A colorimetric sensing method based on enzyme-induced generation of inorganic nanoparticles

An inorganic nanoparticle, colorimetric sensing technology, applied in the field of colorimetric biosensing, can solve the problems of difficulty in detection, limit the application of PB, etc., and achieve the effects of low cost, fast color change time, and high yield

Active Publication Date: 2020-08-11
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is still difficult to detect targets that are not reactive with PB precursors, limiting the application of PB as a colorimetric method

Method used

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  • A colorimetric sensing method based on enzyme-induced generation of inorganic nanoparticles
  • A colorimetric sensing method based on enzyme-induced generation of inorganic nanoparticles
  • A colorimetric sensing method based on enzyme-induced generation of inorganic nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] First prepare a 50 mM MES buffer solution with ultrapure water, and adjust the pH to 3.0 with dilute hydrochloric acid and potassium hydroxide. Use 50mM MES buffer solution to prepare 0.5mM, 1.5mM ferric chloride solution and 0.5mM, 1.5mM potassium ferricyanide solution, 0.5mM ferrous chloride solution, 1.1mg / mL glucose oxidase solution, 22mM glucose solution and 21mM H 2 o 2 solution.

[0045] First to verify that the method as figure 1 There are two pathways that co-enhance the effect. Take 50 μL of 0.5mM ferric chloride solution and 50 μL of 0.5mM potassium ferricyanide solution respectively, and prepare 4 parts, respectively without adding (sample 1), adding 5 μL of 22mM glucose solution (sample 2), adding 5 μL of 1.1mg / mL glucose oxidase solution (Sample 3), 5 μL of 22 mM glucose solution and 5 μL of 1.1 mg / mL glucose oxidase solution were added (sample 4). After reacting for 10 minutes, measure the UV-Vis absorption spectrum, such as figure 2 shown. From ...

Embodiment 2

[0051] To buy semi-sweet red wine from the supermarket, first filter the red wine through a filter membrane with a 0.2 μm pore size for 3 times, and then dilute it by 1 time with 50 mMMES buffer solution. Take 50μL of 1.5mM ferric chloride, 50μL of 1.5mM potassium ferricyanide and 5μL of 1.1mg / mL glucose oxidase, add 5μL of diluted red wine solution to it, and measure the UV-vis absorbance value after 10 minutes of reaction, as Figure 5 . Take the absorbance value at 706nm, and calculate the glucose content as 27.52±0.04mM according to the equation in Example 1 and the dilution factor.

Embodiment 3

[0053] To buy sweet red wine from the supermarket, first filter the red wine 3 times through a filter membrane with a pore size of 0.2 μm, and then dilute it by 1 time with 50 mMMES buffer solution. Take 50μL of 1.5mM ferric chloride, 50μL of 1.5mM potassium ferricyanide and 5μL of 1.1mg / mL glucose oxidase, add 5μL of diluted red wine solution to it, and measure the UV-vis absorbance value after 10 minutes of reaction, as Figure 5 . Taking the absorbance at 706nm, according to the equation in Example 1 and the dilution factor, the glucose content was calculated to be 34.62±0.07mM.

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Abstract

The invention discloses a colorimetric sensing method for generating inorganic nanoparticles based on enzyme catalytic induction. The method comprises the steps: taking a frerric chloride solution, apotassium ferricyanide solution and an enzymatic solution, mixing, respectively adding the mixed solution into target solutions with known different concentrations, carrying out a reaction, quantitatively detecting by ultraviolet-visible spectroscopy to obtain the corresponding ultraviolet-visible absorbance values in all the concentrations, and thus obtaining a linear relationship between the target concentrations and the absorbance values; and adding a to-be-measured target solution to the mixed solution, detecting the absorbance value, and calculating to obtain the target concentration of the to-be-measured target solution. The method directly adopts Prussian blue as a colorimetric probe, and the constructed colorimetric sensing method has the advantages of being fast, simple, low in cost, high in sensitivity, good in selectivity and the like, has satisfactory results in application and has broad application prospects.

Description

technical field [0001] The invention belongs to a colorimetric biosensing technology for quantitative detection, in particular to a colorimetric sensing method based on enzyme-catalyzed induction to generate inorganic nanoparticles. Background technique [0002] Glucose, as the main reducing sugar present in most beverages, is directly related to the total sugar content. Therefore, detection of glucose content in the food industry is of great significance to ensure good food quality. Typically, glucose is detected by conventional methods such as titration and chromatography. However, the end point of the titration method is difficult to judge, especially for beginners. The standard method used to detect glucose in food is chromatography, such as liquid chromatography / high performance liquid chromatography, which has high sensitivity, specificity and reproducibility, but is time-consuming and requires expensive instruments and highly skilled personnel. For simple, rapid an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/3103G01N21/78
Inventor 傅迎春戴煌李延斌
Owner ZHEJIANG UNIV
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