batch No.: 43221005

[0211]

[0212] Certain exemplary embodiments of the present application have been described above by way of illustration, and it is to be understood that various modifications can be made without departing from the spirit and scope of the present application. Therefore, the above-described drawings and descriptions are to be regarded as illustrative in nature and not as restrictive.

Claims

1. A sugar-free spleen-strengthening and kidney-tonifying granule, characterized in that, It is made from the following raw materials in parts by weight: Codonopsis pilosula 180-240 parts, Lycium barbarum 180-240 parts, Ligustrum lucidum 180-240 parts, Atractylodes macrocephala 100-150 parts, Cuscuta chinensis 60-80 parts, Psoralea corylifolia 35-50 parts, Polygonatum sibiricum 60-100 parts, Dioscorea opposita 80-120 parts, and Salvia miltiorrhiza 30-60 parts.

2. The sugar-free spleen-strengthening and kidney-tonifying granules according to claim 1, characterized in that, It also includes the following excipients by weight: 200-400 parts dextrin, 8-12 parts steviol glycosides, 30-60 parts microcrystalline cellulose, and 2-5 parts silicon dioxide.

3. A sugar-free spleen-strengthening and kidney-tonifying granule according to claim 1 or 2, characterized in that, The preferred ratio of raw materials is as follows: Codonopsis pilosula 210 parts, Lycium barbarum 210 parts, Ligustrum lucidum 210 parts, Atractylodes macrocephala 126 parts, Cuscuta chinensis 70 parts, Psoralea corylifolia 42 parts, Polygonatum sibiricum 80 parts, Dioscorea opposita 100 parts, and Salvia miltiorrhiza 45 parts.

4. The method for preparing a sugar-free spleen-strengthening and kidney-tonifying granule as described in any one of claims 1-3, characterized in that, Includes the following steps: Step 1: Raw material pretreatment and group extraction: Mix Codonopsis pilosula, Atractylodes macrocephala, Polygonatum sibiricum, and Dioscorea opposita, add 8 to 12 times their total weight of 60 to 70% ethanol solution, heat and reflux to extract 2 to 3 times, 1 to 2 hours each time, combine the ethanol extracts, filter, recover the ethanol, and concentrate to ethanol extract A with a relative density of 1.10 to 1.15 at 60℃ for later use; Mix wolfberry, privet fruit, dodder seed, salt-processed psoralea, and salvia miltiorrhiza, add 8 to 10 times their total weight of water, decoct 2 to 3 times, 1 to 1.5 hours each time, combine the decoctions, filter, and concentrate to a water extract B with a relative density of 1.10 to 1.15 at 60°C, and set aside. Step 2, Compound Enzymatic Hydrolysis: Cool the water extract B obtained in step 1 to 40-50℃, adjust the pH to 4.5-5.5, add a compound enzyme for enzymatic hydrolysis for 1-2 hours, then heat to 90℃ for sterilization for 10 minutes, filter to obtain enzymatic hydrolysate B'. Step 3, Mixing and Concentration: Combine the alcohol extract A and the enzymatic hydrolysis extract B', stir evenly, and continue to concentrate to a thick paste C with a relative density of 1.25 to 1.30 at 60°C; Step 4: Vacuum belt drying and pulverization: The thick paste C was dried using a vacuum belt dryer until the moisture content was ≤5%; the dried paste was then pulverized into a fine powder that passed through an 80-100 mesh sieve to obtain the dry paste powder D. Step 5, Granulation: Mix the dry extract powder D with the prescribed amount of dextrin, microcrystalline cellulose, silicon dioxide and steviol glycosides thoroughly for 20 minutes until uniform. Use 85% (v / v) ethanol as a wetting agent. Add 1-2% of the dry extract powder weight of povidone K30 to the wetting agent as a binder to make a soft material and granulate it with a 16-mesh sieve. Step 6, Drying and Granulation: The wet granules were placed in a fluidized bed dryer and dried at 65-75°C until the moisture content was ≤6.0%. The granules were then sieved through a 16-mesh sieve and qualified granules were collected. Step 7, Mixing and Packaging: The granulated granules are placed in a three-dimensional mixer and mixed for 20-30 minutes to ensure uniform content; then packaged according to specifications to obtain the sugar-free spleen-strengthening and kidney-tonifying granules.

5. The method for preparing a sugar-free spleen-strengthening and kidney-tonifying granule according to claim 4, characterized in that: In step 2, the compound enzyme is a mixture of cellulase and pectinase in a mass ratio of 1:1 to 1:

2.

6. The preparation method of a sugar-free spleen-strengthening and kidney-tonifying granule according to claim 4, characterized in that: In step 4, the drying temperature is 70-85℃ and the vacuum degree is -0.08 to -0.10MPa.

7. The method for preparing a sugar-free spleen-strengthening and kidney-tonifying granule according to claim 4, characterized in that: In step 5, the ratio of dry extract powder D to dextrin is 1:

2.

8. The method for preparing a sugar-free spleen-strengthening and kidney-tonifying granule according to claim 4, characterized in that: In step 7, the packaging specification is 10g per bag.

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