Rapid glucose detection method

A detection method, glucose technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, color/spectral characteristic measurement, etc., can solve the problems of high price of glucose oxidase, hindering the repeated use of enzymes, and high detection costs, and achieve Low cost, high enzyme stability, and sensitive detection effect

Inactive Publication Date: 2020-05-08
SHANDONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, the conditions for the survival of glucose oxidase and peroxidase are relatively harsh, and they need to be stored in a certain concentration of PBS solution at low temperature. If they are exposed to room temperature or higher temperature for a long time, the enzyme will be deactivated and decomposed at a faster rate. The repeated use of the enzyme is hindered, and the glucose oxidase is expensive and complicated to prepare. The GOX method uses a large amount of enzyme to measure glucose, and the detection cost is relatively high.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Synthesis of GOx&AuNCs@ZIF-8

[0055] (1) Synthesis of AuNCs:

[0056] 5 mL of HAuCl with a solution temperature of 37 °C and a concentration of 10 mmol / L 4 The aqueous solution was added to 5 mL of BSA solution with a concentration of 50 mg / mL and a temperature of 37 ° C under vigorous stirring. After 5 minutes, 0.5 mL of NaOH solution with a concentration of 1.0 mol / L was added, and the mixture solution was stabilized at 37° C. and continued to be vigorously stirred for 24 hours. Stirring was stopped when the color of the solution changed from light yellow to dark brown, and the solution was dialyzed against ultrapure water for 48 hours to remove unreacted impurities.

[0057] (2) Synthesis of GOx&AuNCs@ZIF-8:

[0058] At room temperature, 5 mL of 0.04 mmol / L zinc acetate solution, 5 mL of 0.5 mg / mL GOx solution, 4 mL of 0.8 mg / mL AuNCs solution and 5 mL of 0.16 mmol / L 2-methylimidazole solution Mixed, the mixed solution was stirred for 1 minute and the...

Embodiment 2

[0059] Embodiment 2: Establishment of the linear equation for glucose detection

[0060] (1) GOx&AuNCs@ZIF-8 (prepared in Example 1) solutions with a concentration of 40 mg / mL were prepared respectively, and the concentrations were 0 μmol / L, 0.2 μmol / L, 0.5 μmol / L, 1 μmol / L, 5 μmol / L, 10 μmol / L L, 15μmol / L, 20μmol / L, 25μmol / L, 30μmol / L, 35μmol / L, 40μmol / L glucose solution.

[0061] (2) Mix 500 μL of GOx&AuNCs@ZIF-8 solution obtained in step (1) with 1 mL of different gradient glucose solutions prepared in step (1), and bathe in 37°C water for 30 minutes.

[0062] (3) Add 300 mL of acetic acid buffer solution (pH=3.0) with a concentration of 0.1 mmol / L and 100 μL of a TMB solution with a concentration of 8 mmol / L to the solution obtained in step (2) in sequence. After 10 minutes, measure the UV- Vis spectrum, and obtain the ultraviolet-visible absorption intensity of different concentrations of glucose at 652nm.

[0063] (4) According to the data that step (3) obtains, draw t...

Embodiment 3

[0065] Embodiment 3: the detection of glucose content in the sample

[0066] (1) Prepare a solution of GOx&AuNCs@ZIF-8 (prepared in Example 1) with a concentration of 40 mg / mL.

[0067] (2) Mix 500 μL of the GOx&AuNCs@ZIF-8 solution obtained in step (1) with 1.0 mL of the sample solution to be tested, and bathe in a 37°C water bath for 30 minutes.

[0068] (3) Add 300 mL of acetic acid buffer solution (pH=3.0) with a concentration of 0.1 mmol / L and 100 μL of a TMB solution with a concentration of 8 mmol / L to the solution obtained in step (2) in sequence. After 10 minutes, measure the UV- Vis spectrum, obtain A652nm ultraviolet-visible absorption intensity.

[0069] (4) Substitute the ultraviolet-visible absorption intensity obtained in step (3) into the linear equation y=0.015x+0.167 (obtained in Example 2) to calculate the glucose content of the sample to be tested.

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Abstract

The invention discloses a rapid glucose detection method. According to the method disclosed by the invention, a nanoscale GOx&AuNCs@ZIF-8 compound with high catalytic capability is prepared by utilizing a mild and environment-friendly mode. Compared with a uniform mixture of two free enzymes, namely AuNCs and GOX, the GOx&AuNCs@ZIF-8 compound has the advantages that the biocatalytic cascade of theGOx&AuNCs@ZIF-8 compound is enhanced by about 19.3 times, enzyme stability is high, enzyme dosage is small, cost is low, efficient and sensitive detection of glucose is realized, detection conditionsare mild, operation is simple and convenient, and a strong targeting property is realized. The rapid glucose detection method of the invention overcomes the problems of complicated sample pretreatment procedures and expensive instruments in chromatography, the problems of poor targeting property, poor sensor stability and demands for special electrodes and electrochemical equipment in an electrochemical method, and the problems of high cost of colorimetric sensors, large enzyme dosage, easy enzyme inactivation and the like in traditional enzyme method.

Description

technical field [0001] The invention relates to the technical field of carbohydrate analysis and detection, in particular to a rapid detection method for glucose. Background technique [0002] Glucose content in serum is an important indicator to measure the health status of human body. Glucose content in soil is a common indicator to measure soil characteristics. The glucose index in sewage plays an important role in sewage purification. Glucose content in food and medicine is a commonly used indicator for component analysis. Therefore, glucose detection technology is widely used in the fields of medicine, biotechnology, soil detection, industrial wastewater treatment, food and pharmaceutical processing. The development of easy-to-operate and highly sensitive glucose detection technology has become the focus of this field. [0003] At present, the methods for detecting glucose mainly include colorimetric method, electrochemical method, high-performance liquid chromatograph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/54C12Q1/26G01N21/31G01N21/33
CPCC12Q1/26C12Q1/54G01N21/31G01N21/33
Inventor 李向阳李宗益迟静甜郭曼莉
Owner SHANDONG AGRICULTURAL UNIVERSITY
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