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. -

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

Example Embodiment

[0066] Example 1: Preparation of probe BNBD

[0067] A dual response detection 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, stir for 30min at room temperature to obtain a brown precipitate, filter, and wash the filter cake with ethyl acetate , Dry to obtain 0.29g brown product, obtain product 1 (HNBD), the yield is 97%;

[0069] (2) Dissolve product 1 in 20ml ethanol with a concentration of 0.0195g / mL, add benzaldehyde to a concentration of 1mol / L, add glacial acetic acid to a concentration of 0.1mol / L, reflux for 3h, cool, and precipitate a red-black solid, solid Elute with 1:1 ethyl acetate-petroleum ether eluent, collect the first product, and dry it to obtain a red solid, which is the dual response detection ClO based on hydrogen ion activation - / H 2 S fluores...

Example Embodiment

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

[0078] Three groups of rabbits were cultivated, one group of rabbits were fed with chlorine water; the second group of rabbits were fed with cysteine; the three groups of rabbits were normal rabbits. After half a month of feeding, the gastric juices of three groups of rabbits were measured and tested.

[0079] Prepare solution

[0080] Prepare probe BNBD stock solution: accurately weigh out ClO based on hydrogen ion activation dual response detection - / H 2 S fluorescent molecular probe is dissolved in anhydrous acetonitrile, and the preparation concentration is 2×10 -4 M probe BNBD stock solution;

[0081] Prepare the target substance ClO - Stock solution: accurately weigh the target CIO - Dissolved in water, the preparation concentration is 1×10 -3 M's ClO - Stock sol...

Example Embodiment

[0091] Example 3: Detection of sample ClO in isolated gastric mucosa - And H 2 S content (using the hydrogen ion activated dual response detection sample ClO prepared in Example 1 - / H 2 S fluorescent molecular probe)

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

[0093] To prepare 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, and dilute to 1000ml with water.

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

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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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