Capillary column resistant to protein adsorption and preparation method thereof

An anti-protein adsorption and capillary column technology, which is applied in separation methods, chemical instruments and methods, solid adsorbent liquid separation, etc., can solve the problem of high toxicity in the production process and poor quality of anti-protein adsorption capillary electrophoresis covalently bonded coating columns Excellent, complex preparation process, etc., to achieve excellent anti-protein adsorption ability, excellent anti-protein adsorption performance, simple and fast preparation process

Inactive Publication Date: 2017-11-28
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] For this reason, the technical problem to be solved by the present invention is that the existing preparation process is complex, the operating conditions are harsh, the production process is highly toxic, the production efficiency is low, the production cost is high, and the prepared anti-protein adsorption capillary electrophoresis covalently bonded coating column In order to solve the problem of poor quality, and provide an anti-protein adsorption capillary column and its preparation method, the process flow is simple, the production process is environmentally friendly, the production efficiency is high, the production cost is low, and the prepared anti-protein adsorption capillary electrophoresis covalently bonded coating Layer column is of good quality

Method used

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  • Capillary column resistant to protein adsorption and preparation method thereof

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preparation example Construction

[0028] A preparation method of a capillary column resistant to protein adsorption, comprising the steps of:

[0029] S1, the concentration is 2-6mg·ml -1 Photosensitive diazo resin solution, water, concentration 2-6mg·ml -1 The cyclodextrin aqueous solution and water are slowly injected into the activated capillary successively; the mass ratio of the photosensitive diazo resin to the cyclodextrin is 1:(1-1.3);

[0030] S2. Dry the capillary prepared in step S1, and expose it to an ultraviolet lamp with a wavelength of 248nm-365nm for 10min-30min to construct a protein-adsorption-resistant capillary column with a diazo resin-cyclodextrin composite structure.

[0031] The weight-average molecular weight Mw of the photosensitive diazo resin is 700-5500, and the weight-average molecular weight of the diazo resin is too small, and the degree of crosslinking is not good enough to affect the coating effect; when the weight-average molecular weight is too large, the solubility of the...

Embodiment 1

[0043] This implementation provides a kind of preparation method of the anti-protein adsorption capillary column, comprises the following steps:

[0044] S0, capillary internal activation pretreatment

[0045] successively with a concentration of 0.1mol·L -1 Sodium hydroxide solution, distilled water, the concentration is 0.1mol L -1 Flush the capillary with hydrochloric acid solution, distilled water and methanol for 30min, 10min, 30min, 10min and 10min respectively, and then use N 2 The activated capillary can be obtained by drying the capillary.

[0046] S1. At a temperature of 10°C, use a concentration of 2mg·ml - The photosensitive diazo resin solution (Mw=2500) washes the capillary column for 10 minutes, and the distilled water washes the capillary column for 2 minutes. Then use a concentration of 2mg·ml -1 The cyclodextrin aqueous solution washes the capillary column for 10 minutes, and then washes the capillary column with distilled water for 1 minute, so that an ...

Embodiment 2

[0052] This implementation provides a kind of preparation method of the anti-protein adsorption capillary column, comprises the following steps:

[0053] S0, capillary internal activation pretreatment

[0054] successively with a concentration of 0.09mol·L -1 Sodium hydroxide solution, distilled water, the concentration is 0.11mol L -1 Flush the capillary with hydrochloric acid solution, distilled water and methanol for 35min, 8min, 35min, 8min and 8min respectively, and then use N 2 The activated capillary can be obtained by drying the capillary.

[0055] S1. At a temperature of 20°C, use a concentration of 3mg·ml - The photosensitive diazo resin solution (Mw=750) was used to wash the capillary column for 10 minutes, and the distilled water was used to wash the capillary column for 2 minutes. Then use a concentration of 2mg·ml -1 The cyclodextrin aqueous solution washes the capillary column for 10 minutes, and then washes the capillary column with distilled water for 1 m...

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Abstract

The invention relates to a capillary column resistant to protein adsorption. The capillary column is characterized in that a protein adsorption-resistant covalent bonding coating is modified on an inner wall of the capillary and is formed by performing ultraviolet irradiation on a light-sensitive diazoresin and cyclodextrin so as to carry out a photocuring cross-linking reaction, and the thickness of the protein adsorption-resistant covalent bonding coating is 1-10nm. The preparation method comprises the following step: constructing the protein adsorption-resistant covalent bonding coating on the inner wall of a quartz capillary column by utilizing electrostatic self-assembly and photocuring cross-linking reaction of the cyclodextrin and the light-sensitive diazoresin. The preparation of the coating can be performed in an aqueous phase in a layer-by-layer self-assembly manner, the preparation process is simple, rapid, non-toxic and low in cost, the chemical bonding of the coating is realized by utilizing the photocuring cross-linking reaction of the light-sensitive diazoresin and the cyclodextrin, and the coating has excellent hydrophilic property and excellent protein adsorption resistance. Therefore, the quality problems that the capillary is easily blocked and the like when the capillary column is applied to the coating on the inner wall of the capillary are solved.

Description

technical field [0001] The invention relates to the technical field of capillary column separation, in particular to a protein adsorption-resistant capillary column and a preparation method of the capillary column. Background technique [0002] Capillary electrophoresis not only has the characteristics of high sensitivity, high resolution, less sample injection, low solvent consumption, high speed, and various detection methods, it can also separate a series of compounds from ions to neutral molecules, from small molecules to biological macromolecules, Especially for the separation and analysis of biomacromolecules such as peptides, proteins, and DNA, capillary electrophoresis has developed rapidly in recent years. However, the interaction between the unmodified inner wall of the capillary and the analyzed sample, especially the biomacromolecular sample, causes non-specific adsorption of proteins on the inner wall of the capillary, which often leads to peak tailing, peak mis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/20B01D15/10
CPCB01D15/206B01D15/10
Inventor 于冰丛海林王明红
Owner QINGDAO UNIV
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