CN115626967B

CN115626967B - Dual-functional adsorption resin and preparation method and application thereof

Info

Publication number: CN115626967B
Application number: CN202211073623.2A
Authority: CN
Inventors: 龚波林; 欧俊杰; 韩速; 黄超
Original assignee: Ningxia Bethel Active Carbon Co ltd; North Minzu University
Current assignee: Ningxia Bethel Active Carbon Co ltd; North Minzu University
Priority date: 2022-09-02
Filing date: 2022-09-02
Publication date: 2023-08-29
Anticipated expiration: 2042-09-02
Also published as: CN115626967A

Abstract

The application belongs to the technical field of adsorption resins, and discloses a dual-function adsorption resin, a preparation method and application thereof. The resin matrix is macroporous resin microsphere with epoxy group on the surface, the particle diameter of the microsphere is 5-6 mu m, the surface of the microsphere is grafted with a polymer brush with olefin, and L-cysteine is also introduced into the surface of the microsphere. The preparation method specifically adopts a seed swelling polymerization method, uses 3, 4-epoxy cyclohexyl methacrylate as a monomer and tetraethyleneglycol diacrylate as a cross-linking agent to prepare macroporous resin microspheres with epoxy groups on the surfaces, then utilizes an atom transfer radical polymerization technology to graft polymer brushes with olefin on the surfaces of the microspheres, and finally introduces L-cysteine into the surfaces of the microspheres through photoinitiated 'mercapto-ene' click chemistry reaction to prepare the bifunctional adsorption resin capable of being used for metal ion adsorption and/or glycopeptide enrichment.

Description

Dual-functional adsorption resin and preparation method and application thereof

Technical Field

The application belongs to the technical field of adsorption resins, and particularly relates to a dual-function adsorption resin, a preparation method and application thereof.

Background

Along with the acceleration of the industrialization process, the pollution of heavy metal ions to water bodies is increasingly serious. Copper is a typical heavy metal and is widely applied to the manufacturing industries of printing and dyeing, metallurgy, electronics, nuclear power and the like. Cu (II) ions (Cu) generated during production and manufacture ²⁺ ) Is difficult to be degraded by microorganisms, and can be remained in water for a long time to pollute a water source if the water is directly discharged into the environment without being treated. Once excessive Cu is ingested into human body ²⁺ Gene mutations can be caused due to the characteristics of difficult biodegradation, easy bioaccumulation and the like, and serious health hazards such as Wilson disease, alzheimer disease and the like (documents 1.S.Bolisetty,M.Peydayesh,R.Mezzenga,Sustainable technologies for water purification from heavy metals:review and analysis,Chem.Soc.Rev.48 (2019) 463-487) are caused; thus, cu is treated before the wastewater is discharged ²⁺ And (5) performing adsorption removal.

At present, various Cu removal technologies such as adsorption, precipitation, ion exchange and membrane technology are researched ²⁺ Is a method of (2). Among them, the adsorption method is widely used in water treatment because of its simple operation, low cost and high efficiency (document 2.A.Abidli,Y.Huang,Z.Ben Rejeb,A.Zaoui,C.B.Park,Sustainable and efficient technologies for removal and recovery oftoxic and valuable metals from wastewater:recent progress,challenges,and future perspectives,Chemosphere 292 (2022) 133102).

Proteins are the main contributors and executives of various vital activities. Glycosylation is one of the important post-translational modifications of proteins, and plays an important regulatory role in complex biological processes such as signal transduction, cell proliferation and homeostasis. Aberrant glycosylation is often associated with certain diseases including cancer (document 3.M.Rosato,S.Stringer,T.Gebuis,I.Paliukhovich,K.W.Li,D.Posthuma,P.F.Sullivan,A.B.Smit,R.E.van Kesteren,Combined cellomics and proteomics analysis reveals shared neuronal morphology and molecular pathway phenotypes for multiple schizophrenia risk genes,Mol.Psychiatr.26 (2021) 784-799). Thus, glycopeptide analysis is critical for diagnosis and treatment of diseases. Mass Spectrometry (MS) has become a key tool in proteomics research. However, due to the low abundance of glycopeptides in biological samples, the interference is strong, making direct mass spectrometry of glycopeptides difficult (documents 4.L.Zhang,S.Ma,Y.Chen,Y.Wang,J.Ou,H.Uyama,M.Ye,Facile fabrication of biomimetic chitosan membrane with honeycomb-like structure for enrichment of glycosylated peptides, anal. Chem.91 (2019) 2985-2993). Thus, enrichment of low abundance glycopeptides prior to mass spectrometry has become an essential step.

Up to now, the hydrophilic interaction chromatography (hydroic interaction chromatography, HILIC) strategy of exploring hydrophilic interactions between glycopeptides and adsorbents has been an ideal strategy of enriching glycopeptides due to its good selectivity, unbiased properties and high compatibility with MS (documents 5.Y.Tian,R.Tang,X.Wang,J.Zhou,X.Li,S.Ma,B.Gong,J.Ou,Bioinspired dandelion-like silica nanoparticles modified with L-glutathione for highly efficient enrichment ofN-glycopeptides inbiological samples, anal.Chim. Acta 1173 (2021) 338694), based on which applicant has proposed a bifunctional adsorption resin with good hydrophilic interactions in the present application.

Disclosure of Invention

The application relates to a double-function adsorption resin, in particular to a macroporous resin microsphere with epoxy groups on the surface, the particle diameter of the microsphere is 5-6 mu m, a polymer brush with olefin is grafted on the surface of the microsphere, and L-cysteine is also introduced on the surface of the microsphere.

The application relates to a preparation method of a bifunctional adsorption resin, which specifically adopts a seed swelling polymerization method, uses 3, 4-epoxy cyclohexyl methacrylate as a monomer and tetraethylene glycol diacrylate as a cross-linking agent to prepare macroporous resin microspheres with epoxy groups on the surfaces, then utilizes an atom transfer radical polymerization technology to graft a polymer brush with olefin on the surfaces of the microspheres, and finally introduces L-cysteine into the surfaces of the microspheres through photoinitiated 'mercapto-olefin' click chemical reaction to prepare the bifunctional adsorption resin capable of being used for metal ion adsorption and/or obtaining glycopeptide enrichment.

The preparation process comprises the following steps:

s1, preparing macroporous resin microspheres with epoxy groups on surfaces

Mixing emulsified polyglycidyl methacrylate, 3, 4-epoxycyclohexylmethacrylate, tetraethyleneglycol diacrylate, n-propanol, 1, 4-butanediol, azobisisobutyronitrile and aqueous phase in a ratio of 9-11 mL/6-8 mL/0.4-0.6 g/150-170 mL;

reacting for 12-18 h at room temperature, then heating to 60-80 ℃ and continuing to react for 12-18 h;

and (3) carrying out soxhlet extraction for 24-48 h after the reaction, then respectively washing for 3-5 times by using absolute ethyl alcohol and deionized water, and finally carrying out vacuum drying at 60 ℃ for 12-20 h to obtain the macroporous resin microsphere with epoxy groups on the surface.

S2, grafting a polymer brush with olefin on the surface of the microsphere

Dispersing macroporous resin microspheres with epoxy groups on the surfaces in sulfuric acid solution, reacting for 10-12 hours at 40-60 ℃, washing the reacted product to be neutral, and vacuum drying;

dissolving the dried microspheres in dichloromethane, then adding triethylamine, 2-bromoisobutyryl bromide and 4-dimethylaminopyridine under ice bath condition, reacting at room temperature, and washing to obtain a macromolecular initiator;

dispersing a macromolecular initiator, 2' -bipyridine and allyl methacrylate in a mixed solvent of ethanol and deionized water according to the proportion of 1.0-1.2 g/50-60 mg/0.4-0.6 mL/8-12 mL, wherein the volume ratio of ethanol to deionized water in the mixed solvent of ethanol and deionized water is 4/1;

removing oxygen in the solvent through freezing, vacuumizing and nitrogen filling, adding a cuprous bromide catalyst into the solvent in a proportion of 8-12 mL/25-30 mg, carrying out catalytic reaction for 20-24 h at 50-70 ℃ in a nitrogen atmosphere, and washing with ethanol, deionized water and disodium ethylenediamine tetraacetate aqueous solution for 3-5 times sequentially after the reaction to obtain the microspheres with the surface grafted with the polymer brush containing olefin.

S3, preparing the dual-function adsorption resin

Dissolving a photoinitiator benzoin dimethyl ether in an ethanol-deionized water solution, wherein the volume ratio of ethanol to deionized water in the ethanol-deionized water solution is 1/1;

adding resin matrix of polymer brush with alkene grafted on the microsphere surface, performing ultrasonic dispersion, and then adding L-cysteine; wherein, the mixing proportion of benzoin dimethyl ether, ethanol-deionized water solution, resin matrix of polymer brush grafted with olefin on the microsphere surface and L-cysteine is 20-30 mg/20-30 mL/300-400 mg;

irradiating for 20-40 min under 365nm ultraviolet light, respectively washing for 3-5 times with ethanol and deionized water, and drying at room temperature to obtain the dual-function adsorption resin.

Compared with the prior art, the application has the following beneficial effects:

1. the double-function adsorption resin takes macroporous resin MAR microspheres with epoxy groups on the surfaces as a matrix, adopts an atom transfer radical polymerization technology to graft polymer brushes with olefin on the surfaces of the microspheres, and finally introduces L-cysteine into the surfaces of the microspheres through photoinitiated 'mercapto-olefin' click chemical reaction, wherein the L-cysteine has amino, carboxyl and other functional groups, so that the whole resin microspheres keep good hydrophilicity, can effectively enrich glycopeptides, and can provide rich binding sites for enrichment of metal ions.

2. The preparation method is simple, the raw materials are cheap and easy to obtain, and the prepared product is pure, monodisperse and uniform in pore diameter.

Drawings

FIG. 1 is a flow chart of the preparation of a dual function adsorption resin of the present application;

FIG. 2 is a helium ion electron microscope image of macroporous resin microspheres (a) and bifunctional adsorption resin (b);

FIG. 3 is an infrared signature of macroporous resin microspheres, microspheres with olefin-containing polymer brushes grafted on the surface, dual function adsorption resins;

FIG. 4 is an X-ray diffraction photoelectron spectrum: (a) a full spectrum of microspheres with olefin-containing polymer brushes grafted on the surface, (b) a full spectrum of a bifunctional adsorption resin, (c) an N-1S high resolution spectrum of a bifunctional adsorption resin, and (d) an S-2p high resolution spectrum of a bifunctional adsorption resin;

FIG. 5 is a graph of analysis of bifunctional adsorption resins before and after enrichment of IgG trypsin digest: (a) pre-enrichment analysis, (b) post-enrichment analysis of 10 μg IgG trypsin digest, (c) post-deglycosylation analysis, (d) post-enrichment analysis of 10fmol IgG trypsin digest, (e) signal intensity analysis of 4 selected N-glycopeptides enriched in different amounts; in the figure, the N-glycopeptides are shown;

FIG. 6 shows a dual function adsorption resin for Cu ²⁺ (a) adsorption isotherms and (d) kinetics, and are fitted by (b) Langmuir model, (c) Freundlich model, (e) quasi-primary model and (f) quasi-secondary model.

Detailed Description

The following description of the embodiments of the present application will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.

Example 1

1. Preparation of bifunctional adsorption resin

S1, preparing macroporous resin microspheres with epoxy groups on surfaces

8mL of 3, 4-Epoxycyclohexylmethacrylate (EMA), 8mL of tetraethyleneglycol diacrylate (TEGDA), 8mL of n-propanol, 8mL of 1, 4-butanediol, 0.5g of azobisisobutyronitrile, and 160mL of an aqueous phase comprising Sodium Dodecyl Sulfate (SDS) at a concentration of 0.2% and polyvinyl alcohol (PVA) at a concentration of 0.5% were mixed with 10mL of a polyglycidyl methacrylate (seed) solution after ultrasonic disruption emulsification;

reacting for 16h at room temperature, and then heating to 70 ℃ to continue the reaction for 16h;

and performing soxhlet extraction for 24 hours after the reaction, then respectively washing with absolute ethyl alcohol and deionized water for 3 times, and finally performing vacuum drying at 60 ℃ for 12 hours to obtain macroporous resin Microspheres (MAR) with epoxy groups on the surfaces.

S2, grafting a polymer brush with olefin on the surface of the microsphere

Dispersing macroporous resin microspheres with epoxy groups on the surfaces in sulfuric acid solution, reacting for 12 hours at 60 ℃, washing the reacted product to be neutral, and drying in vacuum;

1.0g of a macroinitiator, 50mg of 2,2' -bipyridine and 0.5mL of Allyl Methacrylate (AMA) are dispersed in 10mL of mixed solvent of ethanol and deionized water, wherein the volume ratio of ethanol to deionized water in the mixed solvent of ethanol and deionized water is 4/1;

performing the operations of freezing, vacuumizing and nitrogen filling for 3 times circularly to remove oxygen in the solvent, and then adding 25mg of cuprous bromide catalyst;

the operations of freezing, vacuumizing and nitrogen filling are performed for 3 times in a recycling way, and the catalytic reaction is carried out for 24 hours at 60 ℃ under the nitrogen atmosphere, and ethanol, deionized water and disodium ethylenediamine tetraacetate aqueous solution are sequentially used for washing 3 times after the reaction, so that the microsphere (poly (AMA) @ MAR microsphere) with the polymer brush containing olefin grafted on the surface is obtained.

S3, preparing the dual-function adsorption resin

30mg of the photoinitiator benzoin dimethyl ether is dissolved in 25mL of ethanol-deionized water, wherein the volume ratio of ethanol to deionized water in the ethanol-deionized water solution is 1/1;

300mg of poly (AMA) @ MAR microspheres were added and dispersed by ultrasound, then 400mg of L-cysteine (L-Cys) was added, irradiated for 30min under 365nm ultraviolet light, washed 3 times with ethanol and deionized water, respectively, and dried at room temperature to obtain a bifunctional adsorption resin (Cys@poly (AMA) @ MAR).

2. Characterization of materials

FIG. 2 is a helium electron microscope image of macroporous resin Microspheres (MARs) and bifunctional adsorption resins (Cys@poly (AMA) @ MARs). From the figure, monodisperse MAR microspheres with a diameter of about 5 μm were successfully prepared by seed swelling polymerization (FIG. 2 a), while the morphology of the modified microspheres (FIG. 2 b) was similar to the original MAR (FIG. 2 a) and the particle surface was slightly rough, indicating that Cys@poly (AMA) @MAR was successfully prepared.

Fig. 3: macroporous resin Microspheres (MAR), microspheres with olefin-containing polymer brushes grafted on the surface (poly (AMA) @ MAR microspheres), and bifunctional adsorption resins (cys @ poly (AMA) @ MAR) were characterized by fourier transform attenuated total reflection infrared spectroscopy (ATR-FTIR), as shown in fig. 3:

in the MAR spectrum, 910cm-1 is a characteristic peak of the epoxide group, which is not present in the other spectra, indicating that the epoxide group on the modified MAR surface has been depleted.

In the poly (AMA) @ MAR spectrum, the characteristic peak at 3531cm-1 corresponds to the stretching vibration of-OH, the weak peak at 1634cm-1 corresponds to the stretching vibration of C=C, and these two characteristic peaks do not appear in the Cys @ poly (AMA) @ MAR spectrum.

In the Cys@poly (AMA) @ MAR spectra, the two characteristic peaks of 3540cm-1 and 1580cm-1 were-NH 2 respectively corresponding to stretching and deforming vibrations, and the weak peak of 1625cm-1 was-COOH corresponding to stretching vibrations, thus indicating successful grafting of L-cysteine (L-Cys) on the MAR surface.

FIG. 4 shows XPS spectrum, specifically:

there was no N, S peak in the spectrum (4 a) of the microsphere (poly (AMA) @ MAR microsphere) with the olefin-containing polymer brush grafted on the surface, and N, S peak in the spectrum (4 b) of the bifunctional adsorbent resin (cys @ poly (AMA) @ MAR) with which the surface cys @ poly (AMA) @ MAR was successfully prepared.

In the high resolution N-1s spectrum of Cys@poly (AMA) @ MAR (FIG. 4C), the characteristic peaks of 399.1eV and 401eV are assigned to C-N and N-H, respectively; in the high resolution S-2p spectrum of Cys@poly (AMA) @MAR (FIG. 4 d), the characteristic peaks of 163.4eV and 164.6eV belong to the 3/2p and 1/2p orbitals of sulfur atoms, based on which it is further shown that Cys@poly (AMA) @MAR was successfully prepared.

3. Glycopeptide enrichment

Taking 5mg of bifunctional adsorption resin (Cys@poly (AMA) @MAR), placing in a centrifuge tube, firstly activating with 200 mu L of loading liquid (acetonitrile ACN/water H2O/trifluoroacetic acid TFA=87/12/1, volume ratio), and then placing in 200 mu L of loading liquid containing a certain amount of protein digestion liquid for incubation for 30min at room temperature;

oscillating and centrifuging at room temperature, and removing supernatant; the shaking is sufficient and is typically 8 hours.

Washing three times with a sample solution to remove the non-glycopeptides adsorbed on the material;

finally, the glycopeptides enriched on the material were eluted with an eluent (acetonitrile ACN/water H2O/trifluoroacetic acid tfa=30/69/1, volume ratio), and the resulting eluent was analyzed by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).

4. Adsorption of metal ions

Kinetic adsorption: 10mg of the bifunctional adsorption resin (Cys@poly (AMA) @MAR) was dispersed to 10mL (6 mmol/L) of Cu ²⁺ In an aqueous solution. After incubation for 20, 40, 60, 90, 120, 150, 180, 240, 300min, the supernatant was analyzed by atomic absorption spectrophotometry through a membrane.

Static adsorption: 10mg of the bifunctional adsorption resin (Cys@poly (AMA) @MAR) was dispersed in 10mL of Cu at different concentrations (1.0, 3.0, 5.0, 6.0, 7.0, 8.0, 9.0 mmol/L) ²⁺ In aqueous solution, incubation was performed at 25℃for 12 hours, then filtration was performed with a 0.22 μm filter membrane, and the collected filtrate was analyzed with an atomic absorption spectrophotometer.

Example 2

Preparation of bifunctional adsorption resin

S1, preparing macroporous resin microspheres with epoxy groups on surfaces

6mL of 3, 4-Epoxycyclohexylmethacrylate (EMA), 6mL of tetraethyleneglycol diacrylate (TEGDA), 6mL of n-propanol, 6mL of 1, 4-butanediol, 0.4g of azobisisobutyronitrile, and 150mL of an aqueous phase comprising Sodium Dodecyl Sulfate (SDS) at a concentration of 0.2% and polyvinyl alcohol (PVA) at a concentration of 0.5% were mixed with 9mL of a polyglycidyl methacrylate (seed) solution after ultrasonic disruption emulsification;

reacting for 12 hours at room temperature, and then heating to 60 ℃ to continue reacting for 12 hours;

S2, grafting a polymer brush with olefin on the surface of the microsphere

S3, preparing the dual-function adsorption resin

Glycopeptide enrichment and metal ion adsorption were performed with the bifunctional adsorption resin (cys@poly (AMA) @mar) of this example, and the procedure was the same as in example 1.

Material application

(1) Application of bifunctional adsorption resin (Cys@poly (AMA) @MAR) in glycopeptide enrichment

The IgG trypsin digestion solution is taken as a sample, and the bifunctional adsorption resin (Cys@poly (AMA) @MAR) prepared in the embodiment 1 is used for enrichment, and due to the introduction of L-cysteine (L-Cys), a large number of polar groups (such as amino groups and carboxyl groups) are arranged on the surface of the bifunctional adsorption resin, so that the Cys@poly (AMA) @MAR has stronger hydrophilicity and can be used as a hydrophilic interaction chromatography (HILIC) adsorbent for enriching glycopeptides. The enriched glycopeptides were determined by MALDI-TOF-MS and the results are shown in FIG. 5:

the non-glycopeptide signal severely masks the N-glycopeptide signal prior to enrichment (fig. 5 a);

after enrichment of 10. Mu.g of IgG trypsin digest with Cys@poly (AMA) @MAR prepared in example 1, the glycopeptide signal increased significantly (signal intensity 1018.3) with weaker non-glycopeptide signal interference as shown in FIG. 5 b;

deglycosylating the eluted glycopeptides with a loading solution (acetonitrile ACN/water H2O/trifluoroacetic acid tfa=87/12/1, volume ratio) PNGase-F, as shown in fig. 5c, the glycopeptide signal disappeared and only two deamidated peptides (1158 m/z and 1190 m/z) were detected, thereby indicating that all disappeared glycopeptide signals belong to the N-glycopeptides;

as shown in FIG. 5d, the 7 typical N-glycopeptides (2601 m/z, 2633m/z, 2764m/z, 2795m/z, 2805m/z, 2925m/z and 2957 m/z) after enrichment were all clearly identified, and the glycopeptide signal intensity was 402.4 when the sample size of the IgG trypsin digest was reduced to 10fmol, thus indicating that Cys@poly (AMA) @MAR had higher sensitivity to N-glycopeptides.

Different levels of IgG trypsin digestion solution were enriched with Cys@poly (AMA) @ MAR, and N-glycopeptides composed of four different sugar chains 2601, 2633, 2763 and 2795 were selected as labels, as shown in FIG. 5e, as the IgG trypsin digestion solution increased, the intensities of the four labels increased correspondingly, and when the IgG trypsin digestion solution increased to 45. Mu.g, the intensity of the label did not increase any more, thus indicating that the maximum adsorption amount of Cys@poly (AMA) @ MAR was 9mg/g.

(2) Bifunctional adsorption resin (Cys@poly (AMA) @MAR) in Cu ²⁺ Application in adsorption

The maximum adsorption amount is a key index for evaluating the adsorption performance of the material.

To evaluate the adsorption capacity of the bifunctional adsorption resin (cys@poly (AMA) @mar) for metal ions, the adsorption capacity for metal ions of different concentrations was tested and the isothermal adsorption curve thereof was plotted (fig. 6 a), from which it is known: when Cu is ²⁺ At a concentration of less than 6mmol/L, the adsorption amount of Cys@poly (AMA) @MAR follows Cu ²⁺ The concentration increases rapidly; when Cu is ²⁺ At concentrations above 6mmol/L, the adsorption isotherm reached the plateau, from which Cys@poly (AMA) @MAR vs Cu was calculated ²⁺ The maximum adsorption amount of (C) was 63mg/g.

To study Cu ²⁺ The adsorption mechanism on Cys@poly (AMA) @ MAR was fitted to adsorption isotherms using Langmuir model and Freundlich model, and the results are shown in FIGS. 6b and 6c, where the linear correlation coefficients of Langmuir model and Freundlich model are 0.9981 and 0.9594, respectively. Obviously Cys@poly (AMA) @MAR vs Cu ²⁺ The adsorption type of (2) is more in accordance with Langmuir model, and is a monomolecular layer adsorption mode.

The rate at which the material reaches adsorption equilibrium is also an important indicator for the study of the material as an adsorbent, and thus the kinetic adsorption process of cys@poly (AMA) @mar is also studied. As shown in FIG. 6d, cys@poly (AMA) @MAR vs Cu ²⁺ The adsorption quantity of the material is rapidly increased in the first 120min, and the adsorption quantity is basically unchanged after the adsorption quantity exceeds 120min, so that the adsorption sites on the surface of the material are completely occupied, and the adsorption balance is achieved. Fitting a quasi-primary and a quasi-secondary kinetic model according to the adsorption curve, as shown in fig. 6e and 6 f: the linear fit correlation coefficient of the quasi-first-order model is 0.8837, and the linear fit correlation coefficient of the quasi-second-order model is 0.9974. Obviously, the pseudo-second-order kinetic model is more suitable for describing Cu ²⁺ Kinetic adsorption process on Cys@poly (AMA) @ MAR.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present application, and are not limiting; although the application has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced with equivalents; such modifications and substitutions do not depart from the spirit of the technical solutions according to the embodiments of the present application.

Claims

1. The preparation method of the bifunctional adsorption resin is characterized by comprising the following steps:

s1, preparing macroporous resin microspheres with epoxy groups on the surfaces, taking the macroporous resin microspheres as a resin matrix,

the preparation method of the macroporous resin microsphere with the epoxy group on the surface comprises the following steps:

mixing emulsified polyglycidyl methacrylate, 3, 4-epoxycyclohexyl methacrylate, tetraethylene glycol diacrylate, n-propanol, 1, 4-butanediol, azodiisobutyronitrile and water phase in a ratio of 9-11 mL/6-8 mL/0.4-0.6 g/150-170 mL, reacting for 12-18 h at room temperature, and then heating to 60-80 ℃ to continue reacting for 12-18 h; after the reaction, carrying out soxhlet extraction for 24-48 hours, then respectively washing with absolute ethyl alcohol and deionized water for 3-5 times, and finally carrying out vacuum drying at 60 ℃ for 12-20 hours to obtain macroporous resin microspheres with epoxy groups on the surfaces;

s2, grafting a polymer brush with olefin on the surface of the microsphere by utilizing an atom transfer radical polymerization technology, wherein the polymer brush specifically comprises the following components:

removing oxygen in the solvent through freezing, vacuumizing and nitrogen filling, adding a cuprous bromide catalyst into the solvent according to the proportion of 8-12 mL/25-30 mg, carrying out catalytic reaction for 20-24 h at 50-70 ℃ in a nitrogen atmosphere, and washing with ethanol, deionized water and disodium ethylenediamine tetraacetate aqueous solution for 3-5 times after the reaction to obtain microspheres with the surface grafted with the polymer brush containing olefin;

s3, introducing L-cysteine into the surface of the microsphere through photoinitiation of 'mercapto-ene' click chemical reaction to obtain the bifunctional adsorption resin, wherein the bifunctional adsorption resin specifically comprises:

adding resin matrix with olefin polymer brush grafted on the microsphere surface, performing ultrasonic dispersion, and then adding L-cysteine, wherein the mixing ratio of benzoin dimethyl ether, ethanol-deionized water solution, resin matrix with olefin polymer brush grafted on the microsphere surface and L-cysteine is 20-30 mg/20-30 mL/300-400 mg;

2.A bifunctional adsorbent resin obtained by the preparation method of claim 1, characterized in that:

the resin matrix is macroporous resin microsphere with epoxy group on the surface;

the microsphere surface is grafted with a polymer brush with olefin, and L-cysteine is also introduced into the microsphere surface.

3. The dual function adsorbent resin of claim 2, wherein: the particle size of the macroporous resin microsphere with the epoxy group on the surface is 5-6 mu m.

4. Use of a bifunctional adsorption resin of claim 2 or 3 for adsorbing metal ions.

5. Use of a bifunctional adsorption resin as claimed in claim 2 or 3 for the separation, enrichment and purification of glycopeptides.