Preparation method and application of hesperidin phospholipid complex

Abstract

The invention provides a preparation method and application of a hesperidin phospholipid complex, and relates to the technical field of bulk drug industry. The preparation method comprises the following steps: adsorbing amorphous hesperidin by using an adsorbent, then desorbing the amorphous hesperidin on the adsorbent by using an organic solvent to obtain a desorption solution, and removing the organic solvent in the desorption solution to obtain amorphous hesperidin powder; mixing the hesperidin amorphous powder with phospholipid to obtain a hesperidin phospholipid complex; the amorphous hesperidin is obtained by dissolving crystalline hesperidin in hot water and converting the crystalline hesperidin into the amorphous hesperidin. The preparation method relieves the problem of low yield of the hesperidin phospholipid complex in the prior art.

Description

Technical Field

[0001] The present invention relates to the technical field of bulk pharmaceutical chemicals industry, and particularly relates to a preparation method and application of hesperidin phospholipid complex. Background Art

[0002] Hesperidin, also known as naringin and naringin, is a glycoside formed by hesperetin and rutinose, and is a dihydroflavonoid derivative. Its chemical formula is shown as follows:

[0003]

[0004] Hesperidin widely exists in plants of Leguminosae, Labiatae, Papilionaceae, and Rutaceae citrus genus, and has the effects of maintaining normal osmotic pressure of blood vessels, reducing vascular fragility, enhancing capillary toughness, and shortening bleeding time. Hesperidin not only has the effect of reducing cholesterol in the human body, but also has the effects of antiviral, improving immunity, and has pharmacological activities such as anti-inflammatory, antioxidant, antibacterial, anticancer, radiation protection, and protecting the cardiovascular system.

[0005] The water solubility and lipid solubility of hesperidin are extremely poor, which greatly limits the product development and application starting from hesperidin. Using traditional process development methods to prepare products with hesperidin as the raw material, due to its characteristics of slightly soluble in water and solvents, its process development is extremely difficult.

[0006] Previous research results show that hesperidin has the best dissolution effect in three solvents: dimethyl sulfoxide (DMSO), pyridine, and N,N-dimethylformamide (DMF). The existing process yield of hesperidin phospholipid complex is low. There is a published literature (Zhang Qing, Guan Haiyan, Jin Zhu, etc. Determination of hesperidin content in hesperidin phospholipid complex by high performance liquid chromatography [J]. China Pharmaceuticals, 2015, 24(13):) that uses hesperidin and soybean lecithin as raw materials and a dimethyl sulfoxide-methanol mixed solution as the reaction solvent to prepare hesperidin phospholipid complex, and its average content is only 282.5 μg / mg. Moreover, the solvent system has high polarity, high boiling point, good thermal stability, and the serious defect that the solvent cannot be completely removed. In addition, DMSO, DMF, and pyridine are highly toxic, interact with the hydrophobic groups of proteins, resulting in protein denaturation, and have vascular toxicity and liver and kidney toxicity. It is not suitable for the preparation process of drug development. Therefore, how to improve the preparation process of hesperidin phospholipid complex is an urgent problem to be solved at present.

[0007] In view of this, the present invention is specifically proposed. Summary of the Invention

[0008] The purpose of the present invention is to provide a preparation method of hesperidin phospholipid complex to alleviate the problem of low yield of hesperidin phospholipid complex existing in the prior art.

[0009] Another object of the present invention is to provide the application of the above hesperidin phospholipid complex.

[0010] To solve the above technical problems, the present invention specifically adopts the following technical solutions:

[0011] In the first aspect, a preparation method of a hesperidin phospholipid complex is provided, which includes using an adsorbent to adsorb amorphous hesperidin, then using an organic solvent to desorb the amorphous hesperidin on the adsorbent to obtain a desorption solution, removing the organic solvent in the desorption solution to obtain an amorphous powder of hesperidin; and then mixing the amorphous powder of hesperidin with phospholipids to obtain the hesperidin phospholipid complex;

[0012] The amorphous hesperidin is obtained by dissolving crystalline hesperidin in water at 60-95°C to convert it into amorphous hesperidin.

[0013] In an optional embodiment, crystalline hesperidin is dissolved in water at 60-95°C and stirred at 300-500 RPM for 1-6 h to obtain an aqueous solution containing amorphous hesperidin.

[0014] In an optional embodiment, the adsorbent is selected from chromatographic column packings.

[0015] In an optional embodiment, the adsorbent is selected from polar or weakly polar chromatographic column packings, and further preferably includes macroporous resin and / or polyamide.

[0016] In an optional embodiment, the adsorbent is added to the aqueous solution containing amorphous hesperidin, and the system temperature is maintained at 60-95°C for adsorption for 1-6 h

[0017] In an optional embodiment, the organic solvent includes methanol or an ethanol aqueous solution with a concentration of at least 90% v / v.

[0018] In an optional embodiment, the phospholipids include one or more of soybean phospholipids and lecithin.

[0019] In an optional embodiment, the amorphous powder of hesperidin and phospholipids are reacted in an ethanol aqueous solution with a concentration of at least 95% v / v to obtain the hesperidin phospholipid complex.

[0020] In the second aspect, a hesperidin phospholipid complex prepared by the preparation method of the first aspect is further provided.

[0021] In the third aspect, an application of the preparation method of the hesperidin phospholipid complex of the first aspect, or the hesperidin phospholipid complex of the second aspect in the preparation of drugs is further provided.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The preparation method of hesperidin phospholipid complex provided by the present invention utilizes the characteristic that hesperidin transforms from crystalline form to amorphous form in aqueous solution under high-temperature conditions, combines the adsorption characteristics of adsorbent for hesperidin and the desorption effect of organic solvent, not only maintains the high solubility of hesperidin but also enables the subsequent preparation process to be carried out at low temperature. The hesperidin phospholipid complex prepared by this process has low toxicity, can be used to prepare oral preparations, and has a high yield, with the highest content reaching 447 μg / mg. The purity of the hesperidin phospholipid complex can reach over 95% calculated by the area normalization method of high-performance liquid chromatography.

[0024] The preparation method of hesperidin phospholipid complex provided by the present invention also has the following advantages:

[0025] (1) This preparation method is simple to operate and has good safety, and can be mastered by general technical personnel.

[0026] (2) The sample treatment process of this preparation method is simple, alleviating the defects of small content of existing hesperidin phospholipid complex and large solvent hazards; in the preferred scheme, all waste liquids generated in this preparation method can be recycled, are green and environmentally friendly, and are suitable for industrial application.

[0027] (3) During the adsorption and desorption processes of this preparation method, the molecular structure of hesperidin is not damaged, and no ring-opening reaction, hydrolysis reaction or esterification or salt formation reaction occurs. Specific embodiments

[0028] Next, the technical solutions of the present invention will be clearly and completely described in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0029] In the first aspect, a preparation method of hesperidin phospholipid complex is provided. This preparation method includes using an adsorbent to adsorb amorphous hesperidin, then using an organic solvent to desorb the amorphous hesperidin on the adsorbent to obtain a desorption solution, removing the solvent in the desorption solution to obtain amorphous powder of hesperidin; then mixing the amorphous powder of hesperidin with phospholipids, and combining the amorphous powder of hesperidin with phospholipids through physical action to prepare a hesperidin phospholipid complex.

[0030] The above-mentioned adsorbent adsorbs the hesperidin transformed from crystalline form to amorphous form through bonding action. Using bonding action to make the adsorbent adsorb amorphous hesperidin has the advantages of simplicity, quickness and low cost, alleviating the problem of a large amount of solvent used in traditional complexes. Since all processes involved are physical actions and do not involve chemical changes, it also avoids the generation of hesperidin by-products caused by unconventional technical means.

[0031] Solubilization using an adsorbent system can greatly increase the solubility of hesperidin in organic solvents without damaging it, and an amorphous powder of hesperidin with high solubility can be obtained. The content of the hesperidin phospholipid complex prepared using the amorphous powder of hesperidin was determined by the area normalization method of high performance liquid chromatography, and the purity of the prepared hesperidin complex can be as high as over 95%.

[0032] Using the aqueous solution heating method, crystalline hesperidin is converted into amorphous hesperidin, and the temperature of the aqueous solution is 60 - 95°C, for example, it can be but not limited to 60, 65, 70, 75, 80, 85, 90 or 95°C.

[0033] In an alternative embodiment, the starting material for the preparation method is a commercially available crude hesperidin raw material.

[0034] In an alternative embodiment, crystalline hesperidin is dissolved in water at 60 - 95°C and stirred at 300 - 500 RPM for 1 - 6 h to obtain an aqueous solution containing amorphous hesperidin. The stirring speed can be, for example, but not limited to 300, 350, 400, 450 or 500 RPM; the stirring time can be, for example, but not limited to 1, 2, 3, 4, 5 or 6 h.

[0035] In an alternative embodiment, the adsorbent is selected from column chromatography packing materials, preferably including polar or weakly polar column chromatography packing materials.

[0036] In an alternative embodiment, the adsorbent includes macroporous resin and / or polyamide.

[0037] In an alternative embodiment, the mass ratio of the charged crystalline hesperidin to the adsorbent is 1:(1 - 9), for example, it can be but not limited to 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8 or 1:9.

[0038] In an alternative embodiment, the adsorption is to saturate the adsorbent with amorphous hesperidin.

[0039] In an alternative embodiment, hesperidin and the adsorbent are adsorbed in an aqueous solution system under the conditions of heating and stirring to prepare an adsorbent adsorbed with hesperidin converted from crystalline form to amorphous form. Specifically, the adsorbent is added to the aqueous solution in which amorphous hesperidin is dissolved, and the system temperature is maintained at 60 - 95°C for adsorption for 1 - 6 h; the system temperature can be, for example, but not limited to 60, 65, 70, 75, 80, 85, 90 or 95°C, preferably 70 - 95°C; the adsorption time can be, for example, but not limited to 1, 2, 3, 4, 5 or 6 h.

[0040] In an alternative embodiment, stirring is carried out during the adsorption process, and the stirring speed is 300 - 500 RPM, for example, it can be, but is not limited to, 300, 350, 400, 450, or 500 RPM.

[0041] In an alternative embodiment, after the adsorption is completed, the adsorbent adsorbed with amorphous hesperidin is separated and dried. The drying temperature can optionally be 40 - 80 °C, for example, it can be, but is not limited to, 40, 45, 50, 55, 60, 65, 70, 75, or 80 °C.

[0042] In an alternative embodiment, the adsorbent saturated with hesperidin is placed in an oven for drying to obtain a dried adsorbent saturated with hesperidin.

[0043] In an alternative embodiment, a 60 - 80 mesh filter cloth is used to separate the adsorbent from the liquid.

[0044] In an alternative embodiment, the separated adsorbent is washed with water at least once to remove the unadsorbed hesperidin.

[0045] In an alternative embodiment, the filtrate and the washing liquid are recovered and added with the adsorbent again for cyclic adsorption.

[0046] In an alternative embodiment, the organic solvents used to desorb the amorphous hesperidin on the adsorbent include methanol or an ethanol aqueous solution with a concentration of at least 90% v / v, preferably 95% v / v ethanol aqueous solution. The desorption solution is an organic solution containing the amorphous hesperidin desorbed from the adsorbent, for example, 90% v / v ethanol containing the amorphous hesperidin desorbed from the adsorbent.

[0047] In an alternative embodiment, the amorphous hesperidin on the adsorbent is desorbed under ultrasonic conditions.

[0048] In an alternative embodiment, the ultrasonic time is 20 - 50 min, for example, it can be, but is not limited to, 20, 25, 30, 35, 40, 45, or 50 min.

[0049] In an alternative embodiment, the removal of the solvent in the desorption solution is carried out at a low temperature not higher than 50 °C, for example, it can be carried out under temperature conditions not higher than 50 °C, not higher than 45 °C, not higher than 40 °C, not higher than 35 °C, or not higher than 30 °C.

[0050] In an alternative embodiment, the solvent in the desorption solution is removed by rotary evaporation under reduced pressure to obtain amorphous powder of hesperidin.

[0051] In an alternative embodiment, the phospholipids include one or more of soy phospholipids and lecithin.

[0052] In an alternative embodiment, the mass ratio of the amorphous powder of hesperidin to phospholipids is 1:(1-9), for example, but not limited to, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, or 1:9.

[0053] In an alternative embodiment, the amorphous powder of hesperidin and phospholipids are reacted in an aqueous ethanol solution with a concentration of at least 95% v / v to obtain a hesperidin phospholipid complex.

[0054] In an alternative embodiment, the amorphous powder of hesperidin and phospholipids are stirred and reacted at a temperature of 30-50 °C for 1-6 h; the reaction temperature can be, for example, but not limited to, 30, 35, 40, 45, or 50 °C, and the reaction time can be, for example, but not limited to, 1, 2, 3, 4, 5, or 6 h.

[0055] An exemplary method for preparing a hesperidin phospholipid complex includes the following steps:

[0056] 1. Preparation of amorphous hesperidin:

[0057] (1) Turn on the constant temperature electromagnetic stirring water bath. After the instrument self-checks, enter the preheating stage and adjust the parameter settings as follows: set the water bath temperature to 60-95 °C.

[0058] (2) Weigh an appropriate amount of hesperidin raw material and place it in a 200 ml round-bottom flask. Add purified water and a magnetic stirring rotor in a ratio of 1:10 to 1:20 by material-liquid ratio; connect the round-bottom flask to a water-cooled reflux device; place it in the water bath and stir at a speed range of 300-500 RPM; stir for 1-6 hours to obtain an amorphous hesperidin solution.

[0059] 2. Adsorption of amorphous hesperidin:

[0060] (1) Weigh an appropriate amount of adsorbent and add it to the round-bottom flask containing the amorphous hesperidin solution, and continue heating and stirring for 1-6 hours.

[0061] (2) After stirring, take out the round-bottom flask, separate the adsorbent from the filtrate through a 60-80 mesh filter cloth, and wash away the unadsorbed hesperidin with purified water. The adsorbent is dried at a temperature of 40 °C - 80 °C.

[0062] 3. Desorption of amorphous hesperidin:

[0063] (1) Add ethanol to the dried adsorbent and adjust the ethanol concentration in the system to ≥90% v / v for desorption of amorphous hesperidin. The resulting solution is the desorbed solution of amorphous hesperidin.

[0064] (2) Remove the organic solvent from the desorbed solution of amorphous hesperidin by rotary evaporation under reduced pressure. The resulting solid is the powder of amorphous hesperidin.

[0065] 4. Preparation of hesperidin phospholipid complex:

[0066] (1) Take an appropriate amount of hesperidin powder obtained in the above steps, add phospholipids, and use 95% ethanol as the reaction solvent. Stir at 30°C to 50°C for 1 to 6 hours. The obtained solution is dried under reduced pressure, and the obtained solid is the hesperidin phospholipid complex.

[0067] In the second aspect, a hesperidin phospholipid complex prepared by the above preparation method is also provided.

[0068] In the third aspect, an application of the preparation method of the hesperidin phospholipid complex in the first aspect or the hesperidin phospholipid complex in the second aspect in the preparation of drugs is also provided.

[0069] In an optional embodiment, the drug is an oral preparation.

[0070] In an optional embodiment, the drug includes drugs for antiviral, immune enhancement, anti-inflammatory, antioxidant, antibacterial, anticancer, radiation protection or cardiovascular system protection.

[0071] The present invention will be further described below through specific examples. However, it should be understood that these examples are only for more detailed description and should not be construed as limiting the present invention in any way.

[0072] Example 1

[0073] 1. A preparation method of hesperidin phospholipid complex, comprising the following steps:

[0074] (1) Turn on the constant temperature electromagnetic stirring water bath. After the instrument self-checks, enter the preheating stage and adjust the parameter settings as follows: set the water bath temperature to 60°C.

[0075] (2) Weigh 60 mg of hesperidin raw material and place it in a 200 ml round-bottom flask. Add purified water and a magnetic stirring rotor at a material-liquid ratio of 1:20; connect the round-bottom flask to a water-cooled reflux device; place it in the water bath and set the stirring speed range to 500 RPM; stir for 1 hour.

[0076] (3) Weigh 400 mg of macroporous resin D101 type and add it to the previously prepared round-bottom flask containing hesperidin, and continue heating and stirring for 1 hour.

[0077] (4) After stirring, take out the round-bottom flask, separate the unadsorbed hesperidin and macroporous resin D101 type through a filter cloth, and wash the macroporous resin D101 type three times with purified water to remove the unadsorbed hesperidin. Dry the macroporous resin D101 type at a temperature of 80°C.

[0078] (5) Add 100 ml of 90% v / v ethanol to the dried macroporous resin D101 for desorption of hesperidin, and the resulting solution is the hesperidin desorption solution.

[0079] (6) Remove ethanol from the hesperidin desorption solution by rotary evaporation under reduced pressure, and the resulting solid is the powder of hesperidin.

[0080] (7) Take 30 mg of the hesperidin powder obtained in the above steps, add 30 mg of egg yolk phospholipid, and use 100 ml of 95% v / v ethanol as the reaction solvent. Stir at 50 °C for 1 hour. The resulting solution is dried under reduced pressure, and the resulting solid is the hesperidin phospholipid complex.

[0081] 2. Preparation of hesperidin phospholipid complex by traditional method:

[0082] Take 30 mg of hesperidin and 30 mg of egg yolk phospholipid, add 100 ml of dimethyl sulfoxide (DMSO)-methanol mixed solution as the reaction solvent, stir at 50 °C for 1 h, and remove the solvent by drying under reduced pressure at low temperature to obtain the hesperidin phospholipid complex.

[0083] Dissolve 10 mg of the samples obtained by the preparation method in the first part of Example 1 and the traditional method in a 10 ml volumetric flask and make up the volume. The solution is filtered through a microporous membrane filter, and the resulting filtrate is calculated by the external standard method using HPLC to compare the content differences.

[0084] The test results show that the content of the hesperidin phospholipid complex prepared by the traditional method is 0.051 mg / mg, and the content of the hesperidin phospholipid complex prepared by the preparation method in the first part is 0.447 mg / mg. Its purity is 97.14 calculated by the area normalization method using HPLC. Compared with the traditional content detection pretreatment method, the content has increased.

[0085] Example 2

[0086] (1) Turn on the constant temperature electromagnetic stirring water bath. After the instrument self-checks, enter the preheating stage and adjust the parameter settings as follows: set the water bath temperature to 95 °C.

[0087] (2) Weigh 60 mg of hesperidin raw material and place it in a 200 ml round-bottom flask. Add purified water and a magnetic stirring rotor at a material-liquid ratio of 1:10; connect the round-bottom flask to a water-cooled reflux device; place it in the water bath and the stirring speed range is 300 RPM; stir for 6 hours to obtain hesperidin.

[0088] (3) Weigh 540 mg of polyamide and add it to the round-bottom flask containing hesperidin before, and continue to heat and stir for 6 hours.

[0089] (4) After the stirring is completed, take out the round-bottom flask, separate the unadsorbed hesperidin from the polyamide through a filter cloth, and wash the polyamide three times with purified water to remove the unadsorbed hesperidin. The polyamide is dried at a temperature of 40 °C.

[0090] (5) Add 100 ml of 95% v / v ethanol to the dried polyamide and ultrasonicate for 50 min for the desorption of hesperidin. The resulting solution is the hesperidin desorption solution.

[0091] (6) Remove the organic solvent from the hesperidin desorption solution by rotary evaporation under reduced pressure. The resulting solid is the powder of hesperidin.

[0092] (7) Take 30 mg of hesperidin powder and add 30 mg of soybean phospholipid. Using 100 ml of 95% v / v ethanol as the reaction solution, stir at 30 °C for 6 hours, and remove the solvent by drying under reduced pressure. The resulting solid is the hesperidin phospholipid complex.

[0093] (8) Take 10 mg of the hesperidin phospholipid complex prepared in the above step into a 10 ml volumetric flask, dissolve it with 95% v / v ethanol and make up to the mark. After filtering through a microporous membrane, the content is detected by the external standard method of HPLC.

[0094] The test results show that: the content of the hesperidin phospholipid complex is 0.317 mg / mg. Compared with the traditional complex preparation method, the content of hesperidin in the hesperidin complex sample treated by the technical solution of the present invention has been significantly improved.

[0095] Example 3

[0096] To further explore the influence of different crystal forms of hesperidin on its process development products, the solubility comparison of hesperidin phospholipid mixture, hesperidin (crystalline) phospholipid complex, and hesperidin (amorphous) phospholipid complex was carried out under the same process conditions. The specific steps are as follows:

[0097] 1. Preparation of hesperidin phospholipid mixture:

[0098] Weigh 500 mg of hesperidin raw material drug and 500 mg of egg yolk phospholipid, add 10 ml of 95% v / v ethanol to prepare a 1 mg per milliliter hesperidin phospholipid mixed solution.

[0099] 2. Preparation of hesperidin (crystalline) phospholipid complex:

[0100] Weigh 500 mg of hesperidin raw material drug and 500 mg of egg yolk phospholipid, add 100 ml of 95% v / v ethanol, heat under reflux and stir at 50 °C for 3 hours, take out and cool to room temperature, remove the solvent at low temperature, dissolve the residue with 95% v / v ethanol and transfer it to a 10 ml volumetric flask, and make up to the mark with 95% v / v ethanol to prepare a 1 mg per milliliter hesperidin (crystalline) phospholipid complex solution.

[0101] 3. Hesperidin (amorphous) phospholipid complex

[0102] (1) Turn on the constant-temperature electromagnetic stirring water bath. After the instrument self-checks, enter the preheating stage and adjust the parameter settings as follows: set the water bath temperature to 95 °C.

[0103] (2) Weigh 60 mg of hesperidin raw material and place it in a 200 ml round-bottom flask. Add 100 ml of purified water and a magnetic stirring rotor; connect the round-bottom flask to a water-cooled reflux device; place it in the water bath and set the stirring speed range to 450 RPM; stir for 3 hours to obtain (amorphous) hesperidin.

[0104] (3) Weigh 400 mg of macroporous resin D101 and add it to the previous round-bottom flask containing (amorphous) hesperidin, and continue heating and stirring for 3 hours.

[0105] (4) After stirring, take out the round-bottom flask, separate the unadsorbed hesperidin and macroporous resin D101 through a filter cloth, and wash the macroporous resin three times with purified water to remove the unadsorbed hesperidin. Dry the macroporous resin at a temperature of 75 °C.

[0106] (5) Weigh 3.3 g of macroporous resin D101 (calculated as 0.153 mg / mg, that is, 3.3 g of macroporous resin D101 can desorb 504.9 mg of hesperidin), add 100 ml of 95% v / v ethanol for desorption, add 500 mg of phospholipid, heat and reflux and stir at 50 °C for 3 hours, take out and cool to room temperature, remove the solvent at low temperature, dissolve the residue in 95% v / v ethanol and transfer it to a 10 ml volumetric flask, and make up to the mark with 95% v / v ethanol to prepare a 1 mg per milliliter hesperidin (amorphous) phospholipid complex solution.

[0107] Filter the above-prepared hesperidin phospholipid mixed solution, hesperidin (crystalline) phospholipid complex solution and hesperidin (amorphous) phospholipid complex solution using a microporous filter membrane, and calculate the filtrate by the external standard method using HPLC.

[0108] The test results show that the average concentration of hesperidin in the hesperidin phospholipid mixture is 0.035 mg / mg, the average concentration of hesperidin in the hesperidin (crystalline) phospholipid complex is 0.041 mg / mg, and the concentration of hesperidin in the hesperidin (amorphous) phospholipid complex is 0.447 mg / mg.

[0109] Example 4

[0110] A preparation method of a hesperidin phospholipid complex, comprising the following steps:

[0111] (1) Turn on the constant-temperature electromagnetic stirring water bath. After the instrument self-checks, enter the preheating stage and adjust the parameter settings as follows: set the water bath temperature to 60 °C.

[0112] (2) Weigh 300 mg of hesperidin raw material and place it in a 200 ml round-bottom flask. Add purified water and a magnetic stirring rotor in a ratio of 1:15 of solid to liquid; connect the round-bottom flask to a water-cooled reflux device; place it in a water bath and stir at a speed range of 500 RPM; stir for 1 hour.

[0113] (3) Weigh 300 mg of macroporous resin D101 type and add it to the previously prepared round-bottom flask containing hesperidin, and continue heating and stirring for 1 hour.

[0114] (4) After stirring, take out the round-bottom flask, separate the unadsorbed hesperidin and macroporous resin D101 type through a filter cloth, and wash the macroporous resin D101 type three times with purified water to remove the unadsorbed hesperidin. Dry the macroporous resin D101 type at a temperature of 80 °C.

[0115] (5) Add 100 ml of 95% v / v ethanol to the dried macroporous resin D101 type for desorption of hesperidin, and the resulting solution is the hesperidin desorption solution.

[0116] (6) Remove ethanol from the hesperidin desorption solution by rotary evaporation under reduced pressure, and the resulting solid is the powder of hesperidin.

[0117] (7) Take 30 mg of the hesperidin powder obtained in the above steps, add 270 mg of soybean phospholipid, and use 100 ml of 95% v / v ethanol as the reaction solvent, stir at 50 °C for 1 hour. Dry the resulting solution under reduced pressure, and the resulting solid is the hesperidin phospholipid complex.

[0118] The test results show that the hesperidin complex rate of the hesperidin phospholipid complex treated by the technical solution of the present invention is calculated by the external standard method to be 75.17%. ， Its purity is calculated by the area normalization method of HPLC to be 96.53.

[0119] According to the test results, it can be clearly concluded that the phospholipid complex of hesperidin (amorphous) has a higher content compared to the phospholipid complex of hesperidin (crystalline), and at the same time, using ethanol as the reaction system, it has higher industrial development value.

[0120] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A preparation method of hesperidin phospholipid complex, characterized in that, It includes adsorbing amorphous hesperidin using an adsorbent, then desorbing the amorphous hesperidin on the adsorbent using an organic solvent to obtain a desorption solution, and removing the organic solvent in the desorption solution to obtain an amorphous powder of hesperidin; then mixing the amorphous powder of hesperidin with phospholipids to obtain the hesperidin phospholipid complex; The amorphous hesperidin is obtained by dissolving crystalline hesperidin in water at 60 - 95 °C to convert it into amorphous hesperidin.

2. The preparation method according to claim 1, characterized in that, Dissolve crystalline hesperidin in water at 60 - 95 °C and stir at 300 - 500 RPM for 1 - 6 h to obtain an aqueous solution containing amorphous hesperidin.

3. The preparation method according to claim 1, characterized in that, The adsorbent is selected from chromatographic column packings; Preferably, the adsorbent is selected from polar or weakly polar chromatographic column packings, and further preferably includes macroporous resin and / or polyamide; Preferably, the mass ratio of crystalline hesperidin to the adsorbent in the feed is 1:(1 - 9).

4. The preparation method according to claim 1, wherein Add the adsorbent to the aqueous solution containing amorphous hesperidin, maintain the system temperature at 60 - 95 °C, and adsorb for 1 - 6 h; Preferably, stir during the adsorption process, and the stirring speed is 300 - 500 RPM; Preferably, after adsorption, separate the adsorbent adsorbed with amorphous hesperidin and dry it; Preferably, the drying temperature is 40 - 80 °C.

5. The preparation method according to claim 1, wherein The organic solvent includes methanol or an ethanol aqueous solution with a concentration of at least 90% v / v; Preferably, the organic solvent is a 95% v / v ethanol aqueous solution; Preferably, desorb the amorphous hesperidin on the adsorbent under ultrasonic conditions; Preferably, the removal of the solvent in the desorption solution is carried out at a temperature not higher than 50 °C; Preferably, remove the solvent in the desorption solution by rotary evaporation under reduced pressure to obtain an amorphous powder of hesperidin.

6. The preparation method according to claim 1, wherein, The phospholipids include one or more of soybean phospholipids and lecithin; Preferably, the mass ratio of the amorphous powder of hesperidin to phospholipids in the feed is 1:(1 - 9).

7. The preparation method according to claim 1, characterized in that, React the amorphous powder of hesperidin and phospholipids in an ethanol aqueous solution with a concentration of at least 95% v / v to obtain a hesperidin phospholipid complex; Preferably, stir and react at a temperature of 30 - 50 °C for 1 - 6 h.

8. The preparation method according to any one of claims 1 to 7, characterized in that, It includes the following steps: (a) Maintain the water bath temperature at 60 - 95 °C, add crystalline hesperidin, water and a magnetic stirring rotor to the reaction vessel; the material ratio of the magnetic stirring rotor to water is 1:(10 - 20); the rotation speed is 300 - 500 RPM; the stirring time is 1 - 6 h to obtain an amorphous hesperidin solution. (b) Add an adsorbent to the system in step (a), and the mass ratio of crystalline hesperidin to the adsorbent in the feed is 1:(1 - 9), continue to maintain the temperature at 60 - 95 °C, and stir for 1 - 6 h; (c) Separate the adsorbent from the liquid through a 60 - 80 - mesh filter cloth, and wash with water to remove the unadsorbed hesperidin, and dry the adsorbent at a temperature of 40 °C - 80 °C; (d) Add ethanol to the dried adsorbent, and adjust the ethanol concentration in the system to ≥90% v / v to desorb the amorphous hesperidin, and the resulting solution is a desorption solution containing amorphous hesperidin; (e) Remove the organic solvent from the desorption solution of amorphous hesperidin by rotary evaporation under reduced pressure, and the resulting solid is an amorphous powder of hesperidin; (f) Take the amorphous hesperidin powder and phospholipids from step (e), use an ethanol aqueous solution at 95% v / v as the reaction solvent, stir at 30 - 50 °C for 1 - 6 hours, and then dry under reduced pressure. The resulting solid is the hesperidin phospholipid complex.

9. The hesperidin phospholipid complex prepared by the preparation method according to any one of claims 1 - 8.

10. The preparation method of the hesperidin phospholipid complex according to any one of claims 1 - 8, or the application of the hesperidin phospholipid complex according to claim 9 in the preparation of drugs.

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