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Preparation method and application of enzyme electrode sensor for SAM detection

A technology of electrode sensor and glassy carbon electrode is applied in the field of preparation of enzyme electrode sensor to achieve the effects of low cost, solving difficulty in detection and simple synthesis method

Inactive Publication Date: 2015-09-23
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The bio-enzyme electrode sensor is currently developed with the characteristics of specificity, stability, fast detection speed, good selectivity, and high sensitivity. It is widely used in the fields of clinical medicine, food, environment, and biological samples. The detection of SAM on the electrode has not been reported

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0026] (1) Electrode activation treatment: the glassy carbon electrode was treated with 0.3 μm and 0.05 μm Al 2 o 3 powder polishing, and ultrasonically cleaned with ethanol and water for 10 min in an ultrasonic cleaner, and the cleaned glassy carbon electrode was used as the working electrode. -1 Scanning in the sulfuric acid solution in the potential range of -0.3 to +1.5 V until it is stable, an activated glassy carbon electrode is obtained;

[0027] (2) Preparation of poly-3,7-diaminophenothiazine-5-one modified electrode: 0.14 mol L -1 The pH value of 3,7-diaminophenothiazine-5-onium was 7.1, and the phosphate buffer solution was passed through nitrogen to remove oxygen for 10 minutes. After the oxygen was removed, the activated glassy carbon electrode was put in, and the voltage was 1.5 V by chronoamperometry. Under pre-positive excitation for 250 s, and then cyclic voltammetry was used to scan 80 cycles to obtain a poly-3,7-diaminophenothiazine-5-onium modified electr...

Embodiment 2

[0031] (1) Electrode activation treatment: the glassy carbon electrode was treated with 0.3 μm and 0.05 μm Al 2 o 3 powder polishing, and ultrasonically cleaned with ethanol and water for 10 min in an ultrasonic cleaner, and the cleaned glassy carbon electrode was used as the working electrode. -1 Scanning in the sulfuric acid solution in the potential range of -0.3 to +1.5 V until it is stable, an activated glassy carbon electrode is obtained;

[0032] (2) Preparation of poly-3,7-diaminophenothiazine-5-one modified electrode: 0.12 mol L -1 The pH value of 3,7-diaminophenothiazine-5-onium was 7.2, and the phosphate buffer solution was passed through nitrogen for deoxygenation for 10 minutes. After deoxygenation, the activated glassy carbon electrode was put in, and the voltage was 1.5 V by chronoamperometry. Under the pre-cationic excitation for 260 s, and then 80 cycles of cyclic voltammetry scanning, the poly-3,7-diaminophenothiazine-5-onium modified electrode was obtained...

Embodiment 3

[0036] (1) Electrode activation treatment: the glassy carbon electrode was treated with 0.3 μm and 0.05 μm Al 2 o 3 powder polishing, and ultrasonically cleaned with ethanol and water for 10 min in an ultrasonic cleaner, and the cleaned glassy carbon electrode was used as the working electrode. -1 Scanning in the sulfuric acid solution in the potential range of -0.3 to +1.5 V until it is stable, an activated glassy carbon electrode is obtained;

[0037] (2) Preparation of poly-3,7-diaminophenothiazine-5-one modified electrode: 0.11 mol L -1 The pH value of 3,7-diaminophenothiazine-5-onium was 7.0, and the pH value of the phosphate buffer solution was passed through nitrogen to remove oxygen for 10 minutes. After the oxygen was removed, the activated glassy carbon electrode was put in, and the voltage was 1.5 V by chronoamperometry. Under the pre-positive excitation for 270 s, and then cyclic voltammetry scanning for 80 cycles, the poly-3,7-diaminophenothiazine-5-onium modifi...

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Abstract

The invention discloses a preparation method and application of an enzyme electrode sensor for SAM detection. The preparation method is characterized by comprising the steps that 1, a glassy carbon electrode is polished with Al2O3 powder and cleaned by ethyl alcohol and water ultrasonic successively in an ultrasonic cleaner to obtain an activated glassy carbon electrode; 2, an electrochemical polymerization method is adopted on the activated glassy carbon electrode to obtain a poly-3,7-diamido-phenothiazine-5-onium modified electrode; 3, transmethylase is fixed to the poly-3,7-diamido-phenothiazine-5-onium modified electrode in a crosslinking mode to obtain a fixed transmethylase electrode sensor. SAM in a sample can be rapidly detected through the electrode, the method is high in sensitivity, good in selectivity, short in response time, low in interference and better than other detection methods, and is an SAM determination method which is simple, rapid, convenient and easy to implement. The obtained fixed transmethylase electrode sensor is low in cost, simple in preparation technology and good in specificity and has the potential for achieving automatic in-situ measurement.

Description

technical field [0001] The invention relates to a preparation method of an enzyme electrode sensor and the technical field of rapid detection application, in particular to a preparation method of an enzyme electrode sensor for detection of S-adenosylmethionine (SAM), which is used for detection of drugs , S-adenosylmethionine technology in biological samples. Background technique [0002] S-adenosylmethionine (SAM), SAM contains active methyl groups, and almost all the methyl groups used for methylation modification in cells come from SAM methyl-thio high-energy bonds. Due to the extensiveness of the methylation reaction, it can be said that SAM is a coenzyme that is second only to ATP in the importance of participating in the reaction in the cell. A small change in the concentration of SAM in the cell will have a significant impact on the growth, differentiation and function of the cell. influences. SAM is mainly synthesized in bacteria by SAM synthase (MetK) through meth...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48
Inventor 李慧芝李冬梅魏琴
Owner UNIV OF JINAN
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