Preparation method and application of pH fluorescence probe

A fluorescent probe and fluorescent molecular probe technology, applied in the field of analytical chemistry, can solve the problems of cell apoptosis, cell dysfunction, organelle damage, etc., and achieve the effects of sensitive detection, novel principle and fast detection process

Active Publication Date: 2017-11-24
QUFU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, abnormal changes in pH, such as acidification, can cause cellular dysfu

Method used

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  • Preparation method and application of pH fluorescence probe
  • Preparation method and application of pH fluorescence probe
  • Preparation method and application of pH fluorescence probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1: Preparation of PH fluorescent probe

[0070] A preparation method of PH fluorescent probe, the steps include:

[0071] (1) Dissolve NBD-Cl (7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole) in chloroform at a concentration of 0.004g / ml, and then add the same volume of chloroform solution and chloroform solution Contain 1% hydrazine hydrate-methanol solution (in the hydrazine hydrate-methanol solution, the volume concentration of hydrazine hydrate is 1%), mix well, stir at room temperature for 2h to obtain brown precipitate, the precipitate is completely precipitated, filtered, the filter cake is passed through ethyl acetate Ester washing and drying to obtain NBD hydrazine;

[0072] (2) Add pyridinium chlorochromate to freshly distilled anhydrous dichloromethane, add 0.083 g / mL concentration, dissolve ultrasonically, and then add 4-(chloromethyl)benzyl alcohol with a concentration of 0.05 g / mL, The reaction was stirred at room temperature for 3 hours under nitrogen protect...

Embodiment 2

[0080] Example 2: Preparation of PH fluorescent probe

[0081] A preparation method of PH fluorescent probe, the steps include:

[0082] (1) Dissolve NBD-Cl (7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole) in chloroform at a concentration of 0.008g / mL, and then add the same volume of chloroform solution and chloroform solution Contain 1% hydrazine hydrate-methanol solution (in the hydrazine hydrate-methanol solution, the volume concentration of hydrazine hydrate is 1.2%), mix well, stir at room temperature for 2 h to obtain brown precipitate, filter, and wash the filter cake with ethyl acetate. Dry it to get NBD hydrazine;

[0083] (2) Add pyridinium chlorochromate to freshly distilled anhydrous dichloromethane, add 0.12g / ml concentration, dissolve by ultrasonic, then add 4-(chloromethyl)benzyl alcohol with a concentration of 0.08 g / mL, The reaction was stirred at room temperature under nitrogen protection for 4 h. After the reaction was completed, 3 times the volume of dichloromethane w...

Embodiment 3

[0085] Example 3: Preparation of PH fluorescent probe

[0086] A preparation method of PH fluorescent probe, the steps include:

[0087] (1) Dissolve NBD-Cl (7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole) in chloroform at a concentration of 0.001g / mL, and then add the same volume of chloroform solution to the chloroform solution Containing 0.39% hydrazine hydrate-methanol solution (in the hydrazine hydrate-methanol solution, the volume concentration of hydrazine hydrate is 0.2%), mix well, stir at room temperature for 2 h to obtain a brown precipitate, filter, and wash the filter cake with ethyl acetate. Dry it to get NBD hydrazine;

[0088] (2) Add pyridinium chlorochromate to dichloromethane, add concentration of 0.041g / ml, dissolve ultrasonically, then add 4-(chloromethyl)benzyl alcohol with a concentration of 0.02 g / mL, room temperature under nitrogen protection Stir the reaction for 3 hours. After the reaction is completed, add 4 times the volume of dichloromethane anhydrous diethy...

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Abstract

The invention provides a preparation method and application of a pH fluorescence probe. A novel fluorescence probe CM-BHNBD is designed, and is from NBD(4-nitrobenzene-2-oxalic acid-13-diazole); the fluorophore intracellular mitochondria/membrane acidification monitoring can be realized; when the method is applied to pH detection in a biological sample, the advantages of good response performance, high data accuracy, good reproducibility, high precision and the like are realized; the implementation feasibility is high.

Description

Technical field [0001] The invention belongs to the field of analytical chemistry, and relates to a method for preparing a pH fluorescent probe and a method for real-time detection of pH value in a biological sample, specifically the acidification monitoring of intracellular mitochondria or mitochondrial membranes. Background technique [0002] Currently, for intracellular mitochondria / intramembrane H + Monitoring analysis, chemical methods mainly include high performance liquid chromatography and mass spectrometry, gas phase, ultraviolet detectors, etc. The number of fluorescent molecular probe detection methods is very small. The gas chromatography detection method has been applied in many fields, but it is not the best choice for the analysis field and the rapid detection of clinical medical biological samples. First of all, the gas chromatography detection method has complex technical steps, numerous sample processing steps and a large workload. , Long operating time, low eff...

Claims

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Application Information

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IPC IPC(8): C07D271/12C09K11/06G01N21/64
CPCC07D271/12C09K11/06C09K2211/1048G01N21/6486
Inventor 陈光窦昆付强李震姜翱刘玉霞尤进茂
Owner QUFU NORMAL UNIV
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