阅读说明：本技术 一种苯并咪唑大孔吸附树脂的制备方法及其应用 (Preparation method and application of benzimidazole macroporous adsorption resin ) 是由 薛兆能 田文辉 于 2021-10-26 设计创作，主要内容包括：本发明公开了一种苯并咪唑大孔吸附树脂的制备方法及其应用,涉及大孔吸附树脂技术领域,本发明先通过悬浮聚合法制备苯乙烯-二乙烯基苯微球,然后采用氯甲醚对苯乙烯-二乙烯基苯微球进行氯甲基化,制得氯甲基化苯乙烯-二乙烯苯微球,最后利用2-甲巯基苯并咪唑与氯甲基化苯乙烯-二乙烯苯微球发生取代反应,制得苯并咪唑大孔吸附树脂,对黄酮类化合物显示出良好的吸附选择性。(The invention discloses a preparation method and application of benzimidazole macroporous adsorption resin, and relates to the technical field of macroporous adsorption resin.)

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

(1) adding a dispersing agent, sodium chloride and methylene blue into water, heating and stirring to completely dissolve the dispersing agent, and cooling to obtain a water phase; adding an oil phase consisting of styrene, divinyl benzene, a pore-forming agent and an initiator into the water phase, heating for reaction, stopping the reaction, alternately washing the obtained white balls with hot water and ethanol until the liquid is clear, and drying to obtain styrene-divinyl benzene microspheres;

(2) swelling the styrene-divinylbenzene microspheres obtained in the step (1) in chloromethyl ether, adding a catalyst, heating to carry out a chloromethylation reaction, washing with water, and drying to obtain chloromethylated styrene-divinylbenzene microspheres;

(3) swelling the chloromethylated styrene-divinylbenzene microspheres prepared in the step (2) in N, N-dimethylformamide, then adding 2-methylmercaptobenzimidazole and anhydrous potassium carbonate, heating for substitution reaction, washing with water, and drying to obtain the benzimidazole macroporous adsorption resin.

2. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the dispersant is at least one of polyvinyl alcohol, gelatin and hydroxyethyl cellulose.

3. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the pore-forming agent is isobutanol, and the initiator is benzoyl peroxide.

4. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the dosage of the dispersant is 0.5-1.5% of the mass of the water phase, the dosage of the sodium chloride is 0.05-0.1% of the mass of the water phase, and the dosage of the methylene blue is 0.01-0.05% of the mass of the water phase.

5. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the amount of the divinyl benzene is 20-30% of the mass of the styrene, the amount of the pore-foaming agent is 50-100% of the total mass of the styrene and the divinyl benzene, the amount of the initiator is 0.5-1% of the total mass of the styrene and the divinyl benzene, and the volume ratio of the water phase to the oil phase is (1-3): 1.

6. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the catalyst is at least one of anhydrous ferric chloride and anhydrous zinc chloride.

7. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the dosage of the catalyst is 20-50% of the mass of the styrene-divinylbenzene microspheres.

8. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the dosage of the 2-methylmercaptobenzimidazole is 15-30% of the mass of the chloromethylated styrene-divinylbenzene microsphere.

9. The preparation method of the benzimidazole macroporous adsorption resin according to claim 1, wherein the benzimidazole macroporous adsorption resin comprises the following steps: the molar ratio of the 2-methylmercaptobenzimidazole to the anhydrous potassium carbonate is 1 (1-1.2).

10. Use of the benzimidazole macroporous adsorbent resin prepared according to any one of claims 1-9 for the separation and purification of flavonoids.

The technical field is as follows:

the invention relates to the technical field of macroporous adsorption resin, in particular to a preparation method and application of benzimidazole macroporous adsorption resin.

Background art:

the macroporous adsorption resin is a polymer adsorption resin which does not contain exchange groups and has a macroporous structure, and can be divided into non-polar, weak-polar and polar resins according to the surface properties of the resin, wherein the non-polar resin is suitable for adsorbing non-polar substances from a polar solvent, and the polar resin is suitable for adsorbing polar substances from a non-polar solvent. The resin has adsorption property due to the action of hydrogen bond or van der waals force, and simultaneously has screening performance due to the net structure and the larger specific surface area, so the macroporous adsorption resin is a material with adsorption and screening capacities.

In recent years, macroporous adsorption resin is widely applied to extraction of effective components of Chinese herbal medicines, so that the separation operation of the effective components can be simplified, and the separated effective components have high purity and low loss. At present, the macroporous adsorption resin which is most applied is polymerized by taking styrene as a polymerization monomer and divinylbenzene as a cross-linking agent and adding a certain proportion of pore-foaming agent.

The extraction method of the effective components of the Chinese herbal medicines mainly comprises a solvent extraction method, a steam distillation method, a squeezing method and a sublimation method, and a person skilled in the art knows that the obtained extract is in a mixture form by adopting any extraction method, the content of the target effective components is very low, and separation and purification operations are required to separate the target effective components with high purity from the extract.

The macroporous adsorption resin adsorption method is a simple separation and purification method, but needs to improve the adsorption selectivity of the macroporous adsorption resin, and uses the macroporous adsorption resin with good selectivity on target active ingredients. However, the macroporous adsorption resin sold in the market at present cannot meet the single use requirement at all, so the invention synthesizes the macroporous adsorption resin with a special structure to be suitable for separating and purifying the effective components of the specific Chinese herbal medicines.

The invention content is as follows:

the invention aims to solve the technical problem of providing a preparation method of a benzimidazole macroporous adsorption resin, wherein the benzimidazole macroporous adsorption resin is prepared by functionally modifying styrene-divinylbenzene microspheres, and is applied to separation and purification of flavonoid compounds, so that the technical effect of remarkably improving the content of the flavonoid compounds in the product is achieved.

The technical problem to be solved by the invention is realized by adopting the following technical scheme:

the invention aims to provide a preparation method of a benzimidazole macroporous adsorption resin, which comprises the following preparation steps:

(1) adding a dispersing agent, sodium chloride and methylene blue into water, heating and stirring to completely dissolve the dispersing agent, and cooling to obtain a water phase; adding an oil phase consisting of styrene, divinyl benzene, a pore-forming agent and an initiator into the water phase, heating for reaction, stopping the reaction, alternately washing the obtained white balls with hot water and ethanol until the liquid is clear, and drying to obtain styrene-divinyl benzene microspheres;

(2) swelling the styrene-divinylbenzene microspheres obtained in the step (1) in chloromethyl ether, adding a catalyst, heating to carry out a chloromethylation reaction, washing with water, and drying to obtain chloromethylated styrene-divinylbenzene microspheres;

(3) swelling the chloromethylated styrene-divinylbenzene microspheres prepared in the step (2) in N, N-dimethylformamide, then adding 2-methylmercaptobenzimidazole and anhydrous potassium carbonate, heating for substitution reaction, washing with water, and drying to obtain the benzimidazole macroporous adsorption resin.

The dispersant is at least one of polyvinyl alcohol, gelatin and hydroxyethyl cellulose.

The pore-forming agent is isobutanol, and the initiator is benzoyl peroxide.

The dosage of the dispersant is 0.5-1.5% of the mass of the water phase, the dosage of the sodium chloride is 0.05-0.1% of the mass of the water phase, and the dosage of the methylene blue is 0.01-0.05% of the mass of the water phase.

The amount of the divinyl benzene is 20-30% of the mass of the styrene, the amount of the pore-foaming agent is 50-100% of the total mass of the styrene and the divinyl benzene, the amount of the initiator is 0.5-1% of the total mass of the styrene and the divinyl benzene, and the volume ratio of the water phase to the oil phase is (1-3): 1.

The styrene-divinylbenzene microspheres prepared in the step (1) belong to conventional macroporous adsorption resin in the field, and although the adsorption property is strong, the adsorption selectivity of the styrene-divinylbenzene microspheres to flavonoids compounds is poor, so that the high-efficiency separation and purification of the flavonoids compounds cannot be realized.

The catalyst is at least one of anhydrous ferric chloride and anhydrous zinc chloride.

The dosage of the catalyst is 20-50% of the mass of the styrene-divinylbenzene microspheres.

Chloromethyl ether is adopted to perform chloromethylation on the styrene-divinylbenzene microspheres in the step (2), chloromethyl is introduced to the styrene-divinylbenzene microspheres, and chloromethyl is taken as an active group, so that subsequent substitution reaction is facilitated.

The dosage of the 2-methylmercaptobenzimidazole is 15-30% of the mass of the chloromethylated styrene-divinylbenzene microsphere.

The molar ratio of the 2-methylmercaptobenzimidazole to the anhydrous potassium carbonate is 1 (1-1.2). Anhydrous potassium carbonate is adopted to neutralize hydrogen chloride generated in the reaction, so that the reaction rate is improved.

In the step (3), 2-methylmercaptobenzimidazole and chloromethyl are subjected to substitution reaction to prepare the benzimidazole macroporous adsorption resin, and the macroporous adsorption resin can show good adsorption selectivity on flavonoids, so that the flavonoids are efficiently separated and purified.

The invention also aims to provide the application of the benzimidazole macroporous adsorption resin in separation and purification of flavonoids compounds. The benzimidazole macroporous adsorption resin prepared by the invention is used for separating and purifying the Chinese herbal medicine extract containing the flavonoid compounds, and the high-content flavonoid compounds are obtained after adsorption-desorption.

The invention has the beneficial effects that:

(1) the preparation method comprises the steps of preparing styrene-divinylbenzene microspheres by a suspension polymerization method, carrying out chloromethylation on the styrene-divinylbenzene microspheres by using chloromethyl ether to prepare chloromethylated styrene-divinylbenzene microspheres, and carrying out substitution reaction on 2-methylmercaptobenzimidazole and the chloromethylated styrene-divinylbenzene microspheres to prepare the benzimidazole macroporous adsorption resin;

(2) the benzimidazole macroporous adsorption resin prepared by the invention has good adsorption selectivity on flavonoid compounds, is easy to desorb, can obtain high-content flavonoid compounds after desorption, and is suitable for separation and purification of the flavonoid compounds, so that the separation and purification operation of the flavonoid compounds is simplified, the loss of the flavonoid compounds in the separation and purification process is reduced, and the purity of the flavonoid compounds in the product is improved.

The specific implementation mode is as follows:

in order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the specific embodiments.

Example 1

(1) Adding gelatin, sodium chloride and methylene blue into water, wherein the dosage of the gelatin is 1% of the mass of the water phase, the dosage of the sodium chloride is 0.1% of the mass of the water phase, and the dosage of the methylene blue is 0.02% of the mass of the water phase, heating and stirring to completely dissolve the dispersing agent, and cooling to obtain a water phase; adding an oil phase consisting of styrene, divinyl benzene, isobutanol and benzoyl peroxide into the water phase, wherein the dosage of the divinyl benzene is 25% of the mass of the styrene, the dosage of the isobutanol is 100% of the total mass of the styrene and the divinyl benzene, the dosage of the benzoyl peroxide is 0.5% of the total mass of the styrene and the divinyl benzene, the volume ratio of the water phase to the oil phase is 2:1, heating to 80 ℃, reacting for 5 hours, stopping the reaction, alternately washing the obtained white balls with hot water and ethanol until the liquid is clear, and drying to obtain the styrene-divinyl benzene microspheres;

(2) swelling the styrene-divinylbenzene microspheres obtained in the step (1) in chloromethyl ether, adding anhydrous zinc chloride accounting for 25% of the mass of the styrene-divinylbenzene microspheres, heating to 40 ℃, reacting for 12 hours, washing with water, and drying to obtain chloromethylated styrene-divinylbenzene microspheres;

(3) swelling the chloromethylated styrene-divinylbenzene microspheres prepared in the step (2) in N, N-dimethylformamide, adding 2-methylmercaptobenzimidazole and anhydrous potassium carbonate which account for 20% of the mass of the chloromethylated styrene-divinylbenzene microspheres, wherein the molar ratio of the 2-methylmercaptobenzimidazole to the anhydrous potassium carbonate is 1:1, heating to 60 ℃, reacting for 5 hours, washing with water, and drying to obtain the benzimidazole macroporous adsorption resin.

Example 2

(1) Adding hydroxyethyl cellulose, sodium chloride and methylene blue into water, wherein the dosage of a dispersing agent is 1.5 percent of the mass of the water phase, the dosage of the sodium chloride is 0.05 percent of the mass of the water phase, and the dosage of the methylene blue is 0.01 percent of the mass of the water phase, heating and stirring to completely dissolve the dispersing agent, and cooling to obtain the water phase; adding an oil phase consisting of styrene, divinyl benzene, isobutanol and benzoyl peroxide into the water phase, wherein the dosage of the divinyl benzene is 25% of the mass of the styrene, the dosage of the isobutanol is 100% of the total mass of the styrene and the divinyl benzene, the dosage of the benzoyl peroxide is 1% of the total mass of the styrene and the divinyl benzene, the volume ratio of the water phase to the oil phase is 1:1, heating to 80 ℃, reacting for 5 hours, stopping the reaction, alternately washing the obtained white spheres with hot water and ethanol until the liquid is clear, and drying to obtain the styrene-divinyl benzene microspheres;

(2) swelling the styrene-divinylbenzene microspheres obtained in the step (1) in chloromethyl ether, adding anhydrous ferric chloride with the mass of 30% of that of the styrene-divinylbenzene microspheres, heating to 40 ℃, reacting for 12 hours, washing with water, and drying to obtain chloromethylated styrene-divinylbenzene microspheres;

(3) swelling the chloromethylated styrene-divinylbenzene microspheres prepared in the step (2) in N, N-dimethylformamide, adding 2-methylmercaptobenzimidazole and anhydrous potassium carbonate which account for 15% of the mass of the chloromethylated styrene-divinylbenzene microspheres, wherein the molar ratio of the 2-methylmercaptobenzimidazole to the anhydrous potassium carbonate is 1:1.1, heating to 60 ℃, reacting for 5 hours, washing with water, and drying to obtain the benzimidazole macroporous adsorption resin.

Example 3

(1) Adding polyvinyl alcohol, sodium chloride and methylene blue into water, wherein the dosage of a dispersing agent is 0.5 percent of the mass of the water phase, the dosage of the sodium chloride is 0.1 percent of the mass of the water phase, and the dosage of the methylene blue is 0.05 percent of the mass of the water phase, heating and stirring to completely dissolve the dispersing agent, and cooling to obtain the water phase; adding an oil phase consisting of styrene, divinyl benzene, isobutanol and benzoyl peroxide into the water phase, wherein the dosage of the divinyl benzene is 20% of the mass of the styrene, the dosage of the isobutanol is 50% of the total mass of the styrene and the divinyl benzene, the dosage of the benzoyl peroxide is 0.8% of the total mass of the styrene and the divinyl benzene, the volume ratio of the water phase to the oil phase is 3:1, heating to 80 ℃, reacting for 5 hours, stopping the reaction, alternately washing the obtained white balls with hot water and ethanol until the liquid is clear, and drying to obtain the styrene-divinyl benzene microspheres;

(2) swelling the styrene-divinylbenzene microspheres obtained in the step (1) in chloromethyl ether, adding anhydrous zinc chloride accounting for 35% of the mass of the styrene-divinylbenzene microspheres, heating to 40 ℃, reacting for 12 hours, washing with water, and drying to obtain chloromethylated styrene-divinylbenzene microspheres;

(3) swelling the chloromethylated styrene-divinylbenzene microspheres prepared in the step (2) in N, N-dimethylformamide, adding 2-methylmercaptobenzimidazole and anhydrous potassium carbonate which account for 30% of the mass of the chloromethylated styrene-divinylbenzene microspheres, wherein the molar ratio of the 2-methylmercaptobenzimidazole to the anhydrous potassium carbonate is 1:1.2, heating to 60 ℃, reacting for 5 hours, washing with water, and drying to obtain the benzimidazole macroporous adsorption resin.

Example 4

(1) Adding polyvinyl alcohol, sodium chloride and methylene blue into water, wherein the dosage of a dispersing agent is 1% of the mass of the water phase, the dosage of the sodium chloride is 0.1% of the mass of the water phase, and the dosage of the methylene blue is 0.03% of the mass of the water phase, heating and stirring to completely dissolve the dispersing agent, and cooling to obtain the water phase; adding an oil phase consisting of styrene, divinyl benzene, isobutanol and benzoyl peroxide into the water phase, wherein the dosage of the divinyl benzene is 30% of the mass of the styrene, the dosage of the isobutanol is 100% of the total mass of the styrene and the divinyl benzene, the dosage of the benzoyl peroxide is 1% of the total mass of the styrene and the divinyl benzene, the volume ratio of the water phase to the oil phase is 1.5:1, heating to 80 ℃, reacting for 5 hours, stopping the reaction, alternately washing the obtained white balls with hot water and ethanol until the liquid is clear, and drying to obtain the styrene-divinyl benzene microspheres;

(2) swelling the styrene-divinylbenzene microspheres obtained in the step (1) in chloromethyl ether, adding anhydrous zinc chloride accounting for 30% of the mass of the styrene-divinylbenzene microspheres, heating to 40 ℃, reacting for 12 hours, washing with water, and drying to obtain chloromethylated styrene-divinylbenzene microspheres;

(3) swelling the chloromethylated styrene-divinylbenzene microspheres prepared in the step (2) in N, N-dimethylformamide, adding 2-methylmercaptobenzimidazole and anhydrous potassium carbonate which account for 25% of the mass of the chloromethylated styrene-divinylbenzene microspheres, wherein the molar ratio of the 2-methylmercaptobenzimidazole to the anhydrous potassium carbonate is 1:1.1, heating to 60 ℃, reacting for 5 hours, washing with water, and drying to obtain the benzimidazole macroporous adsorption resin.

Comparative example 1

Comparative example 1 was conducted by replacing 2-methylmercaptobenzimidazole with benzimidazole in step (3) of example 4, and the rest was not changed.

Comparative example 2

Comparative example 2 in example 4, 2-methylmercaptobenzimidazole was replaced with dimethylamine in step (3), and the rest of the procedure was not changed.

Comparative example 3

Comparative example 3 the procedure of example 4, step (2) and step (3), was deleted and the rest of the procedure was unchanged.

Loading 100g of camellia seed hulls into an extraction tank, adding 1000g of 70% ethanol, performing reflux extraction at 50 ℃ for three times, performing first extraction for 2 hours, performing second extraction for 1 hour, performing third extraction for 1 hour, filtering, collecting extract, combining, concentrating and recovering ethanol to obtain a water phase; filling the benzimidazole macroporous adsorption resin into a column, wherein the mass ratio of the oil tea seed shells to the benzimidazole macroporous adsorption resin is 1:1, enabling the water phase to pass through the resin column at the flow rate of 0.5BV/h, then sequentially eluting with water, 30% ethanol and 50% ethanol at the flow rate of 2BV/h, collecting 50% ethanol eluent, concentrating and drying to obtain the product.

The content of flavonoids in the product was determined by HPLC method and the results are shown in the following table.

	Content of flavonoids/%
		Example 1	59.15
Example 2	54.26
		Example 3	65.38
Example 4	63.52
		Comparative example 1	50.23
Comparative example 2	24.60
		Comparative example 3	18.57

The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.