阅读说明：本技术 一种单分散交联聚苯乙烯-二乙烯基苯微球的制备方法 (Preparation method of monodisperse crosslinked polystyrene-divinylbenzene microspheres ) 是由 刘德云 曾彬 任兴发 屠炳芳 于 2021-09-03 设计创作，主要内容包括：本发明提供一种单分散交联聚苯乙烯-二乙烯基苯微球的制备方法,包括：溶液A乳化后在惰性气体保护下原位聚合；然后轮流加入乳化后的溶液B和乳化后的溶液C进行原位沉积和交联获得所述单分散交联聚苯乙烯-二乙烯基苯微球；溶液A包括2wt％～40wt％苯乙烯、0.5wt％～30wt％二乙烯基苯、0.5wt％～5wt％引发剂、0.5wt％～15wt％稳定剂,余量为水；溶液B包括0.5wt％～5wt％引发剂,还包括5wt％～40wt％苯乙烯和/或1wt％～30wt％二乙烯基苯,余量为水；溶液C包括0.5wt％～15wt％稳定剂,余量为水。其在保证产品质量性能的同时采用水作为反应溶剂,聚合反应体系稳定可控。(The invention provides a preparation method of monodisperse crosslinked polystyrene-divinylbenzene microspheres, which comprises the following steps: emulsifying the solution A and then carrying out in-situ polymerization under the protection of inert gas; then adding the emulsified solution B and the emulsified solution C in turn for in-situ deposition and crosslinking to obtain the monodisperse crosslinked polystyrene-divinylbenzene microspheres; the solution A comprises 2 to 40 weight percent of styrene, 0.5 to 30 weight percent of divinylbenzene, 0.5 to 5 weight percent of initiator, 0.5 to 15 weight percent of stabilizer and the balance of water; the solution B comprises 0.5 to 5 weight percent of initiator, 5 to 40 weight percent of styrene and/or 1 to 30 weight percent of divinylbenzene, and the balance of water; the solution C comprises 0.5 to 15 weight percent of stabilizer and the balance of water. The method adopts water as a reaction solvent while ensuring the quality and performance of the product, and a polymerization reaction system is stable and controllable.)

1. A preparation method of monodisperse cross-linked polystyrene-divinylbenzene microspheres comprises the following steps:

emulsifying the solution A and then carrying out in-situ polymerization under the protection of inert gas; then adding the emulsified solution B and the emulsified solution C in turn for in-situ deposition and crosslinking to obtain the monodisperse crosslinked polystyrene-divinylbenzene microspheres;

the solution A comprises 2 to 40 weight percent of styrene, 0.5 to 30 weight percent of divinylbenzene, 0.5 to 5 weight percent of initiator, 0.5 to 15 weight percent of stabilizer and the balance of water; the solution B comprises 0.5 to 5 weight percent of initiator, 5 to 40 weight percent of styrene and/or 1 to 30 weight percent of divinylbenzene, and the balance of water; the solution C comprises 0.5 to 15 weight percent of stabilizer and the balance of water.

2. The preparation method according to claim 1, wherein the initiator is one or more selected from the group consisting of azobisisobutyronitrile, dimethyl azobisisobutyrate, benzoyl peroxide tert-butyl ester, potassium persulfate, sodium persulfate, and ammonium persulfate.

3. The method according to claim 1, wherein the stabilizer is selected from one or more of polyvinyl alcohol, polyethylene glycol, sodium dodecyl sulfate, sodium dodecyl benzene sulfonate, polyvinylpyrrolidone, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, β -cyclodextrin, gelatin, lignin, tween and span.

4. The method according to claim 1, wherein the weight ratio of the solution A to the solution B is (1-3): (1-55).

5. The method according to claim 1, wherein the weight ratio of the solution B to the solution C is (1-6): (1-20).

6. The method according to claim 1, wherein the solution B and the solution C are added dropwise to the solution A.

7. The method according to claim 1, wherein the solution A is reacted for 1 to 4 hours, and then the solution B is added.

8. The process according to claim 1, wherein the solution C is further added dropwise after the addition of B is completed for at least 30 min.

9. The method of claim 1, wherein the in-situ polymerization temperature, the in-situ deposition temperature, and the crosslinking temperature are 65 to 95 ℃.

10. The method according to claim 1, wherein water is a good solvent for the initiator in the solution A, and water is a poor solvent for the initiator in the solution B.

Technical Field

The invention relates to the field of chromatographic packing, in particular to a preparation method of monodisperse crosslinked polystyrene-divinylbenzene microspheres.

Background

The saccharides are basic substances capable of expressing various biological activities in the bodies of animals and plants, are not only nutrients of organisms, but also participate in various vital activities by forming complexes with proteins, lipids and the like. Saccharides can be classified into monosaccharides, disaccharides, oligosaccharides and polysaccharides according to the degree of polymerization and condensation. The polysaccharide has complex biological activity and function in many aspects, especially the function on the body immunity function, and has become an important component in the research and development of natural medicines and health care products. At present, part of natural polysaccharide compounds are used clinically and show good curative effect. The analysis of the saccharides and sugar chains in the complex carbohydrates can provide valuable information for industries such as medical treatment, scientific research, food and the like. At present, the purification and analysis method of saccharides mainly takes various High Performance Liquid Chromatography (HPLC) modes as main modes, internal fillers mainly take monodisperse bonded silica gel stationary phases, and because the bonded phases of the silica gel fillers are easy to drop in the using process, and the silica gel has poor alkali resistance and is continuously corroded and decomposed under the condition of high pH value to limit the application, the monodisperse sulfonated polystyrene-divinylbenzene ion exclusion fillers with the characteristics of acid resistance, alkali resistance and high temperature resistance are increasingly valued by users. These characteristics of monodisperse sulfonated polystyrene-divinylbenzene fillers greatly improve the analytical efficiency thereof, and are widely used for the separation of sugars, sugar alcohols, organic acids, polypeptides and nucleic acids.

The preparation of the monodisperse polystyrene-divinylbenzene filler is mainly based on a seed swelling method and a dispersion polymerization method.

The seed swelling method is proposed in the beginning of 90 s by Ugelstad J. mainly using particles prepared by emulsion polymerization as seeds, and then carrying out multi-step swelling and polymerization by using monomers, a crosslinking agent, a swelling agent and the like to obtain polymer particles with larger particle size. Patent CN1412554A adopts a seed swelling method to synthesize chloromethylated polystyrene-divinylbenzene microspheres, and prepares sulfonated monodisperse cationic filler by amination and surface deposition on the basis of the chloromethylated polystyrene-divinylbenzene microspheres. However, polystyrene-divinylbenzene particles tend to absorb biological macromolecules irreversibly due to their strong hydrophobicity, resulting in a decrease in separation efficiency. Similarly, patent CN1132213A adopts a one-step seed swelling method to prepare polystyrene crosslinked microspheres and polyvinyl pyridine crosslinked microspheres, but also fails to avoid the problem of irreversible adsorption. The patent CN110314664A and the patent CN1785526A both adopt a seed swelling method to prepare the monodisperse chromatographic packing which can be used for glycosylated hemoglobin analysis, and the monodisperse chromatographic packing with hydrophilic surface is prepared by mainly utilizing hydrophilic monomers and the like to carry out swelling polymerization on seed particles. Although the irreversible adsorption of protein molecules by the filler is reduced to a certain extent, the preparation process of the filler is still complex. In a word, the seed swelling method has the advantages that the prepared filler particles are uniform, and the defects of the whole preparation process are complex operation, harsh conditions, overlong period, overhigh cost, excessive generated waste liquid and the like.

The dispersion polymerization is mainly characterized in that monomers, organic solvents such as methanol or ethanol and the like, oil-soluble initiators and the like are mixed into a uniform system before reaction, after the reaction is initiated by heating, polymer monomers begin to polymerize and reach a certain molecular weight, and then are separated out and wound to finally form polymer particles with a certain size. In the process of preparing the polystyrene-divinylbenzene crosslinked microspheres by using a dispersion polymerization method, when the crosslinked polystyrene microspheres are prepared by adopting one-time feeding, the proportion of a crosslinking agent divinylbenzene cannot be more than 1%, otherwise, the prepared particles are reduced in uniformity and are in a polydisperse state, and the materials are seriously agglomerated to cause complete failure of the reaction.

Disclosure of Invention

In view of the above-mentioned shortcomings of the prior art, the present invention aims to provide a simple, efficient, low-cost, and environmentally friendly method for preparing a monodisperse cross-linked polystyrene-divinylbenzene chromatographic packing for separation and analysis of saccharides, which is used to solve the problems in the prior art.

To achieve the above objects and other related objects, the present invention is achieved by the following technical solutions.

The invention provides a preparation method of monodisperse crosslinked polystyrene-divinylbenzene microspheres, which comprises the following steps:

emulsifying the solution A and then carrying out in-situ polymerization under the protection of inert gas; then adding the emulsified solution B and the emulsified solution C in turn for in-situ deposition and crosslinking to obtain the monodisperse crosslinked polystyrene-divinylbenzene microspheres;

the solution A comprises 2 to 40 weight percent of styrene, 0.5 to 30 weight percent of divinylbenzene, 0.5 to 5 weight percent of initiator, 0.5 to 15 weight percent of stabilizer and the balance of water; the solution B comprises 0.5 to 5 weight percent of initiator, 5 to 40 weight percent of styrene and/or 1 to 30 weight percent of divinylbenzene, and the balance of water; the solution C comprises 0.5 to 15 weight percent of stabilizer and the balance of water.

Preferably, the content of the styrene in the solution A is 8-25 wt%. More preferably, the content of styrene in the solution A is 10-20 wt%.

Preferably, the content of the divinylbenzene in the solution A is 2 to 15 wt%. More preferably, the content of the divinylbenzene in the solution A is 2 to 5 wt%.

Preferably, the content of the initiator in the solution A is 0.5-2 wt%. More preferably, the content of the initiator in the solution A is 0.5-1.5 wt%.

Preferably, the content of the stabilizer in the solution A is 1-8 wt%. More preferably, the content of the stabilizer in the solution A is 1-3 wt%.

Preferably, the content of the styrene in the solution B is 15-40 wt%. More preferably, the content of the styrene in the solution B is 30-40 wt%.

Preferably, the content of the divinylbenzene in the solution B is 10 to 25 wt%. More preferably, the content of the divinylbenzene in the solution B is 18 to 25 wt%.

Preferably, the content of the initiator in the solution B is 1-3 wt%. More preferably, the content of the initiator in the solution B is 1.5-2.5 wt%.

Preferably, the content of the initiator in the solution C is 2-10 wt%. More preferably, the content of the initiator in the solution C is 2-5 wt%.

Preferably, the initiator is selected from one or more of benzoyl peroxide, diisobutyronitrile, azobisisobutyronitrile, dimethyl azobisisobutyrate, benzoyl peroxide tert-butyl ester, potassium persulfate, sodium persulfate and ammonium persulfate.

Preferably, the stabilizer is selected from one or more of polyvinyl alcohol, polyethylene glycol, sodium dodecyl sulfate, sodium dodecyl benzene sulfonate, polyvinylpyrrolidone, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, beta-cyclodextrin, gelatin, lignin, tween and span.

Preferably, the weight ratio of the solution A to the solution B is (1-3): (1-55).

Preferably, the weight ratio of the solution B to the solution C is (1-6): (1-20).

Preferably, the solution A is added into the solution B after reacting for 1-4 hours.

Preferably, the initiator in solution A uses water as a good solvent, and the initiator in solution B uses water as a poor solvent. If the initiator in the solution A is one or more selected from potassium persulfate, sodium persulfate and ammonium persulfate; the initiator in the solution B is one or more of azodiisobutyronitrile, dimethyl azodiisobutyrate and benzoyl peroxide tert-butyl ester. Preferably, solution B and solution C are added dropwise to solution a.

Preferably, solution C is added dropwise at least 30min after addition of B is complete.

Preferably, the temperature for in-situ polymerization, in-situ deposition and crosslinking is 65-95 ℃.

Preferably, the reaction also comprises separation and purification steps after the reaction is finished. More preferably, the separation may be by centrifugation. More preferably, the purification is a wash, such as with one or both of water and alcohol.

The preparation method realizes the one-step preparation of the monodisperse polystyrene-divinylbenzene microspheres with different particle diameters and different crosslinking degrees by adopting the styrene in-situ polymerization and the method of continuously dripping the divinylbenzene crosslinking agent for in-situ deposition and crosslinking, and is used for chromatographic packing for separation and analysis of saccharides.

The monodisperse crosslinked polystyrene-divinylbenzene microsphere obtained by the method has the particle size of 3-10 mu m, good monodispersity, stable and controllable polymerization reaction system and no caking phenomenon in the reaction process.

The technical scheme of the invention has the beneficial effects that:

the preparation method adopts an emulsion polymerization method, ensures the quality and performance of the product, adopts water as a reaction solvent, has no caking phenomenon in the reaction process, is stable and controllable in a polymerization reaction system, and can finally synthesize the microspheres with good monodispersity of 3-10 mu m. In addition, the invention can effectively shorten the production period, simplify the production flow, and reduce the use of organic reagents and the trouble of subsequent recovery treatment, thereby reducing waste and cost.

Drawings

FIG. 1 shows a scanning electron micrograph of crosslinked polystyrene-divinylbenzene microspheres prepared in example 1 of the present invention.

FIG. 2 shows a scanning electron micrograph of crosslinked polystyrene-divinylbenzene microspheres prepared in example 2 of the present invention.

Detailed Description

The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the respective manufacturers.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

Example 1

This example is the preparation of 3 μm crosslinked polystyrene-divinylbenzene microspheres:

in a 250mL beaker, 18.39 wt% styrene, 4.77 wt% divinylbenzene, 72.66 wt% deionized water, 1.36 wt% potassium persulfate, 2.82 wt% hydroxypropyl cellulose were weighed, dissolved with stirring at 300rpm to obtain an emulsified solution A and transferred to a 500mL three-necked flask.

Another 250mL beaker was weighed 31.75 wt% styrene, 19.05 wt% divinylbenzene, 47.62 wt% water, 1.58 wt% azobisisobutyronitrile, and stirred uniformly at 300rpm to obtain emulsified solution B.

2.44 wt% sodium dodecyl sulfate solution in 97.56 wt% deionized water to obtain solution C.

Placing the three-neck flask in an oil bath kettle, starting stirring, heating to 85 ℃, starting to slowly dropwise add 30mL of solution B into the flask after reacting for 2 hours under the protection of inert gas, and dropwise adding 20mL of solution C after 30 minutes. The process is circulated until the solution B, C is completely added. The reaction was continued for 4 hours.

The reaction was then turned off, and the reaction was washed 3 times each with ethanol and deionized water, then dried under vacuum at 70 ℃. The results are shown in FIG. 1.

Example 2

In this example, the preparation of 10 μm crosslinked polystyrene-divinylbenzene microspheres:

10.01 wt% of styrene, 85.31 wt% of deionized water, 0.67 wt% of sodium persulfate and 1.78 wt% of polyvinylpyrrolidone were weighed in a 500mL beaker, and dissolved with stirring at 300rpm to obtain solution A which was transferred to a 500mL three-necked flask.

Another 250mL beaker was weighed to obtain 37.57 wt% styrene, 24.42 wt% divinylbenzene, 35.78 wt% water, 2.23 wt% azobisisobutyronitrile, and stirred at 300rpm to obtain solution B.

4.76 wt% polyvinylpyrrolidone was dissolved in 95.24 wt% n-butanol to obtain solution C.

Placing the three-neck flask in an oil bath, starting stirring and heating to 75 ℃, starting to slowly dropwise add 30mL of solution B into the flask after reacting for 2 hours under the protection of inert gas, dropwise adding 20mL of solution C into the flask after 30 minutes, and circulating until the solution B, C is completely dropwise added. The reaction was continued for 6 hours.

The reaction was then turned off, and the reaction was washed 3 times each with ethanol and deionized water, then dried under vacuum at 70 ℃. The results are shown in FIG. 2.

Comparative example 1

Except that solution a did not contain hydroxypropylcellulose and the amount of deionized water was 75.48 wt%, the same as in example 1.

In the comparative example, no stabilizer is added in the stage of forming the polymerization seeds by reaction, so that the particle size of the seeds formed by in-situ polymerization is uncontrollable, and finally formed microspheres have inconsistent particle sizes and poor monodispersity and do not meet the use requirement of chromatography.

Comparative example 2

The difference from example 1 is that solution A and solution C do not contain a stabilizer, and the stabilizer content in solution A and solution C is replaced by water. Therefore, agglomeration is generated in the reaction process, and the microspheres with good monodispersity cannot be effectively synthesized.

The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.