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A method for quantitative analysis of alcohol based on dual enzyme-inorganic nanoflower composite material

An inorganic nano-composite material technology, applied in the field of quantitative analysis, can solve the problems of low operational stability, narrow linear operating range, low sensitivity, etc., to improve the activity and stability of enzymes, special material morphology, high specific surface area. Effect

Active Publication Date: 2021-11-02
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 through biological enzymes
[0004] As multifunctional tools, enzymes have been extensively studied due to their unique properties, such as high reaction specificity, high catalytic activity, and low toxicity, but natural free enzymes have some inherent disadvantages, such as high preparation and purification costs, and low operational stability , catalytic activity is sensitive to environmental conditions and difficult to recycle

Method used

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  • A method for quantitative analysis of alcohol based on dual enzyme-inorganic nanoflower composite material
  • A method for quantitative analysis of alcohol based on dual enzyme-inorganic nanoflower composite material
  • A method for quantitative analysis of alcohol based on dual 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 dual-enzyme-inorganic nanoflower composite material. The method comprises alcohol oxidase (AOX), horseradish peroxidase (HRP), CaCl 2 The solution was added into a phosphate buffer solution, and left to stand at room temperature for reaction to obtain a dual enzyme-inorganic nanoflower composite material (AOHNF). The reaction conditions are simple, and the enzyme activity and stability of the obtained material are significantly enhanced. Then use AOX to catalyze the oxidation of alcohol in the solution to generate H 2 o 2 , HRP catalyzes the interaction of TMB with H 2 o 2 The reaction makes the solution appear blue, and the absorbance of the absorption wavelength at 650nm is measured to obtain the alcohol concentration in the solution. The analysis method is simple to operate, and the detection result has high sensitivity. The alcohol quantitative analysis method based on the dual enzyme-inorganic nanoflower composite material of the present invention has the characteristics of simple operation, high sensitivity, good stability, etc., 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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Patent Type & Authority Patents(China)
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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