Chitosan microspheres, preparation method thereof and application of chitosan microspheres in vigor-preserving oral liquid preparation

Abstract

The invention relates to the field of traditional Chinese medicine oral liquid preparations, in particular to chitosan microspheres, a preparation method thereof and application of the chitosan microspheres in preparation of a vitality-preserving oral liquid preparation. The chitosan microspheres are prepared from the following components in parts by weight: 80 to 120 parts of chitosan, 2 to 5 parts of an emulsifier and 30 to 40 parts of an oil phase, the chitosan microspheres are obtained by dissolving the chitosan microspheres in an acidic solution and then reacting the chitosan microspheres with an emulsifier and an oil phase. The preparation method comprises the following steps: adding a clarifying agent, namely a chitosan microsphere mixture with different particle sizes, carrying out flocculation adsorption on the mixture, and adding a stabilizer sodium carboxymethyl cellulose CMC-Na; through the synergistic effect of the two components, the quality and safety of the oral liquid are remarkably improved. The colloid adsorbed by the clarifying agent can be more stably dissolved in the liquid medicine under the synergy of the stabilizer sodium carboxymethyl cellulose, so that the problem of preparation precipitation is solved, and the prepared vigor-preserving oral liquid has good stability and immunomodulatory activity.

Description

Technical Field

[0001] The invention relates to the field of Chinese medicine oral liquid preparations, in particular to chitosan microspheres and a preparation method thereof and application thereof in Baoyuan oral liquid preparations. Background Art

[0002] The immune system is the most effective weapon to defend against the invasion of pathogens. However, due to personal genetics and disease, patients' own immune capabilities vary greatly. Baoyuan Decoction is a famous prescription for replenishing qi. Its important mechanism of action is to enhance the body's immune function by replenishing the body's positive qi.

[0003] The composition of Baoyuan Decoction in Wei Zhi's "Love Mirror" in the Ming Dynasty is: ginseng, astragalus, licorice, cinnamon, add a piece of ginger, boiled in water, and taken at any time. It is mainly used to treat weakness, fatigue, and insufficient vitality. Symptoms include fatigue, shortness of breath, fear of cold, and smallpox in children, yang deficiency, sunken head, blood deficiency, clear pulp, difficulty in astringency, pale tongue coating, and weak pulse. Literature research shows that Baoyuan Decoction can increase the cellular immune function of infants and young children after ventricular septal defect repair, T lymphocyte colony count, CD 3+ , CD 4+ / CD 8+ The expression rate was significantly increased and cellular immunity was significantly enhanced.

[0004] Baoyuan Decoction is a classic Chinese prescription with a wide range of applications and definite efficacy. Since ancient times, the history of medication has been based on traditional water decoctions. However, the ingredients of Baoyuan oral liquid are complex. In addition to active ingredients, traditional water decoctions also contain some high-molecular impurities, such as starch, gum, mucus, tannins, pigments, etc. These ingredients are highly soluble when heated, and gradually precipitate after cooling, or precipitate in the oral liquid due to changes in temperature, oxidation, pH value, etc. Therefore, the precipitation problem that occurs after the development of Baoyuan Decoction into an oral liquid has always been a key bottleneck that plagues the development of formulations. Summary of the invention

[0005] The object of the present invention is to provide a chitosan microsphere and a preparation method thereof, and use thereof in the preparation of a stable Baoyuan oral liquid preparation, so as to completely retain the efficacy of the classic prescription.

[0006] In order to achieve the above-mentioned invention object, the technical solution provided by the present invention is as follows:

[0007] A chitosan microsphere comprises, by weight, 80-120 parts of chitosan, 2-5 parts of emulsifier and 30-40 parts of oil phase. The chitosan microsphere is obtained by dissolving in an acidic solution and then reacting with the emulsifier and the oil phase.

[0008] In a preferred embodiment, the pH value of the acidic solution is 2-4.

[0009] The chitosan microspheres prepared in an acidic solution have cations on the outside, are highly positively charged, and have a strong adsorption capacity for anionic suspended matter in the Baoyuan oral solution.

[0010] In a preferred embodiment, the acidic solution includes one or both of acetic acid solution and phosphoric acid solution.

[0011] In a preferred embodiment, the emulsifier includes one or both of sodium lauryl sulfate and Tween.

[0012] In a preferred embodiment, the Tween comprises Tween 80.

[0013] In a preferred embodiment, the oil phase is one or more of soybean oil, olive oil or sunflower oil.

[0014] In a preferred embodiment, the chitosan microspheres include 10-20wt% chitosan microspheres with a particle size less than 0.1 μm, 20-40wt% chitosan microspheres with a particle size of 0.1-40 μm, and 45%-70% chitosan microspheres with a particle size of 45-105 μm.

[0015] Chitosan microspheres with a particle size of 45-105μm are used to absorb and remove macromolecular impurities such as starch and pectin in the liquid medicine, improve the clarity of the liquid medicine, and are filtered out during the preparation of the oral liquid. Chitosan microspheres with a particle size of less than 0.1μm still exist in the oral liquid after preparation, and are used to maintain the stability of the oral liquid. They can effectively retain the effective ingredients in the oral liquid, such as some polysaccharides, so that the efficacy of the oral liquid is retained to a large extent; they can also improve the penetration performance of the drug. Chitosan microspheres with a particle size of 0.1-40μm can absorb phenolic acid and tannin components in the liquid medicine to improve the stability of the liquid medicine.

[0016] The preparation method of chitosan microspheres comprises the following steps: dissolving chitosan in an acidic solution, adding an emulsifier, stirring, then dropping an oil phase, heating the reaction system to 70-90°C after the dropping is completed, reacting for 30-50 minutes, and purifying to obtain chitosan microspheres; the emulsifier is dropped several times, and each drop is 20-40 minutes apart from the last drop.

[0017] The reaction temperature of the system will affect the solubility of chitosan and the viscosity of the solution. If the temperature is too high, the viscosity of the chitosan solution will decrease, while if the temperature is too low, it may lead to incomplete cross-linking, affecting the stability and performance of the microspheres.

[0018] In one preferred embodiment, the emulsifier is added dropwise three times, 50-60wt% of the total emulsifier is added dropwise for the first time, 30-39wt% of the total emulsifier is added dropwise for the second time, and 1-20wt% of the total emulsifier is added dropwise for the third time, and each addition is 20-40 minutes apart from the last addition.

[0019] The addition process and amount of the emulsifier added each time have a significant effect on the particle size and morphology of the microspheres. The multiple drops of the emulsifier and the amount of addition ensure that the chitosan microspheres have a certain weight range and a variety of particle sizes, thereby ensuring the implementation of the function.

[0020] The particle size has an important influence on the application and performance of chitosan. If the particle size is too large, the solubility will be reduced, and if the particle size is too small, the adsorption performance will be reduced.

[0021] The chitosan microspheres prepared by the method of the present invention have stronger adsorption performance than the prior art and are applicable in a wide pH range. The addition of chitosan microspheres needs to be able to play an adsorption role and strengthen the retention of active ingredients on the one hand, and on the other hand, it should not cause the Baoyuan oral liquid to precipitate. The chitosan microspheres synthesized in the prior art that can be used to adsorb tetracycline, Cr or phosphorus elements will obviously cause the Baoyuan oral liquid to precipitate, and are not suitable for the system of the present invention.

[0022] The present invention also claims to protect the use of the chitosan microspheres in preparing Baoyuan oral liquid preparations.

[0023] Based on the same inventive concept, the present invention also claims to protect a method for preparing the Baoyuan oral liquid preparation, comprising the following steps:

[0024] S1, 140-160 parts of ginseng, 300-320 parts of astragalus, 70-85 parts of licorice, 28-35 parts of cinnamon bark, and 110-130 parts of ginger are mixed and baked, and then 12-18 times of water is added, soaked for 40-70 minutes, boiled, and then kept at a slight boil for 1.2-1.5 hours, filtered, and filtrate 1 and dregs are obtained;

[0025] S2. Add 10-14 times the amount of water to the residue and boil it, then keep it at a slight boil for 35-65 minutes, filter it, combine the filtrate, concentrate it, and adjust the pH value to 4.3-5.8;

[0026] S3, adding 2-25 parts of a clarifier and 0.2-1.2 parts of a stabilizer, mixing evenly, filtering, and obtaining the Baoyuan oral liquid preparation;

[0027] The clarifier is the chitosan microspheres.

[0028] In a preferred embodiment, the filtration is performed using a ceramic inorganic membrane.

[0029] In a preferred embodiment, the filtration is carried out by using a ceramic inorganic membrane with a pore size of 0.1-0.45 μm.

[0030] In a preferred embodiment, the concentration is performed by vacuum concentration to 350-450 ml at 60-80°C.

[0031] In step S2, the pH value is adjusted to be within the range of 4.3-5.8, and the Baoyuan oral liquid is more stable and has better clarity. If the acid or base is too high, the stability of the Baoyuan oral liquid will deteriorate. As a preferred embodiment of the present invention, the concentration of the stabilizer sodium carboxymethyl cellulose CMC-Na is 0.06-0.15%.

[0032] Adding too much clarifier will make the sample solution too viscous, affecting operability and subsequent processing. Adding too little clarifier will result in poor adsorption performance and precipitation.

[0033] In a preferred embodiment, the stabilizer is one or both of sodium carboxymethyl cellulose and xanthan gum. Considering the effect of improving texture, sodium carboxymethyl cellulose is preferred.

[0034] In a preferred embodiment, the ginseng has a growth period of 5-8 years.

[0035] In a preferred embodiment, the astragalus cultivation method is simulated wild cultivation.

[0036] In a preferred embodiment, the cinnamon bark is selected from the bark of a cassia tree with an age of 10-15 years.

[0037] In a preferred embodiment, the ginger variety is small yellow ginger.

[0038] In a preferred embodiment, the Baoyuan oral liquid preparation further includes 25-35 parts of flavoring agent, which is added after the stabilizer is added.

[0039] In a preferred embodiment, the flavoring agent includes one or both of steviol glycoside and xylitol.

[0040] In a preferred embodiment, the flavoring agent includes 0.1-0.5 parts of stevioside and 25-35 parts of xylitol.

[0041] In a preferred embodiment, the flavoring agent is one or both of steviol glycoside and xylitol.

[0042] Based on the same inventive concept, the present invention also claims protection for the Baoyuan oral liquid preparation prepared by the above preparation method.

[0043] The beneficial effects of the present invention are:

[0044] (1) The present invention provides a preparation method of Baoyuan oral liquid, which avoids the loss of active substances caused by the drying process, reduces the dosage of traditional water decoctions, and completely retains the active substance basis of traditional decoctions, thereby improving the bioavailability of traditional Chinese medicine oral liquids.

[0045] (2) The composition of Chinese medicine oral liquid is quite complex, belonging to the mixture of solution, colloid, suspension and emulsion. When colloid and suspension are left standing, there are sedimentation and aggregation of particles, and emulsion is prone to creaming and rupture when the conditions change. At the same time, because colloid, suspension and emulsion are the same, the particles that need to be added in the present invention have different charges and will attract each other, resulting in a large amount of precipitation after standing. Therefore, the present invention adds chitosan microspheres as a clarifier, performs flocculation and adsorption, and adds sodium carboxymethyl cellulose CMC-Na as a stabilizer. Stabilizers can promote the effect of clarifiers, and similarly, clarifiers can also promote the effect of stabilizers. The main principle may be: chitosan microspheres with a particle size of less than 0.1μm can improve the permeability of drugs; chitosan microspheres with a particle size of 0.1-40μm can adsorb phenolic acid and tannin components in the drug solution to improve the stability of the drug solution; chitosan microspheres with a particle size of 45-105μm can adsorb macromolecular impurities such as starch and pectin in the drug solution to improve the clarity of the drug solution. Clarifiers adsorb impurities and stabilizers buffer water quality in two aspects: clarifiers mainly remove impurities in oral liquids through adsorption, and stabilizers are mainly used to buffer and stabilize water quality, balance environmental factors such as pH, salinity, and osmotic pressure in water. Clarifiers remove impurities through adsorption, while stabilizers maintain the stability of oral liquids by buffering water quality. The synergistic effect of the two significantly improves the quality and safety of oral liquids. The colloid after adsorption of the clarifier can be more stably dissolved in the medicinal solution with the cooperation of the stabilizer sodium carboxymethyl cellulose, which solves the problem of preparation precipitation and makes the prepared Baoyuan oral solution have both good stability and immunomodulatory activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The Baoyuan oral liquid prepared in Example 1 is in a state after being left to stand for 30 days at a temperature of 25 and a humidity of 60%;

[0047] Figure 2 The state of the Baoyuan oral liquid prepared in Examples 1 and 2 after being left to stand for 30 days at a temperature of 25 and a humidity of 60%;

[0048] Figure 3 The state of the Baoyuan oral liquid prepared in Comparative Example 1 after being left to stand for 30 days at a temperature of 25 and a humidity of 60%;

[0049] Figure 4 The state of the Baoyuan oral liquid prepared in Comparative Example 2 after being left to stand for 30 days at a temperature of 25 and a humidity of 60%;

[0050] Figure 5 This is the state of the Baoyuan oral liquid prepared in Comparative Example 12 after being left to stand at a temperature of 25 and a humidity of 60% for 30 days. DETAILED DESCRIPTION

[0051] The present invention is further described below in conjunction with specific examples, but the content of the present invention is not limited thereto. The molecular weight of the chitosan used in the examples and comparative examples is 161.2.

[0052] Example 1

[0053] The preparation method of chitosan microspheres is as follows: first, 100g chitosan is dissolved in an acetic acid solution with a pH value of 3.2, and the pH value of the system is maintained at 3.2-3.3. Then, an emulsifier (sodium dodecyl sulfate) is added, and the amount of the emulsifier is 2.5% of the mass of chitosan. The emulsifier is added dropwise three times, 50wt% of the total amount of the emulsifier is added dropwise for the first time, 35wt% of the total amount of the emulsifier is added dropwise for the second time, and 15wt% of the total amount of the emulsifier is added dropwise for the third time, and each dropwise addition is 20-40min apart from the last dropwise addition; and the emulsifier is fully stirred to be evenly distributed in the solution. Then, 35g of oil phase (soybean oil) is slowly added dropwise to the solution, while constantly stirring to form tiny oil droplets. The above mixture is then heated to 85°C and maintained for a period of 35-40min to evaporate the water inside the oil droplets to form a hollow structure. Finally, unreacted substances and impurities are removed by filtering, washing and other steps to obtain pure chitosan microspheres. The chitosan microspheres include 20wt% of chitosan microspheres with a particle size less than 0.1μm, 30wt% of chitosan microspheres with a particle size of 0.1-40μm and 50% of chitosan microspheres with a particle size of 45-105μm.

[0054] Example 2

[0055] On the basis of Example 1, the total amount of the emulsifier added was adjusted to 60wt% for the first drop, 39wt% for the second drop, and 1wt% for the third drop, and the interval between each drop and the last drop was 20-40min, so that the obtained chitosan microspheres included 10wt% of chitosan microspheres with a particle size of less than 0.1μm, 35wt% of chitosan microspheres with a particle size of 0.1-40μm, and 55% of chitosan microspheres with a particle size of 45-105μm.

[0056] Example 3

[0057] On the basis of Example 1, the total amount of the emulsifier added was adjusted to 60wt% for the first drop, 30wt% for the second drop, and 10wt% for the third drop, and the interval between each drop and the last drop was 20-40min, so that the obtained chitosan microspheres included 15wt% of chitosan microspheres with a particle size of less than 0.1μm, 25wt% of chitosan microspheres with a particle size of 0.1-40μm, and 60% of chitosan microspheres with a particle size of 45-105μm.

[0058] Comparative Example 1

[0059] On the basis of Example 1, the amount of emulsifier was adjusted to 0.375% of the mass of chitosan and added dropwise at one time, so that the average particle size of the chitosan microspheres was less than 0.1 μm.

[0060] Comparative Example 2

[0061] On the basis of Example 1, the amount of emulsifier was adjusted to 0.875% of the mass of chitosan and added dropwise at one time, so that the particle size of chitosan microspheres was 0.1-40 μm.

[0062] Comparative Example 3

[0063] On the basis of Example 1, the amount of emulsifier was adjusted to 1.25% of the mass of chitosan and added dropwise at one time, so that the particle size of chitosan microspheres was 45-105 μm.

[0064] Example 4

[0065] Based on Example 1, the mixture was heated to 70° C. to obtain chitosan microspheres.

[0066] Example 5

[0067] Based on Example 1, the mixture was heated to 90° C. to obtain chitosan microspheres.

[0068] Comparative Example 3

[0069] On the basis of Example 1, the mixture was heated to 60° C. to obtain chitosan microspheres. The viscosity of the obtained chitosan microspheres decreased, and the adsorption performance was reduced.

[0070] Comparative Example 4

[0071] On the basis of Example 1, the mixture was heated to 100° C. to obtain chitosan microspheres. The chitosan microspheres were not completely cross-linked, which affected the stability and performance of the microspheres.

[0072] Example 6

[0073] On the basis of Example 1, the amount of emulsifier was adjusted to 1% of the mass of chitosan to obtain chitosan microspheres.

[0074] Example 7

[0075] On the basis of Example 1, the amount of emulsifier was adjusted to 5% of the mass of chitosan to obtain chitosan microspheres.

[0076] Comparative Example 5

[0077] On the basis of Example 1, the total amount of emulsifier is adjusted to 2.125% of the mass of chitosan, and the emulsifier is added dropwise twice, the first and second emulsifier amounts are the same as those in Example 1, and each drop is separated from the last drop by 20-40 minutes; chitosan microspheres are obtained. The chitosan microspheres include 37.5wt% of chitosan microspheres with a particle size of 0.1-40μm and 62.5% of chitosan microspheres with a particle size of 45-105μm.

[0078] Comparative Example 6

[0079] On the basis of Example 1, the total amount of emulsifier is adjusted to 1.75% of the mass of chitosan, and the emulsifier is added dropwise twice, the first dropwise addition is the same as that of Example 1, the second dropwise addition amount of emulsifier is the same as that of the third dropwise addition of Example 1, and each dropwise addition is separated from the last dropwise addition by 20-40 minutes; chitosan microspheres are obtained. The chitosan microspheres include 28.6wt% of chitosan microspheres with a particle size of less than 0.1μm and 71.4% of chitosan microspheres with a particle size of 45-105μm.

[0080] Comparative Example 7

[0081] On the basis of Example 1, the total amount of emulsifier is adjusted to 1.25% of the mass of chitosan, and the emulsifier is added dropwise twice, the first dropwise addition is the same as the second dropwise addition amount of Example 1, the second dropwise addition is the same as the third dropwise addition amount of Example 1, and each dropwise addition is separated from the last dropwise addition by 20-40 minutes; chitosan microspheres are obtained. The chitosan microspheres include 40wt% of chitosan microspheres with a particle size of less than 0.1μm and 60wt% of chitosan microspheres with a particle size of 0.1-40μm.

[0082] Comparative Example 8

[0083] First, chitosan and polyethyleneimine (PEI) (mass ratio of chitosan to polyethyleneimine 1:3) were dissolved in 15 mL, 15 wt% acetic acid solution as the water phase system. 90 mL of liquid paraffin and 6 mL of Span 80 were evenly mixed in a 250 mL three-necked flask to form an oil phase system. The three-necked flask was equipped with a condenser tube and mechanical stirring. The water bath temperature was adjusted to 35 ° C, and the mechanical stirring was controlled at 300 r min -1。Add the aqueous solution drop by drop into the oil phase system and emulsify for 1 hour until a white emulsion is obtained. Then add 1.5 mL of 50 wt% glutaraldehyde solution to crosslink the aqueous monomers encapsulated in the oil phase for 2.5 hours. The reactant gradually changes from milky white to brown. The product is centrifuged and washed with petroleum ether, 0.5 M NaOH solution, deionized water, and anhydrous ethanol in sequence. The washed product is placed in an 80°C drying oven and dried for 12 hours. The product (CP) is collected and placed in a dryer for later use.

[0084] The chitosan microspheres prepared in the above examples and comparative examples were used to prepare Baoyuan oral liquid, and the specific process was as follows:

[0085] 149.2 parts of ginseng (five-year-old), 298.4 parts of astragalus (simulated wild), 74.8 parts of licorice, 30.0 parts of cinnamon (fifteen-year-old), and 120.0 parts of ginger (small yellow ginger); after cleaning, steaming, cutting, and baking, the above five Chinese medicines are soaked in 12 times the amount of ultrapure water for 55 minutes, boiled over high heat, then turned to low heat and kept slightly boiling for 1.2 hours, the liquid medicine is filtered with a 0.45um ceramic inorganic membrane, and the residue is added with 11 times the amount of ultrapure After boiling the water over high heat, turn to low heat and keep it at a slight boil for 45 minutes. The liquid medicine is filtered through a 0.45um ceramic inorganic membrane, the filtrate is combined, and vacuum concentrated to 400ml at 65℃, the pH value of the system is adjusted to 4.5, and 0.2g of stevioside and 32g of xylitol as flavoring agents, chitosan microspheres as clarifiers, and sodium carboxymethyl cellulose CMC-Na as stabilizer are added, filtered, adjusted to a total volume of 500ml, stirred evenly, filled, sterilized, and inspected by light.

[0086] The amount of chitosan microspheres added accounted for 0.75% of the sample solution (400 ml).

[0087] The stabilizer sodium carboxymethyl cellulose CMC-Na accounts for 0.09% of the sample solution (400 ml).

[0088] The transmittance, soluble solids and anti-immune antibody level of Baoyuan oral liquid were tested. The test method was as follows: the absorbance of the clarified liquid was measured at a wavelength of 623 nm using an ultraviolet spectrophotometer, and the transmittance was calculated according to the conversion formula of absorbance and transmittance. References: Lu Ming, Zhang Hongxia, He Fu Jin, et al. Relationship between carbon black surface energy and transmittance of toluene extract [J. Rubber Industry, 2020, 67(9): 709-712.

[0089] The test method for the level of anti-immune antibodies is as follows: take a 24-well cell culture plate, add 950 μL of mouse spleen lymphocyte (LP) suspension to each well, and set up 3 duplicate wells for each group. Pre-incubate for 1 hour, add anti-mouse CD3 antibody (1.25 μg / mL, purchased from Biopsies) to stimulate spleen lymphocytes (the control group uses 10 μL PBS), and incubate at 37°C and 5% CO 2Culture in an incubator, shake gently once every 30 minutes to remove non-adherent cells. After 3 hours, wash twice with culture medium to remove non-adherent cells, replace with culture medium containing Baoyuan oral solution (the concentration of Baoyuan oral solution is 3 mg / mL), continue to culture for 48 hours, centrifuge at 3000r / min, collect the supernatant, dilute it with culture medium at a ratio of 1:5, measure the OD value at 450nm on a microplate reader, and calculate the immune antibody level according to the standard curve.

[0090] The results are as follows:

[0091] Table 1 Performance of Baoyuan oral liquid prepared from chitosan microspheres prepared in Examples 1-7 and Comparative Examples 1-8

[0092] Sample Light transmittance (%) Soluble solids (%) Anti-CD3 antibody (μg / mL) Example 1 92 4.6 21.2 Example 2 91 4.3 21.1 Example 3 91 4.5 20.9 Example 4 90 4.2 21.1 Example 5 92 4.3 20.8 Example 6 91 4.2 20.7 Example 7 90 4.3 21.1 Comparative Example 1 84 3.6 18.2 Comparative Example 2 82 3.4 18.1 Comparative Example 3 83 3.3 17.9 Comparative Example 4 85 3.2 17.8 Comparative Example 5 82 3.4 18.3 Comparative Example 6 84 3.3 18.2 Comparative Example 7 83 3.2 17.9 Comparative Example 8 82 3.5 17.6

[0093] The states of the Baoyuan oral liquid prepared in Examples 1 and 2 after being left to stand for 30 days at a temperature of 25 and a humidity of 60% are as follows: Figure 1 and Figure 2 The conditions of the Baoyuan oral liquid prepared in Comparative Examples 1 and 2 after standing for 30 days at a temperature of 25 and a humidity of 60% are as follows: Figure 3 and Figure 4 As shown in Figure 2, it can be seen that there are obvious differences in the stability and clarity of oral liquids.

[0094] Example 8

[0095] An immune function regulating oral solution and a preparation method thereof, comprising the following raw materials in parts by weight: 134.3 parts of ginseng (five-year-old), 268.6 parts of astragalus (simulated wild), 67.3 parts of licorice, 27.0 parts of cinnamon (fifteen-year-old), and 108.0 parts of ginger (small yellow ginger); the above five flavors are cleaned, steamed, cut, and baked, then soaked in 11 times the amount of ultrapure water for 50 minutes, boiled over high heat, then turned to low heat and kept slightly boiling for 1.3 hours, and the liquid medicine is decocted with 0.45um ceramic inorganic Membrane filtration, add 10 times the amount of ultrapure water to the residue, boil it on high heat, then turn to low heat and keep it at a slight boil for 55 minutes, filter the liquid through a 0.45um ceramic inorganic membrane, combine the filtrate, and vacuum concentrate it to 400ml at 65℃, adjust the pH value of the system to 4.5, add 0.2g of stevioside as a flavoring agent, 29g of xylitol, acid chitosan microspheres as a clarifier, and sodium carboxymethyl cellulose CMC-Na as a stabilizer, filter, adjust the total amount to 500ml, stir evenly, fill, sterilize, and inspect by light, and the product is ready;

[0096] The preparation method of chitosan microspheres is as follows: first, 100g chitosan is dissolved in an acetic acid solution with a pH value of 3.8, and the pH value of the system is maintained at 3.2-3.3. Then, an emulsifier (sodium dodecyl sulfate) is added, and the amount of the emulsifier is 3.5% of the mass of chitosan. The emulsifier is added dropwise three times, 50wt% of the total amount of the emulsifier is added dropwise for the first time, 35wt% of the total amount of the emulsifier is added dropwise for the second time, and 15wt% of the total amount of the emulsifier is added dropwise for the third time, and each dropwise addition is 20-40min apart from the last dropwise addition. And fully stir to make the emulsifier uniformly distributed in the solution. Then, 35g of oil phase (soybean oil) is slowly added dropwise to the aqueous phase, and continuously stirred to form tiny oil droplets. Then, the above mixture is heated to 80°C and maintained for 35-40min to evaporate the water inside the oil droplets to form a hollow structure. Finally, unreacted substances and impurities are removed by filtering, washing and other steps to obtain pure chitosan microspheres. The chitosan microspheres include 20wt% of chitosan microspheres with a particle size less than 0.1μm, 20wt% of chitosan microspheres with a particle size of 0.1-40μm and 60% of chitosan microspheres with a particle size of 45-105μm, wherein most of the chitosan microspheres with a particle size of 45-105μm have a particle size of more than 80μm.

[0097] The amount of chitosan microspheres added accounted for 0.15% of the sample solution (400 ml).

[0098] The amount of stabilizer sodium carboxymethyl cellulose CMC-Na added is 0.12% of the sample solution (400 ml).

[0099] Comparative Example 9 (diatomaceous earth, compared with Example 8)

[0100] An immune function regulating oral Baoyuan liquid and a preparation method thereof, comprising the following raw materials in parts by weight: 134.3 parts of ginseng (five-year-old), 268.6 parts of astragalus (simulated wild), 67.3 parts of licorice, 27.0 parts of cinnamon (fifteen-year-old), and 108.0 parts of ginger (small yellow ginger); the above five flavors are cleaned, steamed, cut, and baked, then soaked in 11 times the amount of ultrapure water for 50 minutes, boiled over high heat, then turned to low heat and kept slightly boiling for 1.3 hours, the liquid medicine is filtered through a 0.45um ceramic inorganic membrane, and the residue is added Boil 10 times the amount of ultrapure water over high heat, then turn to low heat and keep it at a slight boil for 55 minutes. Filter the drug solution through a 0.45um ceramic inorganic membrane, combine the filtrate, and vacuum concentrate to 400ml at 65°C. Adjust the pH value of the system to 4.5, add 0.2g of stevioside as a flavoring agent, 29g of xylitol, 0.6% of diatomaceous earth as a clarifier (purchased from Hebei Yousheng Refractory Materials), and sodium carboxymethyl cellulose (CMC-Na) as a stabilizer, filter, adjust the total amount to 500ml, stir evenly, fill, sterilize, and inspect by light.

[0101] Comparative Example 10 (ZTC1+1 Clarifier)

[0102] An immune function regulating Baoyuan oral liquid and a preparation method thereof, comprising the following raw materials in parts by weight: 134.3 parts of ginseng (five-year-old), 268.6 parts of astragalus (simulated wild), 67.3 parts of licorice, 27.0 parts of cinnamon (fifteen-year-old), and 108.0 parts of ginger (small yellow ginger); the above five flavors are cleaned, steamed, cut, and baked, then soaked in 11 times the amount of ultrapure water for 50 minutes, boiled over high heat, then turned to low heat and kept slightly boiling for 1.3 hours, the liquid medicine is filtered with a 0.45um ceramic inorganic membrane, and the residue is added with 10 times the amount of ultrapure water. After boiling ultrapure water over high heat, turn to low heat and keep it at a slight boil for 55 minutes. The drug solution is filtered through a 0.45um ceramic inorganic membrane, the filtrate is combined, and vacuum concentrated to 400ml at 65°C. The pH value of the system is adjusted to 4.5, and 0.2g of stevioside, 29g of xylitol, 0.6% of ZTC1+1 clarifier (purchased from Tianjin Zhengtiancheng Clarification Technology Co., Ltd.), and stabilizer sodium carboxymethyl cellulose CMC-Na are added. Filter, adjust to a total amount of 500ml, stir evenly, fill, sterilize, and inspect by light.

[0103] Comparative Example 11 (Conventional chitosan gel, no acidification and microspheroidization operation)

[0104] An immune function regulating oral Baoyuan liquid and a preparation method thereof, comprising the following raw materials in parts by weight: 134.3 parts of ginseng (five-year-old), 268.6 parts of astragalus (simulated wild), 67.3 parts of licorice, 27.0 parts of cinnamon (fifteen-year-old), and 108.0 parts of ginger (small yellow ginger); the above five flavors are cleaned, steamed, cut, and baked, then soaked in 11 times the amount of ultrapure water for 50 minutes, boiled over high heat, then turned to low heat and kept slightly boiling for 1.3 hours, the liquid medicine is filtered through a 0.45um ceramic inorganic membrane, and the residue is added with 10 Bring to a boil twice the amount of ultrapure water over high heat, then turn to low heat and keep at a slight boil for 55 minutes. Filter the drug solution through a 0.45um ceramic inorganic membrane, combine the filtrate, and concentrate in a vacuum at 65°C to 400ml. Adjust the pH value of the system to 4.5, add 0.2g of stevioside as a flavoring agent, 29g of xylitol, 0.6% of chitosan gel as a clarifier (Chengdu Wanxiang Hongrun Biotechnology Co., Ltd.), and sodium carboxymethyl cellulose (CMC-Na) as a stabilizer, filter, adjust the total amount to 500ml, stir evenly, fill, sterilize, and inspect by light.

[0105] Comparative Example 12 (no stabilizer added)

[0106] Compared with Example 8, the only difference is that the stabilizer sodium carboxymethyl cellulose CMC-Na is not added, and the others are the same.

[0107] Comparative Example 13 (no clarifier added)

[0108] Compared with Example 8, the only difference is that the clarifier acid chitosan microspheres are not added, and the other aspects are the same. The state of the Baoyuan oral liquid prepared in Comparative Example 12 after being left at a temperature of 25 and a humidity of 60% for 30 days is as follows Figure 5 shown.

[0109] The performance test was carried out on Example 8 and Comparative Examples 9-13, and the test method was as above. The test results are shown in the following table:

[0110] Table 2 Performance test of Baoyuan oral liquid of Example 8 and Comparative Examples 9-13

[0111] Sample Light transmittance (%) Soluble solids (%) Anti-CD3 antibody (μg / mL) Example 1 92 4.6 21.2 Example 8 90 4.3 21.3 Comparative Example 9 82 3.5 17.9 Comparative Example 10 88 3.2 17.8 Comparative Example 11 84 3.7 17.6 Comparative Example 12 87 3.9 18.1 Comparative Example 13 82 3.2 17.7

[0112] As can be seen from the above table, the performance of the implementation case is better than that of the comparative example. The main reasons may be: Chinese medicine decoctions are mostly compound preparations, and water is often used as a solvent during extraction, so some high molecular substances such as starch, gum, protein, and sugar are also extracted. In essence, Chinese medicine decoctions are a mixture of solutions, colloids, suspensions, and emulsions. When colloids and suspensions are left standing, there is sedimentation and aggregation of particles, and emulsions are prone to creaming and rupture when conditions change. At the same time, due to the different charges carried by the particles of colloids, suspensions, and emulsions, they will also attract each other, resulting in a large amount of precipitation after standing. Although there are so many reasons for the formation of precipitation, it involves the acid-base combination of Chinese medicine chemical components, tannin precipitation, hydrolysis degradation, and polymerization reactions. But in general, the treatment principle is: to remove precipitation to the maximum extent without affecting the clinical efficacy. The selection of oral liquid clarifiers must first meet safety, and secondly, the retention rate of active ingredients, clarity, etc. Common clarifiers include diatomite, ZTC1+1 clarifier, water-soluble chitosan, etc. Diatomite is not suitable for filtering oral liquid, and the oral liquid concentration is too high, and it is difficult to filter using diatomite. ZTC1+1 has a good filtering effect on oral liquid, but ZTC1+1 is relatively expensive. Compared with acid-soluble chitosan, water-soluble chitosan is looser in flocculation, and the effect is not as good as acid-soluble chitosan. Therefore, the present invention obtains acid-soluble chitosan by acidifying chitosan, and microspheroidizes acid-soluble chitosan to strengthen its porous adsorption function, so that the clarity effect is better. In addition, stabilizer sodium carboxymethyl cellulose CMC-Na is added to the preparation to ensure the subsequent clarity and stability of the preparation. The present invention can not only properly solve the precipitation problem of Baoyuan oral liquid through new pharmaceutical technology and more reasonable preparation process, but also can improve the efficacy of the drug, and this will also provide new ideas for the application and development of Chinese medicine oral liquid dosage forms.

Claims

1. A chitosan microsphere, characterized in that: By weight, it comprises: 80-120 parts of chitosan, 2-5 parts of emulsifier, and 30-40 parts of oil phase; the chitosan microspheres are obtained by dissolving in an acidic solution and then reacting with the emulsifier and the oil phase.

2. The chitosan microspheres according to claim 1, characterized in that The pH value of the acidic solution is 2-4.

3. The chitosan microspheres according to claim 1, characterized in that The acidic solution includes one or both of acetic acid solution and phosphoric acid solution; the emulsifier includes one or both of sodium dodecyl sulfate and Tween; and the oil phase is one or more of soybean oil, olive oil or sunflower oil.

4. The chitosan microspheres according to any one of claims 1 to 3, characterized in that In the chitosan microspheres, The invention comprises 10-20wt% of chitosan microspheres with a particle size less than 0.1μm, 20-40wt% of chitosan microspheres with a particle size of 0.1-40μm and 45%-70% of chitosan microspheres with a particle size of 45-105μm.

5. The method for preparing chitosan microspheres according to any one of claims 1 to 4, characterized in that: The following steps are involved: Dissolve chitosan in an acidic solution, add an emulsifier, stir, then drop the oil phase, heat the reaction system to 70-90°C after the dropwise addition, react for 30-50 minutes, and purify to obtain chitosan microspheres; The emulsifier is added dropwise for several times, with each addition being 20-40 minutes apart from the last addition.

6. The preparation method according to claim 5, characterized in that: The emulsifier is added dropwise three times, 50-60 wt % of the total emulsifier is added dropwise for the first time, 30-39 wt % of the total emulsifier is added dropwise for the second time, and 1-20 wt % of the total emulsifier is added dropwise for the third time, and each addition is 20-40 minutes apart from the last addition.

7. Use of the chitosan microspheres according to any one of claims 1 to 4 in the preparation of Baoyuan oral liquid preparations.

8. A method for preparing a Baoyuan oral liquid preparation, characterized in that: The following steps are involved: S1. 140-160 parts of ginseng, 300-320 parts of astragalus, 70-85 parts of liquorice, and 28-35 parts of cinnamon bark. Mix 110-130 parts of ginger and bake, then add 12-18 times the amount of water, soak for 40-70 minutes, and boil. Then keep it at a slight boil for 1.2-1.5h, filter it, and obtain filtrate 1 and medicinal residue; S2. Add 10-14 times the amount of water to the residue and boil it, then keep it at a slight boil for 35-65 minutes, filter it, and combine the filtrate. Concentrate and adjust pH to 4.3-5.8; S3, adding 2-25 parts of a clarifier and 0.2-1.2 parts of a stabilizer, mixing evenly, filtering, and obtaining the Baoyuan oral liquid preparation; The clarifier is the chitosan microspheres according to any one of claims 1 to 4.

9. The preparation method according to claim 8, characterized in that: The filtration is carried out by using a ceramic inorganic membrane; preferably, the filtration is carried out by using a ceramic inorganic membrane with a membrane pore size of 0.1-0.45 μm.

10. The Baoyuan oral liquid preparation prepared according to the preparation method according to claim 8 or 9.

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