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Preparation method and application of nano-enzyme for colorimetric detection of glucose

A glucose and nano-enzyme technology, applied in the field of analytical chemistry, can solve the problems of poor stability, time-consuming and laborious, and high cost of natural enzyme purification, and achieve the effects of low cost, simple preparation method, good anti-interference and reliability

Active Publication Date: 2021-08-24
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the cost of preparation and purification of natural enzymes is high, their stability is poor, and they are easily digested by proteases. More stable and highly active artificial enzymes are needed to replace them.
At the same time, since the optimal activity conditions of natural enzymes and nanozymes are often different, multi-step reactions are required to realize the detection of glucose, which is time-consuming and laborious.

Method used

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  • Preparation method and application of nano-enzyme for colorimetric detection of glucose
  • Preparation method and application of nano-enzyme for colorimetric detection of glucose
  • Preparation method and application of nano-enzyme for colorimetric detection of glucose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A method for preparing a nanozyme for colorimetric detection of glucose, the steps are as follows:

[0032] Disperse 620μL of chloroauric acid (HAuCl4) aqueous solution (30mM), 4.5mg of sodium tetrachloropalladate (Na2PdCl4) and 5mg of norepinephrine (NE) in 30mL of ethanol solution, and add 2 mL of sodium borohydride (NaBH4) (150 mM), in a 60° C. water bath for 1 h, washed with ethanol and deionized water three times each, and freeze-dried to obtain AuPd-NE nanozyme.

[0033] Such as figure 1 As shown, the principle of the present invention is that glucose is oxidized and decomposed into gluconic acid and H in the presence of AuPd-NE nanozyme 2 o 2 , then H 2 o 2 Continue to decompose into OH and H 2 O, · OH can change the color of the substrate TMB and have absorbance at 652nm. The cascade reaction of AuPd-NE nanozyme is used to realize the highly sensitive and selective detection of glucose.

[0034] figure 2 It is the transmission electron microscope image ...

Embodiment 2

[0037] An AuPd-NE nanozyme is applied to a glucose detection method in human serum, the steps comprising:

[0038] (1) Drawing of working curve

[0039] Add 400 μL of glucose solutions of different concentrations to 1 mL of AuPd-NE nanozyme solution, incubate at 50°C for 4 hours, add 100 μL of TMB solution, record the change of absorbance at 652 nm, and arrange and plot the obtained data to obtain a linear relationship , the linear relationship is: Abs=1.1133[Glu]+0.1143.

[0040] (2) Detection of glucose concentration in human serum

[0041] The human serum of serum samples 1, 2, and 3 were diluted 20 times respectively, and 400 μL of human serum was added to 1 mL of AuPd-NE nanozyme solution. After incubation at 50 °C for 4 h, 100 μL of TMB solution was added. Then record the absorbance change at 652nm, and substitute the absorbance intensity value into the linear relational formula to determine the concentration value of glucose in human serum, and compare it with the blo...

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Abstract

The invention discloses a preparation method and application of a nano enzyme for colorimetric detection of glucose. The preparation method comprises the following steps that HAuCl4, Na2PdCl4 and noradrenaline are : uniformly dispersed in an ethanol solution to obtain a mixed solution; NaBH4 is added into the mixed solution under the stirring condition, reduction is conducted under the water bath condition, then centrifugation, washing and freeze drying are conducted, and the AuPd-NE nano-enzyme is obtained. On the basis, a novel method for detecting the content of glucose in human serum is established by utilizing the catalytic performance of the nano enzyme. Under the optimal condition, the detection range of the method for detecting the glucose is 30-250 muM, and the detection limit is 10 muM. The synthesis method is simple, the selectivity is high, the detection limit is low, and sensitive detection of glucose is realized.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to a preparation method and application of a nanozyme for colorimetrically detecting glucose. Background technique [0002] Glucose can be decomposed into gluconic acid and hydrogen peroxide (H2O2) under the action of natural glucose oxidase, so the cascade colorimetric reaction is used (glucose is decomposed into H2O2 by GOx, and H2O2 can make the substrate under the action of peroxidase Discoloration) can indirectly measure the concentration of glucose. However, the cost of preparation and purification of natural enzymes is high, their stability is poor, and they are easily digested by proteases. More stable and highly active artificial enzymes are needed to replace them. At the same time, since the optimal activity conditions of natural enzymes and nanozymes are often different, multi-step reactions are required to realize the detection of glucose, which is time-consuming and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/78G01N21/3103Y02A50/30
Inventor 徐荣鑫刘思全李贺谭啸峰李涛李琰
Owner UNIV OF JINAN
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