Cation type chitosan bonded and modified capillary electrochromatography open tubular column and manufacturing method thereof

A technology of capillary electrochromatography and cationic chitosan, which is applied in the field of capillary electrochromatography, can solve problems such as poor stability, achieve the effects of avoiding adsorption, good reproducibility, and broad development and application prospects

Inactive Publication Date: 2011-05-25
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to overcome the shortcomings of poor stability existing in existing physical adsorption or electrostatic charge adsorption modification, the present invention provides a cationic chitosan chemically bonded capillary electrochromatographic open-t

Method used

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  • Cation type chitosan bonded and modified capillary electrochromatography open tubular column and manufacturing method thereof
  • Cation type chitosan bonded and modified capillary electrochromatography open tubular column and manufacturing method thereof

Examples

Experimental program
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Embodiment 1

[0013] Embodiment 1: chemical bonding method prepares the open-tubular column modified by carboxymethyl chitosan

[0014] Pour the toluene solution of 30% γ-aminopropyltriethoxysilane into the activated capillary, seal both ends, heat in a gas chromatography furnace at 110°C for 24 hours, take it out and cool it to room temperature, rinse it with toluene and methanol respectively After 0.5 hours, blow dry with nitrogen. The silane-siliconized capillary column was washed with 10% glutaraldehyde (prepared in 50 mM phosphate buffer at pH 8.0) for 1 hour, and then washed with deionized water until neutral to complete the aldehyde group modification of the capillary. Continue to inject 0.2%-0.8% carboxymethyl chitosan solution into the aldehyde-modified capillary, seal both ends, bathe in 60°C water for 1 hour, and blow out the excess solution with nitrogen; finally use 10% glutaraldehyde (pH 8.0 50mM Phosphate buffer) was reacted at room temperature for 1 hour, washed with deioni...

Embodiment 2

[0015] Example 2: Separation of basic proteins by carboxymethyl chitosan modified open-tubular column under anodic electroosmotic flow mode

[0016] Carboxymethyl chitosan was used to modify the open-tubular column (inner diameter 50mm, total length 60.0cm, effective length 40.0cm), with pH 3.020mM phosphate buffer as the mobile phase, separation voltage +15kV, under the CEC anode electroosmotic flow mode , the four basic proteins of trypsin, ribonuclease, cytochrome and lysozyme were separated by point chromatography-ultraviolet detection, and the proteins were completely separated under the above conditions, as shown in figure 2 As shown, the elution peaks are: 1. Trypsin, 2. Ribonuclease, 3. Cytochrome C, 4. Lysozyme, which reduces the tailing phenomenon caused by protein adsorption.

Embodiment 4

[0017] Example 4: Separation of alkaloids by carboxymethyl chitosan modified open-tubular column under cathodic electroosmotic flow mode

[0018] Carboxymethyl chitosan was used to modify the open-tubular column (inner diameter 50mm, total length 60.0cm, effective length 40.0cm), with pH 6.050mM phosphate buffer as the mobile phase, separation voltage +15kV, under the CEC cathodic electroosmotic flow mode , the four basic proteins of morphine, crowbarine, narcotin and papaverine were separated, as shown in Figure 3. The elution peaks are as follows: 1. Morphine, 2. Cybaine, 3. Nacodin, 4. Papaverine. Alkaloids were baseline separated under the above conditions.

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Abstract

The invention discloses a cation type chitosan bonded and modified capillary electrochromatography open tubular column and a manufacturing method thereof. The open tubular column is manufactured by adopting the modifying technologies of silanizing the inner surface of a capillary, modifying aldehyde groups of the capillary wall, polycondensating and bonding chitosan amine aldehyde, crosslinking chitosan molecules, and the like. In the invention, cation type chitosan is bonded in the manufactured capillary open tubular column, and the used cation type chitosan is carboxymethyl chitosan. The cation type chitosan bonded and modified capillary electrochromatography open tubular column has the advantages of firm functional coating chemical bonding, stable quality, double action modes of cathode/anode electroosmotic flows, good separation effect on alkaline protein molecules and active alkaloid and good repeatability, and is suitable for separating and analyzing capillary electrochromatography of positive charge type substances.

Description

Technical field: [0001] The invention relates to a capillary electrochromatographic open-tube column modified by cationic chitosan bonding and a preparation method thereof, belonging to the technical field of capillary electrochromatography. Background technique: [0002] Capillary electrochromatography (capillary electrochromatography) is an efficient and fast new chromatographic micro-separation technology developed in recent years. As one of the main types of capillary chromatographic columns, capillary open-tubular column is an important component of capillary electrochromatographic separation and analysis technology. Compared with capillary packed columns and capillary monolithic columns, capillary open-tubular columns have the characteristics of simple preparation, no need to plug terminals, high column efficiency, and rapid separation, and are more and more valued by chromatographic workers. [0003] In the analysis of basic proteins or alkaloids, the inner wall of t...

Claims

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Application Information

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IPC IPC(8): G01N30/60
Inventor 林旭聪周孙英谢增鸿
Owner FUZHOU UNIV
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