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In situ rapid detection method of hypochlorous acid in cells

A detection method and technology for hypochlorous acid, applied in the field of biological analysis technology or chemical analysis, can solve the problems of weak targeting, difficult to obtain, rare surface-enhanced Raman spectroscopy detection research, and low Raman activity of hypochlorous acid, etc. Achieve the effect of small water interference, fast detection and analysis speed, and high sensitivity

Inactive Publication Date: 2020-04-17
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Due to the low Raman activity or weak targeting effect of hypochlorous acid in cells, it is difficult to obtain characteristic surface-enhanced Raman spectroscopy signals, and related surface-enhanced Raman spectroscopy detection research is relatively rare.

Method used

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  • In situ rapid detection method of hypochlorous acid in cells
  • In situ rapid detection method of hypochlorous acid in cells
  • In situ rapid detection method of hypochlorous acid in cells

Examples

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Embodiment 1

[0046] A kind of in situ rapid detection method of hypochlorous acid in cells, comprising the following steps (see figure 1 ):

[0047] (1) Synthesis of hypochlorous acid recognition molecules

[0048] Mix 5-methoxyl-2-oxo-1,3-benzoxathiolene and potassium hydroxide solution at an equivalent ratio of 1.5, and stir the mixed solution for 6 hours under nitrogen protection , and then added 2M hydrogen chloride solution, the product was extracted with ethyl acetate and washed, and the product was enriched in vacuum to obtain the hypochlorous acid recognition molecule.

[0049](2) Preparation of high-concentration surface-enhanced Raman spectroscopy active substrates

[0050] Take 4.8mL of chloroauric acid solution with a mass ratio of 1%, add it to 100mL of ultrapure water, heat to 100°C (boiling), add 10mL of sodium citrate solution with a mass ratio of 0.1% dropwise, and continue to boil and stirred for 20 minutes and then cooled to 25°C to obtain a stable red surface-enhance...

Embodiment 2

[0060] A method for in-situ rapid detection of hypochlorous acid in cells, comprising the following steps:

[0061] (1) Synthesis of hypochlorous acid recognition molecules

[0062] Mix 5-methoxyl-2-oxo-1,3-benzoxathiolene and sodium hydroxide solution at an equivalent ratio of 1.5, and stir the mixed solution for 7 hours under nitrogen protection , and then added 3M hydrogen chloride solution, the product was extracted with ethyl acetate and washed, and the product was enriched in vacuum to obtain the hypochlorous acid recognition molecule.

[0063] (2) Preparation of high-concentration surface-enhanced Raman spectroscopy active substrates

[0064] Take 4.5mL of chloroauric acid solution with a mass ratio of 2%, add it to 90mL of ultrapure water, heat to 95°C (slight boiling), add 10mL of sodium citrate solution with a mass ratio of 0.2% dropwise, and continue Boil and stir for 20 minutes, then cool to 25°C to obtain a stable red surface-enhanced Raman spectroscopy active s...

Embodiment 3

[0072] A method for in-situ rapid detection of hypochlorous acid in cells, comprising the following steps:

[0073] (1) Synthesis of hypochlorous acid recognition molecules (same as Example 1).

[0074] (2) Preparation of high-concentration surface-enhanced Raman spectroscopy active substrates

[0075] Dissolve silver nitrate in pure water, prepare a silver nitrate solution with a mass ratio of 0.01%, heat it to 90°C (slightly boil), add 2mL of a 0.01% sodium citrate solution drop by drop, continue to boil and stir for 20 minutes, then cool to 25°C, a stable gray surface-enhanced Raman spectroscopy active substrate—silver colloid (see Figure 10 ); put it into a centrifuge tube and centrifuge at high speed for 5 minutes, and remove the layer of high-concentration silver colloid for later use.

[0076] (3) Preparation of nanoprobes

[0077] Take the concentration as 10 -3 M's hypochlorous acid recognition molecule solution, mix it with the silver colloid obtained in step (2...

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Abstract

The invention relates to an in situ rapid detection method for hypochlorous acid in a cell. The in situ rapid detection method comprises the following steps: (1) synthesizing a hypochlorous acid identifying molecule; (2) preparing a high-concentration surface-enhanced Raman spectrum active substrate; (3) preparing a nanoprobe; (4) establishing a characteristic peak intensity-concentration standard curve graph; (5) obtaining a surface enhanced Ramanspectrogramfor the hypochlorous acid in the cell; and (6) carrying out contrast and detection. The invention provides a new method for carrying out in situ rapid detection on the hypochlorous acid in the cell by utilizing a surface enhanced Raman spectrum technology, the method has the advantages of rapid detection, high analysis speed, high sensitivity, low sample amount, strong qualitative capability, photobleaching resistance, small water interference and the like, in situ rapid qualitative and quantitative detection can be carried out on hypochlorous acid in the cell, and detection sensitivity can reach 1MuM or less.

Description

technical field [0001] The invention relates to the field of biological analysis technology or chemical analysis technology, and is a technology for in-situ rapid detection of cells by using the surface-enhanced Raman spectrum effect, specifically, a method for in-situ rapid detection of hypochlorous acid in cells method. Background technique [0002] Reactive oxygen species play key biological effects in living organisms, which can cause DNA damage, affect signal transduction, regulate protein activity, and so on. Studies have shown that hypochlorous acid is closely related to many physiological or pathological processes such as cardiovascular diseases, inflammation, cancer, diabetes, and nervous system lesions. However, many biological functions of hypochlorous acid are still unclear due to the lack of powerful analysis and detection methods. Therefore, the sensitive and accurate in situ detection technology of intracellular hypochlorous acid has important application va...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 李大伟崔景孙佳佳甘振飞郭丹陈华英
Owner EAST CHINA UNIV OF SCI & TECH