Adsorbent for removing myoglobin and method for preparing the same

Abstract

The application provides an adsorbent for removing myoglobin and a preparation method thereof, and the preparation method of the adsorbent for removing myoglobin comprises the following steps: mutual solubility of styrene monomers, polyvinyl cross-linking agents, halogen-containing monomers, initiators and pore-forming agents to obtain an oil phase, dissolving a dispersing agent in water to obtain an aqueous phase, mutual solubility of the aqueous phase and the oil phase to prepare an oil-in-water suspension, suspension polymerization reaction of the oil-in-water suspension to prepare a styrene macroporous resin containing halogenated hydrocarbons; and nucleophilic substitution reaction of the styrene macroporous resin containing halogenated hydrocarbons and an amino acid containing a benzene ring to graft the amino acid to the styrene macroporous resin to prepare the adsorbent. The removal rate and removal speed of the adsorbent prepared by the application to myoglobin are obviously improved, and when the adsorbent is applied to whole blood perfusion treatment, the adsorbent can quickly reduce the myoglobin concentration in the body of a patient in the early stage, and the whole blood perfusion treatment effect on the patient is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of blood purification, in particular to an adsorbent for removing myoglobin and a preparation method thereof. BACKGROUND

[0002] Myoglobin is a binding protein composed of a peptide chain and a heme prosthetic group, which exists in large quantities in heme proteins in striated muscle (skeletal muscle and cardiac muscle) cells. When the heart muscle or skeletal muscle is damaged, myoglobin is released into the vascular system due to the rupture of the cell membrane and can be detected in the blood. And when the muscle damage is more serious, such as acute rhabdomyolysis, it will cause myoglobin to rise, and myoglobin induces the apoptosis of renal tubular epithelial cells through endoplasmic reticulum stress, thereby causing acute kidney injury.

[0003] In the prior art, although myoglobin can be removed by hemodialysis or hemofiltration, there is a lack of adsorbents for effectively adsorbing myoglobin in the clinical treatment process, which not only makes the treatment of patients with high myoglobin inconvenient, but also cannot achieve good treatment effect. SUMMARY

[0004] The present application aims to solve the above-mentioned defects of the prior art and provide an adsorbent capable of effectively removing myoglobin.

[0005] To solve the above-mentioned problems, the present application provides, in a first aspect, a preparation method of an adsorbent for removing myoglobin, comprising the following steps:

[0006] The styrene monomer, the polyvinyl crosslinking agent, the halogen-containing monomer, the initiator and the pore-forming agent are mutually soluble to obtain an oil phase, a dispersant is dissolved in water to obtain an aqueous phase, the aqueous phase and the oil phase are mutually soluble to prepare an oil-in-water suspension, and the oil-in-water suspension is subjected to suspension polymerization to prepare a halogen-containing styrene macroporous resin;

[0007] The halogen-containing styrene macroporous resin and the amino acid containing a benzene ring are subjected to a nucleophilic substitution reaction to graft the amino acid onto the styrene macroporous resin to prepare the adsorbent.

[0008] Further, the nucleophilic substitution reaction of the halogen-containing styrene macroporous resin and the amino acid containing a benzene ring to graft the amino acid onto the styrene macroporous resin to prepare the adsorbent comprises:

[0009] An aqueous solution of the amino acid containing a benzene ring is prepared;

[0010] The aqueous solution of the amino acid containing benzene ring, the styrene macroporous resin containing halogenated hydrocarbon and the promoter are reacted in an alkaline environment at 60-90℃ for 4-24h to make nucleophilic substitution reaction between the amine group in the amino acid containing benzene ring and the halogen in the styrene macroporous resin containing halogenated hydrocarbon, and graft the amino acid to the styrene macroporous resin to obtain the adsorbent.

[0011] Further, the volume ratio of the aqueous solution of the amino acid containing benzene ring and the styrene macroporous resin containing halogenated hydrocarbon is 2:1-1:1, and the mass concentration of the aqueous solution of the amino acid containing benzene ring is 1%-10%.

[0012] Further, the amino acid containing benzene ring is at least one of phenylalanine, tryptophan and tyrosine, and the promoter is potassium iodide.

[0013] Further, the temperature of the suspension polymerization reaction is 60-90℃, and the time of the suspension polymerization reaction is 8-24h.

[0014] Further, the mass ratio of the pore-forming agent and the reaction monomer is 2:1-1:2, the pore-forming agent is a combination of aromatic hydrocarbon, higher alcohol and higher ketone, the mass ratio of the aromatic hydrocarbon and the higher alcohol is 2:1-1:1, the mass ratio of the aromatic hydrocarbon and the higher ketone is 2:1-1:1, the aromatic hydrocarbon is at least one of toluene, ethylbenzene, xylene and n-propylbenzene, the higher alcohol includes at least one of n-octanol, isooctanol and 4-methyl-2-pentanol, and the higher ketone includes 4-methyl-2-pentanone and / or cyclohexanone.

[0015] The reaction monomer is the styrene monomer, the multi-vinyl crosslinking agent and the halogen-containing monomer.

[0016] Further, the styrene monomer is at least one of styrene, methylstyrene and ethylstyrene, and the amount of the styrene monomer is 15%-30% of the mass of the reaction monomer.

[0017] The multi-vinyl crosslinking agent is at least one of divinylbenzene, trivinylbenzene, divinyltoluene and divinyl ethylbenzene, and the amount of the multi-vinyl crosslinking agent is 30%-80% of the mass of the reaction monomer.

[0018] The halogen-containing monomer is at least one of 2-chloromethyl acrylate, 2-bromomethyl methacrylate and chloropropene, and the amount of the halogen-containing monomer is 5%-30% of the mass of the reaction monomer.

[0019] The initiator is at least one of t-butyl peroxy-2-ethylhexanoate, azobisisobutyronitrile and benzoyl peroxide, and the amount of the initiator is 0.1% to 10% of the mass of the reaction monomer;

[0020] Further, the dispersing agent is at least one of polyvinyl alcohol, polyacrylic acid, hydroxymethyl cellulose, gelatin and sodium alginate, and the mass concentration of the dispersing agent in the water phase is 0.05% to 0.15%.

[0021] The second aspect of the present application provides an adsorbent prepared by the preparation method of any one of the first aspect.

[0022] Further, the grafting amount of the amino acid in the adsorbent is 0.05 mmol / mL to 0.8 mmol / mL.

[0023] The adsorbent for removing myoglobin and the preparation method thereof provided by the present application first prepare a styrene macroporous resin through a suspension polymerization reaction, and add a halogenated hydrocarbon-containing monomer before the suspension polymerization reaction to introduce halogen into the styrene macroporous resin, thereby preparing a styrene macroporous adsorption resin containing halogenated hydrocarbons. Then, through a nucleophilic substitution reaction between the amine group in the amino acid and the halogenated hydrocarbon, an amino acid containing a benzene ring is modified to the surface of the styrene macroporous resin, thereby obtaining a styrene macroporous resin grafted and modified with an amino acid. On the one hand, the amino acid grafted on the surface of the styrene macroporous adsorption resin has an amine group and a carboxyl group, which can form an electrostatic structure with the amino acid on the surface of the myoglobin molecule through the adsorption of positive and negative charges, thereby achieving the effect of quickly capturing myoglobin. On the other hand, the amino acid grafted on the surface of the styrene macroporous adsorption resin has a benzene ring structure, which has a hydrophobic interaction with the hydrophobic chain inside the myoglobin, and can be adsorbed more firmly. Therefore, by combining the adsorption of positive and negative charges and the hydrophobic interaction, the adsorbent can not only adsorb myoglobin more firmly, but also has a higher adsorption efficiency for myoglobin. In addition, grafting the amino acid on the surface of the styrene macroporous adsorption resin also increases the content of the hydrophilic group, thereby improving the hydrophilicity of the adsorbent and the blood compatibility of the adsorbent. The adsorbent for effectively adsorbing myoglobin is first prepared by the method of the present application, and the removal rate and removal rate of the adsorbent for myoglobin are both significantly improved. When the adsorbent is applied to whole blood perfusion treatment, it can quickly reduce the concentration of myoglobin in the body of a patient in the early stage, thereby improving the effect of whole blood perfusion treatment on the patient. The adsorbent prepared by the method of the present application has a higher application range and clinical treatment practical value. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 The process flow chart for preparing the adsorbent for removing myoglobin is provided for the embodiments of the present application. DETAILED DESCRIPTION

[0025] In order to make the above objectives, characteristics and advantages of the present application more apparent, concrete embodiments of the present application will be described in detail below with reference to the accompanying drawings.

[0026] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict.

[0027] In addition, the terms "comprising", "including", "containing", "having" are non-limiting, i.e. other steps and other components can be added without affecting the results. Unless otherwise specified, the materials, devices, reagents are commercially available.

[0028] In addition, although the present application describes each step in the preparation in the form of S110, S120, etc., this description is only for the convenience of understanding, and the form of S110, S120 does not represent a limitation on the sequence of each step.

[0029] Figure 1 A process flow chart for preparing the adsorbent for removing myoglobin in the embodiments of the present application is provided. In combination with the process flow chart shown in the figure, the first aspect of the embodiments of the present application provides a preparation method of an adsorbent for removing myoglobin, comprising the following steps: Figure 1

[0030] Step S110, the styrene monomer, the multi-vinyl crosslinking agent, the halogen-containing monomer, the initiator and the pore-forming agent are mutually soluble to obtain an oil phase, the dispersant is dissolved in water to obtain an aqueous phase, the aqueous phase and the oil phase are mutually soluble to prepare an oil-in-water suspension, and the oil-in-water suspension is subjected to a suspension polymerization reaction to prepare a styrene macroporous resin containing halogenated hydrocarbons.

[0031] Specifically, the styrene monomer, the multi-vinyl crosslinking agent, the halogen-containing monomer, the initiator and the pore-forming agent are mutually soluble to obtain an oil phase, the dispersant is dissolved in water to obtain an aqueous phase, the aqueous phase and the oil phase are mixed and stirred uniformly to prepare an oil-in-water suspension, the oil-in-water suspension is subjected to a suspension polymerization reaction at 60-90°C, and after 8-24h of reaction, a styrene macroporous resin containing halogenated hydrocarbons is prepared.

[0032] Wherein, when the aqueous phase and the oil phase are stirred and mixed, the stirring speed is 100-200rpm.

[0033] ​In the embodiment, the styrene macroporous resin is prepared by a suspension polymerization reaction, and the monomer containing halogenated hydrocarbon is added before the suspension polymerization reaction, so that the halogen is more easily introduced into the styrene macroporous resin, and the styrene macroporous resin containing halogenated hydrocarbon is prepared. The styrene macroporous resin containing halogenated hydrocarbon has a better stability, a large crosslinking density, a high strength, a rich pore structure, and a large specific surface area, and is beneficial to improving the adsorption capacity of the styrene macroporous resin containing halogenated hydrocarbon. In addition, by adding the monomer containing halogenated hydrocarbon before the suspension polymerization reaction, the content of the halogen grafted on the surface of the styrene macroporous resin can be adjusted, which is convenient for subsequent grafting of amino acids on the styrene macroporous resin, and improves the preparation efficiency and convenience of the adsorbent.

[0034] As an optional implementation manner, the styrene monomer is at least one of styrene, methylstyrene and ethylstyrene, and the amount of the styrene monomer is 15% to 30% of the mass of the reaction monomer, wherein the reaction monomer refers to the styrene monomer, the multi-vinyl crosslinking agent and the monomer containing halogen, and the mass of the reaction monomer is the sum of the masses of the styrene monomer, the multi-vinyl crosslinking agent and the monomer containing halogen, that is, the amount of the styrene monomer is 15% to 30% of the sum of the masses of the styrene monomer, the multi-vinyl crosslinking agent and the monomer containing halogen, and the same below, which will not be further explained. Therefore, by selecting the above-mentioned substances as the styrene monomer and limiting the mass ratio of the styrene monomer and the reaction monomer in the above range, the crosslinking degree of the styrene macroporous resin containing halogenated hydrocarbon can be adjusted, so as to avoid that the crosslinking degree is too low, which is not conducive to the balling of the styrene macroporous resin containing halogenated hydrocarbon, and avoid that the crosslinking degree is too high, which causes the rigidity of the styrene macroporous resin containing halogenated hydrocarbon to be large and the cracking problem to be prone to occur.

[0035] As an optional implementation manner, the multi-vinyl crosslinking agent is at least one of divinylbenzene, trivinylbenzene, divinyltoluene and divinyl ethylbenzene, and the amount of the multi-vinyl crosslinking agent is 30% to 80% of the mass of the reaction monomer. Therefore, by selecting the above-mentioned substances as the multi-vinyl crosslinking agent and limiting the mass ratio of the multi-vinyl crosslinking agent and the reaction monomer in the above range, the crosslinking degree of the styrene macroporous resin containing halogenated hydrocarbon can be adjusted, so as to avoid that the crosslinking degree is too low, which is not conducive to the balling of the styrene macroporous resin containing halogenated hydrocarbon, and avoid that the crosslinking degree is too high, which causes the rigidity of the styrene macroporous resin containing halogenated hydrocarbon to be large and the cracking problem to be prone to occur.

[0036] As an optional implementation, the halogen-containing monomer is at least one selected from ethyl 2-chloromethacrylate, 2-bromoethyl methacrylate, and allyl chloride, and the amount of the halogen-containing monomer is 5% to 30% of the mass of the reactant monomer. Therefore, by selecting the above-mentioned substances as the halogen-containing monomer and limiting the mass ratio of the halogen-containing monomer to the reactant monomer within the above range, it is not only beneficial to adjust the crosslinking degree of the styrene macroporous resin containing halogenated hydrocarbons, but also beneficial to introduce halogenated hydrocarbons onto the styrene macroporous resin, and to graft an appropriate amount of halogenated hydrocarbons onto the styrene macroporous resin, so as to facilitate the subsequent grafting of an appropriate amount of amino acids and improve the adsorption effect of the adsorbent.

[0037] As an optional implementation, the initiator is at least one selected from tert-butyl peroxide-2-ethylhexanoate, azobisisobutyronitrile, and benzoyl peroxide, and the amount of initiator is 0.1% to 10% of the mass of the reactant monomer. Therefore, selecting the above-mentioned substances as initiators and limiting the mass ratio of initiator to reactant monomer within the above range is beneficial for initiating the suspension polymerization reaction of the oil-in-water suspension.

[0038] As an optional embodiment, the porogen is a composition of aromatic hydrocarbons, higher alcohols, and higher ketones, wherein the mass ratio of aromatic hydrocarbons to higher alcohols is 2:1 to 1:1, and the mass ratio of aromatic hydrocarbons to higher ketones is 2:1 to 1:1. The aromatic hydrocarbon is at least one selected from toluene, ethylbenzene, xylene, and n-propylbenzene; the higher alcohol includes at least one selected from n-octanol, isooctanol, and 4-methyl-2-pentanol; the higher ketone includes 4-methyl-2-pentanone and / or cyclohexanone; and the mass ratio of the porogen to the reactant monomer is 2:1 to 1:2. Therefore, by selecting a composition of aromatic hydrocarbons, higher alcohols, and higher ketones as the porogen, where aromatic hydrocarbons are good solvents and higher alcohols and higher ketones are poor solvents, and by limiting the mass ratio of aromatic hydrocarbons, higher alcohols, and higher ketones within the above-mentioned ranges, and by limiting the mass ratio of the porogen to the reactant monomer within the above-mentioned ranges, it is beneficial to adjust the pore structure of the styrene macroporous resin containing halogenated hydrocarbons, thereby adjusting the pore volume and pore size of the styrene macroporous resin containing halogenated hydrocarbons.

[0039] As an alternative embodiment, the dispersant is at least one selected from polyvinyl alcohol, polyacrylic acid, hydroxymethyl cellulose, gelatin, and sodium alginate, and the mass concentration of the dispersant in the aqueous phase is 0.05% to 0.15%. Therefore, by selecting the above-mentioned substances as dispersants and limiting the amount of dispersant within the above range, it is beneficial to adjust the particle size of the styrene macroporous resin containing halogenated hydrocarbons.

[0040] The styrene macroporous resin containing halogenated hydrocarbons prepared by the method of this embodiment has a particle size range of 0.3 mm to 0.8 mm and a specific surface area (measured by BET method) range of 400 m². 2 / g to 600m 2 / g, pore volume range is 0.8cm³ 30.5 to 1.5 cm 3 / g, and the average pore size ranges from 8 nm to 20 nm. In this way, the subsequent prepared adsorbent can have good adsorption capacity, which is conducive to adsorbing and removing myoglobin in blood. The content of halogen in the halogen-containing styrene macroporous resin prepared by the method of the embodiment ranges from 2% to 10%, which is conducive to subsequent grafting of an appropriate amount of amino acid and improving the adsorption of the adsorbent.

[0041] In step S120, the halogen-containing styrene macroporous resin and the amino acid containing a benzene ring are subjected to a nucleophilic substitution reaction, so that the amino acid is grafted onto the styrene macroporous resin to prepare the adsorbent.

[0042] Specifically, an aqueous solution of the amino acid containing a benzene ring is prepared, and the aqueous solution of the amino acid containing a benzene ring, the halogen-containing styrene macroporous resin, and a promoter are reacted in an alkaline environment at 60°C to 90°C for 4 h to 24 h, so that the amine group in the amino acid containing a benzene ring and the halogen in the halogen-containing styrene macroporous resin are subjected to a nucleophilic substitution reaction, the amino acid is grafted onto the styrene macroporous resin, and the adsorbent is prepared.

[0043] In the embodiment, the amino acid containing a benzene ring is modified to the surface of the styrene macroporous resin through the nucleophilic substitution reaction of the amine group in the amino acid and the halogen, and the styrene macroporous resin grafted with the amino acid is obtained, i.e., the adsorbent in the embodiment. The amino acid grafted on the surface of the styrene macroporous adsorbent resin has an amine group and a carboxyl group, and the amino acid on the surface of the myoglobin molecule exists. The amine group and the carboxyl group in the grafted amino acid and the amino acid on the surface of the myoglobin molecule form an electrostatic structure through the adsorption of positive and negative charges, so as to quickly capture myoglobin. On the other hand, the grafted amino acid has a benzene ring structure, which has a hydrophobic interaction with the hydrophobic chain inside the myoglobin, and can be adsorbed more firmly. In the embodiment, the amino acid with a benzene ring structure is grafted on the surface of the styrene macroporous adsorbent resin, and the adsorbent can more firmly adsorb myoglobin through the combination of the adsorption of positive and negative charges and the hydrophobic interaction.

[0044] As an optional implementation, the volume ratio of the aqueous solution of the amino acid containing a benzene ring to the halogen-containing styrene macroporous resin ranges from 2:1 to 1:1, and the mass concentration of the aqueous solution of the amino acid containing a benzene ring ranges from 1% to 10%.

[0045] In the embodiment, the concentration of the amino acid, the nucleophilic substitution reaction temperature, and the reaction time are adjusted to adjust the grafting amount of the amino acid on the styrene macroporous resin, so that the adsorbent has good adsorbability.

[0046] As an optional embodiment, the amino acid containing benzene ring is at least one of phenylalanine, tryptophan and tyrosine. Thus, by selecting the amino acid with benzene ring structure, the amino acid containing benzene ring and the styrene macroporous resin containing halogenated hydrocarbon are grafted through the π-π interaction between benzene rings, so that the amino acid grafted on the styrene macroporous resin is closer to the skeleton of the styrene macroporous resin, thereby when adsorption is performed, the myoglobin captured by the amino acid can be quickly adsorbed by the styrene macroporous resin.

[0047] As an optional embodiment, the promoter is potassium iodide, and the amount of the promoter added is 1% to 10% of the mass of the amino acid containing benzene ring.

[0048] As an optional embodiment, the alkaline environment is a reaction environment with a pH value of 8 to 11, and the type of the alkaline solution used to adjust the pH value is not limited in the embodiment, and optionally, the pH value of the reaction environment is adjusted to 8 to 11 by adding sodium hydroxide. Thus, the amine group of the amino acid has a positive charge, and in the pH value range, the nucleophilicity of the amine group in the amino acid is strong, which is good for the nucleophilic substitution reaction.

[0049] The amount of the amino acid grafted on the amino acid grafted and modified styrene macroporous resin (i.e. adsorbent) prepared by the method of the embodiment is 0.05 mmol / mL to 0.8 mmol / mL, that is, 0.05 mmol to 0.5 mmol of amino acid is grafted on 1 ml of solid amino acid grafted and modified styrene macroporous resin.

[0050] In this embodiment, the styrene macroporous resin is prepared by a suspension polymerization reaction, and the halogen-containing monomer is added before the suspension polymerization reaction to introduce halogen into the styrene macroporous resin to obtain the styrene macroporous adsorption resin containing halogenated hydrocarbon. Then, the amino acid containing benzene ring is modified to the surface of the styrene macroporous resin through the nucleophilic substitution reaction between the amine group in the amino acid and the halogenated hydrocarbon to obtain the styrene macroporous resin grafted with the amino acid. On the one hand, the amino acid grafted on the surface of the styrene macroporous adsorption resin has amine group and carboxyl group, which can form electrostatic structure with the amino acid on the surface of myoglobin molecule through the adsorption of positive and negative charges to achieve the effect of quickly capturing myoglobin. On the other hand, the amino acid grafted on the surface of the styrene macroporous adsorption resin has benzene ring structure, which has hydrophobic interaction with the hydrophobic chain inside the myoglobin, and can be adsorbed more firmly. Therefore, by combining the adsorption of positive and negative charges and the hydrophobic interaction, the adsorbent can not only adsorb myoglobin more firmly, but also has higher adsorption efficiency for myoglobin. In addition, the grafting of the amino acid on the surface of the styrene macroporous adsorption resin also increases the content of hydrophilic groups and improves the hydrophilicity of the adsorbent, which is beneficial to improve the blood compatibility of the adsorbent. The adsorbent for effectively adsorbing myoglobin is prepared by the method of this embodiment, and the clearance rate and clearance rate of the adsorbent for myoglobin are obviously improved. When the adsorbent is applied to whole blood perfusion treatment, it can quickly reduce the concentration of myoglobin in the patient's body in the early stage, improve the effect of whole blood perfusion treatment on the patient, and has higher application range and clinical treatment practical value.

[0051] The second aspect of this embodiment provides an adsorbent prepared by the preparation method of the first aspect, and the grafting amount of the amino acid in the adsorbent is 0.05 mmol / mL to 0.8 mmol / mL.

[0052] In order to further illustrate the present application, the present application will be further illustrated by combining specific examples. The experimental methods used in the examples in the present application are conventional methods unless otherwise specified; the materials, reagents and the like used in the examples in the present application are commercially available unless otherwise specified.

[0053] Example 1

[0054] The present embodiment provides a preparation method of an adsorbent for removing myoglobin, comprising the following steps:

[0055] (1) Preparation of styrene macroporous resin containing halogenated hydrocarbon: In a 1000 mL three-necked flask, 400 mL of an aqueous solution containing 0.1%wt of polyvinyl alcohol was added as the water phase; in a beaker, 10 g of styrene as a styrene-based monomer, 40 g of 80% divinylbenzene (containing 32 g of divinylbenzene and 8 g of ethyl styrene) as a polyvinyl-based crosslinking agent, 10 g of 2-chloromethyl acrylate as a halogen-containing monomer, 0.06 g of azobisisobutyronitrile as an initiator, 40 g of toluene as a porogen, and 20 g of isooctanol and 20 g of cyclohexanone were mixed uniformly to obtain an oil phase; the prepared oil phase was added to the water phase, the mechanical stirring speed was adjusted to 140 rpm, and after the liquid droplet size was stabilized, an oil-in-water suspension was prepared; the oil-in-water suspension was heated to 80°C, and suspension polymerization was performed for 12 hours; after the reaction was completed, the resin was washed with water to remove excess dispersant, then a Soxhlet extractor was used to extract the resin with ethanol for 12 hours, and then the resin was dried and sieved to obtain a styrene macroporous resin containing halogenated hydrocarbon, with a particle size of 0.4 mm to 0.8 mm; the specific surface area (measured by the BET method) of the styrene macroporous resin containing halogenated hydrocarbon was 550 m 2 / g, the pore volume was 1.2 cm 3 / g, the average pore size was 12.5 nm, and the chlorine content was 3.5% of the mass of the styrene macroporous resin containing halogenated hydrocarbon.

[0056] (2) Graft modification of amino acid: 100 mL of the styrene macroporous resin containing halogenated hydrocarbon prepared in step (1) was taken in a three-necked flask, then 100 mL of a 5% phenylalanine aqueous solution was prepared, 1 g of potassium iodide was added, and the pH of the mixture was adjusted to 10 with sodium hydroxide; the prepared mixture was added to the above flask, and nucleophilic substitution was performed at 80°C for 8 hours; after the reaction was completed, the unreacted phenylalanine and potassium iodide were removed with a large amount of water to obtain a phenylalanine-grafted styrene macroporous resin, i.e., an adsorbent, and the grafting amount of phenylalanine in the adsorbent was 0.3 mmol / mL.

[0057] Example 2

[0058] The present embodiment provides a method for preparing an adsorbent for removing myoglobin, comprising the following steps:

[0059] (1) Preparation of styrene macroporous resin containing halogenated hydrocarbon: In a 1000 mL three-necked flask, 400 mL of an aqueous solution containing 0.05% wt of hydroxymethyl cellulose was added as the water phase; in a beaker, 15 g of methyl styrene was added as the styrene monomer, 45 g of 63% divinyl toluene (containing 28.35 g of divinyl toluene and 16.65 g of ethyl styrene) was added as the polyvinyl crosslinking agent, 10 g of 2-bromoethyl methacrylate was added as the halogen-containing monomer, 0.08 g of benzoyl peroxide was added as the initiator, 36 g of ethylbenzene was added as the porogen, 18 g of 4-methyl-2-pentanol was added, and 24 g of 4-methyl-2-pentanone was added, and the above-mentioned substances were mixed uniformly in the beaker to obtain an oil phase; the prepared oil phase was added to the water phase, the mechanical stirring speed was adjusted to 160 rpm, and after the liquid droplet size was stabilized, an oil-in-water suspension was prepared; the oil-in-water suspension was heated to 70°C, and suspension polymerization was carried out for 16 hours; after the reaction was completed, the resin was washed with water to remove excess dispersant, then a Soxhlet extractor was used to extract the resin with ethanol for 12 hours, and then the resin was dried and sieved, and the resin with a particle size of 0.4 mm to 0.8 mm was selected to obtain the styrene macroporous resin containing halogenated hydrocarbon; the specific surface area (measured by the BET method) of the styrene macroporous resin containing halogenated hydrocarbon was 610 m 2 / g, the pore volume was 1.1 cm 3 / g, the average pore size was 11.2 nm, and the bromine content was 4.6% of the mass of the styrene macroporous resin containing halogenated hydrocarbon.

[0060] (2) Graft modification of amino acid: 100 mL of the styrene macroporous resin containing halogenated hydrocarbon prepared in step (1) was taken in a three-necked flask, and then 100 mL of a tryptophan aqueous solution with a mass fraction of 8% was prepared, 0.5 g of potassium iodide was added, and the pH of the mixture was adjusted to 11 using sodium hydroxide; the prepared mixture was added to the above-mentioned flask, and nucleophilic substitution reaction was carried out at 70°C for 12 hours; after the reaction was completed, a large amount of water was used to remove unreacted tryptophan and potassium iodide, and thus a tryptophan grafted and modified styrene macroporous resin, i.e., an adsorbent, was obtained, and the grafting amount of tryptophan in the adsorbent was 0.42 mmol / mL.

[0061] Example 3

[0062] The present embodiment provides a preparation method of an adsorbent for removing myoglobin, which comprises the following steps:

[0063] (1) Preparation of styrene macroporous resin containing halogenated hydrocarbon: In a 1000 mL three-necked flask, 400 mL of 0.15%wt gelatin aqueous solution was added as the water phase; in a beaker, 10 g of styrene, 8 g of ethylstyrene, 50 g of 80% divinylbenzene (containing 40 g of divinylbenzene and 10 g of ethylstyrene) as the multivinyl crosslinking agent, 20 g of chloropropylene as the halogen-containing monomer, and 0.2 g of t-butyl peroxy-2-ethylhexanoate as the initiator were added, and 50 g of xylene, 36 g of n-octanol, and 32 g of 4-methyl-2-pentanone were added as the porogen to obtain an oil phase; the oil phase was added to the water phase, and the mechanical stirring speed was adjusted to 100 rpm; after the liquid droplet size was stabilized, an oil-in-water suspension was prepared; the oil-in-water suspension was heated to 75°C, and suspension polymerization was performed for 14 hours; after the reaction was completed, the resin was washed with water to remove the excess dispersant, and then the resin was extracted with ethanol for 12 hours using a Soxhlet extractor, and then dried and sieved; the resin with a particle size of 0.4 mm to 0.8 mm was selected to obtain the styrene macroporous resin containing halogenated hydrocarbon; the specific surface area (measured by the BET method) of the styrene macroporous resin containing halogenated hydrocarbon was 580 m 2 / g, the pore volume was 1.16 cm 3 / g, the average pore size was 11.8 nm, and the chlorine content was 9.6% of the mass of the styrene macroporous resin containing halogenated hydrocarbon.

[0064] (2) Graft modification of amino acid: 100 mL of the styrene macroporous resin containing halogenated hydrocarbon prepared in step (1) was taken in a three-necked flask, and then 100 mL of a 10% tyrosine aqueous solution was prepared, 0.6 g of potassium iodide was added, and the pH of the mixture was adjusted to 9 using sodium hydroxide; the prepared mixture was added to the above flask, and nucleophilic substitution reaction was performed at 85°C for 9 hours; after the reaction was completed, the unreacted tyrosine and potassium iodide were removed with a large amount of water to obtain the tyrosine grafted styrene macroporous resin, i.e., the adsorbent; the grafting amount of tyrosine in the adsorbent was 0.72 mmol / mL.

[0065] Comparative Example 1

[0066] The present example provides a preparation method of a styrene macroporous resin containing halogenated hydrocarbon, which is the same as the preparation method in step (1) of Example 1, and will not be further described herein.

[0067] Comparative Example 2

[0068] The present example provides a preparation method of a styrene macroporous resin containing halogenated hydrocarbon, which is the same as the preparation method in step (1) of Example 2, and will not be further described herein.

[0069] Comparative Example 3

[0070] This embodiment provides a method for preparing a styrene macroporous resin containing halogenated hydrocarbons. The method for preparing the styrene macroporous resin containing halogenated hydrocarbons is the same as the method in step (1) of Example 3, and will not be described further here.

[0071] The adsorbents prepared in Examples 1 to 3 were used as experimental groups, and the styrene macroporous resins containing halogenated hydrocarbons prepared in Comparative Examples 1 to 3 were used as control groups. The adsorbents in Examples 1 to 3 and Comparative Examples 1 to 3 were used as blood perfusion adsorbents. 1 mL of each adsorbent was accurately measured and added to 10 mL of plasma with a concentration of 600 ng / mL myoglobin. The plasma was shaken at 37°C. Plasma samples were collected after adsorption at 0.5 hours, 1 hour, 1.5 hours, and 2 hours, and the myoglobin concentration was tested by electrochemiluminescence. The adsorption performance of the adsorbents in each example and the comparative examples for myoglobin is shown in Table 1.

[0072] Table 1 Adsorption performance of myoglobin

[0073]

[0074] As shown in Table 1, in the embodiments of the present invention, styrene macroporous resin is first prepared by suspension polymerization, and a monomer containing halogenated hydrocarbons is added before the suspension polymerization to obtain styrene macroporous adsorption resin containing halogenated hydrocarbons. Then, through the nucleophilic substitution reaction between the amino group in the amino acid and the halogenated hydrocarbon, the amino acid containing the benzene ring is modified onto the surface of the styrene macroporous resin to obtain amino acid grafted styrene macroporous resin. This amino acid grafted styrene macroporous resin not only retains the stability of the styrene-divinylbenzene macroporous resin skeleton structure and the rich pore structure, but also the grafted amino acid with the benzene ring structure can combine through positive and negative charge adsorption and hydrophobic interaction, so that the adsorbent can not only adsorb myoglobin more firmly, but also has a higher adsorption efficiency for myoglobin, thereby significantly improving the adsorption rate and adsorption efficiency of the adsorbent for myoglobin.

[0075] The adsorbent prepared by the method of the present invention has significantly improved the clearance rate and clearance speed of myoglobin. When applied to whole blood perfusion therapy, it can rapidly reduce the concentration of myoglobin in the patient's body in the early stage, thereby improving the therapeutic effect of whole blood perfusion.

[0076] While the disclosure is as stated above, its scope of protection is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of this disclosure, and all such changes and modifications will fall within the protection scope of this invention.

Claims

1. A method for preparing an adsorbent for scavenging myoglobin, characterized in that, Includes the following steps: Styrene monomers, polyvinyl crosslinking agents, halogenated monomers, initiators, and porogens are mutually soluble to obtain an oil phase, and a dispersant is dissolved in water to obtain an aqueous phase. The aqueous phase and the oil are mutually soluble to obtain an oil-in-water suspension. The oil-in-water suspension is subjected to a suspension polymerization reaction to obtain a styrene macroporous resin containing halogenated hydrocarbons. The styrene macroporous resin containing halogenated hydrocarbons and an amino acid containing a benzene ring are subjected to a nucleophilic substitution reaction to graft the amino acid onto the styrene macroporous resin, thereby preparing an adsorbent. The halogen-containing monomer is at least one of ethyl 2-chloromethacrylate, 2-bromoethyl methacrylate, and allyl chloride; The amino acid containing a benzene ring is at least one of phenylalanine, tryptophan, and tyrosine.

2. The method for preparing the adsorbent according to claim 1, characterized in that, The method of preparing an adsorbent by subjecting the styrene macroporous resin containing halogenated hydrocarbons and an amino acid containing a benzene ring to a nucleophilic substitution reaction, thereby grafting the amino acid onto the styrene macroporous resin, comprises: Prepare an aqueous solution of an amino acid containing a benzene ring; An aqueous solution of an amino acid containing a benzene ring, a styrene macroporous resin containing a halogenated hydrocarbon, and an accelerator are reacted in an alkaline environment at 60°C to 90°C for 4 to 24 hours. This allows the amino group in the amino acid containing the benzene ring to undergo a nucleophilic substitution reaction with the halogen in the styrene macroporous resin containing the halogenated hydrocarbon, thereby grafting the amino acid onto the styrene macroporous resin to obtain an adsorbent.

3. The method for preparing the adsorbent according to claim 2, characterized in that, The volume ratio of the aqueous solution containing the benzene ring amino acid to the styrene macroporous resin containing the halogenated hydrocarbon is 2:1 to 1:1, and the mass concentration of the aqueous solution containing the benzene ring amino acid is 1% to 10%.

4. The method for preparing the adsorbent according to claim 2, characterized in that, The accelerator is potassium iodide.

5. The method for preparing the adsorbent according to claim 1, characterized in that, The temperature of the suspension polymerization reaction is 60°C to 90°C, and the time of the suspension polymerization reaction is 8 hours to 24 hours.

6. The method for preparing the adsorbent according to claim 1, characterized in that, The mass ratio of the porogen to the reactant monomer is 2:1 to 1:

2. The porogen is a composition of aromatic hydrocarbons, higher alcohols, and higher ketones, wherein the mass ratio of the aromatic hydrocarbon to the higher alcohol is 2:1 to 1:1, and the mass ratio of the aromatic hydrocarbon to the higher ketone is 2:1 to 1:

1. The aromatic hydrocarbon is at least one selected from toluene, ethylbenzene, xylene, and n-propylbenzene. The higher alcohol includes at least one selected from n-octanol, isooctanol, and 4-methyl-2-pentanol. The higher ketone includes 4-methyl-2-pentanone and / or cyclohexanone. The reaction monomers are the styrene monomers, the polyvinyl crosslinking agents, and the halogen-containing monomers.

7. The method for preparing the adsorbent according to claim 6, characterized in that, The styrene monomer is at least one selected from styrene, methylstyrene, and ethylstyrene, and the amount of the styrene monomer used is 15% to 30% of the mass of the reactant monomer; The polyvinyl crosslinking agent is at least one selected from divinylbenzene, trivinylbenzene, divinyltoluene, and divinylethylbenzene, and the amount of the polyvinyl crosslinking agent is 30% to 80% of the mass of the reactant monomer; The amount of the halogen-containing monomer used is 5% to 30% of the mass of the reactant monomer; The initiator is at least one selected from tert-butyl peroxide-2-ethylhexanoate, azobisisobutyronitrile, and benzoyl peroxide, and the amount of the initiator is 0.1% to 10% of the mass of the reactant monomer.

8. The method for preparing the adsorbent according to claim 1, characterized in that, The dispersant is at least one selected from polyvinyl alcohol, polyacrylic acid, hydroxymethyl cellulose, gelatin and sodium alginate, and the mass concentration of the dispersant in the aqueous phase is 0.05% to 0.15%.

9. An adsorbent, characterized in that, It is prepared by the preparation method described in any one of claims 1 to 8.

10. The adsorbent according to claim 9, characterized in that, The amount of amino acid grafted in the adsorbent is from 0.05 mmol / mL to 0.8 mmol / mL.

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