A compound high-calcium preparation composition and a preparation method thereof

Abstract

The application belongs to the technical field of medicine, and particularly relates to a compound high-calcium preparation composition and a preparation method thereof. The composition raw materials comprise calcium raw materials, zinc raw materials, acylated casein, a co-solvent antifreeze agent, a stabilizer, a sweetening agent, a flavoring essence and water. The calcium raw materials are a combination of calcium gluconate and calcium lactate. The acylated casein, the co-solvent antifreeze agent and the stabilizer can not only improve the solubility of the calcium gluconate and the calcium lactate, but also ensure the stability of calcium gluconate molecules and calcium lactate molecules in an aqueous solution, thereby preventing crystallization under low-temperature conditions. The preparation method is simple, can greatly reduce energy consumption, improve production efficiency and reduce production cost. The composition does not contain monosaccharides and disaccharides, and does not add any preservatives and bacteriostatic agents, thereby avoiding potential safety risks.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a compound high-calcium preparation composition and its preparation method. Background Technology

[0002] Calcium is one of the essential macroelements in the human body, playing a vital physiological role, especially important for infants, children, adolescents, pregnant women, breastfeeding mothers, and the elderly who experience significant calcium loss with age. Currently, there are many calcium supplements on the market, with calcium gluconate being the primary source. Calcium gluconate is a water-soluble organic calcium acid and a commonly used calcium supplement in health foods and pharmaceuticals. Calcium gluconate comes in various dosage forms, such as oral solutions, tablets, granules, and injections. Due to its convenience, lack of disintegration in the body, and rapid absorption, oral solutions of calcium gluconate hold an important position in the calcium supplementation field. Currently, most products on the market contain 50mg-150mg of elemental calcium per 10ml or 15ml, such as compound calcium gluconate oral solutions and calcium gluconate zinc oral solutions. According to the "Chinese Dietary Guidelines (2022)," the recommended daily calcium intake varies across different age groups. Children and adolescents, who experience rapid growth and development, are advised to consume 800mg-1200mg of calcium; pregnant women, lactating mothers, and the elderly are advised to consume 800mg-1000mg. Currently available oral calcium supplements require multiple doses to meet the recommended intake. The low calcium content in calcium gluconate is primarily due to its molecular structure. Its theoretical calcium content is approximately 9.3%, and even 100% pure calcium gluconate powder contains only about 93mg of elemental calcium per gram; this proportion is relatively low compared to other calcium salts. Moreover, calcium gluconate has a solubility of about 3.5% in water. The calcium gluconate in commercially available oral solutions is in a supersaturated state. Simply increasing the concentration of calcium gluconate to increase the calcium content will lead to problems such as calcium ion precipitation in the solution, poor taste, and children refusing to take it. In particular, the system is unstable during long-term storage or under low-temperature conditions, which can easily lead to calcium ion precipitation and crystallization in the solution, affecting the appearance, stability, effectiveness and safety of the product, and thus reducing the calcium supplementation effect. This is the technical challenge currently faced in increasing the calcium content of calcium-containing oral solutions.

[0003] Patent CN119548456A discloses a calcium gluconate zinc oral solution and its preparation method. The solubilizing system used in this patent is a sugar alcohol chelating agent, the stabilizer is sodium chloride or sodium citrate, and the pH is adjusted to 3.0-7.5 using alkaline substances such as sodium hydroxide or calcium hydroxide. However, apart from the pH adjuster, the other substances are acidic or neutral in purified water, and using alkaline substances to adjust the solution pH to acidic is clearly unreasonable. Furthermore, in addition to the antibacterial agent in the formulation, sterilization is required during the process, which is time-consuming and labor-intensive. Patent CN117883555A discloses a compound calcium gluconate oral solution and its preparation method. This patent uses a calcium gluconate promoter to improve the calcium content and the stability of the oral solution. However, the calcium gluconate promoter, polydopamine nanospheres, is a novel excipient, lacking long-term toxicological data, posing safety risks to children or pregnant women. Moreover, the nanosphere preparation process is complex, and industrial scale-up presents certain difficulties.

[0004] In summary, while existing technologies have improved the crystallization phenomenon of calcium gluconate oral solutions, no reports have been found regarding crystallization at high calcium content. Given that these issues remain unresolved, there is an urgent need to provide an oral solution with high calcium content that is highly safe, stable, and easy to prepare. Summary of the Invention

[0005] To address the problem of calcium precipitation in high-calcium preparations stored at low temperatures due to the difficulty in improving stability in existing technologies, this invention provides a compound high-calcium preparation composition and its preparation method. The main components include calcium raw material, zinc raw material, acylated casein, co-solubilizing antifreeze agent, stabilizer, sweetener, flavoring, and water. The calcium raw material is a combination of calcium gluconate and calcium lactate. Since single calcium sources like calcium gluconate or calcium lactate provide insufficient calcium and easily form supersaturated solutions, calcium precipitation occurs during storage. Therefore, this invention uses a combination of calcium gluconate and calcium lactate. The acylated casein, co-solubilizing antifreeze agent, and stabilizer in this formulation improve the solubility of calcium gluconate and calcium lactate while ensuring the stability of calcium gluconate and calcium lactate molecules in aqueous solution, preventing crystallization at low temperatures. The preparation method is simple, significantly reducing energy consumption, improving production efficiency, and lowering production costs. This composition does not contain monosaccharides or disaccharides, and does not contain any preservatives or antibacterial agents, thus avoiding potential safety risks.

[0006] The first aspect of this invention provides a compound high-calcium preparation composition, which includes at least calcium raw material, zinc raw material, acylated casein, co-soluble antifreeze agent, stabilizer and water. Each mL of the compound high-calcium preparation composition includes 5-50 mg of calcium element, 0.12-0.45 mg of zinc element, 10-75 mg of acylated casein, 1.5-20 mg of co-soluble antifreeze agent, and 0.8-1.2 mg of stabilizer. The pH value of the composition is 3.5-4.5. The dosage form of the compound high-calcium preparation composition is an oral solution. The water is purified water with a conductivity of not more than 4.3 μs / cm at 20°C.

[0007] The calcium raw material is a combination of calcium lactate and calcium gluconate, with a calcium element mass ratio of 3:1 between calcium lactate and calcium gluconate; furthermore, the compound high-calcium composition contains 10-45 mg of calcium per mL.

[0008] Since calcium gluconate or calcium lactate, as single calcium sources, provide a small amount of calcium and are prone to forming supersaturated solutions, calcium will precipitate out during storage. Therefore, this invention uses a combination of calcium gluconate and calcium lactate.

[0009] The zinc raw material is one or a combination of two of zinc gluconate and zinc lactate.

[0010] The co-solubilized antifreeze agent is selected from one or more of glycerol, sorbitol, polyethylene glycol 400, propylene glycol, trehalose, and mannitol; preferably, the co-solubilized antifreeze agent is a combination of glycerol and sorbitol, and the weight ratio of glycerol to sorbitol is 1:1 to 3:1.

[0011] Glycerol reduces the polarity of high-calcium formulation solutions through hydrogen bonding interactions, lowering the dielectric constant and weakening the electrostatic attraction between ions, thus promoting easier dissociation of calcium ions in a low-polarity environment. Sorbitol forms a eutectic mixture with water, significantly lowering the freezing point of the solution and inhibiting crystal formation by enhancing hydrogen bonding interference through multiple hydroxyl groups. The combined use of these two agents synergistically enhances the solubilizing effect on calcium gluconate and calcium lactate, prevents the precipitation of calcium gluconate crystals, and improves product stability.

[0012] The stabilizer is one or a combination of two of hydroxypropyl methylcellulose and povidone K30.

[0013] In the above formulation, higher calcium content and lower temperature increase the likelihood of crystallization. Since calcium gluconate in the formulation is a supersaturated solution, preventing calcium gluconate crystallization under low-temperature conditions is the key technology of this invention. This invention improves the solubility of calcium gluconate and calcium lactate and prevents calcium ion precipitation under low-temperature conditions by adding a co-soluble antifreeze agent, acylated casein, and a stabilizer.

[0014] Casein is a milk protein rich in phosphorylated serine residues. The phosphoserine residues in the casein molecule carry a negative charge and bind to calcium ions through electrostatic interactions. Special functional groups in the casein molecule, such as phosphate groups and carboxylic acid groups, can also undergo complexation reactions with calcium ions. When calcium ions bind to casein, the conformation of the casein molecule changes, exposing hydrophobic regions. These exposed hydrophobic regions aggregate to form hydrophobic bonds, further stabilizing the casein-calcium ion complex. When casein is acylated, the acyling agent is linked to the protein peptide chain via an amide bond, thereby altering the charge and conformation of casein, increasing its solubility and enhancing its stability. If too little acylated casein is added, calcium ions cannot be fully chelated or dispersed, which may cause calcium to exist in a free form, leading to protein aggregation or incomplete micelle structure, thereby reducing the thermal and storage stability of the system and causing turbidity, astringency, etc. If too much acylated casein is added, it will over-chelate calcium ions, forming a large number of insoluble complexes, causing flocculation or precipitation in the system, thus inhibiting calcium absorption; it will also destroy the structure of casein micelles and reduce the stability of the system.

[0015] When glycerol and sorbitol are selected as co-solvent antifreeze agents, after mixing with water, glycerol and sorbitol change the solvent polarity through hydrogen bonding or dipole interactions, reducing the dielectric constant and weakening the electrostatic attraction between ions, thus promoting easier dissociation of calcium ions in a low-polarity environment. Glycerol and sorbitol form a eutectic mixture with water, significantly lowering the freezing point of the solution and inhibiting crystallization. Glycerol and sorbitol can also increase the solution viscosity, reduce the diffusion rate of calcium ions and anions, and slow down the crystallization kinetics. This invention, by screening the weight ratio of glycerol to sorbitol in the co-solvent antifreeze agent, determined that a glycerol:sorbitol weight ratio of 1:1 to 3:1 can significantly improve the solubility of calcium gluconate and calcium lactate, and prevent crystallization within 20 days under low-temperature conditions.

[0016] Stabilizers, when mixed with water, increase the viscosity of the solution, preventing small particles in a supersaturated system from coalescing into larger particles, reducing the diffusion rate of calcium ions and anions such as gluconate and lactate, ensuring the stability of the supersaturated system, and delaying calcium ion crystallization. Hydroxypropyl methylcellulose, in addition to increasing the viscosity of the solution, can also use its polymer chains to act as a physical barrier, hindering the aggregation of calcium ions in a supersaturated system, especially at low temperatures, maintaining solution stability, inhibiting phase separation, and preventing calcium ion precipitation. Povidone K30, when mixed with water, binds to calcium ions through hydrogen bonds, forming a soluble complex that inhibits the formation of calcium ion crystal nuclei; povidone K30 can also adsorb onto the surface of crystal nuclei, preventing further crystal growth, especially at low temperatures, maintaining solution stability and reducing calcium ion precipitation caused by phase separation. Stabilizers can work synergistically with co-soluble antifreeze agents to jointly prevent calcium ion precipitation in the solution.

[0017] Furthermore, the compound high-calcium preparation composition may also contain a sweetener and / or flavoring, with each mL of the compound high-calcium preparation containing 1.0-2.5 mg of sweetener and 0.2-0.5 mg of flavoring. The sweetener is steviol glycoside; the flavoring is peach flavoring or strawberry flavoring.

[0018] A second aspect of the present invention also provides a method for preparing the compound high-calcium preparation composition, comprising the following steps: (1) Preparation of acylated casein: Dissolve casein in phosphate buffer solution with pH=8.0-9.0, heat in a water bath at 30-40℃, and stir until completely dissolved; slowly add succinic anhydride powder to the casein solution, with succinic anhydride accounting for 6-10% of the weight of casein, while adjusting and maintaining pH=8.0-9.0; continue to maintain water bath at 30-40℃, turn on stirring at 200-300 rpm for 2-3 hours until the sample is fully acylated; after the reaction is complete, adjust the pH to 4-5; then dialyze with deionized water at 2000-3500D for 24-36 hours to remove free succinic acid and byproducts, and dry to obtain acylated casein powder; There are no special requirements for the solubility ratio of casein to phosphate buffer, as long as it is sufficient to completely dissolve the casein. The ratio of succinic anhydride to casein determines the degree of acylation. A low ratio will result in a low target degree of acylation, affecting the binding with calcium ions; a high ratio will lead to a decrease in lysine utilization or cause excessive protein hydrolysis or cross-linking, reducing product yield. (2) Take 75-80 wt% purified water of the prescription amount, heat to boiling, boil for 5-10 minutes; then slowly add the stabilizer, turn on the stirring until the stabilizer is completely dissolved, then slowly add calcium lactate and calcium gluconate, continue stirring, and observe the dissolution of calcium raw materials until the solution is clear and there is no precipitate. (3) Add 10-15 wt% of the prescribed amount of boiled purified water to the material obtained in step (2). When the temperature drops to 35-50℃, add the prescribed amount of zinc raw material, acylated casein and co-solubilized antifreeze, stir to dissolve, and observe the dissolution until the solution is clear and there is no precipitate. (4) Immerse the ultrasonic probe directly into the material obtained in step (3), start the ultrasonic generator at a frequency of 20-25kHz for 5-30 minutes, observe that the dissolution is complete, and make up the volume after the temperature drops to room temperature; then start the ultrasonic generator again for 8-10 minutes. (5) The material obtained in step (4) is filtered sequentially through filter elements with pore sizes of 0.45μm and 0.22μm, then filled, and sterilized at 100-110℃ for 15-20 minutes to obtain the compound high calcium preparation composition.

[0019] Furthermore, in step (1), the pH of the phosphate buffer solution is 8.5. When adding succinic anhydride powder, a pH adjuster is needed to adjust and maintain the pH at 8.5. Sufficient acylation is required until the target acylation value reaches 72-78%. In a specific embodiment of this invention, the acylation value in the reaction is monitored using the 2,4,6-trinitrobenzenesulfonic acid colorimetric method. This method is a commonly used method in the field for detecting acylation values ​​and can be referenced in the literature (Preparation and Properties Study of Succinylated Modified Fish Collagen [J]. China Leather, 2020, 49(12):9-14+35). If a pH adjuster is required, a commonly used pH adjuster should be selected.

[0020] Furthermore, in step (2), after adding the stabilizer, stir at a speed of 200-300 rpm for 3-5 min; after adding calcium lactate and calcium gluconate, stir at a speed of 200-300 rpm for 3-5 min.

[0021] Furthermore, in step (3), after adding zinc raw material, acylated casein and co-solubilized antifreeze, stir for 3-5 min; sweetener is added together with zinc raw material, acylated casein and co-solubilized antifreeze.

[0022] Furthermore, in step (4), after complete ultrasonic dissolution, the fragrance is added and the volume is adjusted after the temperature drops to room temperature.

[0023] The preparation method employs ultrasonic-assisted dissolution technology to dissolve materials, achieving superior dissolution effect and efficiency compared to ordinary stirrers. Ultrasonic dissolution utilizes the extreme physicochemical effects generated by acoustic cavitation to powerfully break particles at the microscopic scale, promoting dissolution and dispersion with extremely high efficiency. The dispersed materials are uniform and less prone to agglomeration. In contrast, stirrers use mechanical devices (blades, magnetic stir bar, turbine, homogenizer, etc.) to induce macroscopic flow (convection) and relative motion (shear) between different flow velocity layers. While stirrers can also dissolve materials, their efficiency is significantly lower than that of ultrasonic technology. This invention, through a comparative study of ultrasonic-assisted dissolution technology and stirrer dissolution technology, found that ultrasonic-assisted dissolution technology has significantly higher dissolution efficiency than stirring dissolution, and no crystallization occurred within 20 days.

[0024] The present invention has achieved the following beneficial effects: 1) In this invention, the co-solvent is an acylated casein and a co-solvent antifreeze complex, wherein the co-solvent antifreeze is preferably a combination of glycerol and sorbitol, with a weight ratio of 1:1 to 3:1. Glycerol reduces the polarity of the high-calcium formulation solution through hydrogen bonding interactions, lowers the dielectric constant, weakens the electrostatic attraction between ions, and promotes easier dissociation of calcium ions in a low-polarity environment; sorbitol forms a eutectic mixture with water, and significantly lowers the freezing point of the solution and inhibits crystal formation by strengthening hydrogen bonding interference through multiple hydroxyl groups. The combined use of the two has a synergistic effect, significantly improving the solubilizing effect on calcium gluconate and calcium lactate, preventing the precipitation of calcium gluconate crystals, and improving product stability.

[0025] 2) In this invention, the stabilizer is one or a combination of hydroxypropyl methylcellulose and povidone K30, which can improve the stability of calcium gluconate and calcium lactate molecules in aqueous solution, reduce the precipitation of calcium ions caused by solution phase separation, and thus improve the stability of the product.

[0026] 3) This invention can significantly increase calcium content through the synergistic effect of acylated casein, co-soluble antifreeze, and stabilizer.

[0027] 4) The oral liquid in this invention does not contain antibacterial agents or preservatives. Long-term testing has proven that the formulation of this invention has a better antibacterial effect than commercially available products that contain antibacterial agents.

[0028] 5) This invention uses ultrasonic-assisted dissolution technology. This process is simple to operate and can significantly reduce energy consumption, improve production efficiency, and reduce production costs.

[0029] In summary, this invention uses acylated casein, co-solubilizing antifreeze, and stabilizer as a solubilizing system, which can significantly increase the calcium content in compound high-calcium preparations, prevent calcium ion precipitation under low-temperature conditions, and improve the stability of oral solutions. The overall formulation has good stability, contains no monosaccharides or disaccharides, and does not contain preservatives or antibacterial agents, thus ensuring high safety. The preparation process is simple, energy consumption is low, production efficiency is high, and the market prospects are broad. Attached Figure Description

[0030] Figure 1 This is a standard curve for determining the calcium content in the compound high-calcium preparation composition of the present invention. Detailed Implementation

[0031] The technical solution of the present invention will be further described below with reference to specific embodiments. These embodiments are only used to illustrate the technical solution of the present invention in more detail and should not be construed as limiting the scope of protection of the present invention.

[0032] In the following examples and comparative examples, the acylation value in the reaction was monitored using the 2,4,6-trinitrobenzenesulfonic acid colorimetric method. This method is a commonly used method in the art for detecting acylation values ​​and can be referenced in the literature (Preparation and Property Study of Succinylated Modified Fish Collagen [J]. China Leather, 2020, 49(12):9-14+35). The boiled purified water used can be cooled to ambient temperature or can be at any temperature between room temperature and boiling temperature; this is not limited in this invention.

[0033] Example 1: A compound high-calcium preparation composition, the formulation of which per milliliter of the composition is as follows: Calcium lactate (calcium content 11.25mg) 61.48mg Calcium gluconate (calcium content 3.75mg) 40.32mg Zinc lactate (zinc content 0.4 mg) 1.80 mg 40mg of acylated casein 6mg of glycerin Sorbitol 3mg Hydroxypropyl methylcellulose 0.9mg Steviosides 1.5mg Peach flavoring 0.3mg Add purified water to the remaining volume.

[0034] The preparation process of the compound high-calcium preparation composition is as follows: (1) Preparation of acylated casein: Casein was dissolved in 0.1M phosphate buffer (pH=8.5), heated in a water bath (temperature controlled at 35℃), and stirred until completely dissolved; succinic anhydride powder was slowly added to the casein solution (succinic anhydride accounts for 8% of the weight of casein), while maintaining pH=8.5 (adjusted with 1M NaOH solution); the water bath was kept at 35℃, stirring was started at 250 rpm, and the stirring time was 2 hours until the sample was fully acylated (target acylation value 75%); after the reaction was completed, 0.05M dilute hydrochloric acid was added to adjust the pH to 4.5; then the sample was dialyzed with deionized water in a 3500D dialysis bag for 30 hours to remove free succinic acid and byproducts, and freeze-dried (temperature: -40℃, time 24 hours) to obtain acylated casein powder; (2) Take 75 wt% purified water of the prescription amount, heat to boiling, boil for 10 min; then stop heating and immediately and slowly add hydroxypropyl methylcellulose, turn on the electric stirrer (speed 250 rpm), stir for 5 min, after the hydroxypropyl methylcellulose is completely dissolved, slowly add calcium lactate and calcium gluconate, continue stirring at 250 rpm for 5 min, observe the calcium dissolution until the solution is clear and there is no precipitate; (3) Add 15 wt% of the prescribed amount of boiled purified water to the material obtained in step (2). When the temperature drops to 35°C, add the prescribed amount of zinc lactate, acylated casein, glycerol, sorbitol and steviol glycosides. Stir at 250 rpm for 4 min to dissolve. Observe the dissolution until the solution is clear and there is no precipitate. (4) Immerse the ultrasonic probe directly into the material obtained in step (3), start the ultrasonic generator at a frequency of 25kHz for 8 minutes, observe that the dissolution is complete, and add peach flavoring after the temperature drops to room temperature and make up the volume; then start the ultrasonic generator again and sonicate at a frequency of 25kHz for 8 minutes. (5) The material obtained in step (4) is filtered twice through 0.45μm and 0.22μm polyethersulfone filter cartridges, then filled, sterilized at 105℃ for 20min, and packaged to obtain the compound high calcium preparation composition (composition pH=4.0).

[0035] Example 2 A compound high-calcium preparation composition, the formulation of which per milliliter of the composition is as follows: Calcium lactate (calcium content 22.51mg) 123.0mg Calcium gluconate (calcium content 7.50mg) 80.65mg Zinc gluconate (zinc content 0.3mg) 2.10mg 50mg of acylated casein 8mg of glycerin Sorbitol 4mg Povidone K30 1.1mg Steviosides 1.8mg Peach flavoring 0.3mg Add purified water to the remaining volume.

[0036] The preparation process of the compound high-calcium preparation composition is as follows: (1) Preparation of acylated casein: Casein was dissolved in 0.1M phosphate buffer (pH=8.5), heated in a water bath (temperature controlled at 38℃), and stirred until completely dissolved; succinic anhydride powder was slowly added to the casein solution (succinic anhydride accounts for 7% of the weight of casein), while maintaining pH=8.5 (adjusted with 1M NaOH solution); the water bath was kept at 35℃, stirring was started at 250 rpm, and the stirring time was 3 hours until the sample was fully acylated (target acylation value 78%); after the reaction was completed, 0.05M dilute hydrochloric acid was added to adjust the pH to 4.0; then the sample was dialyzed with deionized water in a 2000D dialysis bag for 28 hours to remove free succinic acid and byproducts, and freeze-dried (temperature: -40℃, time 24 hours) to obtain acylated casein powder; (2) Take 75 wt% purified water of the prescription amount, heat to boiling, boil for 10 min; then stop heating and immediately and slowly add povidone K30, turn on the electric stirrer (speed 250 rpm), stir for 5 min, and after the povidone K30 is completely dissolved, slowly add calcium lactate and calcium gluconate, continue stirring at 250 rpm for 5 min, and observe the calcium dissolution until the solution is clear and there is no precipitate; (3) Add 15 wt% of the prescribed amount of boiled purified water to the material obtained in step (2). When the temperature drops to 40°C, add the prescribed amount of zinc gluconate, acylated casein, glycerol, sorbitol and steviol glycosides. Stir at 250 rpm for 5 min to dissolve. Observe the dissolution until the solution is clear and there is no precipitate. (4) Immerse the ultrasonic probe directly into the material obtained in step (3), start the ultrasonic generator at a frequency of 25kHz for 8 minutes, observe that the dissolution is complete, and add peach flavoring after the temperature drops to room temperature and make up the volume; then start the ultrasonic generator again and sonicate at a frequency of 25kHz for 8 minutes. (5) The material obtained in step (4) is filtered twice through 0.45μm and 0.22μm polyethersulfone filter cartridges, then filled, sterilized at 105℃ for 20min, and packaged to obtain the compound high calcium preparation composition (composition pH=4.1).

[0037] Example 3 A compound high-calcium preparation composition, the formulation of each milliliter of the composition is as follows: Calcium lactate (calcium content 33.75mg) 184.43mg Calcium gluconate (calcium content 11.25mg) 120.97mg Zinc lactate (zinc content 0.3 mg) 1.35 mg Acylated casein 60mg 10mg of glycerin Sorbitol 5mg Povidone K30 1.15mg Steviosides 2.0mg Strawberry flavoring 0.5mg Add purified water to the remaining volume.

[0038] The preparation process of the compound high-calcium preparation composition is as follows: (1) Preparation of acylated casein: Casein was dissolved in 0.1M phosphate buffer (pH=8.5), heated in a water bath (temperature controlled at 35℃), and stirred until completely dissolved; succinic anhydride powder was slowly added to the casein solution (succinic anhydride accounts for 9% of the weight of casein), while maintaining pH=8.5 (adjusted with 1M NaOH solution); the water bath was kept at 35℃, stirring was started at 250 rpm, and the stirring time was 3 hours until the sample was fully acylated (target acylation value 75%); after the reaction was completed, 0.05M dilute hydrochloric acid was added to adjust the pH to 4.0; then the sample was dialyzed with deionized water in a 3500D dialysis bag for 28 hours to remove free succinic acid and byproducts, and freeze-dried (temperature: -40℃, time 24 hours) to obtain acylated casein powder; (2) Take 75 wt% purified water of the prescription amount, heat to boiling, boil for 10 min; then stop heating and immediately and slowly add povidone K30, turn on the electric stirrer (speed 250 rpm), stir for 5 min, and after the povidone K30 is completely dissolved, slowly add calcium lactate and calcium gluconate, continue stirring at 250 rpm for 5 min, and observe the calcium dissolution until the solution is clear and there is no precipitate; (3) Add 15 wt% of the prescribed amount of boiled purified water to the material obtained in step (2). When the temperature drops to 40°C, add the prescribed amount of zinc lactate, acylated casein, glycerol, sorbitol and steviol glycosides. Stir at 250 rpm for 5 min to dissolve. Observe the dissolution until the solution is clear and there is no precipitate. (4) Immerse the ultrasonic probe directly into the material obtained in step (3), start the ultrasonic generator at a frequency of 25kHz for 10 minutes, observe that the dissolution is complete, and add strawberry flavoring after the temperature drops to room temperature and make up the volume; then start the ultrasonic generator again and sonicate at a frequency of 25kHz for 8 minutes. (5) The material obtained in step (4) is filtered twice through 0.45μm and 0.22μm polyethersulfone filter cartridges, then filled, sterilized at 105℃ for 20min, and packaged to obtain the compound high calcium preparation composition (composition pH=4.0).

[0039] Example 4 A compound high-calcium preparation composition, the formulation of each milliliter of the composition is as follows: Calcium lactate (calcium content 33.75mg) 184.43mg Calcium gluconate (calcium content 11.25mg) 120.97mg Zinc lactate (zinc content 0.3mg) 1.35mg Acylated casein 85mg 10mg of glycerin Sorbitol 5mg Povidone K30 1.15mg Steviosides 2.0mg Strawberry flavoring 0.5mg Add purified water to the remaining volume.

[0040] (1) Preparation of acylated casein: Casein was dissolved in 0.1M phosphate buffer (pH=8.5), heated in a water bath (temperature controlled at 35℃), and stirred until completely dissolved; succinic anhydride powder was slowly added to the casein solution (succinic anhydride accounts for 9% of the weight of casein), while maintaining pH=8.5 (adjusted with 1M NaOH solution); the water bath was kept at 35℃, stirring was started at 250 rpm, and the stirring time was 3 hours until the sample was fully acylated (target acylation value 75%); after the reaction was completed, 0.05M dilute hydrochloric acid was added to adjust the pH to 4.0; then the sample was dialyzed with deionized water in a 3500D dialysis bag for 28 hours to remove free succinic acid and byproducts, and freeze-dried (temperature: -40℃, time 24 hours) to obtain acylated casein powder; (2) Take 75 wt% purified water of the prescription amount, heat to boiling, boil for 10 min; then stop heating and immediately and slowly add povidone K30, turn on the electric stirrer (speed 250 rpm), stir for 5 min, and after the povidone K30 is completely dissolved, slowly add calcium lactate and calcium gluconate, continue stirring at 250 rpm for 5 min, and observe the calcium dissolution until the solution is clear and there is no precipitate; (3) Add 15 wt% of the prescribed amount of boiled purified water to the material obtained in step (2). When the temperature drops to 40°C, add the prescribed amount of zinc lactate, acylated casein, glycerol, sorbitol and steviol glycosides. Stir at 250 rpm for 5 min to dissolve. Observe the dissolution until the solution is clear and there is no precipitate. (4) Immerse the ultrasonic probe directly into the material obtained in step (3), start the ultrasonic generator at a frequency of 25kHz for 8 minutes, observe that the dissolution is complete, and add strawberry flavoring after the temperature drops to room temperature and make up the volume; then start the ultrasonic generator again and sonicate at a frequency of 25kHz for 8 minutes. (5) The material obtained in step (4) is filtered twice through 0.45μm and 0.22μm polyethersulfone filter cartridges, then filled, sterilized at 105℃ for 20min, and packaged to obtain the compound high calcium preparation composition (pH=4.10).

[0041] Comparative Example 1: A compound high-calcium preparation composition, with the following formulation per milliliter: Calcium lactate (calcium content 33.75mg) 184.43mg Calcium gluconate (calcium content 11.25mg) 120.97mg Zinc lactate (zinc content 0.3mg) 1.35mg 10mg of glycerin Sorbitol 5mg Povidone K30 1.15mg Steviosides 2.0mg Strawberry flavoring 0.5mg Add purified water to the remaining volume.

[0042] (1) Take 75 wt% purified water of the prescription amount, heat to boiling, boil for 10 min; then stop heating and immediately and slowly add povidone K30, turn on the electric stirrer (speed 250 rpm), stir for 5 min, and after the povidone K30 is completely dissolved, slowly add calcium lactate and calcium gluconate, continue stirring at 250 rpm for 5 min, and observe the calcium dissolution until the solution is clear and there is no precipitate; (2) Add 15 wt% of the prescribed amount of boiled purified water to the material obtained in step (1). When the temperature drops to 40°C, add the prescribed amount of zinc lactate, glycerol, sorbitol and steviol glycosides. Stir at 250 rpm for 5 min to dissolve. Observe the dissolution until the solution is clear and there is no precipitate. (3) Immerse the ultrasonic probe directly into the material obtained in step (2), start the ultrasonic generator at a frequency of 20kHz for 10 minutes, observe that the dissolution is complete, and add strawberry flavoring after the temperature drops to room temperature and make up the volume; then start the ultrasonic generator again and sonicate at a frequency of 20kHz for 8 minutes. (4) The material obtained in step (3) is filtered twice through 0.45μm and 0.22μm polyethersulfone filter cartridges, then filled, sterilized at 105℃ for 20min, and packaged to obtain the compound high calcium preparation composition (pH=4.05).

[0043] Comparative Example 2: A compound high-calcium preparation composition, with the following formulation per milliliter: Calcium lactate (calcium content 33.75mg) 184.43mg Calcium gluconate (calcium content 11.25mg) 120.97mg Zinc lactate (zinc content 0.3mg) 1.35mg Casein 60mg 10mg of glycerin Sorbitol 5mg Povidone K30 1.15mg Steviosides 2.0mg Strawberry flavoring 0.5mg Add purified water to the remaining volume.

[0044] (1) Take 78 wt% purified water of the prescription amount, heat to boiling, boil for 10 min; then stop heating and immediately and slowly add povidone K30, turn on the electric stirrer (speed 250 rpm), stir for 5 min, and after the povidone K30 is completely dissolved, slowly add calcium lactate and calcium gluconate, continue stirring at 250 rpm for 5 min, and observe the calcium dissolution until the solution is clear and there is no precipitate; (2) Add 15 wt% of the prescribed amount of boiled purified water to the material obtained in step (1). When the temperature drops to 40°C, add the prescribed amount of zinc lactate, casein, glycerol, sorbitol and steviol glycosides. Stir at 250 rpm for 5 min to dissolve. Observe the dissolution until the solution is clear and there is no precipitate. (3) Immerse the ultrasonic probe directly into the material obtained in step (2), start the ultrasonic generator at a frequency of 25kHz for 10 minutes, observe that the dissolution is complete, and add strawberry flavoring after the temperature drops to room temperature and make up the volume; then start the ultrasonic generator again and sonicate at a frequency of 25kHz for 8 minutes. (4) The material obtained in step (3) is filtered twice through 0.45μm and 0.22μm polyethersulfone filter cartridges, then filled, sterilized at 105℃ for 20min, and packaged to obtain the compound high calcium preparation composition (pH=3.90).

[0045] Experimental Example 1 The products prepared in Examples 1-4, Comparative Examples 1 and 2, and two commercially available oral solutions with different calcium content (hereinafter referred to as S1 and S2) (S1 and S2 have specifications of 15 mg / ml and 12 mg / ml, respectively) were left to stand for 10 days at 4°C and 25°C, and the presence of crystals was observed. The results are shown in Table 1.

[0046] The calcium content determination method referenced the content determination method for compound calcium gluconate oral solution in the 2020 edition of the Pharmacopoeia of the People's Republic of China. Method validation showed a good linear relationship between the calcium gluconate concentration (C) and the EDTA titration volume in the range of 30.26-135.12 mg / ml, i.e., a good linear relationship between the calcium content and the EDTA titration volume in the range of 2.81-12.55 mg / ml. The standard curve equation was y = 0.3741x + 0.0002(R²). 2 =0.9997) Figure 1 As shown in the figure, the precision and accuracy of this method meet the requirements, and the established detection method is accurate and reliable.

[0047] Table 1 Results of static tests under low temperature and normal temperature conditions The results showed that the products prepared in Examples 1-4 did not exhibit crystallization and had better stability than commercially available products S1 and S2, and Comparative Examples 1 and 2.

[0048] In Example 4, the amount of acylated casein added was increased beyond the preferred range of 10-75 mg per milliliter. The results showed that after the amount of acylated casein added was increased to 75 mg per milliliter, further increasing the amount did not significantly improve the performance of the oral solution. Therefore, from a cost perspective, this scheme limits the amount of acylated casein added to 10-75 mg per milliliter.

[0049] Experiment Example 2 The products prepared in Examples 1-4 and Comparative Examples 1 and 2, as well as two commercially available oral solutions with different calcium contents (S1 and S2), were investigated under accelerated testing for 3 months. The accelerated testing followed the "Guidelines for Stability Testing of Raw Materials and Preparations" (9001) of the 2020 edition of the Chinese Pharmacopoeia, Volume IV. Samples were placed at 40℃±2℃ and 75%±5% relative humidity for 3 months, and samples were taken at 0, 1, 2, and 3 months for testing of properties, calcium content, and microorganisms. Microbial limit testing followed the "Microbial Limit Tests for Non-Sterile Products: Microbial Count Method (General Rule 1105) and Control Bacteria Test Method (General Rule 1106)" and the "Microbial Limit Standards for Non-Sterile Drugs (General Rule 1107)" of the 2020 edition of the Chinese Pharmacopoeia, Volume IV. The results are shown in Table 2.

[0050] Table 2 Accelerated Test Results As shown in Table 2, the products obtained under accelerated conditions in Examples 1, 2, 3, and 4 showed no significant changes in appearance or calcium content, and met the microbial limits. Compared with commercially available products, their stability and antibacterial effect were superior. In Comparative Examples 1 and 2, the calcium content decreased to 32% and 40% respectively after one month of accelerated treatment. Compared with commercially available products and Examples 1-4, their stability and antibacterial effect were poor; therefore, no further stability studies were conducted.

[0051] Experimental Example 3 While keeping the total mass (5g) of glycerol and sorbitol constant, 100ml of aqueous solutions with different ratios of the two were prepared, and 20g of calcium gluconate was added to each solution. The solutions were sealed and shaken at room temperature for 24 hours to prepare calcium saturated solutions, which were used as test samples (samples 1-9). Add 20g of calcium gluconate to 100ml of purified water, seal, and shake at room temperature for 24 hours to prepare a calcium saturated solution as a control.

[0052] The calcium content determination method referenced the content determination method for compound calcium gluconate oral solution in the 2020 edition of the Pharmacopoeia of the People's Republic of China. Method validation showed a good linear relationship between the calcium gluconate concentration (C) and the EDTA titration volume in the range of 30.26-135.12 mg / ml, i.e., a good linear relationship between the calcium content and the EDTA titration volume in the range of 2.81-12.55 mg / ml. The standard curve equation was y = 0.3741x + 0.0002(R²). 2 =0.9997) Figure 1 As shown in the figure, the precision and accuracy of this method meet the requirements, and the established detection method is accurate and reliable.

[0053] Table 3. Effects of glycerol and sorbitol on calcium solubilization The results shown in Table 3 indicate that the solubility and crystallization stability of calcium were significantly improved when glycerol and sorbitol were combined in a mass ratio of 1:1 to 3:1. When the ratio of glycerol to sorbitol was not within the 1:1 to 3:1 range (samples 1, 2, 6, 7, 8, and 9), the calcium content was lower and the stability was worse compared to samples 3, 4, and 5.

[0054] Experiment Example 4: Comparative Study of Ultrasonic Dissolution Technology and Stirring Dissolution Technology Prepare materials according to the prescription in Table 4, and follow the preparation process in Table 5. Dissolve the materials using both ultrasonic dissolution and stirrer dissolution methods. Step 4 is the main focus of the study. Samples were taken at 5 min, 8 min, 10 min, and 30 min for testing. Calcium content and stability were used as indicators to examine the two preparation methods. The method for determining calcium content was the same as in Experiment 3.

[0055] Table 4. Prescription information for Experiment Example 4 Table 5. Two preparation processes for Experiment Example 4 Table 6 Results of ultrasonic and stirring dissolution studies The results shown in Table 6 indicate that ultrasonic-assisted dissolution technology is superior to agitator stirring in terms of dissolution efficiency, dissolution state, and stability.

[0056] Experimental Example 5 Monosaccharides and disaccharides were determined in the products obtained in Examples 1-4 and Comparative Examples 1 and 2. The detection method adopted was GB5009.8-2023 National Food Safety Standard for the Determination of Fructose, Glucose, Sucrose, Maltose and Lactose in Food. The results are shown in Table 7.

[0057] Table 7 Sugar content detection The test results showed no monosaccharides or disaccharides, indicating that the formula does not contain sugars that raise blood sugar.

[0058] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any equivalent substitutions or modifications made by those skilled in the art within the spirit and principle of the present invention without any creative effort should be included within the protection scope of the present invention.

Claims

1. A compound high-calcium preparation composition, characterized in that, The compound high-calcium preparation composition includes at least calcium raw material, zinc raw material, acylated casein, co-solubilized antifreeze, stabilizer and water. Each mL of the compound high-calcium preparation composition includes 5-50 mg of calcium element, 0.12-0.45 mg of zinc element, 10-75 mg of acylated casein, 1.5-20 mg of co-solubilized antifreeze, and 0.8-1.2 mg of stabilizer. The pH value of the compound high-calcium preparation composition is 3.5-4.5, and the dosage form is an oral solution. The co-solubilized antifreeze is a combination of glycerol and sorbitol, with a weight ratio of glycerol to sorbitol of 1:1 to 3:

1.

2. The compound high-calcium preparation composition according to claim 1, characterized in that, The preparation method of the acylated casein is as follows: casein is dissolved in phosphate buffer solution with pH=8.0-9.0, and heated in a water bath at 30-40℃, stirring until completely dissolved; succinic anhydride powder is slowly added to the casein solution, with succinic anhydride accounting for 6-10% of the weight of casein, while adjusting and maintaining pH=8.0-9.0; the water bath temperature is maintained at 30-40℃, stirring is started at 200-300 rpm, and the stirring time is 2-3 hours until the sample is fully acylated; after the reaction is completed, the pH is adjusted to 4-5; then, the sample is dialyzed with deionized water at 2000-3500D for 24-36 hours to remove free succinic acid and byproducts, and dried to obtain acylated casein powder; full acylation is achieved when the acylation target value reaches 72-78%.

3. The compound high-calcium preparation composition according to claim 1 or 2, characterized in that, The calcium raw material is a combination of calcium lactate and calcium gluconate, with a calcium elemental mass ratio of 3:1 between calcium lactate and calcium gluconate; the compound high-calcium composition contains 10-45 mg of calcium per mL. The zinc raw material is one or a combination of two of zinc gluconate and zinc lactate; The stabilizer is one or a combination of two of hydroxypropyl methylcellulose and povidone K30.

4. The compound high-calcium preparation composition according to claim 1 or 2, characterized in that, The compound high-calcium preparation composition further comprises a sweetener and / or a flavoring, wherein each mL of the compound high-calcium preparation contains 1.0-2.5 mg of sweetener, 0.2-0.5 mg of flavoring, and the remainder is water; the sweetener is steviol glycoside; the flavoring is peach flavoring or strawberry flavoring; and the water is purified water with a conductivity not exceeding 4.3 μs / cm at 20°C.

5. A method for preparing the compound high-calcium preparation composition according to any one of claims 1-4, characterized in that, Includes the following steps: (1) Preparation of acylated casein: Dissolve casein in phosphate buffer solution with pH=8.0-9.0, heat in a water bath at 30-40℃, and stir until completely dissolved; slowly add succinic anhydride powder to the casein solution, with succinic anhydride accounting for 6-10% of the weight of casein, while adjusting and maintaining pH=8.0-9.0; continue to maintain water bath at 30-40℃, turn on stirring at 200-300 rpm for 2-3 hours until the sample is fully acylated; after the reaction is complete, adjust the pH to 4-5; then dialyze with deionized water at 2000-3500D for 24-36 hours to remove free succinic acid and byproducts, and dry to obtain acylated casein powder; (2) Take 75-80 wt% purified water of the prescription amount, heat to boiling, boil for 5-10 minutes; then slowly add the stabilizer, turn on the electric stirrer and stir until the stabilizer is completely dissolved, then slowly add calcium lactate and calcium gluconate, continue stirring, and observe the dissolution of calcium raw materials until the solution is clear and there is no precipitate. (3) Add 10-15 wt% of the prescribed amount of boiled purified water to the material obtained in step (2). When the temperature drops to 35-50℃, add the prescribed amount of zinc raw material, acylated casein and co-solubilized antifreeze, stir to dissolve, and observe the dissolution until the solution is clear and there is no precipitate. (4) Immerse the ultrasonic probe directly into the material obtained in step (3), start the ultrasonic generator at a frequency of 20-25kHz for 5-30 minutes, observe that the dissolution is complete, and make up the volume after the temperature drops to room temperature; then start the ultrasonic generator again for 8-10 minutes. (5) The material obtained in step (4) is filtered sequentially through filter elements with pore sizes of 0.45μm and 0.22μm, then filled, and sterilized at 100-110℃ for 15-20 minutes to obtain the compound high calcium preparation composition.

6. The method for preparing the compound high-calcium preparation composition according to claim 5, characterized in that, In step (1), the pH of the phosphate buffer is 8.

5. When adding succinic anhydride powder, adjust and maintain the pH at 8.5; fully acylate to achieve the target acylation value of 72-78%.

7. The method for preparing the compound high-calcium preparation composition according to claim 5, characterized in that, In step (2), after adding the stabilizer, stir at 200-300 rpm for 3-5 min; after adding calcium lactate and calcium gluconate, stir at 200-300 rpm for 3-5 min.

8. The method for preparing the compound high-calcium preparation composition according to claim 5, characterized in that, In step (3), zinc raw material, acylated casein and co-solubilized antifreeze are added and stirred for 3-5 min; sweetener is added together with zinc raw material, acylated casein and co-solubilized antifreeze.

9. The method for preparing the compound high-calcium preparation composition according to claim 5, characterized in that, In step (4), after complete ultrasonic dissolution, the fragrance is added and the volume is adjusted after the temperature drops to room temperature.

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