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Method for determining activity of acetylcholin esterase by using bifunctional quantum dot sensor system

A technology of acetylcholinesterase and thioacetylcholine, which is applied in the field of analytical chemistry, can solve problems such as unfavorable quantum dot material properties, limited sensitivity improvement space, and complicated reaction steps, and achieves reduced experimental costs, high sensitivity and responsiveness, and simple methods Effect

Active Publication Date: 2013-11-20
中国人民解放军63975部队
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] These published research works are all good at applying the material properties of quantum dots to the determination of the catalytic activity of acetylcholinesterase, but the quantum dots synthesized in the current published work are all designed based on the response to a single product and synthesis, because these methods only use one product to respond, so they cannot make a breakthrough on the existing detection capability level, which is not conducive to fully utilizing the material characteristics of quantum dots, so there is limited room for sensitivity improvement; in addition, these methods Both need to couple AChE to the surface of QD molecules by covalent bonding or physical adsorption, so the reaction steps are more complicated and the operation is more cumbersome.

Method used

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  • Method for determining activity of acetylcholin esterase by using bifunctional quantum dot sensor system
  • Method for determining activity of acetylcholin esterase by using bifunctional quantum dot sensor system
  • Method for determining activity of acetylcholin esterase by using bifunctional quantum dot sensor system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Synthesis of NaHTe precursor: A 10mL round-bottomed flask is connected to sealed water through a small needle and rubber tube, and 0.2g NaBH 4 And 0.16g Te powder were added to the sealed flask, and N 2 Then inject 3.0 mL of deoxygenated distilled water under an ice bath, and react for 6 hours. The resulting clear liquid is the Te source for the next step of quantum dot synthesis;

[0031] Synthesis of dual-function TGA-CdTe quantum dots: weigh 0.23g CdCl 2 After dissolving in 100mL distilled water, add TGA, Cd 2+ The molar ratio between TGA and TGA is 1:0.75, the pH of the solution is adjusted to 11.2 with 0.1M NaOH, and N is introduced 2 30min, quickly inject the newly synthesized NaHTe solution, Cd 2+ : HTe - The molar ratio is 1:0.5, the solution is in N 2 Reflux at 100°C for 4hr under protection. The obtained quantum dots are filtered through a 0.45μm filter membrane and then passed through N 2 save.

[0032] Prepare 0.2M NaAC-HAC with different pH buffers, pH 3.5, 4.0, ...

Embodiment 2

[0037] Synthesis of NaHTe precursor: A 10mL round-bottomed flask is connected to sealed water through a small needle and rubber tube, and 0.2g NaBH 4 And 0.16g Te powder were added to the sealed flask, and N 2 Then inject 3.0 mL of deoxygenated distilled water under an ice bath, and react for 6 hours. The resulting clear liquid is the Te source for the next step of quantum dot synthesis;

[0038] Synthesis of dual-function TGA-CdTe quantum dots: weigh 0.23g CdCl 2 After dissolving in 100mL distilled water, add TGA, Cd 2+ The molar ratio between TGA and TGA is 1:1.25, use 0.1M NaOH to adjust the pH of the solution to 11.2, and pass N 2 30min, quickly inject the newly synthesized NaHTe solution, Cd 2+ : HTe - The molar ratio is 1:0.5, the solution is in N 2 Reflux at 100°C for 4hr under protection. The obtained quantum dots are filtered through a 0.45μm filter membrane and then passed through N 2 save.

[0039] Prepare 0.2M NaAC-HAC with different pH buffers, pH 3.5, 4.0, 4.5, 5.0, 6....

Embodiment 3

[0044] Synthesis of NaHTe precursor: A 10mL round-bottomed flask is connected to sealed water through a small needle and rubber tube, and 0.2g NaBH 4 And 0.16g Te powder were added to the sealed flask, and N 2 Then inject 3.0 mL of deoxygenated distilled water under an ice bath, and react for 6 hours. The resulting clear liquid is the Te source for the next step of quantum dot synthesis;

[0045] Synthesis of dual-function TGA-CdTe quantum dots: weigh 0.23g CdCl 2 After dissolving in 100mL distilled water, add TGA, Cd 2+ The molar ratio between TGA and TGA is 1:1.75, use 0.1M NaOH to adjust the pH of the solution to 11.2, and pass N 2 30min, quickly inject the newly synthesized NaHTe solution, Cd 2+ : HTe - The molar ratio is 1:0.5, the solution is in N 2 Reflux at 100°C for 4hr under protection. The obtained quantum dots are filtered through a 0.45μm filter membrane and then passed through N 2 save.

[0046] Prepare 0.2M NaAC-HAC with different pH buffers, pH 3.5, 4.0, 4.5, 5.0, 6....

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Abstract

The invention provides a determination method capable of sensitively detecting the activity of acetylcholin esterase by using bifunctional quantum dots which can simultaneously generate sensitive response to two reaction substrates of thio-choline and hydrogen ions in an acetylcholin esterase catalytic system. The bifunctional quantum dots capable of simultaneously responding thio-choline and hydrogen ions are provided; and a method for detecting the activity of acetylcholin esterase is established by combining a catalytic reaction of acetylcholin esterase. The system is composed of mercaptoacetic acid-modified bifunctional cadmium telluride quantum dots, acetylthiocholine and a Tris-HCl buffer solution with a pH value of 7.5. The method comprises the following steps of synthesis of the bifunctional quantum dots, preparation of the reaction solution and detection of the activity of acetylcholin esterase by using the bifunctional quantum dot sensor system. The method has the advantages of novel principle, high sensitivity, simple synthetic method, simple system constitution, no need of biological coupling reaction, etc. The method is suitable for the activity determination of acetylcholin esterase and other detections of target objects related to the activity determination.

Description

Technical field [0001] The invention relates to a dual-function quantum dot sensing system used for acetylcholinesterase (AChE) activity determination, and belongs to the field of analytical chemistry. Background technique [0002] Acetylcholinesterase can catalyze the hydrolysis of the neurotransmitter acetylcholine (actylcholine) to produce choline and hydrogen ions, and acetylcholine is responsible for the transmission of cholinergic impulses, so the main function of acetylcholinesterase in the body is to stop the distribution of different cholinergic nerve terminals Corresponding to the effect of acetylcholine in target organs and postsynaptic sites, the presence of acetylcholinesterase can effectively ensure nerve conduction. Many important pesticides and other toxic and harmful compounds are designed through the principle of inhibiting the catalytic activity of acetylcholinesterase. Although these compounds play a huge role in social production activities, they can also cau...

Claims

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Application Information

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IPC IPC(8): C12Q1/46
Inventor 余涛应天翼宋云扬李艳军孙道鸣吴方晖
Owner 中国人民解放军63975部队
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