Silicon-based rhodamine fluorescent staining reagent and its preparation method and application

A silicon-based rhodamine and fluorescent dyeing technology, used in the fields of immunology, organic chemistry and biochemistry, can solve the problems of small Stokes shift of antibody-labeled dyes and difficult to apply, and achieve small molecular weight, high yield, and detection sensitivity. high effect

Active Publication Date: 2022-03-01
SICHUAN UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a fluorescent dye based on a large Stokes shift of a rhodamine derivative skeleton substituted by a silicon atom, a high fluorescence intensity, a preparation method and an application thereof, so as to solve the problem of Stokes shift of existing antibody labeling dyes. Small problems that are difficult to apply to fluorescence ELISA detection

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
  • Silicon-based rhodamine fluorescent staining reagent and its preparation method and application
  • Silicon-based rhodamine fluorescent staining reagent and its preparation method and application
  • Silicon-based rhodamine fluorescent staining reagent and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] The present embodiment has the preparation method of the silicon-based rhodamine fluorescent dyeing reagent of formula (I), comprising the following steps:

[0107] (1) Synthesis of Intermediate A1

[0108] The synthetic route is as follows:

[0109]

[0110] m-Bromoaniline (1.0 mmol), 1,4-dibromobutane (1.5 mol) and potassium carbonate (3.0 mmol) were mixed and stirred in acetonitrile (5 mL) for 12 hours, then 100 mL of water was added. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (30 mL×3). The organic extracts were washed with brine and treated with Na 2 SO 4 dry. After the solvent was removed and distilled under reduced pressure, it was purified by 200-300 mesh silica gel column chromatography. Eluted with petroleum ether / ethyl acetate (20:1), the intermediate A1 was obtained as a light yellow liquid with a yield of 68%.

[0111] (2) Synthesis of Intermediate A2

[0112] The synthetic route is as follows:

[0113...

Embodiment 2

[0137]This example is basically the same as Example 1, except that the starting material is replaced by the B intermediate obtained by 6-bromoindole, and its synthetic route is shown in Figure 1 (b), which includes the following steps:

[0138] (1) Synthetic Intermediate B1

[0139] The synthetic route is as follows:

[0140]

[0141] 6-Bromoindole (1.0 mmol), iodomethane (1.5 mol) and potassium carbonate (2.0 mmol) were mixed and stirred in acetonitrile (5 mL) for 12 hours, then 100 mL of water was added. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (30 mL×3). The organic extracts were washed with brine and treated with Na 2 SO 4 dry. After the solvent was removed and distilled under reduced pressure, it was purified by 200-300 mesh silica gel column chromatography. Eluted with petroleum ether / ethyl acetate (10:1), the intermediate B1 was obtained as a light yellow liquid with a yield of 75%.

[0142] (2) Synthesis of interm...

Embodiment 3

[0171] This example is basically the same as Example 1, except that the starting material is replaced by the C intermediate prepared by m-bromoaniline, and its synthetic route is shown in Figure 1 (c), which includes the following steps:

[0172] (1) Synthesis of intermediate C1

[0173] The synthetic route is as follows:

[0174]

[0175] m-Bromoaniline (1.0 mmol), acetone (10 mol) and iodine (0.01 mmol) were mixed and stirred in acetonitrile (5 mL) for 12 hours, and then 100 mL of water was added. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (30 mL×3). The organic extracts were washed with brine and treated with Na 2 SO 4 dry. After the solvent was removed and distilled under reduced pressure, it was purified by 200-300 mesh silica gel column chromatography. Eluted with petroleum ether / ethyl acetate (10:1), the intermediate C1 was obtained as a light yellow liquid with a yield of 47%.

[0176] (2) Synthetic intermediate C2 ...

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 a silicon-based rhodamine fluorescent dyeing reagent as well as its preparation method and application, belonging to the fields of organic chemistry and biochemistry. The present invention uses the rhodamine derivatives substituted with silicon atoms as the basic skeleton, and synthesizes a silicon-based rhodamine fluorescent substance with a large Stokes shift and high fluorescence intensity capable of labeling immunoglobulin IgG by modifying different forms of aromatic amines. The staining reagent can be used for in vitro SARS-CoV-2 virus-specific antibody fluorescent ELISA detection. The silicon-based rhodamine fluorescent staining reagent of the present invention has a large Stokes shift (>140nm), can effectively avoid the mutual interference of excitation and emission light, and has high detection sensitivity, and the fluorescent ELISA detection method established based on the fluorescent antibody can be applied Microplate readers with different bandwidths have a wide range of applications.

Description

technical field [0001] The invention relates to the fields of organic chemistry and biochemistry, in particular to the technical field of immunology, and in particular to a silicon-based rhodamine fluorescent staining reagent and its preparation method and application. Background technique [0002] SARS-CoV-2 virus is a novel coronavirus pathogen that caused 2019 Severe Acute Respiratory Syndrome (SARS). The main clinical manifestations of infected patients are respiratory symptoms and pneumonia. In February 2020, the Drug International Virus Taxonomy Committee officially named the virus SARS-CoV-2, and the World Health Organization (WHO) named the disease 2019 coronavirus disease (2019corona virusdisease, COVID-19). The SARS-CoV-2 virus has strong transmission ability, long incubation time, and high incidence rate. Traditional virus defense measures are not effective in controlling the virus. The global pandemic trend is obvious, and the situation of virus prevention and co...

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 Patents(China)
IPC IPC(8): C07D471/04
CPCC09B57/00C07F7/0816C09K11/06G01N33/56983G01N33/533C09K2211/1029C09K2211/1055C09K2211/1096G01N2333/165G01N2469/20
Inventor 李坤张宏刘艳红余孝其
Owner SICHUAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap