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Method and kit for detecting extremely acidic pH

A kit and acidic technology, applied in the field of a method and kit for detecting extremely acidic pH, can solve the problems of strong autofluorescence, short fluorescence signal emission wavelength, high scattering rate, etc., and achieve long fluorescence signal emission wavelength and extremely acidic pH The effect of sensitive response and low autofluorescence

Pending Publication Date: 2022-05-27
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the fluorescence methods for detecting the weakly acidic environment with a pH range of 4-6 include ultraviolet light, visible light and near-infrared fluorescence detection methods, but these methods have many limitations, such as poor acidic responsivity, short fluorescence signal emission wavelength, and penetration Poor depth, strong autofluorescence, high scattering rate, and low signal-to-noise ratio
Higher imaging resolution can be obtained in the wavelength range of the near-infrared second region (900-1700nm, called NIR-II), which further improves the penetration depth, sensitivity and resolution of imaging. However, there is still a lack of detection of pH Near-infrared two-region fluorescence method for extremely acidic pH values ​​less than 1

Method used

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  • Method and kit for detecting extremely acidic pH
  • Method and kit for detecting extremely acidic pH

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Near-infrared second-region fluorescence emission spectra of protonated methylene blue (MBH) under different acidic conditions

[0049] First, use hydrochloric acid to configure the hydrogen ion concentration from 10 -7 A series of solutions from M to 6M, and then adding 50 μg / mL MB, will generate MBH immediately, and the reaction process is as follows figure 1 shown. Excited at 808nm wavelength, the near-infrared second region fluorescence emission spectrum was measured. figure 2 Shown are the near-infrared second region fluorescence emission spectra of MBH under different acidic conditions, figure 2 In a, the abscissa is the wavelength, and the ordinate is the fluorescence intensity, figure 2 In b, the abscissa is the hydrogen ion concentration, and the ordinate is the fluorescence intensity. figure 2 In a, it is shown that the fluorescence emission wavelength of MBH in the second near-infrared region under different acidic conditions is about 950 nm, ...

Embodiment 2A

[0050] Example 2 Azure A and Azure B also have extremely acidic responsive near-infrared second region fluorescence emission properties

[0051] The present inventors also found that MB analogs Azure A (CAS number: 531-53-3, CAS name: 3-Amino-7-(dimethylamino)phenothiazin-5-ium chloride) and Azure B (CAS number: 531-55-5) , CAS name: 3-(Dimethylamino)-7-(methylamino)phenothiazin-5-iumchloride), they also have near-infrared second-region fluorescence emission properties in the environment of pH less than 1, such as image 3 shown, image 3 a and 3b are the structural formulas of Azure A and Azure B, respectively, image 3 c and image 3 d are the near-infrared second region fluorescence emission spectra of Azure A and Azure B under acidic conditions, respectively, image 3 e and image 3 f are the fluorescence intensities of Azure A and Azure B under different hydrogen ion concentrations, respectively. image 3 c and image 3 In d, the abscissa is the wavelength, and the ...

Embodiment 3

[0052] Example 3 The kit of the present invention is used to detect the pH value of the in vitro system

[0053] First, dissolve the phenothiazine salt (which can be any of methylene blue, Azure A and Azure B) in the kit with the solvent in the kit (which can be any of water, dimethyl sulfoxide, ethanol, and tetrahydrofuran). ), configured into a 50-500 μg / mL phenothiazine salt solution. Mix the aqueous solution with the system to be detected, and keep the final concentration of the phenothiazine salt at 50 μg / mL after mixing. After 1 minute, put the mixed system into a fluorescent dish, and use an Edinburgh FLS920 fluorescence spectrometer to detect the fluorescence spectrum. The wavelength is 808nm, the fluorescence signal is collected in the 900-1700nm band, and the intensity of the fluorescence signal is compared with the standard curve (the standard curve of methylene blue is figure 2 b, the standard curve of Azure A is image 3 e, the standard curve of Azure B is im...

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Abstract

The invention relates to a method and a kit for detecting extremely acidic pH, phenothiazine salt is protonated after being combined with hydrogen ions in an acidic environment, a protonated product can emit near-infrared two-region fluorescence, and the pH value is obtained by detecting the fluorescence intensity. Compared with fluorescence detection extremely acidic methods such as ultraviolet light, visible light and near-infrared first region, the method has the advantages of sensitive extremely acidic response, large penetration depth, low autofluorescence, weak scattering, high signal-to-noise ratio and the like, and the kit can be used for detecting the pH value of a biological sample in an extremely acidic environment with the pH value smaller than 1.

Description

technical field [0001] The invention relates to the technical field of biological fluorescence analysis, in particular to a method and a kit for detecting extremely acidic pH. Background technique [0002] Protons, as a familiar cation, play a key role in biology. Fluctuations in pH have pronounced effects on many cellular events, such as cellular metabolism, growth, signal transduction, chemotaxis, apoptosis, and autophagy. Although most organisms cannot survive in a strongly acidic environment, there are still a considerable number of microorganisms such as Helicobacter pylori and "acidophilic bacteria". Like enteric pathogens, it can reach the intestine through the highly acidic mammalian stomach and cause disease infection. Some parts of mammals have very low pH, such as gastric juice, and pH can significantly affect their physiological processes. The precise pH in these cells remains difficult to determine due to the lack of efficient methods to detect the acidic pH ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6439
Inventor 蔡林涛邓冠军张鹏飞龚萍
Owner SHENZHEN INST OF ADVANCED TECH
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