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Fluorescent probe and pesticides residue detection kit based on carboxylesterase inhibition method

A technology of fluorescent probes and carboxylesterases, applied in fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., to achieve fast reaction speed, high sensitivity, and simple post-processing effects

Active Publication Date: 2017-03-15
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are still few reports on the detection of pesticide residues using the principle of carboxylesterase inhibition combined with optical probe methods

Method used

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  • Fluorescent probe and pesticides residue detection kit based on carboxylesterase inhibition method
  • Fluorescent probe and pesticides residue detection kit based on carboxylesterase inhibition method
  • Fluorescent probe and pesticides residue detection kit based on carboxylesterase inhibition method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1: the preparation of fluorescent probe shown in formula (I)

[0058]

[0059] The experimental procedure is as follows: Dissolve the tricarbocyanine IR-780 skeleton (0.41g, 1.0mmol) in N,N-dimethylformamide (10mL), and add 4-(chloromethyl)phenylacetate to it (0.18g, 1.0mmol) and catalyst potassium carbonate (0.21g, 1.5mmol), and the resulting reaction system was reacted at 45°C for 2 hours. After the reaction was completed, it was cooled to room temperature, and the solvent was removed under reduced pressure to obtain a crude product. The obtained crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v)=1 / 1) as the eluent to obtain 0.29 g of a blue-green solid product.

[0060] The structural characterization data results of the fluorescent probe are as follows:

[0061] 1 H NMR (600MHz, CD 3 OD) δ (ppm): 8.67 (d, J = 14.9Hz, 1H), 7.57 (d, J = 7.5Hz, 1H), 7.48-7.40 (m, 4H), 7.40-7.33 (m, 2H), 7.29 (s,1H),7...

Embodiment 2

[0063] Embodiment 2: the spectral properties of fluorescent probe shown in formula (I) reacting with different concentrations of carboxylesterase

[0064] Dissolve fluorescent probe solution b (1mM, 50μL) in phosphate buffer (10mM, 5mL), then add different concentrations of carboxylesterase standard solutions, and then dilute to 10mL with 10mM phosphate buffer. The concentrations of carboxylesterase in different carboxylesterase standard solutions were 0, 0.01, 0.025, 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, 0.6 and 1 U / mL. The volumes of the solutions of the series of carboxylesterase standard products with different concentrations are all 2mL. React at 37°C for 15 minutes, and measure its UV-Vis absorption spectrum and fluorescence emission spectrum. When measuring the fluorescence emission spectrum, the excitation wavelength is 650nm; the slit width for excitation and emission is 10nm; the voltage is 700V. Experimental results such as figure 1 as shown, figure 1 is the fluores...

Embodiment 3

[0068] Embodiment 3: Fluorescent probe shown in formula (I) reacts with other substances (selectivity research)

[0069] Various substances were added to the 10 μM fluorescent probe solution: potassium chloride (150 mM), calcium chloride (2.5 mM), magnesium chloride (2.5 mM), glucose (10 mM), reactive oxygen species (10 μM), glutathione (5mM), cysteine ​​(1mM), homocysteine ​​(1mM), human serum albumin (0.5mg / mL), bovine serum albumin (0.5mg / mL), acetylcholinesterase (0.1μg / L), butyrylcholinesterase (20U / L) and carboxylesterase (1U / mL). After reacting at 37°C for 15 min, the fluorescence emission spectrum was measured using a fluorometer F-4600. When the fluorescence emission spectrum is measured, it is de-excited at 650nm; the slit width for excitation and emission is 10nm; the voltage is 700V. 50 µL of the probe solution (1 mM) was added to 10 mL of the solution in which the above-mentioned substances were mixed, and carboxylesterase was added to make the final concentr...

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Abstract

The invention discloses a fluorescent probe and a pesticide residue detection kit based on a carboxylesterase inhibition method. The pesticide residue detection kit comprises a reagent stock solution a and a reagent stock solution b, wherein the reagent stock solution a is a buffer solution containing phosphate, and the reagent stock solution b is a solution of the fluorescent probe. Experiments find that the strength of the self fluorescence of the probe is very weak, and after carboxylesterase is added, the probe react and release an IR-780 hemicyanine skeleton fluorescent parent body, the fluorescence of the solution is remarkably strengthened, showing that the method can be used for detection of the activity of the carboxylesterase; after adding a tiny amount of pesticides in the carboxylesterase, the fluorescence of the solution is remarkably weakened, showing that a tiny amount of the pesticides can inhibit the activity of the carboxylesterase, and thus the fluorescent probe can be used for detection of pesticide residues. The fluorescent probe has the advantages of being easy to operate, low in cost, fast, efficient and sensitive, and the fluorescent probe is convenient for application and popularization, and has a huge application prospect in the fields of agriculture, food and the like.

Description

technical field [0001] The invention relates to a fluorescent probe, a pesticide residue detection kit based on a carboxylesterase inhibition method and a detection method thereof. Background technique [0002] Excessive pesticide residues in agricultural and sideline products affect the food safety of consumers, so the rapid and accurate detection of pesticide residues in food is an important topic of food safety testing. At present, the commonly used methods for the detection of pesticide residues at home and abroad include spectroscopy, chromatography, and the combination of chromatography and other techniques. These methods have the advantages of accurate detection results and high sensitivity, but the detection cost is high and the measurement time is long. It requires personnel with a certain professional technical level to operate, and it is only suitable for research and analysis in the laboratory. [0003] Enzyme inhibition method is the most mature and widely used...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D405/06C09K11/06G01N21/64
CPCC07D405/06C09K11/06C09K2211/1029C09K2211/1088G01N21/6428
Inventor 李照杨兴斌李东钰
Owner SHAANXI NORMAL UNIV
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