Polysaccharide derivative bonded solvent-resistant chromatographic column and preparation method and application thereof

Abstract

The present invention belongs to the technical field of polymer materials and chromatographic separation, and discloses a polysaccharide derivative bonded solvent-resistant chromatographic column and its preparation method and application. The preparation method is: after the polysaccharide derivative is subjected to an alkalization reaction, an allyl epoxy-terminated polyether is added to carry out an etherification reaction to obtain an allyl polyether-substituted polysaccharide derivative; after the silica microspheres are subjected to an activation treatment, a triethoxysilane modification reaction is used to obtain a silicosilane sphere; the allyl polyether-substituted polysaccharide derivative is subjected to a silicohydrogenation bonding reaction with the silicosilane sphere to obtain a polysaccharide derivative-bonded silicon sphere; and finally, the chromatographic column is filled with the chromatographic column to obtain a polysaccharide derivative-bonded solvent-resistant chromatographic column. The preparation method of the present invention has the advantages of high bonding efficiency and simple preparation process. The obtained chromatographic column has the advantages of good solvent resistance and good separation effect.

Description

Technical Field

[0001] The invention belongs to the technical field of polymer materials and chromatographic separation, and particularly relates to a polysaccharide derivative bonded solvent-resistant chromatographic column and a preparation method and application thereof. Background Art

[0002] Chromatographic separation is an analytical technique based on the difference in the distribution coefficient of a substance between the stationary phase and the mobile phase. Its basic principle is to use the difference in the distribution behavior of each component in the mixture in different phases to achieve separation through the relative movement of the two phases. During the chromatographic separation process, one phase is fixed and is called the stationary phase; the other phase continuously flows through the stationary phase and is called the mobile phase. The mixture is introduced into the chromatographic column, which is filled with the stationary phase, and then the mobile phase is used to move the mixture. Different substances have different distribution coefficients in the stationary phase and the mobile phase, causing them to move along the stationary phase at different speeds, ultimately achieving the effect of separation.

[0003] Chirality is one of the basic properties of nature. From microscopic particles to macroscopic substances, chirality is ubiquitous. For example, most proteins and nucleic acids that make up living organisms are chiral, so chirality research plays an important role in pharmacology, analytical chemistry, life sciences and other fields. The focus of chirality research is mainly on the separation of chiral drugs. Among common drugs, a considerable number exist in the form of racemates. Generally, one of the enantiomers exerts its pharmacological effect, while the other enantiomer usually has no pharmacological effect or even offsets some of the effects. Therefore, exploring technologies and methods that can effectively separate racemates is a major challenge in the development of chiral drugs.

[0004] The separation principle of chiral chromatographic columns is mainly based on the interaction between chiral stationary phases and chiral isomers. This interaction utilizes the stereostructure and chemical properties of chiral stationary phases, so that enantiomers (i.e. chiral isomers) can be effectively separated. The core of chiral chromatographic columns is chiral stationary phases. According to the chemical structure of the stationary phase, chiral chromatographic columns are mainly divided into the following types: 1. Polysaccharide derivatives, 2. Cyclodextrins, 3. Crown ethers, 4. Proteins, and other types (such as macrocycles, ligand exchange types, molecular imprinting polymers, etc.). Each stationary phase has its own specific spatial structure and chemical properties, which determine the way and efficiency of their interaction with chiral isomers.

[0005] At present, the most widely used chiral stationary phase is polysaccharide derivatives. According to the production process, chiral stationary phases of polysaccharide derivatives are divided into two categories: coating type and bonding type. The coating type stationary phase is coated on the substrate by physical action, so some solvents can dissolve and fall off the substrate, resulting in a decrease in its separation effect, so there are restrictions on the solvent of the mobile phase. The bonded stationary phase has fewer restrictions on the use of solvents in the mobile phase, a wider range of mobile phases, and better solvent resistance of the chromatographic column.

[0006] CN 116351399 A discloses a method for preparing a bonded cellulose derivative chiral liquid chromatography column, wherein the cellulose derivative is coated on a macroporous hybrid silica gel modified by amino functionalization, and then the cellulose derivative is bonded to the macroporous hybrid silica gel modified by amino functionalization through the connection of a spacer arm molecule to obtain a target chiral stationary phase. However, the patent adopts the method of first coating and then connecting with a spacer arm molecule having a bifunctional group. In addition to one end of the spacer arm molecule being connected to the amino group on the surface of the macroporous hybrid silica gel modified by amino functionalization and the other end being connected to the hydroxyl group on the cellulose derivative, more often the amino groups on the silica gel surface are connected to the amino groups and the hydroxyl groups on the cellulose derivative are connected to the hydroxyl groups, resulting in low bonding efficiency. At the same time, it requires multi-step functionalization modification, hydroxyl protection and hydroxyl substitution, and the preparation process is complicated.

[0007] Therefore, providing a polysaccharide derivative bonded chromatographic column with higher bonding efficiency and simpler preparation process has broad application prospects. Summary of the invention

[0008] In view of the shortcomings and deficiencies of the above prior art, the primary purpose of the present invention is to provide a method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column.

[0009] Another object of the present invention is to provide a polysaccharide derivative bonded solvent-resistant chromatographic column prepared by the above method.

[0010] Another object of the present invention is to provide the use of the above-mentioned polysaccharide derivative bonded solvent-resistant chromatographic column in drug separation.

[0011] The purpose of the present invention is achieved through the following technical solutions:

[0012] A method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column comprises the following preparation steps:

[0013] (1) mixing a polysaccharide derivative with an alkaline solution for alkalization reaction, then adding an allyl epoxy-terminated polyether, heating to 50-80° C. and stirring for etherification reaction, and separating the product to obtain an allyl polyether-substituted polysaccharide derivative;

[0014] (2) adding the silica microspheres to a hydrochloric acid solution for heating and activation treatment, washing and drying, adding the microspheres to an alcohol solution of triethoxysilane, stirring and refluxing to obtain hydrosilicon spheres;

[0015] (3) adding the allyl polyether substituted polysaccharide derivative of step (1) into an anhydrous solvent to dissolve it, then adding the silicon hydride balls of step (2) to stir and disperse them uniformly, heating to 60 to 90° C. under nitrogen protection, then adding chloroplatinic acid catalyst to stir and react, to obtain silicon balls bonded with the polysaccharide derivative;

[0016] (4) Filling the polysaccharide derivative-bonded silica spheres obtained in step (3) into a chromatographic column to obtain a polysaccharide derivative-bonded solvent-resistant chromatographic column.

[0017] Furthermore, the polysaccharide derivative in step (1) is cellulose or its derivatives (hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, etc.), starch or its derivatives, chitosan or its derivatives, dextran or its derivatives, polysucrose or its derivatives, etc.

[0018] Furthermore, the alkaline solution in step (1) is a sodium hydroxide solution, a potassium hydroxide solution, a sodium ethoxide solution or a potassium ethoxide solution with a mass concentration of 0.5% to 30%; the mass ratio of the polysaccharide derivative to the alkaline solution is 1:0.2 to 5.

[0019] Furthermore, the molecular weight of the allyl epoxy terminated polyether in step (1) is 400 to 1000.

[0020] The present invention uses allyl epoxy-terminated polyether to carry out etherification reaction on the polysaccharide derivative, which can cut off the hydrogen bonding of hydroxyl groups between the polysaccharide derivative molecules and improve the solubility of the subsequent reaction; on the other hand, the connecting arm of appropriate length can promote the extension of the bonded polysaccharide derivative in the mobile phase, improve the specific binding efficiency between the polysaccharide derivative and the mixture to be separated, and effectively improve the separation effect.

[0021] Furthermore, the amount of allyl epoxy-terminated polyether added in step (1) controls the degree of substitution (etherification degree, the average number of hydroxyl groups replaced by the reaction reagent on each dehydrated glucose unit) of the etherification reaction to be 0.05 to 0.2. The present invention can achieve good bonding and separation effects by controlling the degree of substitution of the etherification reaction. If the degree of substitution is too low, its solubility is low, and it is not easy to dissolve in the subsequent bonding reaction process, and the terminal allyl content of the subsequent reaction with the silicon hydride ball is too low, the bonding effect is poor, and the solvent resistance is poor; if the degree of substitution is too high, the bonding density of the subsequent reaction with the silicon hydride ball is too large, and the polysaccharide derivative cannot effectively extend to specifically bind and interact with the mixture to be separated in the mobile phase, and the separation effect becomes poor.

[0022] Furthermore, the separation in step (1) refers to separation by filtration, washing and drying the solid phase to obtain the allyl polyether substituted polysaccharide derivative; or vacuum evaporation of the solvent to obtain the allyl polyether substituted polysaccharide derivative.

[0023] Furthermore, the silica microspheres in step (2) have a particle size of 2 to 10 μm and a pore size of 5 to 200 nm.

[0024] Furthermore, the hydrochloric acid solution in step (2) refers to a hydrochloric acid solution with a concentration of 0.5 to 2 mol / L; and the heating activation treatment refers to stirring the activation treatment at a temperature of 80 to 100° C. for 2 to 4 hours.

[0025] Furthermore, the amount of triethoxysilane added in step (2) is 5% to 15% of the mass of the silica microspheres.

[0026] Furthermore, the anhydrous solvent in step (3) is dimethyl sulfoxide (DMSO), dimethylformamide (DMF), or dimethylacetamide (DMAC).

[0027] Furthermore, in step (3), the mass ratio of the allyl polyether substituted polysaccharide derivative to the silicon hydrosilicon spheres added is 0.1 to 0.6:1.

[0028] A polysaccharide derivative bonded solvent-resistant chromatographic column is prepared by the method.

[0029] The application of the above polysaccharide derivative bonded solvent-resistant chromatographic column in drug separation.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] (1) The present invention uses allyl polyether substituted polysaccharide derivatives to react with hydrosilicon spheres to prepare polysaccharide derivative bonded silicon spheres, and utilizes the high selectivity of hydrosilation reaction to achieve good bonding between the polysaccharide derivatives and the silicon spheres. It has the advantages of high bonding efficiency and simple preparation process.

[0032] (2) The present invention uses allyl polyether to carry out substitution reaction on the polysaccharide derivative in advance, which can improve its solubility in subsequent reactions on the one hand and improve its separation effect on the other hand. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 and Figure 2 The chromatograms of the chromatographic columns obtained in Example 2 and Comparative Example 1 used to separate the sample with structure of Formula I and its corresponding isomers are shown respectively.

[0034] Figure 3 and Figure 4 The chromatograms of the chromatographic columns obtained in Example 2 and Comparative Example 3 used to separate the sample with the structure of Formula II and its corresponding isomers are shown respectively. DETAILED DESCRIPTION

[0035] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto.

[0036] Example 1

[0037] A method for preparing a cellulose-bonded solvent-resistant chromatographic column comprises the following preparation steps:

[0038] (1) 100 g of cellulose was mixed with 150 g of a 10% sodium hydroxide aqueous solution for alkalization reaction for 2 h, and then 35 g of an allyl epoxy-terminated polyether with an average molecular weight of 1000 (Liaoning Kelong Fine Chemical Co., Ltd. KL-91B) was added, and the temperature was raised to 60-70° C. and stirred for etherification reaction for 4 h. The product was sampled and tested for the degree of substitution of allyl polyether, which was 0.051. The product was vacuum filtered, and the solid phase was washed with deionized water, neutralized, washed with ethanol, and vacuum dried at 80-90° C. to obtain cellulose substituted with allyl polyether.

[0039] (2) Add silica microspheres with a particle size of 5 μm and a pore size of 10 to 50 nm to a hydrochloric acid solution with a concentration of 1 mol / L, heat to 85 to 90° C., stir and activate for 2 h, filter, wash with deionized water until neutral, vacuum dry, add to an isopropanol solution of triethoxysilane, stir and reflux for reaction, the amount of triethoxysilane added is 5% of the mass of the silica microspheres, and obtain silicon hydrosilicon spheres.

[0040] (3) The allyl polyether-substituted cellulose of step (1) is added to DMSO and heated to dissolve evenly, and then the silicohydride balls of step (2) are added and stirred to disperse evenly, wherein the mass ratio of the allyl polyether-substituted cellulose to the silicohydride balls is 0.1:1, and after nitrogen is used to remove water, a 100 ppm concentration of chloroplatinic acid catalyst solution is added, and the reaction is stirred at 70-80° C. for 4 h until the allyl reaction is complete, thereby obtaining cellulose-bonded silicohydride balls.

[0041] (4) Filling the cellulose-bonded silica spheres obtained in step (3) into a 250*4.6 mm chromatographic column to obtain a cellulose-bonded solvent-resistant chromatographic column.

[0042] Example 2

[0043] A method for preparing a cellulose-bonded solvent-resistant chromatographic column comprises the following preparation steps:

[0044] (1) 100 g of cellulose was mixed with 80 g of a 20% sodium hydroxide aqueous solution for alkalization reaction for 2 h, and then 40 g of an allyl epoxy-terminated polyether with an average molecular weight of 500 (Liaoning Kelong Fine Chemical Co., Ltd. KL-11B) was added, and the temperature was raised to 70-80° C. and stirred for etherification reaction for 4 h. The product was sampled and tested for a degree of substitution of allyl polyether of 0.12. The solid phase was vacuum filtered, washed with deionized water, neutralized, washed with ethanol, and vacuum dried at 80-90° C. to obtain cellulose substituted with allyl polyether.

[0045] (2) Add silica microspheres with a particle size of 5 μm and a pore size of 10 to 50 nm to a hydrochloric acid solution with a concentration of 1 mol / L, heat to 85 to 90° C., stir and activate for 2 h, filter, wash with deionized water until neutral, vacuum dry, add to an isopropanol solution of triethoxysilane, stir and reflux for reaction, the amount of triethoxysilane added is 10% of the mass of the silica microspheres, and obtain silicon hydrosilicon spheres.

[0046] (3) The allyl polyether-substituted cellulose of step (1) is added to DMSO and heated to dissolve evenly, and then the silicon hydride balls of step (2) are added and stirred to disperse evenly, wherein the mass ratio of the allyl polyether-substituted cellulose to the silicon hydride balls is 0.3:1, and after nitrogen is used to remove water, a 100 ppm concentration of chloroplatinic acid catalyst solution is added, and the reaction is stirred at 70-80° C. for 4 h until the allyl reaction is complete, thereby obtaining cellulose-bonded silicon balls.

[0047] (4) Filling the cellulose-bonded silica spheres obtained in step (3) into a 250*4.6 mm chromatographic column to obtain a cellulose-bonded solvent-resistant chromatographic column.

[0048] Example 3

[0049] A method for preparing a cellulose-bonded solvent-resistant chromatographic column comprises the following preparation steps:

[0050] (1) 100 g of cellulose was mixed with 40 g of a 30% sodium hydroxide aqueous solution for alkalization reaction for 2 h, and then 50 g of an allyl epoxy-terminated polyether (Liaoning Kelong Fine Chemical Co., Ltd. KL-11) with an average molecular weight of 450 was added, and the temperature was raised to 70-80° C. and stirred for etherification reaction for 4 h. The product was sampled and tested for a degree of substitution of allyl polyether of 0.20. The solid phase was vacuum filtered, washed with deionized water, neutralized, washed with ethanol, and vacuum dried at 80-90° C. to obtain cellulose substituted with allyl polyether.

[0051] (2) Add silica microspheres with a particle size of 5 μm and a pore size of 10 to 50 nm to a hydrochloric acid solution with a concentration of 1 mol / L, heat to 85 to 90° C., stir and activate for 2 h, filter, wash with deionized water until neutral, vacuum dry, add to an isopropanol solution of triethoxysilane, stir and reflux for reaction, the amount of triethoxysilane added is 15% of the mass of the silica microspheres, and obtain silicon hydrosilicon spheres.

[0052] (3) The allyl polyether-substituted cellulose of step (1) is added to DMSO and heated to dissolve evenly, and then the silicon hydride balls of step (2) are added and stirred to disperse evenly, wherein the mass ratio of the allyl polyether-substituted cellulose to the silicon hydride balls is 0.6:1, and after nitrogen is used to remove water, a 100 ppm concentration of chloroplatinic acid catalyst solution is added, and the reaction is stirred at 70-80° C. for 4 h until the allyl reaction is complete, thereby obtaining cellulose-bonded silicon balls.

[0053] (4) Filling the cellulose-bonded silica spheres obtained in step (3) into a 250*4.6 mm chromatographic column to obtain a cellulose-bonded solvent-resistant chromatographic column.

[0054] Example 4

[0055] A method for preparing an amylose-bonded solvent-resistant chromatographic column comprises the following preparation steps:

[0056] (1) 100 g of amylose was mixed with 200 g of a 2% sodium hydroxide aqueous solution for alkalization reaction for 1 h, and then 40 g of an allyl epoxy-terminated polyether with an average molecular weight of 500 (Liaoning Kelong Fine Chemical Co., Ltd. KL-11B) was added, and the temperature was raised to 70-80° C. and stirred for etherification reaction for 4 h. The product was sampled and tested for a degree of substitution of allyl polyether of 0.13. The solid phase was vacuum filtered, washed with deionized water, neutralized, washed with ethanol, and vacuum dried at 80-90° C. to obtain amylose substituted with allyl polyether.

[0057] (2) Add silica microspheres with a particle size of 5 μm and a pore size of 10 to 50 nm to a hydrochloric acid solution with a concentration of 1 mol / L, heat to 85 to 90° C., stir and activate for 2 h, filter, wash with deionized water until neutral, vacuum dry, add to an isopropanol solution of triethoxysilane, stir and reflux for reaction, the amount of triethoxysilane added is 10% of the mass of the silica microspheres, and obtain silicon hydrosilicon spheres.

[0058] (3) adding the amylose substituted with allyl polyether of step (1) into DMSO and heating to dissolve evenly, then adding the silicon hydride balls of step (2) and stirring to disperse evenly, wherein the mass ratio of amylose substituted with allyl polyether to silicon hydride balls is 0.3:1, adding 100 ppm chloroplatinic acid catalyst solution after nitrogen removal, and reacting at 70-80° C. with stirring for 4 h until the allyl reaction is complete, thereby obtaining silicon spheres bonded with amylose.

[0059] (4) Filling the amylose-bonded silica spheres obtained in step (3) into a 250*4.6 mm chromatographic column to obtain an amylose-bonded solvent-resistant chromatographic column.

[0060] Example 5

[0061] A method for preparing a dextran-bonded solvent-resistant chromatographic column comprises the following preparation steps:

[0062] (1) 100 g of dextran was mixed with 300 g of a 0.5% sodium hydroxide aqueous solution for alkalization reaction for 0.5 h, and then 40 g of an allyl epoxy-terminated polyether with an average molecular weight of 500 (Liaoning Kelong Fine Chemical Co., Ltd. KL-11B) was added, and the temperature was raised to 70-80° C. and stirred for etherification reaction for 4 h. The product was sampled and tested for the degree of substitution of allyl polyether, which was 0.14. After neutralization, the product was vacuum spray-dried at 80-90° C. to obtain allyl polyether-substituted dextran.

[0063] (2) Add silica microspheres with a particle size of 5 μm and a pore size of 10 to 50 nm to a hydrochloric acid solution with a concentration of 1 mol / L, heat to 85 to 90° C., stir and activate for 2 h, filter, wash with deionized water until neutral, vacuum dry, add to an isopropanol solution of triethoxysilane, stir and reflux for reaction, the amount of triethoxysilane added is 10% of the mass of the silica microspheres, and obtain silicon hydrosilicon spheres.

[0064] (3) The allyl polyether-substituted dextran of step (1) is added to DMF and heated to dissolve evenly, and then the silicon hydride balls of step (2) are added and stirred to disperse evenly, wherein the mass ratio of the allyl polyether-substituted dextran to the silicon hydride balls is 0.3:1, and after nitrogen is removed from the water, a 100 ppm concentration of chloroplatinic acid catalyst solution is added, and the reaction is stirred at 70-80° C. for 4 h until the allyl reaction is complete, thereby obtaining silicon spheres bonded with dextran.

[0065] (4) Filling the dextran-bonded silica spheres obtained in step (3) into a 250*4.6 mm chromatographic column to obtain a dextran-bonded solvent-resistant chromatographic column.

[0066] Comparative Example 1

[0067] A method for preparing a cellulose-bonded solvent-resistant chromatographic column, compared with Example 2, step (1) uses allyl glycidyl ether to replace the allyl epoxy-terminated polyether with an average molecular weight of 500, and the amount of allyl glycidyl ether added is adjusted to make the degree of substitution of the etherification reaction the same as that of Example 2. Steps (2) to (4) are the same as those of Example 2.

[0068] The chromatograms of the chromatographic columns obtained in Example 2 and Comparative Example 1 for separating the sample of formula I and its corresponding isomers are shown as follows: Figure 1 and Figure 2 As shown (mobile phase: tert-butyl dimethyl ether: methanol: ethanolamine = 60:40:0.1 (v / v), 1 ml / min, 25°C).

[0069]

[0070] By comparing the results of this comparative example with those of Example 2, it can be seen that the connecting arm bonded to the cellulose is too short, and the separation effect of the obtained chromatographic column is poor.

[0071] Comparative Example 2

[0072] A method for preparing a cellulose bonded solvent-resistant chromatographic column, compared with Example 2, the amount of allyl epoxy-terminated polyether added in step (1) is reduced to 15 g, and the degree of substitution of the allyl polyether of the product after etherification reaction is tested to be 0.038. The obtained allyl polyether-substituted cellulose is difficult to dissolve in DMSO by heating.

[0073] Comparative Example 3

[0074] A method for preparing a cellulose bonded solvent-resistant chromatographic column. Compared with Example 2, the amount of allyl epoxy-terminated polyether added in step (1) is increased to 90 g, and the degree of substitution of the allyl polyether in the product tested after etherification reaction is 0.25.

[0075] The chromatograms of the chromatographic columns obtained in Example 2 and Comparative Example 3 for separating the sample of formula II structure and its corresponding isomers are shown as follows: Figure 3 and Figure 4 As shown (mobile phase: tert-butyl dimethyl ether: methanol = 80:20 (v / v), 1 ml / min, 25°C).

[0076]

[0077] By comparing the results of this comparative example with those of Example 2, it can be seen that when the substitution degree of the allyl polyether is too high, the separation time of the obtained chromatographic column becomes longer and the separation effect becomes worse.

[0078] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column, characterized in that: The method comprises the following preparation steps: (1) mixing a polysaccharide derivative with an alkaline solution for alkalization reaction, then adding an allyl epoxy-terminated polyether, heating to 50-80° C. and stirring for etherification reaction, and separating the product to obtain an allyl polyether-substituted polysaccharide derivative; (2) adding the silica microspheres to a hydrochloric acid solution for heating and activation treatment, washing and drying, adding the microspheres to an alcohol solution of triethoxysilane, stirring and refluxing to obtain hydrosilicon spheres; (3) adding the allyl polyether substituted polysaccharide derivative of step (1) into an anhydrous solvent to dissolve it, then adding the silicon hydride balls of step (2) to stir and disperse them evenly, heating to 60-90° C. under nitrogen protection, then adding chloroplatinic acid catalyst to stir and react, to obtain silicon balls bonded with the polysaccharide derivative; (4) filling the polysaccharide derivative-bonded silica spheres obtained in step (3) into a chromatographic column to obtain a polysaccharide derivative-bonded solvent-resistant chromatographic column; The polysaccharide derivative in step (1) is cellulose or its derivatives, starch or its derivatives, chitosan or its derivatives, dextran or its derivatives, polysucrose or its derivatives; The alkaline solution is a sodium hydroxide solution, a potassium hydroxide solution, a sodium ethoxide solution or a potassium ethoxide solution with a mass concentration of 0.5% to 30%; the molecular weight of the allyl epoxy-terminated polyether is 400 to 1000; The silica microspheres in step (2) have a particle size of 2-10 μm and a pore size of 5-200 nm; the hydrochloric acid solution refers to a hydrochloric acid solution with a concentration of 0.5-2 mol / L; The heating activation treatment refers to stirring the activation treatment at a temperature of 80-100°C for 2-4 hours; The anhydrous solvent in step (3) is dimethyl sulfoxide, dimethylformamide or dimethylacetamide.

2. The method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column according to claim 1, characterized in that: The mass ratio of the polysaccharide derivative and the alkaline solution mixed in step (1) is 1:0.2~5.

3. The method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column according to claim 1, characterized in that: The amount of allyl epoxy-terminated polyether added in step (1) controls the degree of substitution of the etherification reaction to be 0.05-0.

2.

4. The method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column according to claim 1, characterized in that: The separation in step (1) refers to separation by filtration, washing and drying the solid phase to obtain the allyl polyether substituted polysaccharide derivative; or vacuum evaporation of the solvent to obtain the allyl polyether substituted polysaccharide derivative.

5. The method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column according to claim 1, characterized in that: The amount of triethoxysilane added in step (2) is 5% to 15% of the mass of the silica microspheres.

6. The method for preparing a polysaccharide derivative bonded solvent-resistant chromatographic column according to claim 1, characterized in that: The mass ratio of the allyl polyether substituted polysaccharide derivative to the silicon hydride sphere added in step (3) is 0.1-0.6:

1.

7. A polysaccharide derivative bonded solvent-resistant chromatographic column, characterized in that: It is prepared by the method according to any one of claims 1 to 6.

8. Use of the polysaccharide derivative bonded solvent-resistant chromatographic column according to claim 7 in drug separation.

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