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Polymer monolithic column based on ferrocenyl porphyrin as well as preparation method and application of polymer monolithic column

An iron-based porphyrin and polymer technology, applied in chemical instruments and methods, other chemical processes, chemical/physical processes, etc., can solve the problems of low column efficiency, poor protein selectivity, low sample loading, etc. problems, to achieve good mechanical stability and permeability, less methanol usage, and high separation efficiency

Active Publication Date: 2022-04-08
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although capillary monolithic columns have good separation efficiency for proteins and small molecules, they have inherent defects such as poor repeatability and low sample loading; in addition, traditional size HPLC columns can overcome these limitations of capillary monolithic columns. Disadvantages, but it has poor selectivity for proteins and low column efficiency for small molecules

Method used

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  • Polymer monolithic column based on ferrocenyl porphyrin as well as preparation method and application of polymer monolithic column
  • Polymer monolithic column based on ferrocenyl porphyrin as well as preparation method and application of polymer monolithic column
  • Polymer monolithic column based on ferrocenyl porphyrin as well as preparation method and application of polymer monolithic column

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Use 0.5 mL of N,N-dimethylformamide as a solvent;

[0033] 0.03 mg of ferrocene porphyrin, 0.6 mL of triallyl isocyanurate and 0.5 mL of methyl methacrylate were used as reaction monomers;

[0034] 0.9mL of ethylene glycol dimethacrylate was used as the cross-linking agent;

[0035] 0.8mL of 1,4-butanediol and 1.0mL of dodecanol were used as porogens;

[0036] 0.012 mg of azobisisobutyronitrile was used as initiator.

[0037] S1. Mix the above-mentioned solvent, reaction monomer, crosslinking agent, porogen, and initiator, obtain a prepolymerized mixed solution after ultrasonic dispersion is uniform, and then feed nitrogen to remove the oxygen in the prepolymerized mixed solution;

[0038] S2. Load the prepolymerized mixed solution in step S1 into a 50mm×4.6mmI.D. stainless steel chromatographic column tube, seal both ends, and conduct a water bath polymerization reaction at 60°C for 12h;

[0039] S3. After the reaction is completed, use methanol as a mobile phase in...

Embodiment 2

[0055] Preparation of human plasma samples: 4.0 mL of fresh human plasma was centrifuged at 12000 r / min for 15 min at 4°C to obtain the supernatant, which was stored at 4°C before use.

[0056] The monolithic column prepared in Example 1 was connected to a high-performance liquid chromatograph, the flow rate was 1 mL / min, the wavelength was 280 nm, the temperature was 30 °C, the mobile phase was 0.1% TFA water / 0.1% TFA ACN, and 20 μL was injected. Gradient elution test for elution ability of human plasma proteins, results such as Figure 4 shown.

[0057] from Figure 4It can be seen that when the mobile phase changes linearly from 0%B to 38%B within 60 min, human plasma proteins are adsorbed on the prepared monolithic column, and it can be completely eluted after changing the mobile phase.

Embodiment 3

[0059] Preparation of aromatic homologue samples: All aromatic homologue samples (aniline, benzotriazole, phenol, p-nitroaniline, benzene, α-naphthol, p-nitrochlorobenzene, naphthalene, fluorene, and anthracene) were prepared with methanol Prepared as a 0.01mol / L solution. And these samples were stored at 4°C until use.

[0060] The monolithic column prepared in Example 1 was connected to a high performance liquid chromatograph, and the acetonitrile / water mixed solution with a volume ratio of 45:55 was used as the mobile phase, and the mixed sample of 1 mg / mL was continuously injected, and then methanol was used as the mobile phase. The above 10 samples of aromatic homologues were chromatographically separated with the monolithic column prepared in Example 1, and the results were as follows: Figure 5 shown.

[0061] As can be seen from the figure, the chromatographic separation of the aromatic homologue samples all reached baseline separation within 8 minutes. It can be se...

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Abstract

The invention belongs to the technical field of polymer monolithic columns, and discloses a ferrocenyl porphyrin-based polymer monolithic column and a preparation method and application thereof.The preparation method comprises the steps that S1, N, N-dimethylformamide serves as a solvent, ferrocenyl porphyrin, triallyl isocyanurate, methyl methacrylate, a cross-linking agent, a pore-foaming agent and an initiator are mixed, and a mixture is obtained; carrying out uniform ultrasonic dispersion to obtain a pre-polymerized mixed solution, and introducing nitrogen to remove oxygen in the pre-polymerized mixed solution; s2, filling the pre-polymerized mixed solution into a column tube to carry out polymerization reaction; s3, after the reaction is completed, soluble substances are removed through a chromatographic system, and the ferrocenyl porphyrin-based polymer monolithic column is obtained; in conclusion, the polymer monolithic column with ferrocenyl porphyrin as the monomer is prepared on the basis of a free radical polymerization principle, and the monolithic column has a relatively uniform spatial network structure, relatively low back pressure and good mechanical stability and permeability, and can effectively realize fractional separation of proteins and small molecular substances.

Description

technical field [0001] The invention belongs to the technical field of polymer monolithic columns, and in particular relates to a ferrocene-based porphyrin-based polymer monolithic column and a preparation method and application thereof. Background technique [0002] As active macromolecules in cells, proteins are undoubtedly the main molecules associated with diseases, and changes in protein expression levels are directly related to diseases. Therefore, in the post-genome era, proteomics based on the overall level of proteins plays an important role in human disease research. important role. [0003] The primary content of human proteomics research is the composition of the proteome, which requires researchers to characterize the proteome and achieve the separation, identification and mapping of all proteins. Plasma proteomics can be used to screen disease-related protein disease markers in clinical applications. For example, the expression difference of hydrophobic proteo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/26B01J20/30B01D15/08C08F222/14C08F226/06C08F220/14C08J9/28
Inventor 王全郭圆圆李红亚朱宝成
Owner HEBEI AGRICULTURAL UNIV.
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