CN114133571B

CN114133571B - PMO (SLLTP-POSS) hydrophilic microsphere and preparation method and application thereof

Info

Publication number: CN114133571B
Application number: CN202111210304.7A
Authority: CN
Inventors: 陈桐; 李丙祥; 徐亮; 刘涛; 徐何辰; 肖震; 吴艳玲; 杨敏
Original assignee: Zhenjiang Customs Comprehensive Technology Center
Current assignee: Zhenjiang Customs Comprehensive Technology Center
Priority date: 2021-10-18
Filing date: 2021-10-18
Publication date: 2022-10-04
Anticipated expiration: 2041-10-18
Also published as: CN114133571A

Abstract

The invention discloses a PMO (SLLTP-POSS) hydrophilic microsphere and a preparation method and application thereof. The PMO (SLLTP-POSS) hydrophilic microspheres are used as a hydrophilic stationary phase and have the characteristics of HILIC and PALC, and similar to HILIC, PALC can also retain polar compounds. And the acid and alkali resistance of the PMO (SLLTP-POSS) stationary phase is greatly improved. The stationary phase can separate organic acids, sugar alcohol and amino acid mixture, sweetener and other polar compounds, and has great separation degree and high selectivity. The immobilization is less retention of relatively non-polar and less polar compounds than a conventional C18 column, and more retention of more polar compounds. From the perspective of developing green color spectrum and the harm of ACN to the environment, PALC as a green color spectrum mode is expected to become an alternative mode of HILIC and a complementary mode of RPLC.

Description

PMO (SLLTP-POSS) hydrophilic microsphere and preparation method and application thereof

Technical Field

The invention belongs to the field of new materials, and particularly relates to a PMO (SLLTP-POSS) hydrophilic microsphere as well as a preparation method and an application thereof.

Background

Food safety is always a big concern all over the world. The main separation method of polar compounds in food additives is Hydrophilic interaction chromatography (HILIC) [1,2]. At present, a plurality of hydrophilic chromatographic stationary phases are developed on the market, and most of the stationary phases are prepared by bonding hydrophilic groups on the surface of silica gel. When they are used in the HILIC mode, there are three problems:

(1) The mobile phase needs to use a high percentage of organic solvents, such as Acetonitrile (ACN) and the like [3], and the waste liquid generated after a large amount of use, such as random discharge, can cause serious pollution to the environment.

(2) Although HILIC chromatography columns have a strong ability to separate polar compounds, the column efficiency is usually not high [4].

(3) Such chromatographic columns are not very acid-base resistant and, when used under strong acid or strong base conditions for a long time, the separation capacity and selectivity of the chromatographic column are greatly reduced [5].

Aiming at the problems of low column efficiency and generally low acid and alkali resistance of HILIC chromatographic columns, the organic-inorganic hybrid material-ordered mesoporous organic silicon oxide (PMO), namely (R' O) ₃ Si-R-Si(OR') ₃ [6]And the problem can be solved. PMO can completely incorporate organic groups into the mesoporous framework of the material, rendering the material to exhibit special properties such as greater hydrothermal stability and mechanical stability. More interestingly, the PMO shows good chemical stability in alkaline medium, and is very beneficial to solving the problem that the alkaline compound is easy to form a tailing peak on a silica gel matrix. Meanwhile, various chemical reactions can be carried out through the embedded organic group, so that the purpose of functionally modifying the material is achieved. As the proportion of organic and inorganic components can be regulated and controlled by precursors, the materials show great advantages in the aspects of organic functional group density and high guarantee of column efficiency [7]. Therefore, PMO has been applied to preparative chromatography and high performance liquid chromatography as a new class of chromatographic packing. Li et al [8]Reacting 1, 2-bis (triethoxysilyl) ethane with3-aminopropyl triethoxysilane is copolymerized to prepare the amino-functionalized ethylene bridged hybrid silica gel filler. After the chromatographic column is prepared, the stability of the hybridized amino column in an alkaline medium is obviously superior to that of a bonded amino column. Under the conditions of pH 1.6 and pH 11.8, some release type compounds are separated, and good peak shape and column effect are obtained.

Meanwhile, in a water-rich chromatography (PALC) mode [9], a high percentage of water is usually used as a mobile phase, and the application of the stationary phase to the field of food detection is expected to become a substitute of HILIC and a supplement of reversed-phase chromatography, and can also alleviate the problem that a large amount of organic solvent is required to be used for the mobile phase in the HILIC mode. Currently, the types of commercially available PMO liquid chromatography stationary phases are limited. Therefore, the method has important significance for the preparation and development of the stationary phase, and opens up a new path for developing a new stationary phase with excellent performance.

Reference documents

1.Walker S.H.,Carlisle B.C.,Muddiman D.C.Systematic comparison of reverse phase and hydrophilic interaction liquid chromatography platforms for the analysis of N-Linked glycans.Analytical Chemistry,2012,84(19):8198-8206.

2.Shen Q.,Wu H.M.,Wang H.H.,Zhao Q.L.,Xue J.,Ma J.F.,Wang H.X.Monodisperse microsphere-based immobilized metal affinity chromatography approach for preparing Antarctic krill phospholipids followed by HILIC-MS analysis.Food Chemistry,2021,344:128585-128593.

3.Costa P.P.K.G.,Mendes T.D.,Salum T.F.C.,Pacheco T.F.,Rodrigues C.M.Development and validation of HILIC-UHPLC-ELSD methods for determination of sugar alcohols stereoisomers and its application for bioconversion processes of crude glycerin.Journal of Chromatography A,2019,1589:56-64.

4.LiangT.,Fu Q.,Shen A.J.,Wang H.,Jin Y.,Xin H.X.,Ke Y.X.,Guo Z.M.,Liang X.M.Preparation and chromatographic evaluation of a newly designedsteviol glycoside modified-silica stationary phase in hydrophilicinteraction liquid chromatography and reversed phase liquidchromatography.Journal of Chromatography A,2015,1388:110–118.

5.Spicer V.,Krokhin O.V.Peptide retention time prediction in hydrophilic interaction liquid chromatography and comparison of separation selectivity between bare silica and bonded stationary phases.Journal of Chromatography A,2018,1534:75-84.

6.Huang X.,Zhang M.N.,Wang M.J.,Li W.,Wang C.,Hou X.J.,Luan S.,Wang Q.Gold/periodic mesoporous organosilicas with controllable mesostructure by using compressedCO2.Langmuir,2018,34:3642-3653.

7.Kaczmarek A.M.,Maegawa Y.,Abalymov A.,Skirtach A.G.,Inagaki S.,Voort P.V.D.Lanthanide-grafted bipyridine periodic mesoporous organosilicas(BPy-PMOs)physiological range and wide temperature range luminescence thermometry.ACS Applied Materials&Interfaces,2020,12:13540-13550.

8.Li C.,Di B.,Hao W.,Yan F.,Su M.Aminopropyl-functionalized ethane-bridged periodic mesoporous organosilica spheres:preparation and application in liquid chromatography.Journal of Chromatography A,2016,1218(3):408-415.

9.Dembek M.,Bocian S.Pure water as a mobile phase in liquid chromatography techniques.Trends in Analytical Chemistry,2020,123:115793-115806.

Disclosure of Invention

The invention aims to provide PMO (SLLTP-POSS) hydrophilic microspheres and a preparation method thereof.

The PMO (SLLTP-POSS) hydrophilic microsphere provided by the invention is prepared by the following steps:

1) Synthesis of SLLTP-bridged silane (SLLTPBS)

a. Dissolving Bulbus Lilii polysaccharide (LLTP) in ionic liquid, adding dropwise chlorosulfonic acid diluted with anhydrous pyridine, stirring at constant temperature of 30-70 deg.C for 0.5-3h, and adjusting to neutral with alkali solution to obtain sulfated modified Bulbus Lilii polysaccharide (SLLTP);

b. under the protection of nitrogen, stirring and reacting the N, N-Dimethylformamide (DMF) solution of SLLTP with the THF solution of chloromethyl trimethoxy silane (CMTMS) at the constant temperature of 40-80 ℃ for 3-10h to obtain SLLTP-bridged silane (SLLTPBS);

2) In the presence of NaOH, using C18TACL as template agent to make POSS [ C ₂ H ₄ Si(OEt) ₃ ] ₈ SLLTPBS, C18TACL in a mixed solvent of water and THF; filtering after the reaction is finished, washing the product with ethanol, deionized water and methanol in sequence, and drying in vacuum; then, a solvent extraction method is adopted to remove the template agent, and the PMO (SLLTP-POSS) hydrophilic microspheres are obtained.

In the step a of step 1) of the method, the ionic liquid is an imidazole ionic liquid, and specifically may be any one of the following: 1-butyl-3-methylimidazolium chloride, 1-butylimidazolium chloride, and 1, 3-dimethylimidazolium chloride.

In the step a of step 1) of the method, the ratio of the LLTP to the ionic liquid may be 500mg:10mL-500mg:50mL; specifically, the content is 500mg:30mL.

In the step a of the step 1), the molar amount of the chlorosulfonic acid is 40 to 80 percent, specifically 60 percent, of the molar number of the hydroxyl groups on the LLTP;

in the step a of the step 1), the volume ratio of the chlorosulfonic acid to the anhydrous pyridine is 1:10-1:20.

in step 1) a of the above method, the mixture may be stirred at a constant temperature of 50 ℃ for 1 hour.

In step 1), the alkali may be potassium hydroxide or ammonia.

In the step a of the step 1), after the solution is adjusted to be neutral by the sodium hydroxide solution, the method further comprises the following steps: loading the system adjusted to neutral into dialysis bag with cut-off of 10KDa, dialyzing in ultrapure water for 24-72 hr (specifically 48 hr), concentrating, precipitating with ethanol, and freeze drying at-20 deg.C to obtain SLLTP.

In step b) of the above method, the molar ratio of SLLTP to chloromethyltrimethoxysilane is 1.

In step b) of step 1) of the above method, the volume ratio of chloromethyltrimethoxysilane to THF is 1.

In step b) of the above method, the N, N-dimethylformamide is anhydrous N, N-dimethylformamide; the THF is anhydrous THF.

In the step b) of step 1), the method further comprises the following steps after the reaction is finished: loading the reaction solution into a dialysis bag with cut-off amount of 10KDa, dialyzing in ultrapure water for 24-72 hr (specifically 48 hr), concentrating, precipitating with ethanol, and freeze-drying at-20 deg.C to obtain SLLTPBS.

In the step 2) of the above method, the POSS [ C ] ₂ H ₄ Si(OEt) ₃ ] ₈ And SLLTPBS in a molar ratio of 0.5.

In step 2) of the method, the mass ratio of the C18TACl to the SLLTPBS is 1:17.

in the step 2) of the method, the mass ratio of NaOH to SLLTPBS is 1:37.5.

in the step 2) of the method, the volume ratio of water to THF in the mixed solvent of water and THF may be 30:30-60:30, preferably in a volume ratio of 40:30-50:30.

in step 2) of the above method, the reaction temperature of the reaction may be 50 ℃ to 90 ℃, preferably 60 ℃ to 70 ℃.

In step 2) of the above method, the reaction time of the reaction may be 10 to 20 hours, specifically 15 hours.

In the step 2) of the method, the reaction is carried out in a miniature high-pressure reaction kettle.

In step 2) of the above method, the temperature of the vacuum drying may be 65 ℃.

In the step 2), the specific method for removing the template agent by using the solvent extraction method comprises the following steps: adding the dried solid into 150mL concentrated hydrochloric acid (mass fraction is 36% -38%)/deionized water/ethanol (v/v = 5/45/50) solution per gram, heating and refluxing for 8h to extract the template, repeating twice, filtering, washing and drying.

The method further comprises the step of reaming the obtained PMO (SLLTP-POSS) hydrophilic microspheres, and the specific method comprises the following steps: and (3) reacting the PMO (SLLTP-POSS) hydrophilic microspheres, the DMDA and the DDA in ultrapure water, filtering after the reaction is finished, washing with ultrapure water and methanol in sequence, and drying at 60 ℃ in vacuum to obtain the PMO (SLLTP-POSS) hydrophilic microspheres after pore expansion.

The reaming method further comprises the following steps: and (3) removing the DMDA and the DDA from the pore-enlarged PMO (SLLTP-POSS) hydrophilic microspheres by adopting a solvent extraction method.

Wherein the mass ratio of the PMO (SLLTP-POSS) hydrophilic microspheres to the DMDA and to the DDA can be 4.0: (3.0-6.0): (0.5-1.5), specifically 4.0.

The reaction condition of the reaction is that the mixture is kept stand for 24 to 72 hours at the temperature of 110 ℃.

The invention also provides application of the PMO (SLLTP-POSS) hydrophilic microsphere.

The application of the PMO (SLLTP-POSS) hydrophilic microsphere provided by the invention is the application of the PMO hydrophilic microsphere in preparation of a hydrophilic chromatographic stationary phase.

The invention also provides a hydrophilic chromatographic column.

The stationary phase of the hydrophilic chromatographic column is the PMO (SLLTP-POSS) hydrophilic microsphere.

The invention also protects the application of the hydrophilic chromatographic column in separating and/or detecting polar substances.

Specifically, the polar substance is at least one selected from organic acids, sugar alcohol, amino acid, and sweetener (including artificial and natural sweetener);

the organic acid is at least one selected from oxalic acid, tartaric acid, quinic acid, malic acid, shikimic acid, ascorbic acid, acetic acid and maleic acid;

the sweetener is selected from saccharin sodium, sucralose, sodium cyclamate, aspartame, acesulfame potassium, alitame, neotame, glycyrrhizic acid, glycyrrhetinic acid, stevioside, and steviolbioside.

In the above separation or detection method, the sample to be detected may be food; more specifically, it may be a food containing or suspected of containing the above-mentioned organic acids, monosaccharides, sugar alcohols, amino acids or sweeteners.

The PMO (SLLTP-POSS) filler is prepared as a hydrophilic stationary phase, is characterized by utilizing infrared spectroscopy, elemental analysis, a scanning electron microscope and the like, and is researched for chromatographic behavior. The novel stationary phase has the characteristics of HILIC and PALC. Similar to HILIC, PALC can also retain polar compounds, and the acid and alkali resistance is greatly improved. The stationary phase can separate organic acids, sugar alcohol and amino acid mixture, sweetening agent and other polar compounds, and has great separation degree and high selectivity. The immobilization is less retention of relatively non-polar and less polar compounds than a conventional C18 column, and more retention of more polar compounds. From the perspective of developing green color spectrum and the harm of ACN to the environment, PALC as a green color spectrum mode is expected to become an alternative mode of HILIC and a complementary mode of RPLC.

Drawings

FIG. 1 is a flow chart of the synthetic reaction of PMO (SLLTP-POSS) hydrophilic microspheres.

FIG. 2 is an infrared spectrum of PMO (SLLTP-POSS) hydrophilic microsphere at various stages of synthesis; (A) LLTP; (B) SLLTP; (C) SLLTPBS; (D) non-template-removed PMO (SLLTP-POSS); (E) PMO (SLLTP-POSS) with template removed.

FIG. 3 is a scanning electron micrograph of PMO (SLLTP-POSS) microspheres obtained under different conditions. A and B correspond to experiment 3 in Table 1; c, D and E correspond to runs 8,9 and 10, respectively, in Table 1.

FIG. 4 is a thermogravimetric plot of PMO (SLLTP-POSS) microspheres.

FIG. 5 is a low angle XRD pattern of PMO (SLLTP-POSS) microspheres.

FIG. 6 is a comparison of column pressure/flow rate performance of PMO (SLLTP-POSS) columns prepared with different POSS [ C2H4Si (OEt) 3]8/SLLTPBS molar ratios with commercial C18 columns.

Fig. 7 is a graph of the change in retention factor (a) and number of trays (B) for a PMO (SLLTP-POSS) column versus a commercial C18 column at pH = 11. (mobile phase: aqueous triethylamine solution/ACN =95, flow rate: 1.0 mL/min.); variation of PMO (SLLTP-POSS) column retention factor (C) and number of plates (D) from commercial C18 column at pH = 1.0. (mobile phase: 1% TFA/ACN =92, flow rate: 1.0mL/min.

FIG. 8 is a separation chromatogram of eight organic acids in the PLAC mode. The chromatogram obtained by column separation with PMO (SLLTP-POSS) is shown in (A), the chromatogram obtained by column separation with C18 column is shown in (B), the chromatogram obtained by column separation with PMO (SLLTP-POSS) is shown in (C), and the chromatogram obtained by column separation with C18 column is shown in (D), after column washing with C is shown in (D) for 30 days. Chromatographic peak: (1) oxalic acid; (2) tartaric acid; (3) quinic acid; (4) malic acid; (5) shikimic acid; (6) ascorbic acid; (7) acetic acid; and (8) maleic acid.

Detailed Description

The present invention will be further illustrated with reference to the following examples, but the present invention is not limited to the following examples. The method is a conventional method unless otherwise specified. The starting materials are commercially available from the open literature unless otherwise specified.

The experimental materials used in the following examples are as follows:

the lily polysaccharide is purchased from Shaanxi blue grass Biotechnology Co., ltd. 1-butyl-3-methyl chloride, chlorosulfonic acid, chloromethyltrimethoxysilane, octadecyltrimethylammonium chloride (C18 TACL), octa (triethoxysilylethyl) oligomeric silsesquioxane (POSS [ C18 ] ethyl ester ₂ H ₄ Si(OEt) ₃ ] ₈ ) N, N-dimethyldecylamine (DMDA) and Dodecylamine (DDA) were purchased from Alfa Aesar, germany. Pyridine, tetrahydrofuran, N-dimethylformamide, methanol and ethanol were all purchased from Nanjing chemical reagents, inc. (Nanjing, china).

The compounds adenine, caffeine, clenbuterol, salicylic acid, potassium sorbate, fumaric acid, thiourea, tartrazine, sunset yellow, brilliant blue, new red, vitamin B2 (VB 2) and vitamin B6 (VB 6) for hydrophilic and water-rich chromatographic pattern evaluation were purchased from alatin (beijing, china). 8 organic acid compounds including oxalic acid, tartaric acid, quinic acid, malic acid, shikimic acid, ascorbic acid, acetic acid, maleic acid; ribose, mannitol, sucrose, maltitol, raynaud's sugar, melezitose, phenylalanine, methionine, glutamic acid and histidine were purchased from sigma.

The ultrapure Water used in the experimental procedure was obtained from a BWT ultrapure Water system (Best Water Technology, germany). The mobile phase of the HPLC apparatus was a mixture of acetonitrile (chromatographic grade, merck, germany) and ultrapure water, and both the mobile phase and the test compound were filtered using a 0.22 μm filter before use.

Example 1 preparation of PMO (SLLTP-POSS) hydrophilic microspheres

The process is divided into three steps, and the specific content is as follows:

1. synthesis of SLLTP-bridged silane (SLLTPBS)

Firstly, SLLTP is prepared by adopting a green synthesis technology and an ionic liquid-pyridine chlorosulfonate method. 500mg of lily polysaccharide (LLTP) was dissolved in 30mL of 1-butyl-3-methylimidazolium chloride ([ C4mim ] Cl), and then 5mL of anhydrous pyridine-diluted chlorosulfonic acid (the content of chlorosulfonic acid is 60% of the number of moles of hydroxyl groups on LLTP) was added dropwise thereto, followed by stirring at 50 ℃ for 1 hour and then adjusting to neutrality with a sodium hydroxide solution. Dialyzing with 10kDa dialysis bag in ultrapure water for 48 hr, concentrating, precipitating with ethanol, and freeze-drying at-20 deg.C to obtain SLLTP.

Then, the SLLTP was dissolved in 30mL of anhydrous N, N-Dimethylformamide (DMF). 2mL of Chloromethyltrimethoxysilane (CMTMS) was dissolved in 8mL of anhydrous THF, and added dropwise to the above DMF solution. Stirring at constant temperature of 60 ℃ for 5h under the protection of nitrogen. After the reaction is finished, filling the reaction solution into a dialysis bag with the cut-off amount of 10KDa, dialyzing in ultrapure water for 48 hours, concentrating, precipitating with ethanol, and freeze-drying at the temperature of-20 ℃ to obtain SLLTPBS.

2. Preparation of PMO (SLLTP-POSS) hydrophilic microsphere

A certain amount of C18TACL was weighed into a flask, THF and ultrapure water were sequentially added thereto, and the mixture was sufficiently stirred. Then, naOH was added and dissolved. Reacting POSS [ C ] ₂ H ₄ Si(OEt) ₃ ] ₈ And SLLTPBS were dissolved in THF and an aqueous solution, respectively, and added to the above reaction system. Stirring at room temperature for 5h, and heating in a miniature high-pressure reaction kettle for reaction. After the reaction is finished, filtering, washing the product with ethanol, deionized water and methanol in sequence, and drying in vacuum at 65 ℃.

The optimum reaction conditions were determined by examining the variation of the various reaction parameters in the system, the reaction parameters being set up as shown in table 1. Then, a solvent extraction method is adopted to remove the template agent. The dried solid was added to 150mL concentrated hydrochloric acid (mass fraction 36% -38%)/deionized water/ethanol (v/v = 5/45/50) solution per gram, heated under reflux for 8h of extraction template, and repeated twice. After filtration, washing and drying, the PMO (SLLTP-POSS) hydrophilic microspheres are finally obtained, and the synthetic flow chart is shown in figure 1.

TABLE 1 gradiometer of reaction parameters

Wherein the mass ratio of the C18TACL to the SLLTPBS is 1:17.

3. reaming of PMO (SLLTP-POSS) hydrophilic microspheres

4.0g of PMO (SLLTP-POSS) hydrophilic microspheres, 5.0g of DMDA and 1.2g of DDA were weighed into a flask, 120mL of ultrapure water was added thereto, and stirred at room temperature for 1 hour. Placing into a miniature high-pressure reaction kettle, and placing into an electric heating thermostat for standing for 48h at 110 ℃. And filtering after the reaction is finished, washing with ultrapure water and methanol in sequence, and drying in vacuum at 60 ℃ to obtain the PMO (SLLTP-POSS) hydrophilic microspheres after pore expansion. Then solvent extraction is adopted to remove the template agents DMDA and DDA. The procedure for removing DMDA and DDA was the same as for removing the templating agent in 2 above.

Example 2 characterization of PMO (SLLTP-POSS) hydrophilic microspheres

The change in surface chemical structure of PMO (SLLTP-POSS) hydrophilic microspheres was analyzed by Nicolet is model 10 Fourier Infrared Spectroscopy (Thermo Fisher, USA). C. The results of the variation of the contents of H and S elements were obtained by a Vario EL type element analyzer (Elementar Co., germany). The thermal stability and order of the materials were analyzed on a STA 409PC thermogravimetric analyzer (NETZSCH, germany) and Ultimate type IV X-ray diffractometer (XRD), respectively (Rigaku, japan). JSM-6360LV scanning electron microscope (Japan) observes the surface topography of hydrophilic microspheres and determines particle size. A dynamic light scattering instrument (Brookhaven, USA) of BI-200SM DLS model was used to examine the particle size distribution of PMO (SLLTP-POSS) hydrophilic microspheres prepared under different conditions. Changes in specific surface area, pore diameter and pore volume of PMO (SLLTP-POSS) hydrophilic microspheres were examined using a model ASAP-2460 nitrogen adsorption specific surface area analyzer (Micromeritics Instruments Corporation, USA).

The results are shown below.

1. First, the results of IR spectroscopy and elemental analysis of LLTP, SLLTP, SLLTPBS and PMO (SLLTP-POSS) hydrophilic microspheres are shown in FIG. 2 and Table 2.LLTP is 3400cm ^-1 The broad peak at (A) is O-H stretching vibration, 2910cm ^-1 The absorption peak is C-H stretching vibration, 1380cm ^-1 1020cm of flexural vibration having an absorption peak of C-H ^-1 The absorption peaks at (A) are C-O stretching vibrations, which are characteristic peaks of polysaccharides (FIG. 2A). Compared with LLTP, SLLTP has two new characteristic peaks, one is 1198cm ^-1 Is asymmetric S = O stretching vibration peak, and the other is 820cm ^-1 The peak is a symmetric C-O-S stretching vibration peak (figure 2B). Elemental analysis of table 2 showed that element S (9.02%) was found in SLLTP due to the introduction of sulfonic acid groups. The above results all indicate that the synthesis of the sulfated polysaccharide SLLTP was successful.

FIG. 2C is an IR spectrum of SLLTPBS. As can be seen from the figure, at 1080cm ^-1 A new absorption peak appears, namely a Si-O-Si stretching vibration peak, which indicates the existence of a silicon oxygen group; and a C-O stretching vibration peak (1020 cm) of LLTP ^-1 ) Substantially coincident and become broader and larger absorption peaks. 3400cm ^-1 The absorption peak is significantly reduced because most of the hydroxyl groups on LLTP have been replaced with sulfonic acid groups and silane. The elemental analysis results show that the introduction of silane results in a slight decrease in the C, H and S content of SLLTPBS compared to SLLTP. Compared with SLLTPBS, the PMO (SLLTP-POSS) hydrophilic microspheres without template removal after POSS introduction are at 1080cm ^-1 The stretching vibration peak of Si-O-Si is obviously enhanced; at the same time, the absorption peaks at 2920 and 2818cm-1 were correspondingly enhanced, mainly due to the introduction of the templating agent C18TACL (FIG. 2D). FIG. 2E is an IR spectrum of PMO (SLLTP-POSS) after purification. As can be seen, the template C18TACL (2920 and 2818 cm) ^-1 ) The intensity of the infrared absorption peak of (C18 TACl) was greatly diminished, indicating that most of the C18TACl was removed by solvent extraction. In addition, the infrared spectrum has little change, and the basic chemical structure of the PMO (SLLTP-POSS) hydrophilic microsphere is unchanged after the template is removed. Table 2 shows that compared with PMO (SLLTP-POSS) hydrophilic microspheres without template removal, the C and H contents of the PMO (SLLTP-POSS) without template removal are obviously reduced, and the S content is slightly increased, which also proves thatIt was clear that most of the C18TACL was removed.

TABLE 2 elemental analysis results at various stages of stationary phase synthesis

2. Table 3 and FIG. 3 show the particle size distribution and morphology of PMO (SLLTP-POSS) microspheres obtained under different conditions, respectively. By adopting a controlled variable method, under the condition of no change of other synthesis conditions, POSS [ C ] ₂ H ₄ Si(OEt) ₃ ] ₈ The molar ratio of SLLTPBS was varied from 0.5. It is shown that the average particle size of the microspheres does not change much when the molar ratio is within this range. FIGS. 3A and B show POSS [ C ] ₂ H ₄ Si(OEt) ₃ ] ₈ SEM results at a molar ratio of 0.75/SLLTPBS of 1, and it was found that the obtained microspheres had smooth surfaces, an average particle diameter of about 5 μm and a uniform particle size distribution.

3. In order to explore the influence of the change of the NaOH content on the morphology of the synthesized material, the NaOH content was from 50mg to 400mg under the same conditions, and the results are shown in Table 3. It can be seen that the average particle size of the microspheres is only about 2.1 μm when the NaOH content is 50 mg. The particle size of the microspheres gradually increases with the increase of the NaOH content, and when the NaOH content is increased to 400mg, the average particle size of the finally obtained microspheres reaches about 10.2 mu m.

4. The hydrolysis rate of the silicon source is also greatly affected by the water content, and with a slight increase in the amount of water, the hydrolysis rate of the silicon source is significantly increased. The process of silica particle formation is a complex competing process of hydrolysis, nucleation and particle growth, wherein hydrolysis is the key to the overall reaction, and thus the change in water content is also one of the main factors affecting the morphology of the final particles. When H is present ₂ O/THF =30/30, the resulting material had irregular morphology (fig. 3C); with H ₂ The appearance of the material gradually approaches to regular sphere (FIGS. 3A, B and D) with gradually increasing volume ratio of O/THF (40/30 and 50/30), average particle size of 4.2 μm and 4.9 μm (Table 3), and the particle size distribution of the microspheres is comparedUniformity; when the volume ratio is further increased to 60/30, the average particle size of the microspheres becomes smaller and the particle size distribution becomes broader (fig. 3E). The influence of the water content on the morphology of the material is mainly related to the hydrolytic polycondensation rate of the silicon source. When the water content is low, the relative content of NaOH becomes high, at the moment, the silicon source polycondensation rate is increased and is far greater than the hydrolysis rate, and the obtained material is thick and irregular; with the gradual increase of the water content, the hydrolytic polycondensation rate of the silicon source gradually tends to be balanced, so that the shape of the material gradually tends to be regular and uniform; when the water content is further increased, the hydrolysis rate of the silicon source becomes higher, and the nucleation rate is faster than the growth rate, so that new and old crystal nuclei grow together, resulting in uneven size of finally obtained particles.

5. The reaction temperature also has a great influence on the formation of the mesoporous hybrid silica material morphology. The changes in particle size of the microspheres at 50 deg.C, 60 deg.C, 70 deg.C, 80 deg.C and 90 deg.C, respectively, were investigated and are shown in Table 3. It can be seen that PMO (SLLTP-POSS) microspheres having an average particle size of about 5.9 μm were obtained at a reaction temperature of 50 ℃. The particle size of the microspheres tends to decrease gradually with increasing temperature. When the temperature reaches 90 ℃, the average grain diameter of the microspheres is reduced to about 1.3 μm.

TABLE 3 particle size distribution of PMO (SLLTP-POSS) microspheres obtained under different conditions

6. Physical parameters such as specific surface area and pore size of non-expanded PMO (SLLTP-POSS) and expanded PMO (SLLTP-POSS) microspheres are shown in Table 4. The results showed that the specific surface area, pore diameter and pore volume of the non-pore-enlarged PMO (SLLTP-POSS) were 517m, respectively ² G, 3.7nm and 0.74cm ³ (iv) g. And the pore diameter of the reamed PMO (SLLTP-POSS) is 9.6nm, which is far larger than that of the non-reamed PMO (SLLTP-POSS), and the specific surface area is slightly increased. Due to the fact thatThus, the reamed PMO (SLLTP-POSS) has a high surface area, large pore size and spherical morphology and is suitable as an HPLC packing material.

TABLE 4 specific surface area and pore Structure parameters of microspheres at different reaction stages

7. From an application point of view, the thermal stability of PMO (SLLTP-POSS) microspheres is an important aspect, and its thermogravimetric analysis curve in air is shown in fig. 4. As can be seen from FIG. 4, from 50 ℃ to 320 ℃, there is almost no loss of weight of the PMO (SLLTP-POSS) microspheres, which ensures a satisfactory thermal stability of the material. The temperature for starting pyrolysis is about 330 ℃, the decomposition and combustion of organic components in the hybrid microsphere skeleton are attributed to, and finally, inorganic residues (mainly SiO crosslinked by silane) of the PMO (SLLTP-POSS) microspheres ₂ ) 46 percent, and ensures the stronger mechanical strength of the microsphere. The higher thermal stability of PMO (SLLTP-POSS) microspheres is mainly due to the fact that POSS has a regular cage-shaped structure, and the inorganic silica framework structure of the POSS has obvious reinforcing effects on the thermal stability and the mechanical strength of the microspheres.

8. The structural order of the organic-inorganic hybrid microspheres was evaluated by XRD. FIG. 5 shows the XRD pattern of PMO (SLLTP-POSS) microspheres. In the low-angle spectrum, the main peak observed at 2 θ =0.74 ° can be attributed to the diffraction peak of the (100) plane, and in addition, the two small peaks observed at 2 θ =1.63 ° and 2.02 ° are attributed to the diffraction peaks of the (110) and (200) planes, respectively, which is the structure of a typical hexagonal mesoporous material, indicating that PMO (SLLTP-POSS) microspheres have a highly ordered characteristic.

Example 3 chromatographic evaluation of PMO (SLLTP-POSS) hydrophilic microspheres as a hydrophilic stationary phase

The chromatographic analysis was performed on an agilent 1260 high performance liquid chromatograph (usa).

1. Preparation of PMO (SLLTP-POSS) hydrophilic chromatographic column

A PMO (SLLTP-POSS) hydrophilic chromatographic column is prepared by a homogenization method. Using isopropanol/trichloromethane =1 (v/v) as a homogenate, adding 4.0g of PMO (SLLTP-POSS) hydrophilic microspheres into the homogenate, performing ultrasonic treatment for 10min to uniformly disperse the microspheres, and pouring the mixture into a homogenate tank. Methanol was used as a displacement liquid, and the displacement liquid was packed in a stainless steel column tube (150 mm. Times.4.6 mm) under a pressure of 370bar, to obtain a novel hydrophilic column.

2. Examination of mechanical Strength of PMO (SLLTP-POSS) hydrophilic column

In the PALC mode, whether the mechanical strength of the microspheres meets the packing requirement of a chromatographic column is judged by examining the relationship between the flow rate and the pressure drop of the PMO (SLLTP-POSS) chromatographic column. The column pressure was measured by changing the flow rate under the conditions of 100% methanol as the mobile phase and room temperature as the column temperature, i.e., 0.25mL/min,0.5mL/min,0.75mL/min,1.0mL/min,1.25mL/min,1.5mL/min,1.75mL/min,2.0mL/min,2.5mL/min,3.0mL/min,3.5mL/min and 4.0 mL/min.

The mechanical strength of the chromatographic column is very important for HPLC and UPLC separations. Whether the column pressure is in direct proportion to the flow rate can be observed, and if the column pressure is in direct proportion to the flow rate, the column pressure has better mechanical strength. The PMO (SLLTP-POSS) packings prepared under reaction conditions Nos. 1 to 4 in Table 1 were labeled PMO (SLLTP-POSS) -1, PMO (SLLTP-POSS) -2, PMO (SLLTP-POSS) -3 and PMO (SLLTP-POSS) -4, respectively, and packed into a column. Column pressures were measured at different flow rates and commercial, same-specification C18 columns (5 μm,150 mm. Times.4.6 mm) were selected for comparison. The results are shown in FIG. 6. As can be seen from FIG. 6, the molar ratios of POSS [ C2H4Si (OEt) 3]8/SLLTPBS of 1.5. Under the condition that the flow rate reaches 4mL/min, the relation between the column pressure and the flow rate still does not deviate from linearity, which shows that the three hybrid fillers have good mechanical stability, and the chromatographic column is well filled. However, at a molar ratio of 0.5, corresponding to PMO (SLLTP-POSS) -4, good linearity was exhibited at flow rates from 0.2 to 2.5 mL/min. However, when the flow rate exceeds 2.5mL/min, the column pressure rapidly increases, and the deviation from linearity is severe. It is probably because the POSS content is less, the rigidity of the material is poorer, and when the pressure of a chromatographic column is overlarge, partial microsphere particles collapse.

Thus, PMO (SLLTP-POSS) -1, PMO (SLLTP-POSS) -2 and PMO (SLLTP-POSS) -3 can be used as chromatographic stationary phases. However, PMO (SLLTP-POSS) -3 was selected as a subject of subsequent studies, considering that the higher the organic content, the stronger the separation ability of the column.

3. Investigation of acid and alkali resistance and stability of PMO (SLLTP-POSS) hydrophilic chromatographic column

And (4) investigating acid and alkali resistance of the chromatographic column. Maintaining the flow rate at 1.0mL/min, and adjusting the pH value to 1.0 by adopting ACN and trifluoroacetic acid aqueous solution with the mass fraction of 1% as an acid-proof test mobile phase under the condition that the column temperature is room temperature; the alkali-resistant test mobile phase adopts ACN/50mmol/L triethylamine aqueous solution, and the pH value is adjusted to 11.0. Fumaric acid and thiourea were used as test probes, and the injection was performed every 8h for 15 times for a total of 120h. The acid and base resistance stability of the column was judged by the remaining percentage of fumaric acid and thiourea retention, respectively, relative to the respective initial retention.

3.1 alkaline stability

For silica gel matrix fillers, the generally accepted failure mechanism at high pH is that the silica gel particles will dissolve under base catalysis. The base stability of the column was investigated by successive washings of the column under high pH conditions. FIGS. 7A and B show the retention factor and the percentage of theoretical plate number remaining relative to the initial value for thiourea plotted against the rinse time. The retention factor and the number of plates of thiourea on the chromatographic column were 93.8% and 95.2% respectively of the initial value within 120h, and both remained above 90%, although there was a decrease, but not significant. Comparative C18-SiO ₂ The column, at the same time, showed a faster drop in retention factor and number of plates, 69.2% and 60.1% of the initial values, respectively, probably due to a severe drop in separation capacity caused by partial dissolution of the silica gel particles. Therefore, the PMO (SLLTP-POSS) stationary phase has good alkaline stability.

3.2 acid stability

Under acidic conditions, the degradation mechanism of the silica gel bonded stationary phase is the hydrolysis of the Si-O-Si type bonded phase under acid catalysis, resulting in a decrease in the retention capacity of the chromatographic column. To improve the stability of the packing under acidic conditions, a PMO (SLLTP-POSS) column was used to overcome this difficulty. The stability of the column was tested by continuous washing under acidic conditionsAnd (5) performing qualitative determination. As shown in fig. 7C and D, the retention factor and the percentage remaining of the number of plates relative to the initial value were plotted against the rinsing time. The results show that after 120h of continuous rinsing, commercial C18-SiO ₂ On the column, the retention factor and number of plates of fumaric acid were reduced by 17% and 21%, respectively, compared to the initial values, indicating that the C18-bonded phase was significantly lost during the flushing of the mobile phase at pH = 1.0. On a PMO (SLLTP-POSS) chromatographic column, the retention factor and the number of the fumaric acid plates are respectively 94.2 percent and 97.7 percent of the initial values, and the reduction is less than 5 percent, which shows that the acidity stability of the newly prepared stationary phase is obviously improved. The above results demonstrate that this hybrid chromatography column has good acid resistance.

And (5) examining the stability in water. Under the conditions of mobile phase water/ACN =90 (v/v), detection wavelength of 260nm and flow rate of 1.0mL/min, a mixture of VB2, VB6, adenine and caffeine (each substance concentration is 30 mg/mL) is separated on a chromatographic column, the column is continuously flushed with a mobile phase for three months in the period of 1 time every other day, and the change of a fixed phase relative to the retention factor and the peak area of the compound is considered, so that the stability of the filler under the condition of a long-time water-rich mobile phase is verified by respectively calculating corresponding RSD.

The results show that the retention factor RSD values of the four compounds are respectively 3.7%,3.2%,2.3% and 4.1%, and the peak area RSD values are respectively 3.5%,3.9%,2.3% and 3.9%, and are all less than 5%, which proves that the separation performance of the chromatographic column is still very stable after the chromatographic column is used for three months in the PALC mode.

5. Chromatographic column process repeatability investigation

20 batches of PMO (SLLTP-POSS) -3 packing (product prepared under reaction conditions No. 3 in Table 1) were synthesized as in example 1 and packed into a chromatographic column. 7 of these were randomly sampled and a mixture of four synthetic pigments, lemon yellow, sunset yellow, brilliant blue and new red, was isolated (each substance concentration was 30 mg/mL). The reproducibility of the synthesis process and the column packing technique of this column packing was evaluated using the Relative Standard Deviation (RSD) of five chromatographic parameters of retention time, peak area, peak width, peak asymmetry and retention factor with a mobile phase of water/ACN =80 (v/v), a flow rate of 1.0mL/min and a detection wavelength of 254 nm. As can be seen from Table 5, the chromatographic columns prepared from 7 batches of the packing have the retention time, the peak width and the RSD of the peak asymmetry of less than 7%, the peak area RSD is between 5% and 8%, and the retention factor RSD is between 6% and 9.2%, so that the separation capability and the column efficiency of 7 chromatographic columns are basically consistent, and the stability of the synthetic process and the column packing technology of the packing is better.

TABLE 5 comparison of RSD values of 5 chromatographic parameters for separation of 4 synthetic pigments by PMO (SLLTP-POSS) hydrophilic stationary phase in batches

rt:retention time；pa:peak area；pw:peak width；paf:peak asymmetry factor；rf:retention factor.

6. Reproducibility test

Using VB ₂ And VB ₆ To test the probes, the reproducibility of the columns was evaluated by the RSD of the day and day retention factors and peak areas. In the case where the mobile phase was water/ACN =90 (10 (v/v), the flow rate was 1.0mL/min, and the detection wavelength was 260nm, VB was measured ₂ And VB ₆ The mixture of (2) (each substance concentration is 30 mg/mL) was injected repeatedly 10 times a day for 6 consecutive days, and the daily and daytime RSD values were calculated, respectively. The results show that VB ₂ And VB ₆ The RSD of the retention factors in the day is respectively 2.1 percent and 2.7 percent, the RSD of the peak area is respectively 1.9 percent and 3.1 percent, the RSD of the retention factors in the day is respectively 2.8 percent and 3.5 percent, the RSD of the peak area is respectively 3.9 percent and 4.3 percent, and both are less than 5 percent, which indicates that the prepared chromatographic column has good reproducibility.

Example 4 separation of eight organic acids by PMO (SLLTP-POSS) hydrophilic column

Eight organic acids, namely oxalic acid, tartaric acid, quinic acid, malic acid, shikimic acid, ascorbic acid, acetic acid and maleic acid, are taken as research objects, and the separation capability of a PMO (SLLTP-POSS) hydrophilic stationary phase (PMO (SLLTP-POSS) -3) under the PALC condition is examined.

The mobile phase is formed by the following steps: disodium phosphate-phosphate buffer (0.01 mol/L, pH = 2.2) and ii: and (3) ACN. Gradient elution procedure: 0-2min,95% I → 90% I; 2.1-7min,90% I → 70% I, 7.1-13min,70% I → 95% I, 13.1-15min,95% I. At room temperature, the flow rate was 1.0mL/min, and the detection wavelength was 210nm.

Preparation of PMO (SLLTP-POSS) -3 hydrophilic column preparation was carried out as under 1 in example 3. A C18 column (150 mm. Times.4.6 mm,5 μm) was used for comparison. Then, under the above mobile phase conditions, two kinds of the columns were continuously flushed for 30 days, and eight kinds of organic acids were again separated to evaluate retention behavior.

When organic acids are separated, because of their high polarity, the ionization of the acids often occurs when the proportion of water in the mobile phase is high, thereby affecting the separation effect. It is common practice to select acidic conditions at low pH to reduce the ionization of the target compound, which can improve the peak shape and retention of the acidic compound. As shown in fig. 8A, under the condition of mobile phase pH 2.2, the mixture of eight organic acids on the new stationary phase obtained good separation within 10min, with higher separation degree. Compared to the C18 column, within 12min, oxalic acid, tartaric acid, quinic acid, acetic acid and maleic acid reached baseline separation, but the malic, shikimic and ascorbic acid chromatographic peaks partially overlapped, not reaching baseline separation (fig. 8B). Then, after 30 days of continuous column flushing, the column chromatography of PMO (SLLTP-POSS) showed no significant change in the separation of organic acids as shown in FIG. 8C. However, on the C18 column, the separation efficiency of the eight organic acids was significantly decreased, the chromatographic peaks of tartaric acid and quinic acid partially overlapped, and the chromatographic peaks of malic acid, shikimic acid and ascorbic acid were completely stacked together and could not be separated (fig. 8D). This is mainly because too low a pH value can seriously affect the lifetime of the C18 column. Therefore, the PMO (SLLTP-POSS) chromatographic column has better acid resistance, and has obvious advantages in separating organic acid.

Example 5 separation of monosaccharides, sugar alcohols and amino acid mixtures by PMO (SLLTP-POSS) hydrophilic column

A mixture of 10 monosaccharides, sugar alcohols and amino acids was selected as a test probe, including ribose, mannitol, sucrose, maltitol, raynaud's sugar, melezitose, phenylalanine, methionine, glutamic acid and histidine, and their separation ability by PMO (SLLTP-POSS) hydrophilic stationary phase (PMO (SLLTP-POSS) -3) was studied. Preparation of PMO (SLLTP-POSS) -3 hydrophilic column preparation was carried out as under 1 in example 3.

Meanwhile, a C18 column and a HILIC column were selected as controls, and the specifications were all (150 mm. Times.4.6 mm,5 μm). Mobile phase, i: ammonium formate (200 mM) and II: and (3) ACN. Optimal separation conditions for PMO (SLLTP-POSS) chromatography columns (gradient elution procedure): 0-2min,98% I → 90% I; 2.1-6min,90% I → 65% I, 6.1-11min,65% I, 11.1-12min,65% I → 98% I, 12.1-14min,98% I; optimal separation conditions for a HILIC column: 0-1min,5% I; 1.1-10min,25% I, 10.1-12min,25% I → 5% I, 12.1-14min and 5% I; optimal separation conditions for C18 column: 0-2min,90% I; 2.1-8min,90% I → 75% I, 8.1-12min,75% I, 12.1-13min,75% I → 90% I, 13.1-15min,90% I. The column temperature was room temperature, the flow rate was 1.0mL/min, the drift tube temperature of the evaporative light scattering detector was set to 55 ℃ respectively, and the flow rate of high purity nitrogen was 2.2L/min.

10 polar compounds of ribose, sucrose, reynolds sugar, melezitose, maltitol, mannitol, phenylalanine, methionine, glutamic acid and histidine are taken as research objects, and PMO (SLLTP-POSS) chromatographic columns, HILIC chromatographic columns and C18 chromatographic columns are compared, so that the separation capacities of the three chromatographic columns on the test compounds are respectively and optimally separated.

On a PMO (SLLTP-POSS) chromatographic column, the 10 polar compounds can be subjected to baseline separation within 18min, which is probably because the polysaccharide structure can provide a large number of hydrophilic action sites and ion exchange sites, the column efficiency of the chromatographic column can be greatly improved, and the retention and separation capacity of the polar compounds is strong. On the HILIC column, the chromatographic peaks of ribose and phenylalanine partially overlapped, and the remaining compounds substantially reached baseline separation. The theoretical plate number of the PMO (SLLTP-POSS) column is significantly higher than that of the HILIC column for the above 10 polar compounds. On the C18 chromatographic column, all peaks of 10 compounds appear within 10min, but chromatographic peaks of five compounds of histidine, maltitol, sucrose, mannitol and methionine are stacked together and cannot be separated; at the same time, the chromatographic peaks for melezitose and reynolds sugar did not achieve baseline separation, indicating that commercial C18 columns have poor retention and separation of the test probes. In addition, in consideration of the influence of green chromatography and ACN on the environment, PALC can separate a mixture of the above 10 sugars, sugar alcohols and amino acids instead of the HILIC method.

Claims

1. The preparation method of the ordered mesoporous organic silicon oxide hydrophilic microsphere comprises the following steps:

1) Synthesis of SLLTP-bridged silanes

a. Dissolving lily polysaccharide in ionic liquid, then dropwise adding chlorosulfonic acid diluted by anhydrous pyridine, stirring at constant temperature of 30-70 ℃ for 0.5-3h, and adjusting to neutrality by using an alkali solution to obtain sulfated and modified lily polysaccharide, which is marked as SLLTP;

b. under the protection of nitrogen, stirring and reacting an N, N-dimethylformamide solution of SLLTP and a THF solution of chloromethyltrimethoxysilane at the constant temperature of 40-80 ℃ for 3-10 hours to obtain SLLTP-bridged silane which is marked as SLLTPBS;

2) In the presence of NaOH, using C18TACL as template agent to make POSS [ C ] ₂ H ₄ Si(OEt) ₃ ] ₈ SLLTPBS, C18TACL in a mixed solvent of water and THF; filtering after the reaction is finished, washing the product with ethanol, deionized water and methanol in sequence, and drying in vacuum; then, removing the template agent by adopting a solvent extraction method to obtain the ordered mesoporous organic silicon oxide hydrophilic microspheres.

2. The method of claim 1, wherein:

in the step 1), the ionic liquid is imidazole ionic liquid, and is specifically selected from any one of the following: 1-butyl-3-methylimidazolium chloride salt, 1-butylimidazolium chloride salt, 1, 3-dimethylimidazolium chloride salt;

in the step 1), the ratio of the lily polysaccharide to the ionic liquid is 500mg:10mL-500mg:50mL;

in the step 1), the molar usage amount of the chlorosulfonic acid is 40-80% of the molar number of hydroxyl groups in the lily polysaccharide;

in the step 1), the volume ratio of the chlorosulfonic acid to the anhydrous pyridine is 1:10-1:20;

in the step 1), the alkali is potassium hydroxide or ammonia water;

in the step 1), after the solution is adjusted to be neutral by sodium hydroxide solution, the method further comprises the following steps: loading the system adjusted to neutral into a dialysis bag with cut-off amount of 10KDa, dialyzing in ultrapure water for 24-72h, concentrating, precipitating with ethanol, and lyophilizing at-20 deg.C to obtain SLLTP.

3. The method according to claim 1 or 2, characterized in that:

in b of the step 1), the molar ratio of SLLTP to chloromethyltrimethoxysilane is 1;

in the step 1) b, the volume ratio of the chloromethyltrimethoxysilane to the THF is 1;

in the step 1) b, the N, N-dimethylformamide is anhydrous N, N-dimethylformamide; the THF is anhydrous THF;

in step b) of step 1), the method further comprises the following steps after the reaction is finished: putting the reaction solution into a dialysis bag with cut-off amount of 10KDa, dialyzing in deionized water for 48h, concentrating, precipitating with ethanol, and freeze-drying at-20 deg.C to obtain SLLTPBS.

4. The method of claim 1, wherein:

in the step 2), the POSS [ C ] ₂ H ₄ Si(OEt) ₃ ] ₈ And SLLTPBS in a molar ratio of 0.5;

in the step 2), the mass ratio of the C18TACl to the SLLTPBS is 1:17;

in the step 2), the mass ratio of NaOH to SLLTPBS is 1:37.5;

in the step 2), the volume ratio of water to THF in the mixed solvent of water and THF is 30:30-60:30, of a nitrogen-containing gas;

in the step 2), the reaction temperature of the reaction is 50-90 ℃;

in the step 2), the reaction time is 10-20 h;

in the step 2), the specific method for removing the template agent by adopting a solvent extraction method comprises the following steps: adding the dried solid into 150mL of mixed solution of concentrated hydrochloric acid/deionized water/ethanol per gram, heating and refluxing for 8h to extract the template agent, repeating twice, filtering, washing and drying; wherein the volume ratio of the concentrated hydrochloric acid to the deionized water to the ethanol in the mixed solution is 5:45:50; the mass fraction of the concentrated hydrochloric acid is 36-38%.

5. The method of claim 4, wherein:

in the step 2), the POSS [ C ] ₂ H ₄ Si(OEt) ₃ ] ₈ And SLLTPBS in a molar ratio of 0.75 to 1.5;

in the step 2), the volume ratio of water to THF in the mixed solvent of water and THF is 40:30-50:30, of a nitrogen-containing gas;

in the step 2), the reaction temperature of the reaction is 60-70 ℃.

6. The method of claim 1, wherein: the method also comprises a step of reaming the obtained ordered mesoporous organic silicon oxide hydrophilic microspheres, and the specific method comprises the following steps: reacting the ordered mesoporous organic silicon oxide hydrophilic microspheres, N-dimethyl decylamine and dodecylamine in ultrapure water, filtering after the reaction is finished, washing with ultrapure water and methanol in sequence, and drying in vacuum at 60 ℃ to obtain the expanded ordered mesoporous organic silicon oxide hydrophilic microspheres;

the reaming method further comprises the following steps: and removing N, N-dimethyl decylamine and dodecylamine from the expanded ordered mesoporous organic silicon oxide hydrophilic microspheres by a solvent extraction method.

7. Ordered mesoporous organosilica hydrophilic microspheres prepared by the method of any one of claims 1-6.

8. The use of the ordered mesoporous organosilica hydrophilic microspheres of claim 7 for preparing a hydrophilic chromatographic stationary phase.

9. A hydrophilic chromatographic column, the stationary phase of which is the ordered mesoporous organic silicon oxide hydrophilic microsphere of claim 7.

10. Use of a hydrophilic chromatography column according to claim 9 for separating and/or detecting polar substances.

11. Use according to claim 10, characterized in that: the polar substance is at least one of organic acids, sugar alcohol, amino acid and sweetener;

the sweetener is selected from saccharin sodium, sucralose, sodium cyclamate, aspartame, acesulfame potassium, alitame, neotame, glycyrrhizic acid, glycyrrhetinic acid, stevioside and steviolbioside.