A chitosan-based hemoperfusion adsorbent, its preparation method and application

Abstract

The application relates to the material field and specifically discloses a chitosan-based hemoperfusion adsorbent as well as a preparation method and application thereof. The hemoperfusion adsorbent takes modified chitosan microspheres as a carrier, grafts chicory acid on the surface, and solidifies manganese dioxide nanometer enzymes to form a composite adsorbent. The hemoperfusion adsorbent provided by the application can efficiently remove active oxygen and various free radicals due to the fact that manganese dioxide nanometer enzymes are solidified on the surface, has high adsorption capacity, can effectively adsorb inflammatory factors in blood in cooperation with the chitosan microspheres after being solidified on the surface of the chitosan microspheres, improves the blood purification effect, has blood compatibility and safety, and can be used for blood purification treatment of inflammatory patients.

Description

Technical Field

[0001] This application relates to the field of medical devices, and more specifically, to a chitosan-based blood perfusion adsorbent, its preparation method, and its application. Background Technology

[0002] Hemoperfusion is an extracorporeal blood purification method that involves drawing blood from the patient's body, passing it through a perfusion device containing a specific adsorbent to remove harmful substances, and then returning the purified blood to the patient. This technology has shown great potential in treating a variety of diseases, especially inflammatory diseases, such as severe hepatitis, psoriasis, and sepsis.

[0003] However, existing hemoperfusion adsorbents have limitations in clearing inflammatory factors from the blood. Patients with inflammatory diseases often have high concentrations of inflammatory factors in their blood, such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), which are key mediators of the inflammatory response. Existing adsorbents have limited efficiency in clearing these inflammatory factors, leading to poor treatment outcomes. Furthermore, inflammatory patients often experience increased oxidative stress due to the inflammatory response, resulting in elevated levels of free radicals and reactive oxygen species in the blood. These substances damage cell nuclei and tissues, and existing adsorbents are often ineffective against them.

[0004] Therefore, it is necessary to develop a blood perfusion adsorbent for patients with inflammation in order to improve the blood perfusion purification effect. Summary of the Invention

[0005] To improve the clearance rate of inflammatory factors, free radicals and reactive oxygen species during hemoperfusion while reducing hemolysis and safety, this application provides a chitosan-based hemoperfusion adsorbent, its preparation method and application.

[0006] The technical solution adopted in this application is as follows: In a first aspect, this application provides a chitosan-based hemoperfusion adsorbent, which uses modified chitosan microspheres as a carrier, grafts chicoric acid onto the surface, and then immobilizes manganese dioxide nanozymes to form a composite adsorbent for clearing inflammatory factors from the blood of patients with inflammation.

[0007] The above technical solution optimizes the selection of adsorbent materials, surface modification and alteration, and uses chitosan microspheres as a carrier, which has good biocompatibility and mechanical strength. After surface modification, chicoric acid is grafted onto the surface, and manganese dioxide nanozymes are immobilized on the surface of the adsorbent using chicoric acid as a linker.

[0008] Manganese dioxide nanozymes possess three types of antioxidant enzyme activity, including superoxide dismutase-like, catalase-like, and glutathione peroxidase-like activities, enabling them to efficiently scavenge reactive oxygen species and various free radicals. Furthermore, they exhibit high adsorption capacity; when immobilized on the surface of chitosan microspheres, they synergistically and effectively adsorb inflammatory factors in the blood, enhancing blood purification. Simultaneously, the presence of two adjacent carboxyl groups and four phenolic hydroxyl groups in the chicoric acid molecule provides a stable chemical bond connecting the chitosan microsphere carrier and the manganese dioxide nanozyme ligand, acting as a bridge. Moreover, the modification with chicoric acid enhances the hydrophilicity of the adsorbent surface, improving blood compatibility and reducing the risk of blood coagulation and platelet activation.

[0009] Preferably, in a preferred embodiment of this application, the chitosan microspheres have a particle size of 100-200 μm and a coefficient of variation (CV) < 5%. This type of chitosan microsphere has strong monodispersity, is not prone to aggregation during subsequent chemical modification, and can provide a more consistent adsorption surface area and pore structure during blood perfusion, which helps to improve the efficiency of blood perfusion.

[0010] Furthermore, in a preferred embodiment of this application, the modification method of the above-mentioned modified chitosan microspheres includes: (1) Chitosan microspheres were dispersed in an alkaline solution and sonicated for 20-30 min to obtain a chitosan microsphere dispersion; (2) Add 2,3-epoxypropyltrimethylammonium chloride and ferrous sulfate to the chitosan microsphere dispersion, and react at 40-60℃ for 7-9 hours under stirring, and then wash.

[0011] Preferably, the ultrasound time is 24-26 minutes; Preferably, the reaction is carried out at a temperature of 45-55℃ for 7.5-8.5 hours.

[0012] Further, in a preferred embodiment of this application, the mass ratio of the chitosan microspheres to the 2,3-epoxypropyltrimethylammonium chloride is 20-30:1; preferably, the mass ratio is 23-26:1.

[0013] Secondly, this application provides a method for preparing the above-mentioned blood perfusion adsorbent, comprising: (1) Chitosan microspheres are dispersed in an alkaline solution and sonicated for 20-30 min to obtain a chitosan microsphere dispersion. 2,3-epoxypropyltrimethylammonium chloride and ferrous sulfate are added to the chitosan microsphere dispersion and reacted at 40-60℃ for 7-9 h under stirring. After washing, modified chitosan microspheres are obtained. (2) Add the modified chitosan microspheres to an ethanol aqueous solution containing chicoric acid, sonicate for 40-50 min, add stannous chloride, and react at 70-80℃ for 3-4 h to obtain chicoric acid-modified chitosan microspheres. Stannous chloride acts as a catalyst, which helps to accelerate the reaction rate. Preferably, the reaction temperature is 73-77°C and the reaction time is 3.5-4 hours.

[0014] 1. The chitosan microspheres acidified by chicory are dispersed in an ethylene glycol solution containing manganese nitrate hexahydrate, an alkaline solution is added, and a pyrolysis reaction is carried out. After centrifugation and washing, the blood perfusion adsorbent is obtained.

[0015] Manganese nitrate hexahydrate (Mn(NO3)2·6H2O) decomposes under alkaline conditions upon heating to form MnO2. The free MnO2 in the solution is captured by chitosan microspheres acidified by chicory during the gradual formation process and is immobilized on the chitosan microspheres to form the blood perfusion adsorbent of this application.

[0016] Furthermore, in a preferred embodiment of this application, the mass ratio of the modified chitosan microspheres to chicoric acid is 15-20:1; preferably, the mass ratio is 16-18:1.

[0017] Furthermore, in a preferred embodiment of this application, the mass ratio of the modified chitosan microspheres to manganese nitrate hexahydrate is 6-8:1; preferably, the mass ratio is 6.5-7.5:1.

[0018] Furthermore, in a preferred embodiment of this application, the reaction temperature of the above-mentioned depyrolysis reaction is 50-80°C and the reaction time is 3-6 hours; preferably, the reaction temperature is 60-70°C and the reaction time is 4-5 hours.

[0019] Thirdly, this application provides the application of the above-mentioned blood perfusion adsorbent in the preparation of blood perfusion equipment.

[0020] In summary, this application has the following beneficial effects: 1. The blood perfusion adsorbent provided in this application uses chitosan microspheres as a carrier. It has good biocompatibility and mechanical strength. After surface modification, the structural stability of the chitosan microspheres can be increased, and the chemical reactivity of the microsphere surface can be improved, which is conducive to further grafting chicoric acid.

[0021] 2. The hemoperfusion adsorbent provided in this application uses chicoric acid as a linker. Because the chicoric acid molecule contains two adjacent carboxyl groups and four phenolic hydroxyl groups, it can act as an intermediate bridge, providing a stable chemical bond to connect the chitosan microsphere carrier and the manganese dioxide nanozyme ligand. Furthermore, the modification with chicoric acid enhances the hydrophilicity of the adsorbent surface, which helps improve blood compatibility and reduce the risk of blood coagulation and platelet activation.

[0022] 3. The hemoperfusion adsorbent provided in this application uses manganese dioxide nanozymes as ligands. Manganese dioxide nanozymes possess three types of antioxidant enzyme activity, including superoxide dismutase (SOD)-like, catalase (CAT)-like, and glutathione peroxidase (GPX)-like activities, which can efficiently scavenge reactive oxygen species and various free radicals. Furthermore, they have high adsorption capacity; after being immobilized on the surface of chitosan microspheres, they can synergistically and effectively adsorb inflammatory factors in the blood with the chitosan microspheres, thereby improving the blood purification effect. Detailed Implementation

[0023] The embodiments of the present invention will be described in detail below with reference to the examples. However, those skilled in the art will understand that the following examples are only for illustrating the present invention and should not be regarded as limiting the scope of the present invention. Specific conditions not specified in the examples shall be carried out according to conventional conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0024] The following provides a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit the scope of the invention.

[0025] Example Example

[0026] This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which includes the following steps: 1. Disperse 50g of chitosan microspheres with a particle size of 100-200μm (coefficient of variation CV < 5%, purchased from Shanghai Pengzan Biotechnology Co., Ltd.) in 250ml of 0.05mol / L NaOH solution, and sonicate for 25min to obtain chitosan microsphere dispersion; 2. Add 2,3-epoxypropyltrimethylammonium chloride (2g) to the chitosan microsphere dispersion, stir evenly, add a catalytic amount of ferrous sulfate, stir and react for 8 hours in a water bath at 50℃, and then wash with ethanol and deionized water to obtain modified chitosan microspheres. 3. Dissolve chicoric acid (2g) in 100mL of 70% ethanol solution to obtain chicoric acid solution; add 35g of modified chitosan microspheres obtained in step (2) to chicoric acid solution, sonicate for 45min, add catalytic amount of stannous chloride, place the mixed reaction solution in a water bath at 72℃ for 4h, filter and wash with ethanol and deionized water in sequence to obtain chicoric acidified chitosan microspheres; 4. Dissolve 5.3g of manganese nitrate hexahydrate crystals in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution. Then add chitosan microspheres obtained in step (3), sonicate for 30min, place in a water bath at 60℃, add 4mL of ammonia water while stirring at 250rpm, continue the reaction for 5h, centrifuge, wash with ethanol, wash with deionized water until neutral, and freeze-dry under vacuum.

[0027] Example 2 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (2): 2,3-epoxypropyltrimethylammonium chloride (2.5 g) was added to the chitosan microsphere dispersion and stirred until homogeneous. Then, a catalytic amount of ferrous sulfate was added, and the mixture was stirred in a water bath at 50 °C for 8 h. The mixture was then washed with ethanol and deionized water to obtain modified chitosan microspheres.

[0028] Example 3 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (2): 2,3-epoxypropyltrimethylammonium chloride (1.6 g) was added to the chitosan microsphere dispersion and stirred until homogeneous. Then, a catalytic amount of ferrous sulfate was added, and the mixture was stirred in a water bath at 50 °C for 8 h. The mixture was then washed with ethanol and deionized water to obtain modified chitosan microspheres.

[0029] Example 4 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (2): 2,3-epoxypropyltrimethylammonium chloride (2g) was added to the chitosan microsphere dispersion and stirred until homogeneous. Then, a catalytic amount of ferrous sulfate was added and the mixture was stirred in a water bath at 60℃ for 7 hours. The mixture was then washed with ethanol and deionized water to obtain modified chitosan microspheres.

[0030] Example 5 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (2): 2,3-epoxypropyltrimethylammonium chloride (2g) was added to the chitosan microsphere dispersion and stirred until homogeneous. Then, a catalytic amount of ferrous sulfate was added and the mixture was stirred in a water bath at 40°C for 9 hours. The mixture was then washed with ethanol and deionized water to obtain modified chitosan microspheres.

[0031] Example 6 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (3): Chicoric acid (2g) was dissolved in 100mL of 70% ethanol solution to obtain a chicoric acid solution. 40g of the modified chitosan microspheres obtained in step (2) were added to the chicoric acid solution. After sonication for 45min, a catalytic amount of stannous chloride was added. The mixed reaction solution was heated in a water bath at 72℃ for 4h. After filtration, the mixture was washed with ethanol and deionized water in sequence to obtain chicoric acidified chitosan microspheres.

[0032] Example 7 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (3): Chicoric acid (2g) was dissolved in 100mL of 70% ethanol solution to obtain a chicoric acid solution. 30g of the modified chitosan microspheres obtained in step (2) were added to the chicoric acid solution. After sonication for 45min, a catalytic amount of stannous chloride was added. The mixed reaction solution was heated in a water bath at 72℃ for 4h. After filtration, the mixture was washed with ethanol and deionized water in sequence to obtain chicoric acidified chitosan microspheres.

[0033] Example 8 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (3): Chicoric acid (2g) was dissolved in 100mL of 70% ethanol solution to obtain a chicoric acid solution. 35g of the modified chitosan microspheres obtained in step (2) were added to the chicoric acid solution. After sonication for 45min, a catalytic amount of stannous chloride was added. The mixed reaction solution was heated in a water bath at 80℃ for 3h. After filtration, the mixture was washed with ethanol and deionized water in sequence to obtain chicoric acidified chitosan microspheres.

[0034] Example 9 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (3): Chicoric acid (2g) was dissolved in 100mL of 70% ethanol solution to obtain a chicoric acid solution. 35g of the modified chitosan microspheres obtained in step (2) were added to the chicoric acid solution. After sonication for 45min, a catalytic amount of stannous chloride was added. The mixed reaction solution was heated in a water bath at 70℃ for 4h. After filtration, the mixture was washed with ethanol and deionized water in sequence to obtain chicoric acidified chitosan microspheres.

[0035] Example 10 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (4): 4.3 g of manganese nitrate hexahydrate crystals were dissolved in 200 mL of ethylene glycol to obtain a manganese nitrate hexahydrate solution. Then, chitosan microspheres acidified by chicory obtained in step (3) were added, ultrasonically dispersed for 30 min, placed in a water bath at 60 °C, and 4 mL of ammonia water was added while stirring at 250 rpm. The reaction was continued for 5 h. After centrifugation, the microspheres were washed with ethanol and then washed with deionized water until neutral. Finally, they were freeze-dried.

[0036] Example 11 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (4): 5.8 g of manganese nitrate hexahydrate crystals were dissolved in 200 mL of ethylene glycol to obtain a manganese nitrate hexahydrate solution. Then, chitosan microspheres acidified in step (3) were added, ultrasonically dispersed for 30 min, placed in a water bath at 60 °C, and 4 mL of ammonia water was added while stirring at 250 rpm. The reaction was continued for 5 h. After centrifugation, the microspheres were washed with ethanol and then washed with deionized water until neutral. Finally, they were freeze-dried.

[0037] Example 12 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (4): 5.3 g of manganese nitrate hexahydrate crystals were dissolved in 200 mL of ethylene glycol to obtain a manganese nitrate hexahydrate solution. Then, chitosan microspheres acidified by chicory obtained in step (3) were added, ultrasonically dispersed for 30 min, placed in a water bath at 50 °C, and 4 mL of ammonia water was added while stirring at 250 rpm. The reaction was continued for 6 h. After centrifugation, the microspheres were washed with ethanol and then washed with deionized water until neutral. Finally, they were freeze-dried.

[0038] Example 13 This embodiment provides a chitosan-based blood perfusion adsorbent, the preparation method of which differs from that of Example 1 in step (4): 5.3 g of manganese nitrate hexahydrate crystals were dissolved in 200 mL of ethylene glycol to obtain a manganese nitrate hexahydrate solution. Then, chitosan microspheres acidified by chicory obtained in step (3) were added, ultrasonically dispersed for 30 min, placed in a water bath at 80 °C, and 4 mL of ammonia water was added while stirring at 250 rpm. The reaction was continued for 3 h. After centrifugation, the microspheres were washed with ethanol and then washed with deionized water until neutral. Finally, they were freeze-dried.

[0039] Comparative Example Comparative Example 1 This comparative example provides a chitosan-based adsorbent, the preparation method of which includes: (1) Dissolve chicoric acid (2g) in 100mL of 70% ethanol solution to obtain chicoric acid solution; add 35g of chitosan microspheres (coefficient of variation CV<5%, purchased from Shanghai Pengzan Biotechnology Co., Ltd.) to chicoric acid solution, sonicate for 45min, add catalytic amount of stannous chloride, heat the mixed reaction solution in a water bath at 72℃ for 4h, filter and wash with ethanol and deionized water in sequence to obtain chicoric acidified chitosan microspheres; (2) Dissolve 5.3g of manganese nitrate hexahydrate crystals in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then add the above chitosan microspheres acidified by chicory, sonicate for 30min, add 4mL of ammonia water under stirring, and react in a water bath at 60℃ for 5h. After centrifugation, wash with ethanol, wash with deionized water until neutral, and freeze-dry under vacuum.

[0040] Comparative Example 2 This comparative example provides a chitosan-based adsorbent, the preparation method of which includes: 1. Disperse 50g of chitosan microspheres with a particle size of 100-200μm (coefficient of variation CV < 5%, purchased from Shanghai Pengzan Biotechnology Co., Ltd.) in 250ml of 0.05mol / L NaOH solution, and sonicate for 25min to obtain chitosan microsphere dispersion; 2. Add 2,3-epoxypropyltrimethylammonium chloride (2g) to the chitosan microsphere dispersion, stir evenly, add a catalytic amount of ferrous sulfate, stir and react for 8 hours in a water bath at 50℃, and then wash with ethanol and deionized water to obtain modified chitosan microspheres. 3. Dissolve 5.3g of manganese nitrate hexahydrate crystals in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then add the modified chitosan microspheres obtained in step (2), ultrasonically disperse for 30min, add 4mL of ammonia water while stirring, and react in a 60℃ water bath for 5h. After centrifugation, wash with ethanol, wash with deionized water until neutral, and freeze-dry under vacuum.

[0041] Comparative Example 3 This comparative example provides a chitosan-based adsorbent, the preparation method of which includes: 1. Disperse 50g of chitosan microspheres with a particle size of 100-200μm (coefficient of variation CV < 5%, purchased from Shanghai Pengzan Biotechnology Co., Ltd.) in 250ml of 0.05mol / L NaOH solution, and sonicate for 25min to obtain chitosan microsphere dispersion; 2. Add 2,3-epoxypropyltrimethylammonium chloride (2g) to the chitosan microsphere dispersion, stir evenly, add a catalytic amount of ferrous sulfate, stir and react for 8 hours in a water bath at 50℃, and then wash with ethanol and deionized water to obtain modified chitosan microspheres. 3. Dissolve chicoric acid (2g) in 100mL of 70% ethanol solution to obtain chicoric acid solution; add 35g of modified chitosan microspheres obtained in step (2) to the chicoric acid solution, sonicate for 45min, add catalytic amount of stannous chloride, heat the mixed reaction solution in a water bath at 72℃ for 4h, filter and wash with ethanol and deionized water in sequence to obtain chicoric acidified chitosan microspheres.

[0042] Performance testing Detection methods 1. Blood compatibility test: According to GB / T16886.4-2003 Biological evaluation of medical devices - Part 4 Selection of blood interaction tests, hemolysis tests were performed on the adsorbents provided in Examples 1-13 and Comparative Examples 1-3.

[0043] Calculate the hemolysis rate using the following formula: Hemolysis rate = (AB) / (CB)*100%, where A is the absorbance of the sample group; B is the absorbance of the negative control group; and C is the absorbance of the positive control group.

[0044] Experimental results show that the hemolysis rate of the adsorbents provided in Examples 1-13 and Comparative Examples 1-3 is less than 3%, which meets the national standard (requiring less than 5%). This indicates that the adsorbents provided in this application have good biocompatibility.

[0045] 2. Experiment on the clearance rate of inflammatory factors: The adsorbents provided in Examples 1 and 6, as well as Comparative Examples 1-3, were wet-packed into columns (φ1×10cm). The columns were pre-purged with physiological saline, and then 30mL of bovine plasma with added TNF-α and IL-6 (wherein the concentration of TNF-α was 1ng / mL and the concentration of IL-6 was 0.5ng / mL) was added. The columns were dynamically adsorbed and cyclical for 3h. The levels of inflammatory factors TNF-α and IL-6 in the bovine plasma before and after adsorption were tested, and the clearance rate was calculated. The results are shown in Table 1.

[0046] Table 1

[0047]

[0048] As shown in Table 1, the adsorbents provided in Examples 1-13 of this application can achieve a clearance rate of over 80% for both inflammatory factors TNF-α and IL-6. In contrast, Comparative Examples 1 and 2 show weaker clearance rates for TNF-α and IL-6. This is mainly because the chitosan microspheres were not modified and activated (Comparative Example 1) or because the chitosan microspheres were not grafted with chicoric acid (Comparative Example 2), resulting in only a small amount of manganese dioxide nanozyme being immobilized on the surface of the chitosan microspheres through electrostatic adsorption, thus affecting its adsorption efficiency. In Comparative Example 3, since the adsorbent surface did not have manganese dioxide nanozyme immobilized, it could only rely on the adsorption capacity of the chitosan microspheres themselves, thus resulting in an even weaker clearance rate for both TNF-α and IL-6.

[0049] 3. Free radical scavenging rate experiment: A hydroxyl radical scavenging kit (purchased from Nanjing Jiancheng Biological Institute) was used to determine the scavenging ability of the adsorbents prepared in Example 1 and Comparative Examples 1-3. The assay was performed according to the kit instructions. The absorbance of the reaction solution at 550 nm was measured using a microplate reader, and the scavenging rate of each adsorbent for hydroxyl radicals was calculated. The results are shown in Table 2. Table 2 Example 1 75.23 Comparative Example 1 35.71 Comparative Example 2 37.24 Comparative Example 3 16.84 As shown in Table 2, the adsorbent provided in Example 1 of this application exhibits a very strong scavenging rate for hydroxyl radicals because the manganese dioxide nanozyme is immobilized on the surface of chitosan microspheres through a very stable bonding effect via chicoric acid. In Comparative Examples 1-2, due to the lower immobilization amount of manganese dioxide nanozyme, only a limited scavenging effect on hydroxyl radicals was observed. In Comparative Example 3, since no manganese dioxide nanozyme was immobilized, the scavenging effect on hydroxyl radicals was very weak.

[0050] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A chitosan-based blood perfusion adsorbent, characterized in that, The blood perfusion adsorbent uses modified chitosan microspheres as a carrier, grafts chicoric acid onto the surface, and then immobilizes manganese dioxide nanoenzymes to form a composite adsorbent for clearing inflammatory factors from the blood of patients with inflammation. The blood perfusion adsorbent is prepared by the following method: Chitosan microspheres are dispersed in an alkaline solution and sonicated for 20-30 min to obtain a chitosan microsphere dispersion; 2,3-epoxypropyltrimethylammonium chloride and ferrous sulfate are added to the chitosan microsphere dispersion, and the mixture is reacted at 40-60℃ for 7-9 h under stirring, followed by washing to obtain modified chitosan microspheres; The modified chitosan microspheres are added to an ethanol aqueous solution containing chicoric acid, sonicated for 40-50 min, followed by the addition of stannous chloride, and reacted at 70-80℃ for 3-4 h to obtain chicor-acidified chitosan microspheres; The chicor-acidified chitosan microspheres are dispersed in an ethylene glycol solution containing manganese nitrate hexahydrate, an alkaline solution is added, and the mixture is heated at 50-80℃ for 3-6 h, followed by centrifugation and washing to obtain the blood perfusion adsorbent.

2. The chitosan-based blood perfusion adsorbent according to claim 1, characterized in that, The chitosan microspheres have a particle size of 100-200 μm and a coefficient of variation (CV) < 5%.

3. The chitosan-based blood perfusion adsorbent according to claim 1, characterized in that, The mass ratio of the chitosan microspheres to the 2,3-epoxypropyltrimethylammonium chloride is 20-30:

1.

4. A method for preparing a blood perfusion adsorbent according to any one of claims 1-3, characterized in that, It includes: Chitosan microspheres were dispersed in an alkaline solution and sonicated for 20-30 minutes to obtain a chitosan microsphere dispersion. 2,3-epoxypropyltrimethylammonium chloride and ferrous sulfate were added to the chitosan microsphere dispersion and reacted at 40-60℃ for 7-9 hours under stirring. After washing, modified chitosan microspheres were obtained. The modified chitosan microspheres were added to an ethanol aqueous solution containing chicoric acid, sonicated for 40-50 minutes, and then stannous chloride was added. The mixture was reacted at 70-80℃ for 3-4 hours to obtain chicor-acidified chitosan microspheres. The chicor-acidified chitosan microspheres were dispersed in an ethylene glycol solution containing manganese nitrate hexahydrate, and an alkaline solution was added. The mixture was heated at 50-80℃ for 3-6 hours, and after centrifugation and washing, the blood perfusion adsorbent was obtained.

5. The method for preparing the blood perfusion adsorbent according to claim 4, characterized in that, The mass ratio of the modified chitosan microspheres to chicoric acid is 15-20:

1.

6. The method for preparing the blood perfusion adsorbent according to claim 4, characterized in that, The mass ratio of the modified chitosan microspheres to manganese nitrate hexahydrate is 6-8:

1.

7. The use of a blood perfusion adsorbent according to any one of claims 1-3 in the preparation of a blood perfusion device.

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