阅读说明：本技术 杜仲多糖锶络合物及其制备方法和应用 (Eucommia polysaccharide strontium complex and preparation method and application thereof ) 是由 唐斌 邓雨晴 马粉波 彭颖 贺文琪 李思敬 于 2019-08-02 设计创作，主要内容包括：本发明涉及一种杜仲多糖锶络合物及其制备方法和应用。一种杜仲多糖锶络合物的制备方法,包括如下步骤：在控制pH为9～11的条件下,将杜仲多糖和水溶性锶盐在水中反应,得到反应液；在反应液中加入有机沉淀剂,然后静置反应,再经固液分离,收集沉淀,得到杜仲多糖络合物。上述杜仲多糖锶络合物的毒性较小且具有较好的骨修复和关节病防治效果。(The invention relates to an eucommia polysaccharide strontium complex and a preparation method and application thereof. A preparation method of eucommia polysaccharide strontium complex comprises the following steps: reacting eucommia ulmoides polysaccharide and water-soluble strontium salt in water under the condition that the pH value is controlled to be 9-11 to obtain a reaction solution; adding an organic precipitator into the reaction solution, standing for reaction, performing solid-liquid separation, and collecting precipitate to obtain the eucommia ulmoides polysaccharide complex. The eucommia polysaccharide strontium complex has low toxicity and good bone repair and joint disease prevention and treatment effects.)

1. The preparation method of the eucommia polysaccharide strontium complex is characterized by comprising the following steps:

reacting eucommia ulmoides polysaccharide and water-soluble strontium salt in water under the condition that the pH value is controlled to be 9-11 to obtain a reaction solution; and

and adding an organic precipitator into the reaction solution, standing for reaction, performing solid-liquid separation, and collecting precipitate to obtain the eucommia ulmoides polysaccharide complex.

2. The method for preparing eucommia ulmoides polysaccharide strontium complex according to claim 1, wherein the mass molar ratio of the eucommia ulmoides polysaccharide to strontium element in the water-soluble strontium salt is 60g:1mol to 250g:1 mol.

3. The method for preparing eucommia ulmoides polysaccharide strontium complex according to claim 1, wherein the step of reacting eucommia ulmoides polysaccharide with water-soluble strontium salt in water under the condition of controlling pH to be 9-11 comprises the following steps:

respectively dissolving the eucommia ulmoides polysaccharide and the water-soluble strontium salt in water to obtain an aqueous solution of the eucommia ulmoides polysaccharide and an aqueous solution of the water-soluble strontium salt; adding the water-soluble strontium salt water solution into the eucommia ulmoides polysaccharide water solution under the conditions of continuously stirring and controlling the pH value to be 9-11, and then continuously stirring and reacting for more than 1 hour under the condition of controlling the pH value to be 9-11.

4. The method for preparing eucommia ulmoides polysaccharide strontium complex according to claim 3, wherein the concentration of the eucommia ulmoides polysaccharide aqueous solution is 0.5 g/mL-1.5 g/mL; and/or the concentration of strontium element in the water solution of the water-soluble strontium salt is 1.5-2 mol/L; and/or the volume ratio of the organic precipitator to the reaction liquid is 2: 1-5: 1.

5. The method for preparing eucommia ulmoides polysaccharide strontium complex according to claim 1, wherein the water-soluble strontium salt is strontium chloride; and/or the organic precipitator is anhydrous acetone or anhydrous ethanol.

6. The method for producing a eucommia ulmoides polysaccharide strontium complex according to claim 1, wherein the time for the standing reaction is 24 hours or more.

7. The method for preparing eucommia ulmoides polysaccharide strontium complex according to claim 1, wherein sodium hydroxide is used as a pH regulator in the step of reacting eucommia ulmoides polysaccharide with a water-soluble strontium salt in water under the condition of controlling pH to 9-11.

8. The method for preparing eucommia ulmoides polysaccharide strontium complex according to claim 1, wherein the step of reacting eucommia ulmoides polysaccharide with water-soluble strontium salt in water is performed at 10-40 ℃.

9. The method for producing a eucommia ulmoides polysaccharide strontium complex according to any one of claims 1 to 8, further comprising a step of washing the precipitate with ethanol.

10. The method for preparing eucommia ulmoides polysaccharide strontium complex according to claim 9, wherein the step of washing the precipitate with ethanol comprises: and sequentially washing the precipitate by using ethanol with different concentrations, and drying the precipitate.

11. The eucommia ulmoides polysaccharide strontium complex prepared by the method for preparing eucommia ulmoides polysaccharide strontium complex according to any one of claims 1 to 10.

12. Use of the eucommia ulmoides polysaccharide complex of claim 11 for preparing a bone repair material.

Technical Field

The invention relates to the field of biological medicines, in particular to an eucommia polysaccharide strontium complex and a preparation method and application thereof.

Background

Strontium plays an important role in bone formation, and can promote the proliferation of osteoblasts and multifunctional stem cells, improve bone metabolism and the like, however, free strontium ions have toxicity in vivo and are difficult to develop for the application of strontium.

Disclosure of Invention

Therefore, a preparation method of eucommia polysaccharide strontium complex with low toxicity and good bone repair and joint disease prevention and treatment effects is needed.

In addition, an eucommia polysaccharide strontium complex and application thereof are also provided.

A preparation method of eucommia polysaccharide strontium complex comprises the following steps:

reacting eucommia ulmoides polysaccharide and water-soluble strontium salt in water under the condition that the pH value is controlled to be 9-11 to obtain a reaction solution; and

and adding an organic precipitator into the reaction solution, standing for reaction, performing solid-liquid separation, and collecting precipitate to obtain the eucommia ulmoides polysaccharide complex.

In one embodiment, the mass molar ratio of the eucommia ulmoides polysaccharide to the strontium element in the water-soluble strontium salt is 60g:1mol to 250g:1 mol.

In one embodiment, the step of reacting eucommia ulmoides polysaccharide and a water-soluble strontium salt in water under the condition of controlling the pH to be 9-11 comprises the following steps:

respectively dissolving the eucommia ulmoides polysaccharide and the water-soluble strontium salt in water to obtain an aqueous solution of the eucommia ulmoides polysaccharide and an aqueous solution of the water-soluble strontium salt; adding the water-soluble strontium salt water solution into the eucommia ulmoides polysaccharide water solution under the conditions of continuously stirring and controlling the pH value to be 9-11, and then continuously stirring and reacting for more than 1 hour under the condition of controlling the pH value to be 9-11.

In one embodiment, the concentration of the eucommia ulmoides polysaccharide aqueous solution is 0.5 g/mL-1.5 g/mL; and/or the concentration of strontium element in the water solution of the water-soluble strontium salt is 1.5-2 mol/L; and/or the volume ratio of the organic precipitator to the reaction liquid is 2: 1-5: 1.

In one embodiment, the water soluble strontium salt is strontium chloride; and/or the organic precipitator is anhydrous acetone or anhydrous ethanol.

In one embodiment, the standing reaction time is more than 24 hours.

In one embodiment, sodium hydroxide is used as a pH regulator in the step of reacting the eucommia ulmoides polysaccharide and the water-soluble strontium salt in water under the condition of controlling the pH to be 9-11.

In one embodiment, the step of reacting the eucommia ulmoides polysaccharide and the water-soluble strontium salt in water is performed at 10-40 ℃.

In one embodiment, the method further comprises the step of washing the precipitate with ethanol.

In one embodiment, the washing the precipitate with ethanol comprises: and sequentially washing the precipitate by using ethanol with different concentrations, and drying the precipitate.

The eucommia polysaccharide strontium complex prepared by the preparation method of the eucommia polysaccharide strontium complex.

The application of the eucommia ulmoides polysaccharide complex in preparing bone repair materials.

Experiments prove that: according to the preparation method of the eucommia polysaccharide strontium complex, the eucommia polysaccharide strontium complex with good bone repair and joint disease prevention and treatment effects can be prepared according to the steps, and has low toxicity compared with free strontium ions.

Drawings

FIG. 1 is a flowchart of a method for preparing an eucommia ulmoides polysaccharide complex according to an embodiment;

FIG. 2 is an XRD pattern of the product of example 1 and the eucommia ulmoides polysaccharide of comparative example 3;

FIG. 3 is an infrared spectrum of the product obtained in example 1 and the eucommia ulmoides polysaccharide of comparative example 3;

FIG. 4 is a scanning electron micrograph of the product of example 1;

FIG. 5 is a scanning electron micrograph of a eucommia ulmoides polysaccharide of comparative example 3;

FIG. 6 is a bar graph of macrophages cultured on the culture solution prepared from the eucommia ulmoides polysaccharide strontium complex of example 1 and the control experiment at the third day, respectively.

Detailed Description

In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

As shown in fig. 1, a method for preparing a eucommia ulmoides polysaccharide strontium complex according to an embodiment includes the steps of:

step S110: and reacting the eucommia ulmoides polysaccharide and the water-soluble strontium salt in water under the condition of controlling the pH value to be 9-11 to obtain a reaction solution.

The pH is controlled to 9 to 11 so that the reaction proceeds smoothly, thereby producing a eucommia polysaccharide strontium complex.

Specifically, the mass molar ratio of the eucommia ulmoides polysaccharide to the strontium element in the water-soluble strontium salt is 60g:1 mol-250 g:1 mol. Too much strontium can leave a large amount of strontium ions in the complex, which can cause the material to be harmful in vivo. If the strontium element is less, the eucommia polysaccharide is excessive, and the complex reaction is insufficient.

In one embodiment, step S110 includes: respectively dissolving eucommia ulmoides polysaccharide and water-soluble strontium salt in water to obtain an aqueous solution of eucommia ulmoides polysaccharide and an aqueous solution of water-soluble strontium salt; adding a water-soluble strontium salt aqueous solution into the eucommia ulmoides polysaccharide aqueous solution under the conditions of continuously stirring and controlling the pH value to be 9-11, and then continuously stirring and reacting for more than 1 hour under the condition of controlling the pH value to be 9-11. Preferably, the water used in the step of dissolving the eucommia ulmoides polysaccharide and the water-soluble strontium salt in water is deionized water, so as to reduce impurities as much as possible and improve the purity of the product.

It is understood that, in other embodiments, the step S110 is not limited to the above-mentioned step, and in other embodiments, an aqueous solution of eucommia ulmoides polysaccharide may be added to an aqueous solution of water-soluble strontium salt; or directly adding the eucommia ulmoides polysaccharide and the water-soluble strontium salt into water, and stirring for reaction.

In one embodiment, the concentration of the aqueous solution of the eucommia ulmoides polysaccharide is 0.5 g/mL-1.5 g/mL, the maximum concentration of the aqueous solution of the eucommia ulmoides polysaccharide is 1.5g/mL, and the concentration of the aqueous solution is too low, so that the content of the eucommia ulmoides polysaccharide is too low, and the complex reaction is difficult to occur. The concentration of strontium element in the water solution of the water-soluble strontium salt is 1.5-2 mol/L, the strontium ion concentration is too high, the hydrolysis degree of the salt is reduced, and hydrogen ions in the solution are increased, so that the pH is influenced, and the generated complex is unstable. If the concentration of strontium is too low, the formation of coordinate bonds is less and the reaction is incomplete.

The water-soluble strontium salt can be a strontium salt which is commonly used in the field and can be dissolved in water, such as strontium chloride or strontium nitrate, and preferably, the water-soluble strontium salt is strontium chloride which can enable the prepared eucommia polysaccharide strontium complex to have fewer byproducts, higher purity and almost no toxicity.

Specifically, in step S110, sodium hydroxide or potassium hydroxide is used as a pH adjuster. It is understood that the pH regulator is not limited to sodium hydroxide, and in other embodiments, the pH regulator may be ammonia water, etc., however, the eucommia ulmoides polysaccharide contains sodium and potassium, and the sodium hydroxide or potassium hydroxide is used for regulating the pH regulator, so that redundant impurities are not generated, and the ammonia water may introduce new impurities into the system. In one embodiment, the pH regulator is sodium hydroxide aqueous solution with the concentration of 1.5-2 mol/L or potassium hydroxide aqueous solution with the concentration of 1.5-2 mol/L. The sodium hydroxide and the potassium hydroxide are strong bases, the increase of the concentration can cause local oxidation of the eucommia polysaccharide, the excessively low concentration can cause the overlarge volume of the eucommia polysaccharide, so that the concentration of each substance in a reaction system is reduced too much, and the reaction is not easy to occur.

Specifically, the step of reacting the eucommia ulmoides polysaccharide and the water-soluble strontium salt in water is performed under a room temperature condition. Herein, room temperature means 10 ℃ to 40 ℃.

Step S120: adding an organic precipitator into the reaction solution, standing for reaction, performing solid-liquid separation, and collecting precipitate to obtain the eucommia ulmoides polysaccharide complex.

Adding organic precipitant, and standing for reaction to precipitate the eucommia ulmoides polysaccharide complex in the reaction solution. The organic precipitant may be an organic precipitant commonly used in the art. Specifically, the organic precipitant is anhydrous acetone or anhydrous ethanol. Preferably, the organic precipitator is absolute ethyl alcohol, and the toxicity of the absolute ethyl alcohol is lower compared with that of absolute acetone, so that the prepared eucommia polysaccharide strontium complex has lower toxicity, and the absolute ethyl alcohol is cheap, and is beneficial to reducing the production cost.

Specifically, the reaction time is 24 hours or more so that the eucommia ulmoides polysaccharide complex in the reaction solution can be precipitated as much as possible.

Specifically, the volume ratio of the organic precipitator to the reaction solution is 2: 1-5: 1, so that the purpose of removing impurities by washing is achieved on the premise of not wasting more organic precipitator.

Further, after step S120, a step of washing the precipitate with ethanol is further included to perform a purification process on the eucommia ulmoides polysaccharide complex. Washing with ethanol to remove strontium chloride in the precipitate and improve the purity of the eucommia ulmoides polysaccharide complex.

Specifically, the step of washing the precipitate with ethanol comprises: washing the precipitate with ethanol of different concentrations, and drying the precipitate. In one embodiment, the drying temperature in the step of drying the precipitate is 37 ℃ to 45 ℃.

In one embodiment, the step of sequentially washing the precipitate with ethanol of different concentrations comprises: ethanol with a volume percentage of 80%, ethanol with a volume percentage of 60% and ethanol with a volume percentage of 40% were mixed with the precipitate in sequence, with stirring for at least 30 minutes each time. Specifically, the volume ratio of ethanol with each concentration to the reaction solution obtained in step S110 is 2.5:1 to 5:1, so as to achieve the purpose of effective washing.

Experiments prove that: the eucommia polysaccharide strontium complex prepared by the preparation method of the eucommia polysaccharide strontium complex can avoid the toxicity of free strontium ions through the complexation reaction of strontium and macromolecules of eucommia polysaccharide. Meanwhile, the prepared eucommia polysaccharide strontium complex also has the characteristics of the eucommia polysaccharide strontium complex and enables strontium to be effectively released in vivo, plays the roles of reducing inflammation and proliferating bone cells in strontium and can better assist in treating osteoarthritis. The eucommia polysaccharide strontium complex can change the immune environment of bones and promote the proliferation and differentiation of cells related to bone repair. Can play an important role in bone repair by regulating bone immunity.

The eucommia ulmoides polysaccharide strontium complex prepared by the preparation method of eucommia ulmoides polysaccharide strontium complex of one embodiment. The eucommia polysaccharide strontium complex has good bone repair and joint disease prevention and treatment effects, and has low toxicity.

The eucommia polysaccharide complex can be used for preparing bone repair materials, so that the bone repair materials have good bone repair and joint disease prevention and treatment effects and low toxicity.

The following are specific examples (the following examples, unless otherwise specified, contain no other components not specifically indicated except for unavoidable impurities):