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Fluorescent probe molecule for detecting diaphorase based on rhodamine derivative and preparation method and application thereof

A technology of fluorescent probe and diaphorase, which is applied in fluorescence/phosphorescence, chemical instruments and methods, and material analysis through optical means, can solve the problem of rare diaphorase and achieve good stability of physical and chemical properties , good application prospects and good selectivity

Pending Publication Date: 2020-07-10
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few researches on probes for diaphorase

Method used

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  • Fluorescent probe molecule for detecting diaphorase based on rhodamine derivative and preparation method and application thereof
  • Fluorescent probe molecule for detecting diaphorase based on rhodamine derivative and preparation method and application thereof
  • Fluorescent probe molecule for detecting diaphorase based on rhodamine derivative and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Synthesis of fluorescent probe molecules for detection of diaphorase based on rhodamine derivatives

[0043] For specific synthetic routes, see figure 1 .

[0044] (1) Preparation of Compound 5: Under ice-bath conditions, 14 mL of concentrated sulfuric acid was added to a round-bottomed flask, and cyclohexanone (12.74 mmol, 1.32 mL) was added dropwise with a dropping funnel and stirred evenly. 2-(4-(Diethylamino)-2-hydroxybenzoyl)benzoic acid (6.4 mmol, 2.0 g) was added in portions and stirred. The mixture was heated to reflux at 90°C for 2 hours, cooled to room temperature, the reaction solution was poured into ice water, and 1.4 mL of 70% perchloric acid was added. After the precipitate was precipitated, it was suction-filtered, the filter cake was washed with water, and dried to obtain compound 5 as a black solid with a yield of 90%.

[0045] (2) Preparation of Compound 4: Dissolve Compound 5 (1.72g, 3.6mmol) in glacial acetic acid, add 4-hydroxybenzal...

Embodiment 2

[0050] Example 2: Selectivity of fluorescent probe molecules for diaphorase detection based on rhodamine derivatives

[0051] Using CH 3 OH (methanol): PBS (0.01mol / L, pH=7.4) = 9:1 (v:v) solution controls the experimental conditions.

[0052] The fluorescent probe molecule of the present invention is used CH 3 OH:PBS=9:1 (v:v) was dissolved in a solvent and fixed to a volumetric flask of 100mL, and the molecular concentration of the fluorescent probe was prepared to be 2×10 -5 mol / L solution.

[0053] The sample vials were divided into 17 groups, each group of sample vials were added with 5mL concentration of 2×10 -5 CH of the fluorescent probe molecule of the present invention of mol / L 3 OH:PBS (0.01mol / L, pH=7.4)=9:1 (v:v) solution, the first bottle of solution was used as a blank group, and 50 μL of Co at a concentration of 0.01mol / L was added to the other 16 groups 2+ , Ni 2+ ,Fe 3+ , K + ,Pb 2+ , HSO 3 - ,NO 3 - ,H 2 o 2 ,H 2 S,ClO - , Cys, Hcy, Ala, Cys...

Embodiment 3

[0054] Example 3: Fluorescence titration of diaphorase detection fluorescent probe molecules based on rhodamine derivatives to diaphorase

[0055] Using CH 3 OH (methanol): PBS (0.01mol / L, pH=7.4) = 9:1 (v:v) solution controls the experimental conditions.

[0056] Fluorescent probe molecules of the present invention are dissolved with a solvent of DMSO:PBS=9:1 (v:v) and settled in a volumetric flask of 1000mL, and the concentration of fluorescent probe molecules is prepared as 2×10 -5 mol / L solution.

[0057] 3 mg of diaphorase (DTD) was dissolved in 100 μL of deionized water to prepare an aqueous solution of diaphorase with a concentration of 900 U / mL.

[0058] Divide the sample bottles into 21 groups, add 0μL-40μL diaphorase aqueous solution with a concentration of 900U / mL to each group of sample bottles, and then use a concentration of 2×10 -5 CH of the fluorescent probe molecule of the present invention of mol / L 3 OH:PBS (0.01mol / L, pH=7.4)=9:1 (v:v) solution was adjus...

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Abstract

The invention relates to a fluorescent probe molecule for detecting diaphorase based on a rhodamine derivative and a preparation method and application thereof. The molecular structure of the fluorescent probe is shown in figure 1. The invention also discloses a preparation method of the fluorescent probe molecule for detecting the diaphorase based on the rhodamine derivative, and application of the fluorescent probe molecule in detecting the diaphorase in a water body sample or biological cells. The fluorescent probe molecule synthesis method is easy to synthesize and high in yield.

Description

technical field [0001] The invention belongs to an organic functional material for detecting trace substances, and relates to a fluorescent probe molecule, a preparation method and an application. Background technique [0002] Diaphorase (DTD), a cytoplasmic flavoproteinase widely distributed in the body tissues of almost all animals, is a detoxifying enzyme present at low levels in most healthy tissues, but exposed to the production of reactive oxygen species Substances called quinones (for example, those found in certain foods, pollutants, and cigarette smoke) are often overexpressed. On the one hand, it uses NADH (the reduced state of nicotinamide adenine dinucleotide, reduced coenzyme I) as an electron donor to reduce various quinones, prevent quinone free radicals and prevent quinone free radicals from triggering the generation of super Oxygen anion O 2 - , preventing or reducing the production of free radicals and protecting cells from damage, this effect makes it c...

Claims

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Application Information

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IPC IPC(8): C07D311/88C09K11/06G01N21/64
CPCC07D311/88C09K11/06G01N21/6428C09K2211/1088C09K2211/1007
Inventor 周莹陈秋玲乔怡木王光科
Owner YUNNAN UNIV
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