Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of ratio type fluorescent probe in detection of peroxynitrite anions

A peroxynitroso, fluorescent probe technology, applied in the field of fluorescent probes, can solve the problems of low sensitivity and large background interference, and achieve the effects of high sensitivity, obvious phenomenon and high detection

Active Publication Date: 2020-07-17
QILU UNIV OF TECH
View PDF2 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the defects of low sensitivity and large background interference in the fluorescent probe for detecting peroxynitroso anion in the prior art, the present invention provides an application of a ratiometric fluorescent probe in detecting peroxynitroso anion , it can be found through fluorescence spectrophotometer detection that the fluorescent probe has high sensitivity to the detection of peroxynitroso anion, and the phenomenon is obvious, which is easy to identify

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of ratio type fluorescent probe in detection of peroxynitrite anions
  • Application of ratio type fluorescent probe in detection of peroxynitrite anions
  • Application of ratio type fluorescent probe in detection of peroxynitrite anions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Add 0.276 g of compound 1 (6-bromo-1H,3H-benzo[de]isotryptamine-1,3-dione) and 0.08 g of propargylamine into 20ml of ethanol, and react at 80°C for 12 Hours, TLC monitors the reaction process, after the reaction is over, the reaction solution is rotated to remove the solvent, the solid is dissolved in dichloromethane, and the mixed solvent column chromatography of dichloromethane and methanol with a volume ratio of 5:1 is used for separation to obtain compound 2;

[0035](2) Dissolve 0.156 g of compound 2 and 0.078 g of N-(2-aminoethyl)morpholine in 8ml of ethylene glycol monomethyl ether, and react at 110°C for 12 hours. TLC monitors the reaction progress. Wash with dilute hydrochloric acid with a fraction of 10%, and extract the reaction liquid obtained after the reaction with dichloromethane to remove the water phase, then use rotary distillation to remove the solvent, use a small amount of dichloromethane to dissolve the solid, and use a volume ratio of 20:1. Dic...

Embodiment 2

[0042] Titration Experiment of Fluorescent Probe and Peroxynitrite Anion

[0043] In PBS buffer (pH=7.4), add the fluorescent probe with an initial concentration of 1 mM, so that the concentration of the fluorescent probe in the solution is 10 μM. Then, different amounts of peroxynitroso anions with an initial concentration of 1.00 mM were sequentially added, so that the concentrations of peroxynitroso anions in the solution were 0.5 μM, 1 μM, 2 μM, 3 μM, 4 μM, and 5 μM, respectively. The nitroso anion was used as a control, and it was allowed to stand still for 0.5h to fully react the peroxynitroso anion and the fluorescent probe.

[0044] Fluorescence spectra under different concentrations of peroxysubunit anions were tested with a fluorescence spectrometer. The excitation wavelengths of the fluorescence spectra were 420 nm and 580 nm, the emission wavelengths were 490 nm and 660 nm, and the detection wavelengths were 490 nm and 660 nm. figure 2 As shown, as the concentrat...

Embodiment 3

[0047] (1) MTT detection of cell viability by fluorescent probe

[0048] HeLa cells were cultured in a 37°C incubator for 24 hours, then they were cultured into 96-well plates and incubated with different concentrations of probes for 12 hours. 10 μL of MTT was subsequently added to each well of the plate and further incubated for 4 h before the addition of DMSO (100 μL). The number of cells in each well was measured using a Herm Fisher Scientific plate reader. Cytotoxic structures such as Figure 4 As shown, therefore, the fluorescent probes in this patent are low toxic to cells.

[0049] (2) Selectivity test of fluorescent probes for detection of peroxynitrite anion

[0050] Under the same test conditions, add excessive amounts of other bioactive small molecules to the solution, and test the fluorescence spectrum after adding different bioactive small molecules, the results are as follows: Figure 5 shown. Depend on Figure 5 It can be seen that from 1-14 represent the ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an application of a ratio type fluorescent probe in detection of peroxynitrite anions, and the structural formula of the ratio type fluorescent probe is as follows: when the fluorescent probe does not react with peroxynitrite anions, intramolecular carboxyl can form an intramolecular spiral ring, a conjugated structure of a fluorophore is destroyed so that fluorescence quenching is caused, when the fluorescent probe reacts with peroxynitrite anions, the spiro ring can be opened to recover the conjugated structure of the fluorophore again, and strong fluorescence is displayed under the illumination condition. The preparation method of the fluorescent probe is simple and has a high product yield, the probe is simple in structure and suitable for large-scale promotionand application, peroxynitrite anions can be recognized and detected in a water system and an organic solvent system in a high-selectivity mode, the ratio of two fluorescence signals serves as an output signal through the ratio type fluorescence probe, interference of background factors can be well eliminated, interference of other factors is reduced, and the detection sensitivity and accuracy areimproved.

Description

technical field [0001] The invention belongs to the technical field of fluorescent probes, and in particular relates to the application of a ratio-type fluorescent probe in detecting peroxynitroso anions. Background technique [0002] Peroxynitrite anion (ONOO-) is considered to be one of the most important anions in the human body by excess nitric oxide (NO) and oxygen free radicals ( . o 2 - ) reaction, it is not only a strong oxidant, but also a nitrating agent, which can react with macromolecular substances such as nucleic acid, protein, and lipid in human cells, making cell metabolism difficult and causing the body to consume a large amount of energy, thus causing a series of Biological pathological processes such as tumors, arteriosclerosis, body inflammation, retinal damage, etc. In recent years, there has been a lot of research on the detection of ONOO- in the medical field, but due to the strong oxidation of ONOO-, it is very unstable in the body, which brings in...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07D405/14C09K11/06G01N21/64
CPCC07D405/14C09K11/06G01N21/6428C09K2211/1088C09K2211/1029C09K2211/1059C09K2211/1033G01N2021/6432
Inventor 刘克印包罗张昌炜
Owner QILU UNIV OF TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com