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A method for detecting homocysteine ​​in urine by using an enzyme-modified electrode

A technology of homocysteine ​​and modified electrodes is applied in measurement devices, material analysis by electromagnetic means, instruments, etc., to achieve the effects of rapid detection and simple preparation process

Active Publication Date: 2022-02-08
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the specific detection of Hcy can be achieved by loading an enzyme specific to Hcy on nanoporous gold. The method of detecting homocysteine ​​in urine using an enzyme-modified electrode has not been reported yet.

Method used

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  • A method for detecting homocysteine ​​in urine by using an enzyme-modified electrode
  • A method for detecting homocysteine ​​in urine by using an enzyme-modified electrode
  • A method for detecting homocysteine ​​in urine by using an enzyme-modified electrode

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Example 1: A method for the rapid and accurate detection of Hcy using an enzyme-modified electrode

[0044] (1) Preparation and characterization of nanoporous gold electrodes:

[0045] The gold-silver alloy film with a thickness of 100nm was placed in concentrated nitric acid at 30°C and corroded for 45min to obtain NPG. The prepared NPG was pasted on the surface of a glassy carbon electrode (GCE), and 3 μL of 0.05% Nafion was added dropwise on the surface of the NPG. After drying, it was baked with an infrared lamp for 10 min to obtain a nanoporous gold electrode. (NPG / GCE).

[0046] The prepared NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electrode system. 2 SO 4 The cyclic voltammetry was used to scan 20 circles, the scanning low potential was +0.35V, and the scanning high potential range was +1.55V. The effective area of...

Embodiment 2

[0056] Example 2: A method for rapid and accurate detection of Hcy using an enzyme-modified electrode

[0057] (1) Preparation and characterization of nanoporous gold electrodes:

[0058] The gold-silver alloy film with a thickness of 90nm was placed in concentrated nitric acid at 28°C and corroded for 30min to obtain NPG. The prepared NPG was pasted on the surface of a glassy carbon electrode (GCE), and 2 μL of 0.05% Nafion was added dropwise on the surface of the NPG. After drying, it was baked with an infrared lamp for 10 min to obtain a nanoporous gold electrode ( NPG / GCE).

[0059] The prepared NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electrode system. 2 SO 4 The cyclic voltammetry was used to scan 20 circles, the scanning low potential was +0.3V, and the scanning high potential range was +1.5V. The effective area of ​​the ...

Embodiment 3

[0066] Example 3: A method for rapid and accurate detection of Hcy using an enzyme-modified electrode

[0067] (1) Preparation and characterization of nanoporous gold electrodes:

[0068] The gold-silver alloy film with a thickness of 110nm was placed in concentrated nitric acid at 32°C and corroded for 60min to obtain NPG. The prepared NPG was pasted on the surface of a glassy carbon electrode (GCE), and 5 μL of 0.05% Nafion was added dropwise on the surface of the NPG. After drying, it was baked with an infrared lamp for 10 min to obtain a nanoporous gold electrode. (NPG / GCE).

[0069] The prepared NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electrode system. 2 SO 4 The cyclic voltammetry was used to scan 20 circles, the scanning low potential was +0.4V, and the scanning high potential range was +1.6V. The effective area of ​​the...

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Abstract

The invention discloses a method for detecting homocysteine ​​in urine by using an enzyme-modified electrode. The steps are: 1) preparation and characterization of a nanoporous gold electrode; 2) preparation of an enzyme-modified electrode; 3) lyase with methionine The modified electrode or cystathionine synthase modified electrode detects the current response values ​​generated by different concentrations of Hcy, and makes a linear standard curve of Hcy; 4) Use the methionine lyase modified electrode or cystathionine synthase modified electrode to detect the urine dilution containing Hcy sample. Compared with other detection methods, the method of the invention does not require expensive instruments and reagents, greatly reduces the detection cost, does not require complicated sample pretreatment process, and has simple operation and rapid detection. with broadly application foreground.

Description

technical field [0001] The invention relates to a method for detecting homocysteine ​​(Homocysteine, Hcy), in particular to a method for detecting homocysteine ​​in urine by using an enzyme-modified electrode. It belongs to the field of electrochemical analysis and testing. Background technique [0002] Homocysteine ​​(Hcy) mainly exists in the form of binding in human plasma, and only about 5% of Hcy exists in the form of free. The sum of the amount of bound and free Hcy is the total homocysteine ​​(Total homocysteine, tHcy). The normal content of tHcy in plasma is in the range of 5-15 μM, and the abnormal increase of tHcy level in plasma is hyperhomocysteinemia. According to the level of tHcy in plasma, it can be divided into mild, moderate and severe hyperhomocysteinemia. At present, many clinical studies have shown that abnormal levels of tHcy in plasma are related to cardiovascular diseases, pregnancy complications, diabetes, Alzheimer's disease, renal failure, depre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/3272
Inventor 王霞高岩
Owner SHANDONG UNIV