Method for detecting hydrogen peroxide and lactic acid based on colorimetric principle and application of method

A horseradish peroxidase and ratio technology, applied in the field of medical testing, can solve the problems of lack of biochemical sensing detection system, less biochemical detection, and immature development

Active Publication Date: 2021-09-03
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still few biochemical tests based on smartphones
This may be due to the lack of establishment of a biochemical sensing detection system suit

Method used

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  • Method for detecting hydrogen peroxide and lactic acid based on colorimetric principle and application of method
  • Method for detecting hydrogen peroxide and lactic acid based on colorimetric principle and application of method
  • Method for detecting hydrogen peroxide and lactic acid based on colorimetric principle and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Embodiment 1 A kind of synthetic method of MP-Au NPs

[0111] 1.1 Synthesis of PNAAN-Au NPs

[0112] 4 μL HAuCl 4 (10%, 1.14mM) was injected into 961μL of 10mM sodium hydroxide solution, and then 25μL of 32mg / mL polyvinylpyrrolidone (PVP; M=40000) solution was added, mixed evenly (30s) and stored in a 4℃ refrigerator for 30 Minutes, this is solution 1; N-(3-mimidyl)-aniline (NAAN) is dissolved in water and made into a 400mg / ml solution and placed in a refrigerator at 4°C for 30 minutes, this is solution 2; after 30 minutes, Inject 10 μL of solution 2 into solution 1 (the final concentration of NAAN in the system is 16 mM), mix well, shake at room temperature at 300 rpm for 10 minutes and take it out to obtain poly-N-(3-mimidyl)-aniline-protected gold nanoparticles (PNAAN -Au NPs) solution.

[0113] 1.2 Ligand exchange

[0114] In the present invention, mercapto groups can be used to form tight and stable gold-sulfur bonds with gold, so 100 μL of 1 mM p-mercaptopheno...

Embodiment 2

[0116] Example 2 A detection of H using the SPR optical properties of MP-Au NPs 2 o 2 Methods

[0117] Due to its surface plasmon resonance (SPR) effect, gold nanoparticles (Au NPs) will cause strong light absorption at specific wavelengths. As the particle size of gold nanoparticles increases, the position of its absorption peak shifts red toward the long wavelength direction, and the color of the solution gradually changes from red to dark purple, blue or even blue-white. Such as figure 1 As shown, the present invention utilizes horseradish peroxidase to catalyze hydrogen peroxide to generate hydroxyl radicals, and utilize hydroxyl radicals to cross-link phenol groups to mediate aggregation of gold nanoparticles, providing a method for detecting H 2 o 2 Methods:

[0118] 2.1 MP-Au NPs vs H 2 o 2 the response to

[0119] ① Purify the MP-Au NPs solution, centrifuge the MP-Au NPs prepared in 1.2 of Example 1 at 5000rpm for 8 minutes, then remove the supernatant, add an ...

Embodiment 3

[0127] Embodiment 3 Stability comparative experiment of gold nanoparticles

[0128] In order to further evaluate the stability of the p-mercaptophenol-modified gold nanoparticles prepared by the present invention, this experiment was carried out in different media (such as salt solution with higher concentration, strong acid and strong Alkali) in the stability is compared, in order to assess whether PNAAN can effectively improve the stability of gold nanoparticles as a stabilizing agent in the present invention, concrete steps are as follows:

[0129] 3.1 Materials:

[0130] ①P-mercaptophenol was purchased from Shanghai McLean Biochemical Technology Co., Ltd.; sodium chloride, hydrochloric acid, and sodium hydroxide were purchased from Tianjin Damao Chemical Reagent Factory;

[0131] ② The p-mercaptophenol-modified gold nanoparticles (MP-Au NPs 1 for short) prepared based on carboxyl-protected gold nanoparticles (COOH-Au NPs) refer to the Chinese patent (patent number "201710...

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Abstract

The invention discloses a method for detecting hydrogen peroxide and lactic acid based on a colorimetric principle and application of the method. According to the method, gold nanoparticles protected by polyaniline derivatives are prepared by taking chloroauric acid, polyvinylpyrrolidone and N-(3-amidinyl)-aniline as raw materials, and then the gold nanoparticles are further reacted with p-mercaptophenol to prepare the p-mercaptophenol gold nanoparticles. Experiments show that horse radish peroxidase is utilized to catalyze decomposition of hydrogen peroxide to generate hydroxyl radicals, and the hydroxyl radicals can enable phenol to be subjected to cross-linking polymerization, so that the prepared mercaptophenol gold nanoparticles can be effectively promoted to be aggregated, and the color of a solution is changed; the change is closely related to the concentration of the hydrogen peroxide, so that the p-mercaptophenol gold nanoparticles can be used for detecting the hydrogen peroxide or detecting related substances which can generate the hydrogen peroxide in an enzyme catalytic metabolism process, such as lactic acid, glucose, cholesterol and the like.

Description

technical field [0001] The invention belongs to the field of medical detection, in particular to a method for detecting hydrogen peroxide and lactic acid based on a colorimetric principle and an application thereof. Background technique [0002] Hydrogen peroxide (H 2 o 2 ) as one of the reactive oxygen species (reactive oxygen species, ROS) in the human body, is an important signaling molecule and oxidative stress indicator, excessive H 2 o 2 It can cause DNA damage, cell death, and aging, which can further lead to diseases such as chronic inflammation, cancer, and central nervous system diseases. Therefore, to establish efficient H 2 o 2 Analytical methods are of great importance. At the same time, H 2 o 2 As an intermediate in the biochemical process, it is mainly produced through the oxidative decomposition of substances in the metabolic process, for example, lactate oxidase (Lactate oxidase, LOX) oxidizes lactic acid (Lactate). Therefore, by detecting the H pro...

Claims

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Application Information

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IPC IPC(8): G01N21/33G01N21/78G01N21/84C09K9/00B82Y40/00B82Y30/00
CPCG01N21/33G01N21/78G01N21/84C09K9/00B82Y30/00B82Y40/00
Inventor 李楠张美莹陈凯欣牟宗霞崔鑫周平薛巍
Owner JINAN UNIVERSITY
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