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Magnetic graphene enzyme-mimicking property-based acetylcholine visual-detection method

A technology of acetylcholine and detection method, which is applied in the direction of analyzing by chemical reaction of the material, and analyzing the material by observing the influence on the chemical indicator, so as to achieve the effects of simple and flexible operation, simple equipment and low detection cost.

Inactive Publication Date: 2014-04-09
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But Fe 3 o 4 The characteristics of easy agglomeration limit the application of its properties to a certain extent

Method used

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  • Magnetic graphene enzyme-mimicking property-based acetylcholine visual-detection method
  • Magnetic graphene enzyme-mimicking property-based acetylcholine visual-detection method
  • Magnetic graphene enzyme-mimicking property-based acetylcholine visual-detection method

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Experimental program
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Effect test

Embodiment 1

[0027] Embodiment one: Fe 3 o 4 / rGO Preparation and H 2 O 2 Colorimetric detection of

[0028] (1) Fe 3 o 4 Preparation of / rGO: First, 0.5 mmol FeCl 2 4H 2 O and 1 mmol FeCl 3 ·6H 2 O was added to 10 mL of DEG solution containing 5 mg GO, and stirred at 90 oC for 30 min; then 10 mL of DEG solution containing 0.8 M NaAc was added dropwise, and after stirring for 10 min, the mixture was transferred to the reaction kettle , heated at 200 oC for 6 h; finally the product was cooled to room temperature, washed several times with ethanol and dried in a vacuum oven at 50 °C to obtain Fe 3 o 4 / rGO as figure 1 shown.

[0029] (2)H 2 o 2 Response relationship between concentration and absorbance: First, 965 μL ABS containing 0.5 mg TMB was mixed with 10 μL H 2 o 2 Mix well; then add 25 μL of 2 mg·mL –1 Fe 3 o 4 / rGO, mixed well and incubated at room temperature for 15 min. Finally, the Fe in the reaction system is removed by an external magnetic field (magn...

Embodiment 2

[0030] Embodiment two: detection of acetylcholine and standard curve drawing

[0031] (1) Enzyme-catalyzed reaction of acetylcholine: 200 μL of PBS containing 0.5 units of acetylcholinesterase (AchE) and 0.5 units of choline oxidase (ChOx) were mixed with different amounts (0, 5×10 –5 、10 –4 , 5×10 –4 、10 –3 、10 –2 、10 –1 , 1, 10 μmol) of acetylcholine were mixed evenly, and incubated at 37°C for 30 min.

[0032] (2) Correspondence between acetylcholine concentration and absorbance: 800 μL contains 0.5 mg TMB and 50 μg Fe 3 o 4 The ABS of rGO / rGO was added to the solution incubated in step (3), mixed evenly, and incubated at room temperature for 30 min. Then, the Fe in the reaction system was removed by an external magnetic field (magnet) 3 o 4 / rGO, take pictures to observe the color change, and measure the absorbance of the reaction solution at a wavelength of 652 nm. Finally, establish the corresponding relationship between the concentration of acetylcholine and...

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Abstract

The invention relates to a magnetic graphene enzyme-mimicking property-based acetylcholine visual-detection method, belonging to the technical field of biosensing. A hydrothermal method is used for synthesizing Fe3O4 / rGO by one step, H2O2 (hydrogen peroxide) is catalyzed to oxidize a substrate TMB (tetramethylabenzidine) to generate blue by utilizing the peroxidase-like property of the Fe3O4 / rGO, and the characteristic absorption intensity of an oxidation product of the TMB at the wavelength of 652nm is detected to realize the detection of hydrogen peroxide; furthermore, the H2O2 can be catalyzed to oxidize and convert the colorless TMB into a corresponding blue product based on the Fe3O4 / rGO, and the blue product is combined with acetyl choline to decompose H2O2 in the presence of both acetylcholinesterase and choline oxidase, and a novel acetylcholine visual-sensing method is established. The invention aims to provide an acetylcholine colorimetric detection method, and according to the method, the operation is convenient and flexible, the detection cost is low, the sensitivity is high, and instruments and equipment are simple.

Description

technical field [0001] The invention relates to the construction of a visual detection method for acetylcholine, and technically relates to the fields of material science, optical analytical chemistry, biosensing and the like. Background technique [0002] Acetylcholine (Ach) is a neurotransmitter that transmits information between neurons or between neurons and effectors. It mainly exists in the peripheral nervous system and central nervous system of organisms. There is a large amount of acetylcholine in human brain tissue, but the content of acetylcholine will decline with age. Normal elderly people have a 30% drop compared to their youth, and the decline in senile dementia patients is even more severe, up to 70% to 80%. When the amount of acetylcholine in the brain falls below a certain level, it increases the chances of developing Alzheimer's disease. Alzheimer's disease is a chronic and fatal neurodegenerative disease. Its clinical manifestations are continuous degene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 王坤杨兴旺钱静
Owner JIANGSU UNIV
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