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Method for measuring glucose by fluorescence spectrometry of enzyme-catalyzed oxidation TMB

A technology of catalytic oxidation and fluorescence spectroscopy, applied in the field of glucose determination by enzyme-catalyzed oxidation TMB fluorescence spectroscopy, to achieve the effect of simple method, good selectivity and good reproducibility

Inactive Publication Date: 2019-01-11
GUANGXI NORMAL UNIV
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Problems solved by technology

So far, there have been no reports of glucose oxidase (GOx for short) catalyzing the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) for fluorescent quantitative detection of glucose.

Method used

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  • Method for measuring glucose by fluorescence spectrometry of enzyme-catalyzed oxidation TMB

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Embodiment

[0018] A method for measuring glucose with enzyme-catalyzed oxidation TMB fluorescence spectrum, comprising the steps:

[0019] (1) Prepare glucose standard solution system with known concentration: add 1 μL, 4 μL, 15 μL, 20 μL, 30 μL, 40 μL 500 μmol / L glucose standard solution to 6 different graduated test tubes, and then add 15 μL 0.11 mg / L glucose standard solution in sequence. mL glucose oxidase solution, 150 μL 0.02mol / L pH 7.4 PBS buffer solution and 100 μL 0.5mmol / L TMB solution, shake well, react in a water bath at 40°C for 20min, stop the reaction with ice water, and dilute to 1.5mL with twice distilled water;

[0020] (2) Prepare blank control solution system: adopt the method of step (1) without adding glucose standard solution to prepare blank control solution system;

[0021] (3) Get respectively the glucose standard solution system and the blank control solution system prepared by steps (1), (2) and pour in the quartz cuvette, on the fluorescence spectrometer, se...

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Abstract

The invention discloses a method for measuring glucose by fluorescence spectrometry of enzyme-catalyzed oxidation TMB. The method comprises the following steps: (1) preparing a glucose standard solution system with known concentration, measuring the fluorescence peak intensity value at 380nm as F; (2) preparing a blank control solution system, and measuring the fluorescence peak intensity value asF0; (3) calculating the formula: deltaF=F-F0; (4) making a working curve for the concentration of glucose by deltaF; (5) preparing a measured sample solution, measuring the fluorescence peak intensity value as Fsample, and calculating the formula: deltaFsample=Fsample-F0; and (6) calculating the concentration of glucose in the measured sample according to the working curve. This method uses enzyme to catalyze the oxidation of TMB to produce a product with fluorescence, and uses the reaction product as a fluorescent molecular probe to linearly correlate the fluorescence peak intensity with theconcentration of the object under measurement, thereby realizing the quantitative determination of the target. The method is convenient, good in selectivity, and high in sensitivity.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to a method for measuring glucose by enzyme-catalyzed oxidation of TMB fluorescence spectrum. Background technique [0002] Glucose is one of the most important sources of energy for living cells. It is often used as a nutrient in medicine and is widely used in biomedicine. Therefore, it is of great significance to detect the content of glucose. Currently, glucose detection methods include colorimetric methods, electrochemical methods, fluorescence methods, and chemiluminescence methods. It has been reported in the literature that the prior art has the following methods: a catalytically active multi-element-doped carbon dot (ME-CDs), which is used as a colorimetric sensor for the detection of glucose, and the method is linear The range is 0.20-2.50mmol / L, and the detection limit is 0.06mmol / L. This method has good specificity, but low sensitivity; 2 o 2 Under the condition of ...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N21/78
CPCG01N21/643G01N21/78
Inventor 蒋治良姚东梅李重宁梁爱惠
Owner GUANGXI NORMAL UNIV
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