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Preparation method and application of dual-response fluorescent molecular probe for detecting ClO<->/H2S based on hydrogen ion activation

A fluorescent molecular probe and dual-response technology, applied in the field of analytical chemistry, can solve problems such as limitations and achieve sensitive detection

Inactive Publication Date: 2017-03-22
QUFU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to the best of our knowledge, no probe can detect ClO under acidic conditions. - / H 2 S was detected, which clearly limits the ClO in animals - and H 2 S-Balance Study

Method used

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  • Preparation method and application of dual-response fluorescent molecular probe for detecting ClO&lt;-&gt;/H2S based on hydrogen ion activation
  • Preparation method and application of dual-response fluorescent molecular probe for detecting ClO&lt;-&gt;/H2S based on hydrogen ion activation
  • Preparation method and application of dual-response fluorescent molecular probe for detecting ClO&lt;-&gt;/H2S based on hydrogen ion activation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: Preparation of probe BNBD

[0067] A dual-response detection of ClO based on hydrogen ion activation - / H 2 The preparation method of S fluorescent molecular probe, the steps are:

[0068] (1) Dissolve 0.3g NBD-Cl in 50mL chloroform, then add 50mL hydrazine hydrate-methanol solution with a volume concentration of 0.6%, mix well, and stir at room temperature for 30min to obtain a brown precipitate, filter, and wash the filter cake with ethyl acetate , dried to obtain 0.29g of brown product, to obtain product 1 (HNBD), and the yield was 97%;

[0069] (2) Dissolve product 1 in 20ml of ethanol with a concentration of 0.0195g / mL, add benzaldehyde at a concentration of 1mol / L, add glacial acetic acid at a concentration of 0.1mol / L, reflux for 3 hours, cool, and precipitate a red-black solid, solid Elute with ethyl acetate-petroleum ether eluent with a volume ratio of 1:1, collect the first product, and dry it to obtain a red solid, which is the dual-response det...

Embodiment 2

[0077] Embodiment 2: Detection of ClO in isolated gastric juice - and H 2 The content of S (using the dual response detection ClO based on hydrogen ion activation prepared in Example 1 - / H 2 S fluorescent molecular probes)

[0078] Three groups of rabbits were cultivated respectively. Rabbits in group 1 were fed with chlorine water; rabbits in group 2 were fed with cysteine; rabbits in group 3 were normal rabbits. After half a month of feeding, the gastric juices of the three groups of rabbits were taken for measurement and detection.

[0079] Prepare the solution

[0080] Preparation of probe BNBD stock solution: Accurately weigh ClO based on dual-response detection based on hydrogen ion activation - / H 2 S fluorescent molecular probes were dissolved in anhydrous acetonitrile and prepared at a concentration of 2×10 -4 Probe BNBD stock solution of M;

[0081] Preparation of target ClO - Stock solution: Accurately weigh the target ClO - Soluble in water, the preparat...

Embodiment 3

[0091] Example 3: Detection of sample ClO in isolated gastric mucosa - and H 2 The content of S (using the dual response detection sample ClO based on hydrogen ion activation prepared in Example 1 - / H 2 S fluorescent molecular probes)

[0092] Isolated gastric mucosa: taken from the three groups of rabbits in Example 2, the gastric mucosa was first rinsed with artificial gastric juice to scrape off the impurities outside the mucosa, then a thin layer of about 1mm was taken, soaked and cleaned with normal saline, and placed on a glass slide for use imaged under a microscope.

[0093] Preparation of artificial gastric juice: Take 16.4ml of dilute hydrochloric acid with a mass concentration of 9.5-10.5%, add 800ml of water and 10g of pepsin, shake well, add water to dilute to 1000ml.

[0094]1) Prepare the solution: the method is the same as in Example 2, the blank isolated gastric mucosa sample and anhydrous acetonitrile are mixed at a volume ratio of 5:1 to obtain a serum ...

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Abstract

The present invention relates to a preparation method and an application of a dual-response fluorescent molecular probe for detecting ClO<-> / H2S based on hydrogen ion activation. The method comprises the steps of dissolving NBD-Cl in chloroform, adding a hydrazine hydrate-methanol solution, and performing uniform mixing to obtain a product 1; dissolving the product 1 in ethanol, adding benzaldehyde and glacial acetic acid, and conducting refluxing, cooling, precipitation, etc. to obtain the final product. The fluorescent molecular probe is applicable to qualitative analysis and quantitative analysis of ClO<-> / H2S in biological samples, and detection is sensitive, accurate and fast; the biological samples mainly include gastric juice, gastric mucosa, cells, and so on. The fluorescent molecular probe can be used in the related fields of analytical chemistry, life organic analysis, disease pre-diagnosis and clinical examination.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and relates to a double response detection ClO based on hydrogen ion activation - / H 2 Preparation method and application of S fluorescent molecular probe. Background technique [0002] Redox balance Reactive oxygen species (ROS) and reduced sulfur species (RSS) play key roles in regulating biological processes such as cell proliferation, differentiation, and apoptosis. Disruption of redox balance due to overproduction of reactive oxygen species, overwhelming the cell's protective defense mechanisms will lead to consequences such as peroxidation, DNA damage, and even more deleterious outcomes if genetic variants affect normal repair proteins altered. As a well-known molecule RSS, hydrogen sulfide (H 2 S), has been shown to have anti-apoptotic and anti-inflammatory effects. Due to its protective effect on the gastric mucosa, hydrogen sulfide has been exploited in the design of new medicines ...

Claims

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Application Information

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IPC IPC(8): C07D271/12C09K11/06G01N21/64
CPCC07D271/12C09K11/06C09K2211/1048G01N21/6486
Inventor 刘玉霞付强姜翱胡金莲窦昆陈光尤进茂
Owner QUFU NORMAL UNIV
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