Alcohol quantitative analysis method based on double-enzyme-inorganic nanoflower composite material

A technology of inorganic nano and composite materials, applied in the field of quantitative analysis, can solve problems such as low operational stability, narrow linear operating range, low sensitivity, etc., and achieve improved enzyme activity and stability, special material morphology, and high specific surface area Effect

Active Publication Date: 2019-07-12
NANJING NORMAL UNIVERSITY
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Problems solved by technology

Such sensors have high selectivity, but their associated usage costs are high and cannot adequately address the shortcomings of alcohol detection, such as low sensitivity, low stability, and narrow linear operating range in practical applications
Biological enzymes have low stability, are not easy to store for a long time, and are easy to inactivate, which brings certain limitations to the above method of alcohol detection ...

Method used

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  • Alcohol quantitative analysis method based on double-enzyme-inorganic nanoflower composite material
  • Alcohol quantitative analysis method based on double-enzyme-inorganic nanoflower composite material
  • Alcohol quantitative analysis method based on double-enzyme-inorganic nanoflower composite material

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Embodiment 1

[0025] A method for quantitative analysis of alcohol based on double enzyme-inorganic nano flower composite material, comprising the following steps:

[0026] (1) Preparation of dual enzyme-inorganic nanoflower composite materials: such as figure 1 As indicated, 15 μL of alcohol oxidase (AOX) solution 1.0 mg / mL and horseradish peroxidase (HRP) 1.0 mg / mL were added to 1.41 mL of phosphate buffered saline (PBS), and then 168 μL of 100 mM CaCl 2 Add the solution to the above solution, mix it evenly, let it stand still, incubate at 30°C for 48 hours, take it out, then centrifuge at 10000r / min for 15min, remove the supernatant, then add 150μL of PBS, mix well, and you can get The dual enzyme-inorganic nanoflower composite (AOHNF) required for the experiment.

[0027] Such as figure 2 As shown, the dual enzyme-inorganic nanoflower composite material is characterized by SEM. The dual enzyme-inorganic nanoflower composite material is a uniform nanoflower-shaped ball with an averag...

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Abstract

The invention discloses an alcohol quantitative analysis method based on a double-enzyme-inorganic nanoflower composite material. The method comprises the following steps: adding alcohol oxidase (AOX), horse radish peroxidase (HRP) and CaCl2 solution into a phosphate buffer solution, standing and reacting at room temperature to obtain the double-enzyme-inorganic nanoflower composite material (AOHNF). The reaction conditions are simple, and the enzyme activity and stability of the obtained material are remarkably enhanced. Alcohol in the solution is catalyzed and oxidized by using AOX to generate H2O2, TMB is catalyzed by using HRP to react with H2O2 to enable the solution to be blue, and the absorbance of the absorption wavelength at 650 nm is measured, so that the alcohol concentration inthe solution is obtained. The analysis method is simple to operate and high in detection result sensitivity. Double enzyme-based method disclosed by the invention The alcohol quantitative analysis method based on the double-enzyme-inorganic nanoflower composite material has the characteristics of simplicity in operation, high sensitivity, good stability and the like, and has a good development prospect.

Description

technical field [0001] The invention belongs to the field of quantitative analysis, in particular to an alcohol quantitative analysis method based on a double enzyme-inorganic nano flower composite material. Background technique [0002] With the development of the times, the detection of alcohols with high sensitivity, high selectivity and high accuracy has been widely used in many different fields, and the rapid, sensitive, selective and quantitative detection of alcohols is very important for law enforcement, chemical industry, pharmaceuticals and fermentation It is also of great significance to industries such as , where accurate and rapid determination of ethanol content is very important in clinical and forensic analysis. Furthermore, simple, fast and economical methods of alcohol analysis are also used in the pulp, food and beverage industries to control the quality of fermentation processes and their products. In the processes of traffic law enforcement, forensic me...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/31G01N21/78
Inventor 韦天香徐乔任琳琳
Owner NANJING NORMAL UNIVERSITY
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