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Method for detecting concentration of acetylcholin esterase

A technology of acetylcholinesterase and acetylcholine, which is applied in the field of acetylcholinesterase detection, can solve the problems of complex operation methods, low sensitivity, and long reaction time, and achieve high sensitivity, low detection limit, and simple and convenient detection process

Inactive Publication Date: 2019-07-23
南宁师范大学
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most methods have defects such as complicated operation methods, long reaction time, and low sensitivity.

Method used

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  • Method for detecting concentration of acetylcholin esterase
  • Method for detecting concentration of acetylcholin esterase
  • Method for detecting concentration of acetylcholin esterase

Examples

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Embodiment 1

[0019] Preparation of carbon quantum dots: Take 1g of charcoal and 200mL of HNO with a concentration of 5mmoL 3 Solution, after mixing, reflux with magnetic stirring in an oil bath at 120°C for 8 hours, filter the refluxed reaction solution to obtain a supernatant; use NaOH solution to adjust the pH value of the supernatant to neutral, and use 0.22 μm Filter through a microporous membrane, take the filtrate, and freeze-dry to obtain the obtained product.

Embodiment 2

[0021] A method for detecting the concentration of acetylcholinesterase, comprising:

[0022] Step 1. Dissolve 2g of trypsin in 10ml of ultrapure water. After dispersion, put it into a 20ml reaction kettle, react at 180°C for 10h, take it out, and freeze-dry it to obtain carbon quantum dots. The pH used for the carbon quantum dots is 7.4 and the concentration is 0.05mol / L Tris-HCl buffer solution prepared as a concentration of 5.3 × 10 -6 g / mL carbon quantum dot solution;

[0023] Step 2, first use acetylcholinesterase dry powder and ultrapure water configuration concentration to be the stoste of 1000U / L, described stoste is carried out gradient dilution with described Tris-HCl buffer solution, make the acetylcholinesterase standard solution of different concentrations, its Concentrations are 0U / L, 2U / L, 5U / L, 10U / L, 25U / L in turn, and then add 100μl to 100μl of the acetylcholinesterase standard solution with different concentrations as described above to obtain a concentrati...

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Abstract

The invention discloses a method for detecting the concentration of acetylcholin esterase. The method comprises the following steps that: S1: preparing carbon quantum dots, and preparing the carbon quantum dots into carbon quantum dot solution; S2: preparing acetylcholin esterase standard solutions of different concentrations, and independently adding acetyl choline solution, choline oxidase solution, horseradish peroxidase solution and o-phenylenediamine solution into the carbon quantum dot solution, and obtaining a mixed liquid system; S3: independently adding the carbon quantum dot solutioninto the mixed liquid system to obtain a standard sample system; S4: independently detecting the fluorescence intensities I442 and I573 of the standard sample system on the positions of 442nm and 573nm, and establishing a linear relational expression between I573 / I442 and the concentration of the acetylcholin esterase; and S5: preparing the to-be-detected solution of the acetylcholin esterase, and referring to S2, S3 and S4 to obtain the concentration of the acetylcholin esterase in the to-be-detected solution of the acetylcholin esterase. The method has the advantages of high-sensitivity identification and the like of the acetylcholin esterase in the sample.

Description

technical field [0001] The invention relates to the field of detection of acetylcholinesterase. More specifically, the present invention relates to a method for detecting the concentration of acetylcholinesterase. Background technique [0002] Acetylcholinesterase is mainly distributed in nervous system tissues, which can quickly catalyze the hydrolysis of neurotransmitter acetylcholine and lead to the termination of nerve impulse transmission, thus ensuring the normal physiological functions of the human body. Human brain tissue contains a large amount of acetylcholine, and its content will decrease with age. When the content of acetylcholine in the brain is too low, Alzheimer's disease will occur. At present, the detection methods of acetylcholinesterase at home and abroad mainly include chemiluminescence, fluorescence, spectrophotometry and electrochemical methods. However, most methods have defects such as complex operation methods, long reaction time, and low sensitiv...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432G01N2021/6439
Inventor 黄珊肖琦姚建东刘义
Owner 南宁师范大学
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