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Mesoporous carbon immobilized enzyme biological sensing material doped with nitrogen and preparing method thereof

A nitrogen-doped mesoporous carbon, immobilized enzyme technology, applied in the fields of biocatalysis and biosensing, can solve the problem of poor biocompatibility of mesoporous carbon materials, unsuitable for large biomolecular loads, and small pore size of mesoporous carbons. and other problems, to achieve the effect of environmental friendliness, strong practicability and high thermal stability

Inactive Publication Date: 2009-05-20
SHANGHAI INST OF CERAMIC CHEM & TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the biocompatibility of mesoporous carbon materials is not good, and the activity of biomolecules is seriously lost after combining with them, which largely limits its application in the field of biosensing.
In addition, the mesoporous carbon prepared in the past has a small pore size, which is not suitable for large biomolecular loading, which is also a disadvantage for its use in the preparation of biosensors.

Method used

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  • Mesoporous carbon immobilized enzyme biological sensing material doped with nitrogen and preparing method thereof
  • Mesoporous carbon immobilized enzyme biological sensing material doped with nitrogen and preparing method thereof
  • Mesoporous carbon immobilized enzyme biological sensing material doped with nitrogen and preparing method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Accurately measure 2.4ml of ethylenediamine and 3.4ml of carbon tetrachloride, mix well, add 0.5g of MCF template, reflux at 90°C for 6 hours, then transfer to an oven to dry for 12 hours, the obtained solid is finely ground with a mortar, and then Carbonization was carried out at 600° C. for 5 hours in a nitrogen atmosphere. The resulting carbide was treated with 10% HF solution to remove the silicon template. The nitrogen adsorption curve and pore size distribution of the obtained nitrogen-doped mesoporous carbon are as follows: figure 1 and figure 2 As shown, its specific surface area and pore volume are 474m 2 g- 1 and 0.85cm 3 g -1 , the pore size is about 17nm obtained from the adsorption curve.

[0030] Weigh 80 mg of the obtained nitrogen-doped mesoporous carbon, put it into 4 ml of 20% nitric acid, react at 60° C. for 6 h for modification, filter, wash and dry.

[0031] Weigh 10 mg of the above-mentioned modified nitrogen-doped mesoporous carbon, add to...

Embodiment 2

[0037] The pyrolytic graphite electrode was sanded smooth on sandpaper, sonicated in deionized water for 10 min, and removed to dry. Then take 10 mg of the immobilized enzyme obtained in Example 1, disperse it in 2 ml of 5wt% Nafion phosphate buffer solution, take 0.5 ml, evenly drop-coat it on the pyrolytic graphite electrode, dry it at room temperature to form a film, and form a biosensor for electrochemical determination.

[0038] The electrochemical determination was carried out on a CHI 440 electrochemical workstation, using a three-electrode system, in which the graphite electrode was used as the working electrode, the platinum wire was used as the counter electrode, and Ag / AgCl was used as the reference electrode. The cyclic voltammogram of the prepared enzyme sensor is as follows: Figure 4 As shown, the appearance of a pair of reversible redox peaks indicates that the enzyme sensor can realize direct electron transfer between the enzyme and the electrode, and its ele...

Embodiment 3

[0042] In Example 2, other experimental methods and preparation steps remained unchanged, except that 10 mg of the immobilized enzyme in Example 1 was dispersed in 4 ml of 5 wt% Nafion phosphate buffer solution, and then 0.5 ml was used to prepare the biosensor. The response time of the enzyme sensor is also less than 5 seconds, and the linear range is 1-80 μmol. The curve of its current response value to the concentration of hydrogen peroxide is as follows Figure 5 As shown, the calculated sensitivity of the enzyme sensor is 142mA L mol -1 cm- 2 , all lower than those in Example 2 and Comparative Example 2.

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Abstract

The invention relates to a nitrogen-doped mesoporous carbon immobilized enzyme biosensing material and a preparation method thereof, which belongs to the field of biocatalysis and biosensing. Mesoporous carbon is prepared by taking mesoporous silica as a template and ethylenediamine, acrylonitrile or tripolycyanamide as a carbon-nitrogen source, and then nitric acid is used to modify the surface of the mesoporous carbon to obtain hydrophilically modified nitrogen-doped mesoporous carbon. The material has high specific surface area, large pore volume, large aperture and good biocompatibility, and also has good water and thermal stability and electric conductivity at the same time. The activity of enzyme molecules mobilized by taking the material as a carrier is well maintained. The biosensing based on nitrogen-doped mesoporous carbon immobilized enzyme has quick response time, wide linear range, high sensitivity, and low detection limit. In addition, a preparation process for the immobilized enzyme biosensing material is simple and controllable, and has the advantages of mild conditions, easily-obtained raw materials, and conventional equipment.

Description

technical field [0001] The invention relates to a nitrogen-doped mesoporous carbon-immobilized enzyme biosensing material and a preparation method thereof. It belongs to the field of biocatalysis and biosensing. Background technique [0002] Biosensors are sensors composed of immobilized cells, enzymes, antibodies or other biologically active substances and signal converters. It is a new type of information technology developed by the interpenetration of biomedicine, electronics and engineering in recent years. It has the advantages of strong specificity, high sensitivity, fast response, low cost and long service life. , Biosensors have a wide range of application values ​​in medicine, medicine, agriculture, animal husbandry and other disciplines, food, fermentation, pharmaceutical and other industrial sectors, and environmental monitoring. [0003] The selection of appropriate carrier materials to immobilize biomolecules is the key technology to determine the characterist...

Claims

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Application Information

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IPC IPC(8): G01N27/327
Inventor 高秋明师琳璞
Owner SHANGHAI INST OF CERAMIC CHEM & TECH CHINESE ACAD OF SCI
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