阅读说明：本技术 一种杂化硅胶色谱填料的制备方法 (Preparation method of hybrid silica gel chromatographic packing ) 是由 范大双 韩海峰 张明亮 张大兵 于 2020-06-12 设计创作，主要内容包括：一种杂化硅胶色谱填料的制备方法,以1,2-二(三乙氧基硅基)乙烷和有机硅烷在表面活性剂溶液中溶胶凝胶得到杂化硅球,再后续N,N-二甲基十六胺水热扩孔制备杂化硅胶色谱填料,得到的杂化硅胶填料还可以进行后续改性。本发明制备的杂化硅胶球形度高、粒径均一、金属离子含量低、比表面积大、孔径大,同时水热稳定性、化学稳定性和机械稳定性强,直接共聚不需要商品化硅胶填料不需要甲苯等有毒溶剂不需要氮气保护,后续可改性强。(A method for preparing hybridized silica gel chromatographic packing comprises the steps of carrying out sol-gel on 1, 2-di (triethoxysilyl) ethane and organosilane in a surfactant solution to obtain hybridized silica spheres, carrying out subsequent hydrothermal reaming on N, N-dimethylhexadecylamine to prepare the hybridized silica gel chromatographic packing, and carrying out subsequent modification on the obtained hybridized silica gel chromatographic packing. The hybrid silica gel prepared by the invention has the advantages of high sphericity, uniform particle size, low metal ion content, large specific surface area, large pore diameter, strong hydrothermal stability, chemical stability and mechanical stability, no need of commercial silica gel filler for direct copolymerization, no need of toxic solvents such as toluene and the like, no need of nitrogen protection, and strong subsequent modification.)

1. A preparation method of a hybrid silica gel chromatographic packing is characterized by comprising the following steps:

the method comprises the following steps: preparing a template solution;

step two: carrying out sol on the template solution to obtain a sol mixture;

step three: the sol mixture is gelled to obtain small-hole silicon spheres;

step four: reaming the small-hole silicon ball to obtain silica gel containing a template agent;

step five: and removing the template agent from the silica gel containing the template agent to obtain the hybrid silica gel filler.

2. The method according to claim 1, characterized in that said step one is in particular: weighing a certain amount of surfactant, alkali, alcohol and water in turn, and stirring at 15-35 ℃ until the mixture is clear in a round-bottom flask to obtain a template solution.

3. The method of claim 1, wherein: the second step is specifically as follows: dropwise adding a certain amount of 1, 2-bis (triethoxysilyl) ethane BTEE alcohol solution and organosilane alcohol solution into the template solution obtained in the step one under strong stirring, and continuously stirring for 10-20 hours at 15-35 ℃ to obtain a sol mixture.

4. The method according to claim 1, wherein the third step is specifically: and (3) transferring the sol mixture obtained in the step two into a polypropylene plastic bottle, standing and aging at the temperature of 80-140 ℃ for 10-40 hours under the hydrothermal condition, then filtering, washing with deionized water and ethanol, and drying at the temperature of 50-80 ℃ to obtain the small-hole silica gel.

5. The method according to claim 1, wherein the fourth step is specifically: and (3) adding the small-hole silicon spheres obtained in the third step into a hydrothermal kettle containing an aqueous solution of N, N-dimethylhexadecylamine DMHA and ammonium fluoride NH4F, stirring for 1-5 hours at 15-35 ℃, then placing into an oven for standing for 1-5 days at 100-150 ℃, filtering, and washing with deionized water, ethanol and isopropanol to obtain the silica gel containing the template agent.

6. The method according to claim 1, wherein the step five is specifically: and D, extracting the silica gel containing the template agent obtained in the step four by using an alcoholic solution containing a small amount of acid to remove the template agent, and drying at 50-80 ℃ to obtain the hybrid silica gel filler.

7. The method according to claim 3, wherein the molar ratio of the reaction raw materials of the sol mixture is organosilane to surfactant to alkali to alcohol to water =1:0.8 to 2.0:1.5 to 3.5:40 to 100:250 to 450.

8. The method according to claim 2, characterized in that the surfactant is an alkyltrimethylammonium halide or a combination of alkyltrimethylammonium halide and dodecylamine DDA in a ratio of alkyltrimethylammonium halide/dodecylamine DDA molar ratio of 0.7:0.3 to 1.0:0, preferably the alkyltrimethylammonium halide is cetyltrimethylammonium bromide CTAB, cetyltrimethylammonium chloride CTAC, octadecyltrimethylammonium bromide, octadecyltrimethylammonium chloride.

9. The method of claim 3, wherein the molar ratio of organosilane to 1, 2-bis (triethoxysilyl) ethane is organosilane: 1, 2-bis (triethoxysilyl) ethane =0.05: 0.95-0.35: 0.65.

10. Process according to claim 3, characterized in that the organosilanes comprise phenyltriethoxysilane PhTES, mercaptopropyltriethoxysilane MPTES, vinyltriethoxysilane VTES, methyltriethoxysilane MTES, butyltriethoxysilane BTES, octyltriethoxysilane OTES, chloropropyltriethoxysilane CPTES, aminopropyltriethoxysilane APTES, tetraethylorthosilicate TEOS.

11. The method according to claim 2, characterized in that the base is an alkali metal hydroxide or an ammonium hydroxide.

12. The method according to claim 1, further comprising a sixth step of subjecting the hybrid silica gel obtained in the fifth step to subsequent modification (oxidation, click chemistry, chiral bonding, ionic liquid modification) to obtain reversed phase hybrid silica spheres, ion exchange hybrid silica spheres, hydrophilic reversed phase hybrid silica spheres, chiral hybrid silica spheres, ionic liquid modified hybrid silica spheres.

Technical Field

The invention relates to a preparation method of a chromatographic packing, in particular to a preparation method of a hybrid silica gel chromatographic packing.

Background

The traditional silica gel filler has many defects in the separation and purification process of products, such as poor chemical stability; the usable pH value range is narrow; silica gel itself is unstable and readily soluble under alkaline conditions (pH > 8); under acidic conditions (pH <2), the covalent bond connecting the active group and the silica gel surface can be hydrolyzed and broken, thereby causing the loss of the bonded silica gel stationary phase and becoming a main factor influencing the separation effect of the silica gel matrix liquid chromatography stationary phase. The hybrid silica gel matrix filler has the characteristics of good chemical stability, wide applicable pH range, difficult generation of irreversible nonspecific adsorption and the like, and has the defects of good pressure resistance, swelling in an organic solvent and the like; the commercial chromatographic column packing is prepared by a subsequent grafting method, namely, organosilane is bonded to silicon hydroxyl on spherical silica gel under the anhydrous and oxygen-free conditions, the method has strict operating conditions, a large amount of toxic organic solvents (such as toluene, pyridine and the like) are required, and the specific surface area of the method is strongly limited by silica gel raw materials. The market has a great need for chromatographic packing with strong chemical stability, strong mechanical stability and little pollution in the preparation process.

Disclosure of Invention

The invention provides a preparation method of a hybrid silica gel chromatographic packing, which comprises the following specific technical scheme:

a preparation method of a hybrid silica gel chromatographic packing is characterized by comprising the following steps:

the method comprises the following steps: preparing a template solution;

step two: carrying out sol on the template solution to obtain a sol mixture;

step three: the sol mixture is gelled to obtain small-hole silicon spheres;

step four: reaming the small-hole silicon ball to obtain silica gel containing a template agent;

step five: and removing the template agent from the silica gel containing the template agent to obtain the hybrid silica gel filler.

The first step is specifically as follows: weighing a certain amount of surfactant, alkali, alcohol and water in turn, and stirring at 15-35 ℃ until the mixture is clear in a round-bottom flask to obtain a template solution.

The second step is specifically as follows: dropwise adding a certain amount of 1, 2-bis (triethoxysilyl) ethane BTEE alcohol solution and organosilane alcohol solution into the template solution obtained in the step one under strong stirring, and continuously stirring for 10-20 hours at 15-35 ℃ to obtain a sol mixture.

The third step is specifically as follows: and (3) transferring the sol mixture obtained in the step two into a polypropylene plastic bottle, standing and aging at the temperature of 80-140 ℃ for 10-40 hours under the hydrothermal condition, then filtering, washing with deionized water and ethanol, and drying at the temperature of 50-80 ℃ to obtain the small-hole silica gel.

And step four, specifically, adding the small-pore silicon spheres obtained in the step three into a hydrothermal kettle containing an aqueous solution of N, N-dimethylhexadecylamine DMHA and ammonium fluoride NH4F, stirring for 1-5 hours at 15-35 ℃, then placing into an oven, standing for 1-5 days at 100-150 ℃, filtering, and washing with deionized water, ethanol and isopropanol to obtain the silica gel containing the template agent.

And step five, specifically, extracting the silica gel containing the template agent obtained in the step four by using an alcoholic solution containing a small amount of acid to remove the template agent, and drying at 50-80 ℃ to obtain the hybrid silica gel filler.

Preferably, the molar ratio of the reaction raw materials of the sol mixture is organosilane to surfactant to alkali to alcohol to water =1: 0.8-2.0: 1.5-3.5: 40-100: 250-450.

The surfactant is alkyl trimethyl ammonium halide or the combination of alkyl trimethyl ammonium halide and dodecylamine DDA, the proportion of the combination is that the molar ratio of alkyl trimethyl ammonium halide to dodecylamine DDA is 0.7:0.3-1.0:0, and the alkyl trimethyl ammonium halide is preferably cetyl trimethyl ammonium bromide CTAB, cetyl trimethyl ammonium chloride CTAC, octadecyl trimethyl ammonium bromide and octadecyl trimethyl ammonium chloride.

The molar ratio of the organosilane to the 1, 2-bis (triethoxysilyl) ethane is organosilane: 1, 2-bis (triethoxysilyl) ethane =0.05: 0.95-0.35: 0.65.

The organosilane includes phenyltriethoxysilane PhTES, mercaptopropyltriethoxysilane MPTES, vinyl triethoxysilane VTES, methyl triethoxysilane MTES, butyltriethoxysilane BTES, octyltriethoxysilane OTES, chloropropyltriethoxysilane CPTES, aminopropyltriethoxysilane APTES, tetraethyl orthosilicate TEOS.

The alkali is hydroxide of alkali metal or hydroxide of ammonium.

And step six, performing subsequent modification (oxidation, click chemistry, chiral bonding and ionic liquid modification) on the hybrid silica gel obtained in the step five to obtain reversed phase hybrid silica spheres, ion exchange hybrid silica spheres, hydrophilic reversed phase hybrid silica spheres, chiral hybrid silica spheres and ionic liquid modified hybrid silica spheres.

The invention has the beneficial effects that: (1) the sphericity is high and the particle size is uniform; (2) the content of metal ions is low; (3) large specific surface area (300-600 m)² g^-1) The aperture is large; (4) the hydrothermal stability, the chemical stability and the mechanical stability are strong; (5) direct copolymerization is carried out, commercial silica gel filler is not needed, toxic solvents such as toluene and the like are not needed, and nitrogen protection is not needed; (6) the obtained hybrid silica gel has strong subsequent modification (oxidation, click chemistry, chiral bonding, ionic liquid modification and the like), and different types of fillers (reverse phase, ionic type, amphiprotic type, ionic liquid type, chiral and the like) can be obtained.

Drawings

FIG. 1 microscopic image of phenyl hybrid filler

FIG. 2 pore size distribution curves for phenyl hybrid fillers

FIG. 3 microscopic image of vinyl hybrid fillers

FIG. 4 pore size distribution curves for vinyl hybrid fillers

FIG. 5 microscopic image of mercapto hybrid filler

FIG. 6 microscopic image of methyl hybrid filler

FIG. 7. microscopic image of ethoxy hybrid filler.

Examples

The present invention will be described in detail with reference to examples

Example 1

15.0g of cetyltrimethylammonium bromide, 1.0g of dodecylamine, 250g of deionized water, 80g of methanol and 5.5g of potassium hydroxide were weighed in this order and placed in a 500mL round bottom flask and stirred at 20 ℃ until completely dissolved to give a clear and clear solution. 1, 2-bis (triethoxysilyl) ethane methanol solution (10.66 g BTEE +10g methanol) and phenyltriethoxysilane methanol solution (3.36 g PhTES +10g methanol) were slowly added dropwise simultaneously on both sides with vigorous stirring, stirring was continued at 25 ℃ for 20 hours, and then transferred to a polypropylene plastic bottle, allowed to stand at 90 ℃ for 30 hours, filtered and washed with deionized water, and dried at 50 ℃.

Weighing 1g of the crude product in a polytetrafluoroethylene hydrothermal kettle, adding 0.5g of N, N-dimethylhexadecylamine DMHA, 60mg of ammonium fluoride NH4F and 30g of water, stirring for 1 hour at room temperature, standing for 1.5 days at 120 ℃, and then filtering, washing and drying to obtain a white powder crude product.

Adding the crude product into 200g of ethanol solution containing concentrated hydrochloric acid (5 g of 36 wt%), performing Soxhlet extraction at 60 ℃ for 8 hours, filtering, washing with deionized water and ethanol for several times, repeating the extraction operation, and drying at 50 ℃ to obtain the phenyl hybrid silica gel filler.

The appearance of the phenyl hybrid silica gel filler of the embodiment is observed by using a binocular biomicroscope XSP-8C, and the phenyl hybrid silica gel filler has the advantages that the surface of spherical particles is smooth, the particle size is 3-6 mu m, and the particle size is uniform; the specific surface area and pore parameters were measured by McTriStar II 3020 physical adsorption apparatus, the pore size distribution curve is shown in FIG. 2, and the specific surface area is 457m² g^-1Average pore diameter 13.7 nm.

The metal ion content was determined using a seemer fly inductively coupled plasma emission spectrometer iCAP 7000, see table 1 for details. It can be seen from the table that all the metal ion contents are lower than 50ppm, the contents of Al, Ca, Fe and Na are slightly higher, and the contents of the rest metal ions are lower than 10ppm, so that the requirements of the liquid chromatography packing on the metal ion contents are met.

TABLE 1 Metal ion content of hybrid silica gel Filler

Example 2

15g of cetyltrimethylammonium chloride, 1.5g of dodecylamine, 250g of deionized water, 70g of methanol and 4.0g of sodium hydroxide are weighed in sequence, placed in a 500mL round-bottom flask, and stirred at 30 ℃ until completely dissolved to obtain a transparent clear solution. 1, 2-bis (triethoxysilyl) ethane methanol solution (13.74 g BTEE +15g methanol) and vinyltriethoxysilane methanol solution (1.08 g VTES +15g methanol) were slowly added dropwise simultaneously on both sides with vigorous stirring, stirring was continued at 30 ℃ for 20 hours, and then transferred to a polypropylene plastic bottle, allowed to stand at 95 ℃ for 24 hours, filtered and washed with deionized water, and dried at 60 ℃.

Weighing 1g of the crude product in a polytetrafluoroethylene hydrothermal kettle, adding 0.5g of N, N-dimethylhexadecylamine DMHA, 60mg of ammonium fluoride NH4F and 30g of water, stirring for 2 hours at room temperature, standing for 1 day at 125 ℃, and then filtering, washing and drying to obtain a white powder crude product.

Adding the crude product into 150g of ethanol solution containing concentrated hydrochloric acid (5 g of 36 wt%), performing Soxhlet extraction at 70 ℃ for 8 hours, filtering, washing with deionized water and ethanol for several times, and repeating the extraction operation; drying at 55 ℃ to obtain the vinyl hybrid silica gel filler.

The morphology of the vinyl hybrid silica gel filler of the embodiment was observed with a binocular biomicroscope XSP-8C, which is specifically shown in FIG. 3; the method has the advantages that the surface of the spherical particles is smooth, the particle size is 3-6 mu m, and the particle size is uniform; the specific surface area and pore parameters were measured by a Michelrastar II 3020 physical adsorption apparatus, the pore size distribution curve is shown in FIG. 4, and the specific surface area is 336m² g^-1Average pore diameter is 7.8 nm.

The content of metal ions is measured by a Sammer fly inductively coupled plasma emission spectrometer iCAP 7000, which is specifically shown in Table 1; it can be seen from the table that all the metal ion contents are lower than 50ppm, the contents of Al, Ca, Fe and Na are slightly higher, and the contents of the rest metal ions are lower than 10ppm, so that the requirements of the liquid chromatography packing on the metal ion contents are met.

Example 3

Weighing 16g of octadecyl trimethyl ammonium bromide, 1.5g of dodecylamine, 300g of deionized water, 60g of methanol and 6g of potassium hydroxide in sequence, placing the weighed materials in a 500mL round-bottom flask, and stirring at 35 ℃ until the materials are completely dissolved to obtain a transparent clear solution; 1, 2-bis (triethoxysilyl) ethane methanol solution (13.74 g BTEE +5g methanol) and mercaptopropyltriethoxysilane methanol solution (3.36 g MPTES +5g methanol) were slowly added dropwise simultaneously on both sides with vigorous stirring, stirring was continued at 35 ℃ for 20 hours, and then transferred to a polypropylene plastic bottle, allowed to stand at 95 ℃ for 30 hours, filtered, washed with deionized water, and dried at 50 ℃.

Weighing 1g of the crude product in a polytetrafluoroethylene hydrothermal kettle, adding 0.5g of N, N-dimethylhexadecylamine DMHA, 30mg of ammonium fluoride NH4F and 30g of water, stirring for 2 hours at room temperature, standing for 1 day at 120 ℃, and then filtering, washing and drying to obtain a white powder crude product.

Adding the crude product into 150g of ethanol solution containing concentrated hydrochloric acid (5 g of 36 wt%), performing Soxhlet extraction at 60 ℃ for 8 hours, filtering, washing with deionized water and ethanol for several times, and repeating the extraction operation; drying at 65 ℃ to obtain the sulfhydryl hybridized silica gel filler.

The appearance of the mercapto hybrid silica gel filler of the present example was observed with a binocular biomicroscope XSP-8C, specifically shown in fig. 5; the method has the advantages that the surface of the spherical particles is smooth, the particle size is 3-6.5 mu m, and the particle size is uniform; the specific surface area, pore size, pore distribution, and metal ion content were similar to those of example 1.

Example 5

14g of octadecyl trimethyl ammonium chloride, 250g of deionized water, 80g of methanol and 3.8g of sodium hydroxide are weighed in sequence, placed in a 500mL round bottom flask, and stirred at 25 ℃ until the octadecyl trimethyl ammonium chloride, the deionized water, the methanol and the sodium hydroxide are completely dissolved to obtain a transparent clear solution. While stirring vigorously, a solution of 1, 2-bis (triethoxysilyl) ethane in methanol (11.0 g BTEE +10g methanol) and a solution of methyltriethoxysilane in methanol (2.6 g MTES +10g methanol) were added dropwise slowly on both sides simultaneously, stirring was continued for 20 hours at 25 ℃ and then transferred to a polypropylene plastic bottle, left to stand at 95 ℃ for 30 hours, filtered and washed with deionized water, and dried at 65 ℃.

Weighing 1g of the crude product in a polytetrafluoroethylene hydrothermal kettle, adding 0.15g of N, N-dimethylhexadecylamine DMHA, 30mg of ammonium fluoride NH4F and 30g of water, stirring for 2 hours at room temperature, standing for 1 day at 120 ℃, and then filtering, washing and drying to obtain a white powder crude product.

Adding the crude product into 200g of ethanol solution containing concentrated hydrochloric acid (6 g of 36 wt%), performing Soxhlet extraction at 60 ℃ for 8 hours, filtering, washing with deionized water and ethanol for several times, and repeating the extraction operation; drying at 65 ℃ to obtain the methyl hybrid silica gel filler.

The methyl hybrid silica gel filler of the embodiment has the advantages that the surface of spherical particles is smooth, the particle size is 3.5-6 μm, the particle size is uniform, the appearance is shown in figure 6, and the specific surface area, the pore diameter, the pore distribution and the metal ion content are similar to those of example 1.

Example 6

16g of hexadecyl trimethyl ammonium bromide, 0.5g of dodecylamine, 250g of deionized water, 70g of methanol and 4.0g of sodium hydroxide are weighed in turn and placed in a 500mL round bottom flask, the mixture is stirred at 25 ℃ until the mixture is completely dissolved to obtain a transparent clear solution, a 1, 2-bis (triethoxysilyl) ethane methanol solution (10.5 g of BTEE +15g of methanol) and a tetraethoxysilane methanol solution (2.6 g of TEOS +15g of methanol) are slowly dripped into two sides of the flask simultaneously under vigorous stirring, the mixture is stirred for 20 hours at 25 ℃, and then the mixture is transferred into a polypropylene plastic bottle and is kept stand for 30 hours at 95 ℃, filtered, washed by deionized water and dried at 60 ℃.

Weighing 1g of the crude product in a polytetrafluoroethylene hydrothermal kettle, adding 0.15g of N, N-dimethylhexadecylamine DMHA, 30mg of ammonium fluoride NH4F and 30g of water, stirring for 2 hours at room temperature, standing for 1 day at 120 ℃, and then filtering, washing and drying to obtain a white powder crude product.

The crude product is added into 200g of ethanol solution containing concentrated hydrochloric acid (6 g of 36wt percent), Soxhlet extraction is carried out for 8 hours at the temperature of 60 ℃, filtration is carried out, deionized water and ethanol are used for washing for a plurality of times, the extraction operation is repeated, and the ethoxyl hybrid silica gel filler is obtained after drying at the temperature of 60 ℃.

The ethoxy hybrid silica gel filler of the embodiment has the advantages that the surface of spherical particles is smooth, the particle size is 4-7 μm, the particle size is uniform, the appearance is shown in figure 7, and the specific surface area, the pore diameter, the pore distribution and the metal ion content are similar to those of the example 1.

Example 7

3g of the vinyl hybrid silica gel filler obtained in the example 2 was put in a 100mL three-neck flask, 3g of octadecyl mercaptan, 70mL of toluene and 0.01g of azobisisobutyronitrile were added to the flask, stirred at 80 ℃ for 24 hours under the protection of nitrogen, filtered, washed with toluene, water and ethanol in sequence, and vacuum-dried to obtain a C18 modified vinyl hybrid silica gel filler.

Example 8

3.5g of the mercapto hybrid silica gel filler obtained in example 3 was placed in a 100mL three-necked flask, and 2g of 1-vinyl-3-octyl imidazole bromide, 12mL of chloroform and 0.03g of azobisisobutyronitrile were added to the flask, stirred at 60 ℃ for 24 hours, filtered, washed with chloroform, methanol and ethyl acetate in this order, and vacuum-dried to obtain 1-vinyl-3-octyl imidazole bromide modified hybrid silica gel filler.

TABLE 1 Metal ion content of hybrid silica gel Filler

Name (R)	Al	Ca	Fe	K	Mg	Na	Ti	Zr
									Phenyl hybrids	42.9	41.7	42.9	9.4	7.5	47.3	3.4	2.2
Vinyl hybridization	34.1	25.6	44.1	9.9	3.9	28.3	1.8	1.2

The present invention is illustrated by the above embodiments, but the present invention is not limited to the above detailed process equipment and process flow, i.e. it is not meant that the present invention is implemented only by relying on the above detailed process equipment and process flow; it should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.