Preparation method and application of piperine-rhein-chlorogenic acid self-assembled hydrogel

Abstract

The invention relates to the field of preparation of natural products, and particularly provides a preparation method and application of piperine-rhein-chlorogenic acid self-assembled hydrogel. The method comprises the following steps: dissolving piperine and rhein, mixing, adjusting the pH value, heating for reaction, purifying and dialyzing to obtain piperine-rhein particles; and then, carrying out ultrasonic-assisted reaction with chlorogenic acid, and carrying out vacuum freeze drying, thereby obtaining the chlorogenic acid. The method is simple to operate, and the prepared piperine-rhein-chlorogenic acid can remarkably improve the anti-inflammatory effect of cells and has the advantages of high biocompatibility, small toxicity and adverse reaction and the like.

Description

Technical Field

[0001] The invention belongs to the field of preparation of natural products, and specifically relates to a preparation method and application of a piperine-rhein-chlorogenic acid self-assembled hydrogel. Background Art

[0002] Inflammation is the body's defensive response to various inflammatory factors. Moderate inflammatory response is beneficial to the body's health, while excessive and persistent inflammatory response can cause damage to the body, such as cancer and autoimmune diseases. With the in-depth research on natural compounds in recent years, the anti-inflammatory effects of more and more natural compounds have been recognized. Natural compounds improve inflammation with multiple pathways, multiple levels, and high safety.

[0003] Piperine is the main active ingredient of pepper, with multiple effects such as antibacterial, anti-inflammatory, antioxidant, and anticonvulsant, but its poor water solubility and low bioavailability limit its efficient use. Small molecule compounds are directly assembled into hydrogels, effectively replacing exogenous inactive ingredients with active ingredients, achieving high loading of nutrients, and having the characteristics of high biocompatibility. Designing hydrogels suitable for loading piperine has become an urgent problem to be solved.

[0004] In recent years, the use of small molecule compounds to self-assemble into hydrogels has become a research hotspot. A large number of studies have found that some natural active ingredients such as alkaloids, polyphenols, and flavonoids have the ability to self-assemble or co-assemble to form delivery carriers. Compared with most hydrogels that rely on polymer carriers or incorporate polymer compounds, metal ions, etc. as media, natural product self-assembled hydrogels have the advantages of high biocompatibility, strong synergistic effects, low toxicity and side effects. However, there are few reports on piperine self-assembled hydrogels in the prior art. Summary of the invention

[0005] Based on the structural characteristics of piperine and previous research foundation, the present invention utilizes piperine, rhein and chlorogenic acid to construct a hydrogel system to improve the anti-inflammatory ability of piperine.

[0006] In order to achieve the above object, the present invention provides the following technical solutions: The first aspect of the present invention provides a piperine-rhein-chlorogenic acid self-assembled hydrogel with anti-inflammatory effect, characterized in that the self-assembled hydrogel is self-assembled by piperine, rhein and chlorogenic acid; the preparation method is as follows: 1.00 mmol / L piperine and 10.00 mmol / L rhein are mixed to obtain a mixed solution; the pH of the mixed solution is adjusted to 7.0-7.5 using a 5 mg / mL sodium hydroxide solution, and then an equal volume of phosphate buffer to the mixed solution is added, and the mixture is reacted at 60°C and 180 r / min for 10 min to obtain a reaction mixture; the reaction mixture is subjected to normal phase silica gel column chromatography and eluted with 20% v / v ethanol; the eluate is concentrated to 1 / 2 of the original volume, and dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein particle solution; an equal volume of phosphate buffer containing chlorogenic acid is added to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 2%, and the mixture is ultrasonically treated at 30°C and 100W for 10 min. min, and after standing for 1.0 h, vacuum freeze-drying was performed to obtain the piperine-rhein-chlorogenic acid self-assembled hydrogel dry product; Wherein, the preparation method of piperine is: A1: uniformly mixing citric acid and choline chloride in a molar ratio of 3:1 and heating until the mixture forms a clear and transparent liquid to obtain a citric acid-choline chloride deep eutectic solvent; the heating temperature is 55° C. and the heating time is 30 min; A2: adding white pepper powder to the citric acid-choline chloride low eutectic solvent in step A1 to obtain a mixed solution, heating the mixed solution to 60° C. and then continuously heating at a constant temperature for 30 min, extracting the mixed solution, centrifuging and filtering the extracted mixed solution, collecting the filtrate, removing the filter residue to obtain a clarified piperine extract, wherein the solid-liquid ratio of the white pepper powder to the citric acid-choline chloride is 1 / 35 g / mL; A3: adding the piperine extract described in step A2 to an extraction solvent, oscillating at room temperature and then ultrasonically treating the mixture, standing for phase separation, and obtaining an upper phase and a lower phase; the extraction solvent is composed of isopropanol and water, and the mass ratio of the piperine extract, isopropanol, and water is 2:2:3; the upper phase is an isopropanol phase containing piperine, and the lower phase is a low eutectic solvent phase containing a mixture of the piperine extract; the upper phase is distilled under reduced pressure to 1 / 8 of the original volume, and residual isopropanol is removed to obtain a piperine extract; the piperine extract is subjected to vacuum freeze-drying treatment to obtain high-purity piperine; The rhein and chlorogenic acid were purchased from Shanghai Yuanye Biotechnology Co., Ltd. with a purity of >90%.

[0007] Furthermore, the solvent for the 1.00 mmol / L piperine is a methanol solution; and the solvent for the 10.00 mmol / L rhein is a dimethyl sulfoxide solution.

[0008] Furthermore, the volume ratio of the 1.00 mmol / L piperine and the 10.00 mmol / L rhein is 1:1.

[0009] Furthermore, the pH of the phosphate buffer is 7.4.

[0010] The second aspect of the present invention provides the use of the above-mentioned piperine-rhein-chlorogenic acid self-assembled hydrogel with anti-inflammatory effect in health products, food and feed.

[0011] Beneficial effects of the present invention: The method is simple to operate, and the prepared piperine hydrogel has the advantages of high biocompatibility, low toxicity and adverse reactions, and can significantly improve the anti-inflammatory ability of piperine. DETAILED DESCRIPTION

[0012] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0013] Example 1 Preparation of piperine A1: uniformly mixing citric acid and choline chloride in a molar ratio of 3:1 and heating until the mixture forms a clear and transparent liquid to obtain a citric acid-choline chloride deep eutectic solvent; the heating temperature is 55° C. and the heating time is 30 min; A2: adding white pepper powder to the citric acid-choline chloride low eutectic solvent in step A1 to obtain a mixed solution, heating the mixed solution to 60° C. and then continuously heating at a constant temperature for 30 min, extracting the mixed solution, centrifuging and filtering the extracted mixed solution, collecting the filtrate, removing the filter residue to obtain a clarified piperine extract, wherein the solid-liquid ratio of the white pepper powder to the citric acid-choline chloride is 1 / 35 g / mL; A3: adding the piperine extract described in step A2 to an extraction solvent, oscillating at room temperature and then ultrasonically treating, standing for phase separation, and obtaining an upper phase and a lower phase; the extraction solvent is composed of isopropanol and water, and the mass ratio of the piperine extract, isopropanol, and water is 2:2:3; the upper phase is an isopropanol phase containing piperine, and the lower phase is a low eutectic solvent phase containing a mixture of the piperine extract; the upper phase is distilled under reduced pressure to 1 / 8 of the original volume, and residual isopropanol is removed to obtain a piperine extract; the piperine extract is subjected to vacuum freeze-drying treatment to obtain high-purity piperine.

[0014] Example 2 Rhein was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with a purity of >90%.

[0015] Example 3 Chlorogenic acid was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with a purity of >90%.

[0016] Example 4 Preparation of a piperine-rhein-chlorogenic acid self-assembled hydrogel This embodiment provides a method for preparing a piperine-rhein-chlorogenic acid self-assembled hydrogel, the method comprising the following steps: (1) 1.00 mmol / L piperine and 10.00 mmol / L rhein were mixed to obtain a mixed solution. The pH of the mixed solution was adjusted to 7.0-7.5 using 5 mg / mL NaOH, and an equal volume of phosphate buffer (pH 7.4) was added. The mixture was reacted at 60°C and 180 r / min for 10 min to obtain a reaction mixture. The reaction mixture was subjected to normal phase silica gel column chromatography and eluted with 20% v / v ethanol; the eluate was concentrated to 1 / 2 of the original volume and dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein granular solution. The piperine was obtained in Example 1, and the rhein was obtained in Example 2.

[0017] (2) Add an equal volume of phosphate buffer (pH 7.4) containing chlorogenic acid to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 2%, and ultrasonicate for 10 min at 30°C and 100W, stand for 1.0 h, and then freeze-dry in vacuum to obtain a piperine-rhein-chlorogenic acid self-assembled hydrogel dry product. The chlorogenic acid is obtained by Example 3.

[0018] Example 5 Preparation of piperine-rhein self-assembled particles This embodiment provides a method for preparing piperine-rhein self-assembled particles, the method comprising the following steps: 1.00 mmol / L piperine and 10.00 mmol / L rhein were mixed to obtain a mixed solution. The pH of the mixed solution was adjusted to 7.0-7.5 using 5 mg / mL NaOH, and an equal volume of phosphate buffer (pH 7.4) was added, and the mixture was reacted at 60 °C and 180 r / min for 10 min to obtain a reaction mixture. The reaction mixture was subjected to normal phase silica gel column chromatography and eluted with 20% v / v ethanol; the eluate was concentrated to 1 / 2 of the original volume and dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein granular solution, which was freeze-dried. The piperine was obtained in Example 1, and the rhein was obtained in Example 2.

[0019] Example 6 Preparation method of piperine-rhein-chlorogenic acid self-assembled hydrogel This embodiment provides a method for preparing a piperine-rhein-chlorogenic acid self-assembled hydrogel, the method comprising the following steps: (1) 1.00 mmol / L piperine and 10.00 mmol / L rhein were mixed to obtain a mixed solution. The pH of the mixed solution was adjusted to 7.0-7.5 using 5 mg / mL NaOH, and an equal volume of phosphate buffer (pH 7.4) was added. The mixture was reacted at 50 °C and 180 r / min for 10 min to obtain a reaction mixture. The reaction mixture was dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein particle solution. The piperine was obtained in Example 1, and the rhein was obtained in Example 2.

[0020] (2) Add an equal volume of phosphate buffer (pH 7.4) containing chlorogenic acid to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 2%, and ultrasonicate for 10 min at 30°C and 100W, stand for 1.0 h, and then freeze-dry in vacuum to obtain a piperine-rhein-chlorogenic acid self-assembled hydrogel dry product. The chlorogenic acid is obtained by Example 3.

[0021] Example 7 Preparation method of piperine-rhein-chlorogenic acid self-assembled hydrogel This embodiment provides a method for preparing piperine-rhein self-assembled nanoparticles, the method comprising the following steps: (1) 1.00 mmol / L piperine and 10.00 mmol / L rhein were mixed to obtain a mixed solution. The pH of the mixed solution was adjusted to 7.0-7.5 using 5 mg / mL NaOH, and an equal volume of phosphate buffer (pH 7.4) was added. The mixture was reacted at 60°C and 180 r / min for 10 min to obtain a reaction mixture. The reaction mixture was subjected to normal phase silica gel column chromatography and eluted with 20% v / v ethanol; the eluate was concentrated to 1 / 2 of the original volume and dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein granular solution. The piperine was obtained in Example 1, and the rhein was obtained in Example 2.

[0022] (2) Add an equal volume of phosphate buffer (pH 7.4) containing chlorogenic acid to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 3%, and ultrasonicate for 10 min at 30°C and 100W, stand for 1.0 h, and then freeze-dry in vacuum to obtain a piperine-rhein-chlorogenic acid self-assembled hydrogel dry product. The chlorogenic acid is obtained by Example 3.

[0023] Example 8 Preparation method of piperine-rhein-chlorogenic acid self-assembled hydrogel This embodiment provides a method for preparing a piperine-rhein-chlorogenic acid self-assembled hydrogel, the method comprising the following steps: (1) 1.00 mmol / L piperine and 10.00 mmol / L rhein were mixed to obtain a mixed solution. The pH of the mixed solution was adjusted to 7.0-7.5 using 5 mg / mL NaOH, and an equal volume of phosphate buffer (pH 7.4) was added. The mixture was reacted at 60 °C and 180 r / min for 10 min to obtain a reaction mixture. The reaction mixture was dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein particle solution. The piperine was obtained in Example 1, and the rhein was obtained in Example 2.

[0024] (2) Add an equal volume of phosphate buffer (pH 7.4) containing chlorogenic acid to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 2%, and ultrasonicate for 10 min at 30°C and 100W, stand for 1.0 h, and then freeze-dry in vacuum to obtain a piperine-rhein-chlorogenic acid self-assembled hydrogel dry product. The chlorogenic acid is obtained by Example 3.

[0025] Example 9 Preparation of a piperine-rhein-chlorogenic acid self-assembled hydrogel This embodiment provides a method for preparing a piperine-rhein-chlorogenic acid self-assembled hydrogel, the method comprising the following steps: (1) 1.00 mmol / L piperine and 10.00 mmol / L rhein were mixed to obtain a mixed solution. The pH of the mixed solution was adjusted to 7.0-7.5 using 5 mg / mL NaOH, and an equal volume of phosphate buffer (pH 7.4) was added. The mixture was reacted at 60°C and 180 r / min for 10 min to obtain a reaction mixture. The reaction mixture was subjected to normal phase silica gel column chromatography and eluted with 20% v / v ethanol; the eluate was concentrated to 1 / 2 of the original volume and dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein granular solution. The piperine was obtained in Example 1, and the rhein was obtained in Example 2.

[0026] (2) Add an equal volume of phosphate buffer (pH 7.4) containing chlorogenic acid to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 2%, react at 30°C for 10 min, stand for 1.0 h, and then freeze-dry in vacuum to obtain a piperine-rhein-chlorogenic acid self-assembled hydrogel dry product. The chlorogenic acid is obtained in Example 3.

[0027] Example 10 Preparation of a piperine-rhein-chlorogenic acid self-assembled hydrogel This embodiment provides a method for preparing a piperine-rhein-chlorogenic acid self-assembled hydrogel, the method comprising the following steps: (1) 1.00 mmol / L piperine and 10.00 mmol / L rhein were mixed to obtain a mixed solution. The pH of the mixed solution was adjusted to 7.0-7.5 using 5 mg / mL NaOH, and an equal volume of phosphate buffer (pH 7.4) was added. The mixture was reacted at 60 °C and 180 r / min for 10 min to obtain a reaction mixture. The reaction mixture was subjected to normal phase silica gel column chromatography and eluted with 20% v / v ethanol; the eluate was concentrated to 1 / 2 of the original volume and dialyzed in a 1.0 KDa filter membrane for 16 h to obtain a piperine-rhein granular solution. The piperine was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (purity>90%), and the rhein was obtained in Example 2.

[0028] (2) Add an equal volume of phosphate buffer (pH 7.4) containing chlorogenic acid to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 2%, and ultrasonicate for 10 min at 30°C and 100W, stand for 1.0 h, and then freeze-dry in vacuum to obtain a piperine-rhein-chlorogenic acid self-assembled hydrogel dry product. The chlorogenic acid is obtained by Example 3.

[0029] Experiment 1 Anti-inflammatory effect determination experiment Cell culture: RAW264.7 cells were cultured in DMEM complete medium (containing 10% newborn calf serum and 90% DMEM high glucose medium) in a 37°C, 5% CO2 incubator. Cells in the logarithmic growth phase were selected for the experiment.

[0030] Determination of cell survival rate: Take cells in the logarithmic growth phase, adjust the cell concentration to 1×104 / mL, inoculate them on a 96-well plate, 100 μL per well, and culture them at 37°C, 5% CO2 for 24 h. RAW264.7 cells were divided into a normal group, a model group (lipopolysaccharide) and an experimental group. A blank group containing only 100 μL of complete culture medium was also set up without cells. The normal group and the model group were not intervened by samples and lipopolysaccharide, and an equal volume of complete culture medium was added and cultured for 2 h; the experimental group was sequentially added with a sample solution with a final concentration of 10 mg / L and cultured for 2 h. Then, the model group and the experimental group were cultured with a lipopolysaccharide solution with a final concentration of 0.5 mg / L. After 24 h of culture, the culture medium was discarded, and 90 μL of complete culture medium and 10 μL of 5 mg / mL thiazolyl blue were added to each well. The culture was continued for 4 h, the upper liquid was discarded, and 110 μL of dimethyl sulfoxide was added. The cells were shaken until all the crystals dissolved. The OD value of each well was detected at 490 nm, and the cell survival rate was calculated. Cell survival rate = (OD value of experimental group - OD value of blank group) / (OD value of normal group - OD value of blank group) × 100%.

[0031] Determination of NO level in RAW 264.7 cells. Cells were inoculated in 24-well plates at 1.0×105 / mL, 0.5mL per well, and cultured at 37℃ and 5% CO2 for 24 hours. The old culture medium was discarded and RAW264.7 cells were divided into normal group, model group (lipopolysaccharide) and experimental group. A blank group without cells and only 100 μL complete culture medium was set up. The normal group and model group were not intervened by samples and lipopolysaccharide, and an equal volume of complete culture medium was added and cultured for 2 hours; the experimental group was added with sample solutions with a final concentration of 10 mg / L in turn and cultured for 2 hours. Then, the model group and the experimental group were cultured with lipopolysaccharide solution with a final concentration of 0.5 mg / L, and the culture was continued for 24 hours. The cell supernatant was collected, and the NO level in the cell supernatant was detected using a nitrate reductase (NR) activity detection kit (Griess kit).

[0032] The specific experimental results are shown in Table 1.

[0033] Table 1 Effects of different groups on RAW264.7 cell survival rate and NO release

[0034] Note: Different lowercase letters in the same category indicate significant differences between different groups (p>0.05). From Table 1, we can see that: (1) Compared with Examples 1-3, the cell survival rate of piperine-rhein-chlorogenic acid self-assembled hydrogels in Examples 4 and 6-9 was significantly increased, and the NO concentration was significantly reduced. It can be seen that piperine-rhein-chlorogenic acid self-assembled hydrogels can enhance the anti-inflammatory effect of piperine.

[0035] (2) Compared with Example 5, the cell survival rate of piperine-rhein-chlorogenic acid self-assembled hydrogel in Experimental Examples 4, 6-9 was significantly increased, and the NO concentration was significantly reduced. It can be seen that the anti-inflammatory effect of piperine-rhein-chlorogenic acid self-assembled hydrogel is stronger than that of piperine-rhein self-assembled particles.

[0036] (3) Compared with Examples 6, 7, 8, and 9, the cell survival rate of piperine-rhein-chlorogenic acid self-assembled hydrogel in Experimental Example 4 was significantly increased, and the NO concentration was significantly reduced. It can be seen that the changes in the preparation conditions of piperine-rhein-chlorogenic acid self-assembled hydrogel are not conducive to its anti-inflammatory effect. Among them, the lack of silica gel column chromatography to purify piperine-rhein solution is the most unfavorable to the anti-inflammatory effect of piperine-rhein-chlorogenic acid self-assembled hydrogel, followed by ultrasound-assisted chlorogenic acid reaction, followed by the amount of chlorogenic acid added, and the reaction temperature of piperine-rhein solution has the weakest effect.

[0037] (4) Compared with Example 10, the cell survival rate of the piperine-rhein-chlorogenic acid self-assembled hydrogel prepared by using the piperine extracted by the present invention (Example 1) was significantly increased, and the NO concentration was significantly reduced. It can be seen that the piperine extracted by using the citric acid-choline chloride low eutectic solvent two-phase system can enhance the anti-inflammatory effect of the piperine-rhein-chlorogenic acid self-assembled hydrogel compared with the piperine purchased from the market.

[0038] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A piperine-rhein-chlorogenic acid self-assembled hydrogel with anti-inflammatory effect, characterized in that: The self-assembled hydrogel is self-assembled by piperine, rhein and chlorogenic acid. The preparation method comprises the following steps: mixing 1.00 mmol / L piperine and 10.00 mmol / L rhein to obtain a mixed solution; adjusting the pH value of the mixed solution to 7.0-7.5 using a 5 mg / mL sodium hydroxide solution, adding an equal volume of phosphate buffer to the mixed solution, reacting at 60°C and 180 r / min for 10 min to obtain a reaction mixture; subjecting the reaction mixture to normal phase silica gel column chromatography and eluting with 20% v / v ethanol; concentrating the eluate to 1 / 2 of the original volume, placing the eluate in a 1.0 KDa filter membrane for dialysis for 16 h to obtain a piperine-rhein particle solution; adding an equal volume of phosphate buffer containing chlorogenic acid to the piperine-rhein particle solution, wherein the mass concentration of the chlorogenic acid is 2%, ultrasonicating for 10 min at 30°C and 100W, and standing for 1.0 h later, vacuum freeze drying was performed to obtain the piperine-rhein-chlorogenic acid self-assembled hydrogel dry product; Wherein, the preparation method of piperine is: A1: uniformly mixing citric acid and choline chloride in a molar ratio of 3:1 and heating until the mixture forms a clear and transparent liquid to obtain a citric acid-choline chloride deep eutectic solvent; the heating temperature is 55° C. and the heating time is 30 min; A2: adding white pepper powder to the citric acid-choline chloride low eutectic solvent in step A1 to obtain a mixed solution, heating the mixed solution to 60° C. and then continuously heating at a constant temperature for 30 min, extracting the mixed solution, centrifuging and filtering the extracted mixed solution, collecting the filtrate, removing the filter residue to obtain a clarified piperine extract, wherein the solid-liquid ratio of the white pepper powder to the citric acid-choline chloride is 1 / 35 g / mL; A3: adding the piperine extract described in step A2 to an extraction solvent, oscillating at room temperature and then ultrasonically treating the mixture, standing for phase separation, and obtaining an upper phase and a lower phase; the extraction solvent is composed of isopropanol and water, and the mass ratio of the piperine extract, isopropanol, and water is 2:2:3; the upper phase is an isopropanol phase containing piperine, and the lower phase is a low eutectic solvent phase containing a mixture of the piperine extract; the upper phase is distilled under reduced pressure to 1 / 8 of the original volume, and residual isopropanol is removed to obtain a piperine extract; the piperine extract is subjected to vacuum freeze-drying treatment to obtain high-purity piperine; The rhein and chlorogenic acid are commercially available with a purity of >90%.

2. The piperine-rhein-chlorogenic acid self-assembled hydrogel with anti-inflammatory effect according to claim 1, characterized in that: The solvent for the 1.00 mmol / L piperine is a methanol solution; the solvent for the 10.00 mmol / L rhein is a dimethyl sulfoxide solution.

3. The piperine-rhein-chlorogenic acid self-assembled hydrogel with anti-inflammatory effect according to claim 1, characterized in that: The volume ratio of the 1.00 mmol / L piperine and the 10.00 mmol / L rhein is 1:

1.

4. The piperine-rhein-chlorogenic acid self-assembled hydrogel with anti-inflammatory effect according to claim 1, characterized in that: The pH of the phosphate buffer is 7.

4.

5. Use of the piperine-rhein-chlorogenic acid self-assembled hydrogel with anti-inflammatory effect as claimed in claim 1 in the preparation of anti-inflammatory products, food and feed.

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