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

Hydrogenated dichlorofluorescein diacetamide derivative as well as preparation method and application thereof

An amide derivative, dichlorofluorescein technology, applied in the field of heme detection, can solve the problem of few reports on heme fluorescence measurement

Pending Publication Date: 2020-04-14
GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there are still few reports on the fluorescence measurement of heme

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
  • Hydrogenated dichlorofluorescein diacetamide derivative as well as preparation method and application thereof
  • Hydrogenated dichlorofluorescein diacetamide derivative as well as preparation method and application thereof
  • Hydrogenated dichlorofluorescein diacetamide derivative as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Design and synthesis of embodiment 1 fluorescent reporter molecule

[0054] (1) Design of Hydrogenated Dichlorofluorescein Diacetamide Derivatives

[0055] 2',7'-dichlorofluorescein (DCF) was selected as the dye to prepare the fluorescent reporter molecule, which is a cheap and commercially available dye with green fluorescence suitable for naked-eye observation. In a preliminary study, it was found that the maximum intensity of the spectrum was normalized by setting the molecular concentration of fluorescein, DCF and amide derivative intermediate (DCFA) at 20 μM in BR buffer (pH 12) with excitation at 475 nm is 1, its UV-Vis spectrum and fluorescence spectrum are as follows figure 2 shown. Depend on figure 2 It can be seen that the acylation modification of DCF with diethylamine can lead to a significant red shift in its fluorescence spectrum, with a peak at around 530 nm. Therefore, the emission of DCFA is closer to the eye-sensitive yellow-green (550nm), so the...

Embodiment 2

[0061] The optimization of embodiment 2 detection conditions

[0062] (1) Optimization of detection condition pH value

[0063] Firstly, the pH conditions of the reaction were optimized. DCFHA and corresponding peroxides were prepared in BR buffer (0.04M H 3 BO 3 , 0.04MH 3 PO 4 and 0.04M CH 3 COOH, 1.0MmEDTA, NaOH), and then the reaction was monitored by fluorescence kinetics, and the fluorescence was recorded at intervals of 10 seconds with a microplate reader, and the excitation wavelength was 475nm. All experiments were performed with 50 μM DCFHA and 1 μM heme, and 100 μM H2 o 2 , TBHP at 200 μM and ART at 5 μM. The effect of pH on the detection of hemoglobin is as follows: Figure 6 As shown, it can be seen that increasing the pH from 9 to 12 has no effect on DCFHA / H 2 o 2 The reaction rate of the DCFHA / TBHP system has a great influence, therefore, the optional range of pH is 9-12. And increasing the pH can improve the response of the DCFHA / ART detection system...

Embodiment 3

[0067] Embodiment 3 quantitative measurement of hemoglobin

[0068] (1) Use DCFHA / H 2 o 2 System for quantitative hemoglobin measurement

[0069] DCFHA(50μM) / H when the detected hemoglobin concentration is 0~2μM 2 o 2 (100μM) absorption spectrum and fluorescence spectrum of the solution, and the fluorescence intensity I 530 The titration curve obtained by plotting the hemoglobin concentration, all solutions are excited at 475nm, such as Figure 9 shown.

[0070] Depend on Figure 9 It can be seen that when hemoglobin is 0-20nM, I 530 There is an excellent linear relationship between and hemoglobin concentration. When the hemoglobin is between 0 and 0.1 μM, the fluorescence intensity I 530 The increase showed a clear concentration dependence. Compared with the control without heme, 0.1 μM heme can lead to I 530 Enhancement up to 150-fold; further increases in hemoglobin concentration, I 530 No significant increase. UV-Vis spectra showed similar trends as fluorescen...

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 relates to a hydrogenated dichlorofluorescein diacetamide derivative as well as a preparation method and application thereof, and belongs to the technical field of heme detection. The hydrogenated dichlorofluorescein diacetamide derivative disclosed by the invention can be used as a fluorescence reporter molecule for heme detection, and the detection method disclosed by the inventioncan be used for conveniently and quickly detecting heme; according to the invention, three peroxides H2O2, TBHP and ART are respectively mixed with DCFHA to prepare a detection reagent; the three systems can be used for rapid detection of heme, the detection limit can reach the picomole level, the DCFHA / H2O2 detection method can successfully distinguish bloodstains and coffee stains, and fluorescence can be seen with naked eyes under ultraviolet light; the DCFHA / ART is doped into the filter paper to prepare the detection test paper, and the detection test paper has good effects in heme detection and blood determination, can be used as a portable device, and is used in criminal investigation and other practical applications.

Description

technical field [0001] The invention relates to a hydrogenated dichlorofluorescein diethylamide derivative and a preparation method and application thereof, belonging to the technical field of heme detection. Background technique [0002] Heme is a complex of ferric iron and protoporphyrin [Fe(III)PPIX], which plays an important role in living organisms. Heme and its derivatives mainly include hemin and hematin. The content of heme in the human body is very high, and it participates in the reversible redox reaction of Fe(III) / Fe(II) as an electron transfer medium. In addition to being involved in oxygen transport processes, heme is involved in many other cellular processes and is considered a key regulator of cellular homeostasis. Abnormal hemoglobin concentration is associated with a series of pathophysiological changes, such as secondary brain damage caused by hemorrhagic stroke, induction of neuroglobulin expression in nerve cells, promotion of endogenous carbon monoxid...

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
IPC IPC(8): C07D311/82C09K11/06G01N21/64
CPCC07D311/82C09K11/06G01N21/6486C09K2211/1088C09K2211/1007
Inventor 刘芳邓涛赵靓陈福乾
Owner GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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