CN114832799B - Preparation and application of carbon-point bonded silica gel chromatographic stationary phase based on eutectic solvent - Google Patents
Preparation and application of carbon-point bonded silica gel chromatographic stationary phase based on eutectic solvent Download PDFInfo
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Abstract
The invention discloses a preparation method of a carbon-point bonded silica gel chromatographic stationary phase based on a eutectic solvent, which comprises the steps of mixing choline chloride and lactic acid according to a molar ratio, heating to form the eutectic solvent, reacting at 150-220 ℃ for 2-10 h to form carbon-point DESCDs based on the eutectic solvent, carrying out stirring reflux reaction on silica gel at 90-150 ℃ for 12-48 h by using 3-aminopropyltrimethoxysilane or 3-aminopropyltriethoxysilane to obtain Sil-MPS through modification, mixing the DESCDs with the Sil-MPS, adding an initiator, and carrying out stirring reaction at 50-90 ℃ for 12-48 h to obtain the carbon-point bonded silica gel chromatographic packing Sil-DESCDs based on the eutectic solvent. The prepared eutectic solvent carbon dot bonded silica gel can be used as a reverse phase chromatographic packing, can be used for high-selectivity separation of polycyclic aromatic hydrocarbon, alkylbenzene, aromatic amine, phenols, flavonoids compounds, hydrocortisone and prednisolone, and can be used for quantitative detection of calycosin glucoside, formononetin, calycosin and formononetin in an astragalus medicinal material extracting solution.
Description
Technical Field
The invention relates to a preparation method of a carbon point bonding silica gel chromatographic stationary phase based on a eutectic solvent, which is mainly used for separating polycyclic aromatic hydrocarbon, alkylbenzene, aromatic amine, phenols, flavonoids, hydrocortisone and prednisolone, and belongs to the technical field of chromatographic stationary phases.
Background
High performance liquid chromatography has become the most widely used analytical technique nowadays due to its advantages of good selectivity, fast analysis speed, simple operation, etc. CDs have the characteristics of nano size, rich surface functional groups, high stability, moderate adsorption capacity and the like, can avoid the phenomenon of agglomeration of other carbon materials when used as a chromatographic stationary phase, can uniformly modify the surface of silica gel, and has wide application prospect in a chromatographic separation collar.
Since the eutectic solvent (DES) was discovered in 2003, the eutectic solvent has attracted more and more attention of researchers due to the characteristics of environmental friendliness, high biocompatibility, easy biodegradation, low cost, simple preparation method and the like, and is widely applied to the fields of sample pretreatment, material preparation, electrochemistry and the like. The eutectic solvent is widely applied to the chromatographic field as a green solvent of a new generation, for example, as a reaction solvent in the preparation process of a stationary phase, as an additive of a mobile phase or a mobile phase, as a material of a chromatographic stationary phase and the like. When the eutectic solvent is used as a carbon source to prepare the carbon dots, part of the eutectic solvent contains heteroatoms, so that more abundant components can be added for preparing the carbon dots, and no research report about preparing the carbon dot bonded silica gel stationary phase by using the eutectic solvent as a raw material exists at present.
Disclosure of Invention
The invention aims to provide a preparation method of a silica gel chromatographic stationary phase based on eutectic solvent carbon point bonding;
the invention also aims to research the chromatographic separation performance of the eutectic solvent carbon point bonded silica gel chromatographic packing, and the eutectic solvent carbon point bonded silica gel chromatographic packing is used for separating polycyclic aromatic hydrocarbon, alkylbenzene, aromatic amine, phenols, flavonoids, hydrocortisone and prednisolone.
1. Preparation of eutectic solvent carbon point bonded silica gel chromatographic packing
S1, preparing eutectic solvent carbon point DESCDs: mixing choline chloride and lactic acid, continuously heating and stirring for 1-2h at 80 ℃ to form a uniform eutectic solvent, and placing the eutectic solvent into a reaction kettle to react for 2-10 h at 150-220 ℃; and cooling to room temperature after the reaction is finished, putting the product into a dialysis bag for dialysis, filtering by a filter membrane, performing rotary evaporation concentration, and freeze-drying the concentrated solution to obtain the DESCDs. Wherein the molar ratio of choline chloride to lactic acid is 1 to 2 to 1; the cut-off molecular weight of the dialysis bag is 500 to 1000 Da, and the dialysis time is 24 to 48 hours.
S2, preparation of aminopropyl modified silica gel Sil-APS: firstly, uniformly dispersing spherical silica gel in toluene, adding 3-aminopropyltrimethoxysilane or 3-aminopropyltriethoxysilane, and stirring and refluxing for reaction at 90-150 ℃ for 12-48 h; and after the reaction is finished, centrifuging, washing and drying to obtain Sil-MPS. Wherein the mass volume ratio of the spherical silica gel to the 3-aminopropyltrimethoxysilane or the 3-aminopropyltriethoxysilane is 1 to 2g/mL.
S3, preparing carbon point bonded silica gel chromatographic packing Sil-DESCDs: mixing DESCDs prepared in S1 and Sil-APS prepared in S2, then uniformly dispersing in toluene, adding an initiator, and reacting at 50-90 ℃ for 12-48 h with stirring; and centrifuging, washing and drying after the reaction is finished to obtain carbon-point bonded silica gel chromatographic packing Sil-DESCDs. The mass ratio of DESCDs to Sil-APS is 1; the initiator is diethyl pyrocarbonate and triethylamine, the mass ratio of the DESCDs to the diethyl pyrocarbonate is 1 to 1.
2. Structure and performance of eutectic solvent carbon point bonded silica gel chromatographic packing
1. Elemental analysis
Table 1 is an elemental analysis of the eutectic solvent carbon dot bonded silica gel chromatographic packing. The elemental analysis results include eutectic solvent carbon dots (DESCDs), aminopropyl modified silica gel (Sil-APS), and eutectic solvent carbon dot bonded silica gel (Sil-DESCDs). The results show that the content of C% and H% of Sil-DESCDs is obviously increased compared with the content of Sil-APS, and the successful bonding of the carbon points of the eutectic solvent to the surface of the silica gel is confirmed.
TABLE 1 elemental analysis of eutectic solvent carbon point, aminopropyl modified silica gel and eutectic solvent carbon point-bonded silica gel
2. Infrared spectroscopic analysis (FT-IR)
FIG. 1 is a FT-IR spectrum of Sil-DESCDs, sil-MPS, silica and DESCDs. 3343 cm −1 And 3415 cm −1 The absorption peak at (A) is attributed to the O-H absorption peak on DESCDs and Sil-DESCDs, 3383 cm −1 The absorption peak at (A) is attributed to the tensile vibration absorption peak of N-H on Sil-MPS. 1742 cm of −1 The C = O absorption peak of Sil-DESCDs, corresponding to the characteristic peak C = O in the spectrum of DESCDs, appears at 1722 cm −1 To (3). Compared with Sil-MPS, a new absorption peak 1722 cm appears in the Sil-DESCDs spectrum −1 And 1542 cm −1 This indicates that DESCDs are successfully modified on silica surfaces.
3. Transmission electron microscopy analysis (TEM)
FIG. 2 is a Transmission Electron Microscope (TEM) image of a eutectic solvent carbon dot. The DESCDs can be observed to be in a quasi-spherical shape through a transmission electron microscope.
4. Hydrophobic determination chromatogram analysis
Separating five substances of toluene, ethylbenzene, propylbenzene, butylbenzene and pentylbenzene under the chromatographic conditions of 40-60% of acetonitrile, 60-40% of water, column temperature of 30 ℃, UV detector 254 nm and flow rate of 1 ml/min. The retention time of the five alkylbenzenes is reduced with the increase of the acetonitrile content, which is consistent with the hydrophobicity, indicating that the Sil-DESCDs stationary phase is a hydrophobic chromatographic stationary phase.
As shown in FIG. 3, the compound (1) is toluene, (2) is ethylbenzene, (3) is propylbenzene, (4) is butylbenzene, and (5) is pentylbenzene.
5. Separation chromatogram of polycyclic aromatic hydrocarbons
Chromatographic conditions are as follows: (a) 28% acetonitrile, 60% water, 30 ℃ column temperature, 254 nm UV detector, flow rate of 1 mL/min; (b) 60% acetonitrile, 40% water, column temperature 30 ℃, UV detector 254 nm, flow 1 mL/min.
FIG. 4 is a graph of the chromatographic separation of 11 polycyclic aromatic hydrocarbons using Sil-DESCDs column (a) and commercial ZORBAX SB-C18 column (b). As shown in FIG. 4, the anthracene compound (1) is anthrone, (2) is naphthalene, (3) is 2-methylnaphthalene, (4) is acenaphthylene, (5) is fluorene, (6) is phenanthrene, (7) is anthracene, (8) is pyrene, (9) is triphenylene, (10) is chrysene, (11) is 1,2-benzanthracene. As shown in the figure, the Sil-DESCDs column has high selectivity separation for phenanthrene and anthracene, which are isomers of two benzene rings, pyrene, triphenylene, chrysene, and 1,2-benzanthracene, which are isomers of four benzene rings.
6. Separation chromatogram of aromatic amine
Chromatographic conditions are as follows: (a) 16% acetonitrile, 84% water, 30 ℃ column temperature, 254 nm UV detector, 1 mL/min flow rate; (b) 45% acetonitrile, 55% water, column temperature 30 ℃, UV detector 254 nm, flow 1 mL/min.
FIG. 5 is a chromatogram of the separation of 12 aromatic amines using Sil-DESCDs column (a) and commercial ZORBAX SB-C18 column (b). As shown in FIG. 5, the compound is represented by (1) p-phenylenediamine, (2) p-toluidine, (3) N-methylaniline, (4) m-nitroaniline, (5) dimethylaniline, (6) 4-chloroacetanilide, (7) toluidine, (8) β -naphthylamine, (9) 2,4-dichloroaniline, (10) 2,6-diisopropylaniline, (11) diphenylamine, and (12) tribromoaniline. As shown in FIG. 5, sil-DESCDs column has good separation effect on 12 aromatic amines.
7. Separation chromatogram of phenolic compound
Chromatographic conditions are as follows: (a) 16% acetonitrile, 84% water, 30 ℃ column temperature, 254 nm UV detector, 1 mL/min flow rate; (b) 40% acetonitrile, 60% water, column temperature 30 ℃, UV detector 254 nm, flow 1 mL/min.
FIG. 6 is a chromatogram of the separation of 7 phenols by Sil-DESCDs column (a) and commercial ZORBAX SB-C18 column (b). As shown in FIG. 6, the compound (1) is p-aminophenol, (2) is m-aminophenol, (3) is 2-aminophenol, (4) is phenol, (5) is 3,5-dimethylphenol, and (6) is p-chlorophenol. As shown in FIG. 6, sil-DESCDs column has good separation effect on 7 phenolic compounds.
8. Separation chromatogram of flavonoid compound
Chromatographic conditions are as follows: (a) 22% acetonitrile, 78% 0.1% formic acid water solution, column temperature 30 ℃, UV detector 254 nm, flow rate 1 mL/min; (b) 30% acetonitrile, 70% water, column temperature 30 ℃, UV detector 254 nm, flow rate 1 mL/min.
FIG. 7 is a chromatogram of the separation of 6 flavonoids on Sil-DESCDs column (a) and a commercial ZORBAX SB-C18 column (b). As shown in FIG. 7, the compounds include (1) calycosin glucoside, (2) formononetin, (3) astragalin, (4) calycosin, (5) formononetin, and (6) genistein. As can be seen, the Sil-DESCDs column has good separation effect on 6 flavonoid compounds.
9. Separation chromatogram of hydrocortisone and prednisolone
Chromatographic conditions are as follows: (a) 100% water, column temperature 30 ℃, UV detector 254 nm, flow rate 1 mL/min.
(b) 25% acetonitrile, 75% water, 30 ℃ column temperature, 254 nm UV detector, flow 1 mL/min.
FIG. 8 is a separation chromatogram of hydrocortisone and prednisolone from Sil-DESCDs column (a) and a commercial ZORBAX SB-C18 column (b). As shown in fig. 8, wherein (1) hydrocortisone and (2) prednisolone. As shown in FIG. 8, the Sil-DESCDs column can separate hydrocortisone, which is structurally similar to prednisolone, well under the condition that the mobile phase is pure water.
1. Separation of actual samples of Astragalus membranaceus
Chromatographic conditions are as follows: 22 % acetonitrile, 78% 0.1% aqueous formic acid, column temperature 30 deg.C, UV detector 254 nm.
As shown in FIG. 9 (a), there are (1) calycosin glucoside, (2) formononetin, and (3) calycosin (4) formononetin. The standard solutions of 4 flavonoids are accurately prepared in the concentration range of 0.05-0.8 mg/mL, the obtained standard curve is shown in figure 9 (b), and the linear regression equations are respectively as follows:
calycosin glucoside: y =7212.10x-18.73
Formononetin: y =5444.67x +0.68
Calycosin: y =15336.16x-190.48
Formononetin: y =10935.23x-249.48
Fitting to obtain its correlation coefficient R 2 0.9980, 0.9973, 0.9968 and 0.9945, respectively. Wherein x is the concentration of calycosin glucoside, formononetin, calycosin and formononetin, unit: mg/mL; y is the area of the chromatographic peak, unit: and (7) mAU.
Quantitative analysis is carried out on 4 flavonoid components through a standard curve, and the concentrations of calycosin glucoside, formononetin glucoside, calycosin and formononetin are 0.050 mg/mL, 0.031 mg/mL, 0.023 mg/mL and 0.034 mg/mL respectively.
In conclusion, the method is based on the preparation of the eutectic solvent carbon dot bonded silica gel chromatographic stationary phase, the carbon dots are synthesized by taking the eutectic solvent as the raw material, and the carbon dot bonded silica gel chromatographic stationary phase is further prepared. The stationary phase can be used for separating polycyclic aromatic hydrocarbon, alkylbenzene, aromatic amine, phenols and flavonoids compounds in a high selectivity mode, and can separate prednisolone and hydrocortisone with similar structures under the condition that pure water is used as a mobile phase. And can be used for qualitative and quantitative analysis of calycosin glucoside, formononetin, calycosin and formononetin in radix astragali medicinal material extract.
Drawings
FIG. 1 is an infrared spectrum (FT-IR) of a eutectic solvent carbon dot bonded silica gel chromatographic stationary phase.
FIG. 2 is a Transmission Electron Microscope (TEM) image of a eutectic solvent carbon dot.
FIG. 3 is a diagram of chromatographic separation of 5 alkylbenzenes by Sil-DESCDs column.
FIG. 4 is a separation chromatogram of Sil-DESCDs column (a) and commercial ZORBAX SB-C18 column (b) for 11 polycyclic aromatic hydrocarbons.
FIG. 5 is a chromatogram of the separation of 12 aromatic amines using Sil-DESCDs column (a) and commercial ZORBAX SB-C18 column (b).
FIG. 6 is a chromatogram of the separation of 7 phenols by Sil-DESCDs column (a) and commercial ZORBAX SB-C18 column (b).
FIG. 7 is a chromatogram of the separation of 6 flavonoids on Sil-DESCDs column (a) and a commercial ZORBAX SB-C18 column (b).
FIG. 8 is a separation chromatogram of hydrocortisone and prednisolone from Sil-DESCDs column (a) and a commercial ZORBAX SB-C18 column (b).
FIG. 9 is a chromatogram diagram of the Sil-DESCDs column separating the flavonoid compounds and the flavone standard substance in the radix astragali extract.
Detailed Description
The preparation method of the eutectic solvent carbon dot bonded silica gel chromatographic stationary phase of the invention is further explained by the specific examples.
Example 1
S1, preparing eutectic solvent carbon points DESCDs:
choline chloride and lactic acid are mixed according to a molar ratio (1:2) and placed in a 100 mL round-bottom flask, 1.5 h is continuously heated and stirred at 80 ℃ to form a uniform solution serving as a eutectic solvent DES, 35 mL DES is measured and placed in a reaction kettle to react at 150 ℃ for 3 h. Cooling to room temperature after the reaction is finished, putting the product into a dialysis bag with the molecular weight cutoff of 500 Da for dialysis 24 h, filtering by a filter membrane, performing rotary evaporation concentration, and freeze-drying the concentrated solution to obtain DESCDs;
s2, preparing aminopropyl modified silica gel Sil-APS:
uniformly dispersing 2.8 g spherical silica gel in 20 mL toluene, and then adding 1.4 mL of 3-aminopropyltriethoxysilane; stirring and refluxing the mixture at 100 ℃ to react with 17 h; centrifuging after the reaction is finished, washing by adopting methylbenzene, ethanol water solution and methanol, and drying to obtain Sil-MPS;
s3, bonding:
0.6 g of DESCDs and 2.7 g of Sil-APS were uniformly dispersed in 20 mL toluene, 1.3 g diethylpyrocarbonate and 0.9g triethylamine were added and the reaction was stirred at 50 ℃ for 18 h; and centrifuging after the reaction is finished, washing by using toluene, ethanol and methanol in sequence, and drying to obtain carbon-point bonded silica gel chromatographic packing Sil-DESCDs. The eutectic solvent of the stationary phase has good carbon point bonding amount and good separation effect.
Example 2
S1, preparing eutectic solvent carbon points DESCDs:
choline chloride and lactic acid are mixed according to a molar ratio (1:3) and placed in a 100 mL round-bottom flask, 1.5 h is continuously heated and stirred at 80 ℃ to form a uniform solution serving as a eutectic solvent DES, 35 mL DES is measured and placed in a reaction kettle to react at 180 ℃ for 4 h. Cooling to room temperature after the reaction is finished, putting the product into a dialysis bag with the molecular weight cutoff of 500 Da for dialysis 24 h, filtering by a filter membrane, performing rotary evaporation concentration, and freeze-drying the concentrated solution to obtain DESCDs;
s2, preparing aminopropyl modified silica gel Sil-APS:
3 g spherical silica gel is uniformly dispersed in 20 mL toluene, and 1.5 mL3-aminopropyltrimethoxysilane is added; stirring and refluxing the mixture at 110 ℃ to react with 24 h; after the reaction is finished, centrifuging, washing by adopting methylbenzene, ethanol water solution and methanol, and drying to obtain Sil-MPS;
s3, bonding:
0.8 g of DESCDs and 2.8 g of Sil-APS were uniformly dispersed in 20 mL toluene, 1.5 g diethylpyrocarbonate and 1.1g of triethylamine were added, and 24 h was reacted with stirring at 60 ℃; and centrifuging after the reaction is finished, washing by using toluene, ethanol and methanol in sequence, and drying to obtain carbon-point bonded silica gel chromatographic packing Sil-DESCDs. The eutectic solvent carbon dot bonding amount of the stationary phase is optimal, and the separation effect is optimal.
Example 3
S1, preparing a eutectic solvent carbon point DESCDs:
choline chloride and lactic acid are mixed according to the molar ratio (1:3) and placed in a 100 mL round-bottom flask, the mixture is continuously heated and stirred at 80 ℃ for 1.5 h, a uniform solution is formed and used as a eutectic solvent DES, 35 mL DES is measured and placed in a reaction kettle to react at 200 ℃ for 6 h. Cooling to room temperature after the reaction is finished, putting the product into a dialysis bag with the molecular weight cutoff of 1000 Da for dialysis 48 h, filtering by a filter membrane, performing rotary evaporation concentration, and freeze-drying the concentrated solution to obtain DESCDs;
s2, preparing aminopropyl modified silica gel Sil-APS (ammonium propyl ether) xander:
3.2 g spherical silica gel is evenly dispersed in 20 mL toluene, and then 1.6 mL of 3-aminopropyltriethoxysilane is added; stirring and refluxing the mixture at 130 ℃ to react with 36 h; centrifuging after the reaction is finished, washing by adopting methylbenzene, ethanol water solution and methanol, and drying to obtain Sil-MPS;
s3, bonding:
0.9g of DESCDs and 3 g of Sil-APS were uniformly dispersed in 20 mL toluene, 1.7 g diethylpyrocarbonate and 1.4 g triethylamine were added, and the reaction mixture was stirred at 60 ℃ for 24 h; and centrifuging after the reaction is finished, washing by using toluene, ethanol and methanol in sequence, and drying to obtain carbon-point bonded silica gel chromatographic packing Sil-DESCDs. The eutectic solvent of the stationary phase has good bonding amount of carbon points and good separation effect.
Example 4
S1, preparing eutectic solvent carbon points DESCDs:
choline chloride and lactic acid are mixed according to a molar ratio (1:4) and placed in a 100 mL round-bottom flask, 1.5 h is continuously heated and stirred at 80 ℃ to form a uniform solution serving as a eutectic solvent DES, 35 mL is measured, and the DES is placed in a reaction kettle to react at 200 ℃ for 8 h. Cooling to room temperature after the reaction is finished, putting the product into a dialysis bag with the molecular weight cutoff of 1000 Da for dialysis 48 h, filtering by a filter membrane, performing rotary evaporation concentration, and freeze-drying the concentrated solution to obtain DESCDs;
s2, preparing aminopropyl modified silica gel Sil-APS:
3.4 g spherical silica gel is evenly dispersed in 20 mL toluene, and 1.7 mL3-aminopropyltrimethoxysilane is added; stirring and refluxing at 130 ℃ to react 48 h; centrifuging after the reaction is finished, washing by adopting methylbenzene, ethanol water solution and methanol, and drying to obtain Sil-MPS;
s3, bonding:
0.6 g of DESCDs and 3 g of Sil-APS were uniformly dispersed in 20 mL toluene, 1.6 g diethylpyrocarbonate and 1.2 g triethylamine were added and the reaction was stirred at 75 ℃ for 36 h; and centrifuging after the reaction is finished, washing by using toluene, ethanol and methanol in sequence, and drying to obtain carbon-point bonded silica gel chromatographic packing Sil-DESCDs. The eutectic solvent of the stationary phase has good bonding amount of carbon points and good separation effect.
Example 5
S1, preparing eutectic solvent carbon points DESCDs:
choline chloride and lactic acid are mixed according to a molar ratio (1:4) and placed in a 100 mL round-bottom flask, 1.5 h is continuously heated and stirred at 80 ℃ to form a uniform solution serving as a eutectic solvent DES, 35 mL DES is measured and placed in a reaction kettle to react at 220 ℃ for 10 h. Cooling to room temperature after the reaction is finished, putting the product into a dialysis bag with the molecular weight cutoff of 1000 Da for dialysis 48 h, filtering by a filter membrane, performing rotary evaporation concentration, and performing freeze drying on the concentrated solution to obtain DESCDs;
s2, preparing aminopropyl modified silica gel Sil-APS:
3.4 g spherical silica gel is evenly dispersed in 20 mL toluene, and then 1.9 mL of 3-aminopropyltriethoxysilane is added; stirring and refluxing at 150 ℃ to react 48 h; centrifuging after the reaction is finished, washing by adopting methylbenzene, ethanol water solution and methanol, and drying to obtain Sil-MPS;
s3, bonding:
0.5 g of DESCDs and 3.2 g of Sil-APS were uniformly dispersed in 20 mL toluene, 1.7 g diethylpyrocarbonate and 1.3 g triethylamine were added and the reaction was stirred at 90 ℃ for 48 h; and centrifuging after the reaction is finished, washing by using toluene, ethanol and methanol in sequence, and drying to obtain carbon-point bonded silica gel chromatographic packing Sil-DESCDs. The eutectic solvent of the stationary phase has good bonding amount of carbon points and good separation effect.
Claims (9)
1. A preparation method of a silica gel chromatographic stationary phase based on eutectic solvent carbon point bonding comprises the following steps:
s1, preparing eutectic solvent carbon point DESCDs: mixing choline chloride and lactic acid, continuously heating and stirring for 1-2h at 80 ℃ to form a uniform eutectic solvent, and placing the eutectic solvent into a reaction kettle to react for 2-10 h at 150-220 ℃; cooling to room temperature after the reaction is finished, putting the product into a dialysis bag for dialysis, filtering by a filter membrane, performing rotary evaporation concentration, and freeze-drying the concentrated solution to obtain DESCDs;
s2, preparation of aminopropyl modified silica gel Sil-APS: firstly, uniformly dispersing spherical silica gel in toluene, adding 3-aminopropyltrimethoxysilane or 3-aminopropyltriethoxysilane, and stirring and refluxing for reaction at 90-150 ℃ for 12-48 h; after the reaction is finished, centrifuging, washing and drying to obtain Sil-MPS;
s3, preparing a carbon-point bonded silica gel chromatographic packing Sil-DESCDs: mixing DESCDs prepared in S1 and Sil-APS prepared in S2, then uniformly dispersing in toluene, adding an initiator, and reacting at 50-90 ℃ for 12-48 h with stirring; and centrifuging, washing and drying after the reaction is finished to obtain carbon-point bonded silica gel chromatographic packing Sil-DESCDs.
2. The method for preparing the chromatographic stationary phase based on the eutectic solvent carbon point bonding silica gel, according to claim 1, is characterized in that: in the step S1, the molar ratio of choline chloride to lactic acid is 1 to 2-1.
3. The method for preparing the chromatographic stationary phase based on the eutectic solvent carbon point bonding silica gel, according to claim 1, is characterized in that: in the step S1, the cut-off molecular weight of the dialysis bag is 500 to 1000 Da, and the dialysis time is 24 to 48 h.
4. The method for preparing the chromatographic stationary phase based on the eutectic solvent carbon point bonding silica gel, according to claim 1, is characterized in that: in the step S2, the mass-to-volume ratio of the spherical silica gel to the 3-aminopropyltrimethoxysilane or the 3-aminopropyltriethoxysilane is 1 to 2g/mL.
5. The method for preparing the chromatographic stationary phase based on the eutectic solvent carbon point bonding silica gel, according to claim 1, is characterized in that: in the step S3, the mass ratio of DESCDs to Sil-APS is 1 to 3.
6. The method for preparing the chromatographic stationary phase based on the eutectic solvent carbon point bonding silica gel, according to claim 1, is characterized in that: in the step S3, initiating agents are diethyl pyrocarbonate and triethylamine; the mass ratio of DESCDs to diethylpyrocarbonate is 1 to 1; the mass ratio of the DESCCDs to the triethylamine is 1 to 1.
7. The application of the chromatographic stationary phase based on the eutectic solvent carbon point bonding silica gel prepared by the method of claim 1 as a reversed phase chromatographic packing is characterized in that: the carbon point bonded silica gel chromatographic packing of the eutectic solvent is used for selectively separating polycyclic aromatic hydrocarbon, alkylbenzene, aromatic amine, phenols, flavonoids, hydrocortisone and prednisolone.
8. The use of the chromatographic stationary phase based on eutectic solvent carbon point bonded silica gel as reversed phase chromatographic packing according to claim 7, characterized in that: the polycyclic aromatic hydrocarbon is anthrone, naphthalene, 2-methylnaphthalene, acenaphthene, fluorene, phenanthrene, anthracene, pyrene, triphenylene, chrysene, 1,2-benzanthracene; the alkylbenzene is toluene, ethylbenzene, propyl benzene, butylbenzene or pentylbenzene; the aromatic amine is p-phenylenediamine, p-toluidine, N-methylaniline, m-nitroaniline, dimethylaniline, 4-chloroacetanilide, methylnaphthylamine, beta-naphthylamine, 2,4-dichloroaniline, 2,6-diisopropylaniline, diphenylamine and tribromoaniline; the phenols are p-aminophenol, m-aminophenol, 2-aminophenol, phenol, 3,5-dimethylphenol, p-chlorophenol and p-tert-butylphenol; the flavonoid compounds are calycosin glucoside, formononetin, calycosin, formononetin, astragalin, and genistein.
9. The application of the chromatographic stationary phase based on eutectic solvent carbon point bonding silica gel prepared by the method of claim 1 as a reversed phase chromatographic packing is characterized in that: separating and quantitatively detecting calycosin glucoside, formononetin, calycosin and formononetin in the radix astragali medicinal material extract by using the Sil-DEScDs as reversed phase chromatography packing; the concentration range of calycosin glucoside, formononetin, calycosin and formononetin is 0.05-0.8 mg/mL, and the quantitative linear regression equations are respectively as follows:
calycosin glucoside: y =7212.10x-18.73, R 2 =0.9980；
Formononetin: y =5444.67x +0.68 2 =0.9973；
Calycosin: y =15336.16x-190.48, R 2 =0.9968；
Formononetin: y =10935.23x-249.48 2 =0.9945；
Wherein x is the concentration of calycosin glucoside, formononetin, calycosin and formononetin, unit: mg/mL; y is the area of the chromatographic peak, unit: and (7) mAU.
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| CN106268715A (en) * | 2016-09-12 | 2017-01-04 | 中国科学院兰州化学物理研究所 | A kind of green synthesis method of carbon quantum dot bonded silica gel chromatograph packing material |
| CN108261801A (en) * | 2018-02-09 | 2018-07-10 | 长治学院 | A kind of stationary phase containing eutectic solvent and its preparation method and application |
| CN111250069A (en) * | 2020-01-20 | 2020-06-09 | 浙江月旭材料科技有限公司 | Silica gel chromatographic packing and preparation method and application thereof |
| CN113262770A (en) * | 2021-04-07 | 2021-08-17 | 中国科学院兰州化学物理研究所 | Carbon quantum dot and precursor co-bonded silica gel chromatographic packing and application thereof |
| CN113274993A (en) * | 2021-05-25 | 2021-08-20 | 烟台大学 | Preparation method of silica gel matrix chromatographic packing for separating strong-polarity drugs |
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| US20050106741A1 (en) * | 2003-10-17 | 2005-05-19 | Abraham Dijke | Determination of aromatic hydrocarbons in edible oil products |
| US7557232B2 (en) * | 2007-05-25 | 2009-07-07 | Dionex Corporation | Compositions useful as chromatography stationary phases |
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| CN111250069A (en) * | 2020-01-20 | 2020-06-09 | 浙江月旭材料科技有限公司 | Silica gel chromatographic packing and preparation method and application thereof |
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