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Dual-emission ratio fluorescence sensor, preparation method and application thereof

A fluorescent sensor and ratiometric technology, applied in the field of nanomaterial science and biochemical sensing, can solve the problems of time-consuming, labor-intensive, expensive reagents, etc., and achieve the effect of convenient operation and simple sample processing

Active Publication Date: 2018-06-19
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Traditional pesticide detection methods include mass spectrometry, gas chromatography, high performance liquid chromatography and enzyme-linked immunoreaction, etc. However, these methods are time-consuming and labor-intensive, and require expensive reagents, complex sample processing, sophisticated instruments and skilled operations personnel

Method used

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  • Dual-emission ratio fluorescence sensor, preparation method and application thereof
  • Dual-emission ratio fluorescence sensor, preparation method and application thereof
  • Dual-emission ratio fluorescence sensor, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of dual emission ratio fluorescence sensor

[0043] The preparation method of the dual emission ratio fluorescence sensor includes the following steps:

[0044] Step S1: preparing a solution of glutathione-modified gold nanoparticles;

[0045] Specifically:

[0046] Step S11: Mix concentrated nitric acid and concentrated hydrochloric acid according to the ratio of 1:3 to make aqua regia, let it stand for 30 minutes to maximize its oxidizing ability, then transfer it to the flask and shake it gently. After 30 minutes, transfer the regia Water, wash the flask and other reaction glassware with super water, and dry them for later use;

[0047] Step S12: Add 50 mL of glutathione solution with a concentration of 3 mM to the reaction glassware washed in step S11, add 1500 μL of chloroauric acid solution with a concentration of 100 mM while stirring vigorously, and place the entire device at 95°C. React for 35 minutes in an oil bath to prepare a uniformly dispers...

Embodiment 2

[0059] Example 2: Application of dual emission ratio fluorescence sensor in the detection of acetylcholinesterase

[0060] The method for detecting acetylcholinesterase with a dual emission ratio fluorescent sensor includes the following steps:

[0061] Step S1: Prepare the reaction system and incubate it at 37°C for 25 min;

[0062] Specifically, the preparation method of the reaction system includes:

[0063] Add 10 μL of the substrate thioacetylcholine with a concentration of 1 mM, 3 μL of the dual emission ratio fluorescence sensor prepared in Example 1, the acetylcholinesterase to be measured and an appropriate amount of ultrapure water to prepare a 100 μL reaction system;

[0064] Step S2: Use a fluorescence spectrometer to perform spectrum detection, and realize the detection of acetylcholinesterase through the fluorescence intensity changes of the dual emission ratio fluorescence sensor at 600 nm and 800 nm;

[0065] Specifically, a fluorescence detector is used for detection; ac...

Embodiment 3

[0067] Example 3: Application of dual emission ratio fluorescence sensor in detecting pesticide residues

[0068] The method for detecting pesticide residues with dual emission ratio fluorescent sensors includes the following steps:

[0069] Step S1: Mix the acetylcholinesterase with the pesticide to be tested, and incubate at 37°C for 20 minutes;

[0070] Specifically, adding 8 μL of acetylcholinesterase at a concentration of 1 U / mL and the pesticide to be tested, the incubation temperature is 37° C., and the reaction time is 20 min;

[0071] Step S2: Add the substrate thioacetylcholine and the dual emission ratio fluorescence sensor prepared in Example 1 in proportion to form a reaction system, and incubate at 37°C for 5 minutes;

[0072] Specifically, add 10 μL of 1 mM thioacetylcholine, 3 μL of the dual emission ratio fluorescence sensor prepared in Example 1 and an appropriate amount of water to form a 100 μL reaction system, the incubation temperature is 37° C., and the time is 5 ...

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Abstract

The invention discloses a dual-emission ratio fluorescence sensor. The dual-emission ratio fluorescence sensor is prepared from glutathione modified gold nanoparticles, and has strong fluorescence emission peaks at places 600nm and 800nm respectively. The dual-emission ratio fluorescence sensor can be applied to acetylcholin esterase and pesticide detection, based on Au-S covalent bonding, acetylcholin esterase can hydrolyze acetylthiocholine to generate substances with -SH or enzyme inhibiting activity, and the nature of dual fluorescence emission peaks of glutathione modified gold nanoparticles themselves is combined, the dual-emission ratio fluorescence sensor has the advantages of convenient operation, simple sample processing, high efficiency, high sensitivity detection and the like,and has good application prospects. Based on the dual-emission ratio fluorescence sensor, the invention provides a preparation method of the dual-emission ratio fluorescence sensor and application thereof in detection of acetylcholin esterase and pesticide residue.

Description

Technical field [0001] The invention relates to the fields of nanomaterial science and biochemical sensing technology, in particular to a dual emission ratio fluorescent sensor and a preparation method and application thereof. Background technique [0002] Pesticides, including insecticides, herbicides and fungicides, are widely used in modern agriculture due to their high toxicity. Although they can prevent pests and diseases, they also infiltrate human public health. in. According to the data analysis of the World Health Organization, about 1.5 billion children suffer food poisoning including pesticide residues every year; the typical representatives are carbamates and organophosphorus pesticides. The key mechanism is carbamate and organophosphorus. Pesticides can inhibit the activity of acetylcholinesterase, a catalytically active enzyme in the nervous system, thereby inhibiting neurotransmission, leading to organ failure and death. Therefore, to protect humans from pesticid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6439
Inventor 李旺马欢杨涛林亲录
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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