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Rapid detection method of glutathione

A glutathione and detection method technology, applied in the field of analysis and detection, can solve the problems of cumbersome operation steps, high analysis cost, and long time consumption

Inactive Publication Date: 2020-03-24
HUBEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a fast detection method for glutathione in view of the current glutathione detection method that has cumbersome operation steps, takes a long time, or relies on expensive instruments, and has high analysis costs.

Method used

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  • Rapid detection method of glutathione
  • Rapid detection method of glutathione

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A kind of detection method of glutathione of the present embodiment, get the chitosan solution that 200 μ L concentration is 1.0mg / mL, 50 μ L concentration is the H of 1.0mM 2 o 2 The solution was mixed with 50µL of TMB solution with a concentration of 5.0mM, and reacted at 37ºC for 30min, and 12 experiments were performed in parallel. Then add 100µL of glutathione solutions with initial concentrations of 0µM, 1.25µM, 10µM, 20µM, 30µM, 40µM, 50µM, 60µM, 80µM, 100µM, 125µM and 200µM to the 12 groups of solutions, mix well and store at 37ºC Continue to react for 30min. After the reaction is completed, it can be clearly observed that as the concentration of glutathione solution increases, the blue color of the solution gradually becomes lighter, see figure 2 , indicating that the depth of the color of the solution is related to the concentration of glutathione. Based on this, the rapid colorimetric detection of glutathione solution can be carried out, and the semi-quant...

Embodiment 2

[0030]In order to investigate the chitosan-based catalytic H 2 o 2 For the feasibility of colorimetric analysis of glutathione by oxidized TMB color development, the following three sets of experiments were designed in this embodiment, see Table 1 below. Wherein the chitosan solution initial concentration used is 1.0mg / mL, H 2 o 2 The initial concentration of the solution was 1.0 mM, and the initial concentration of the TMB solution was 5.0 mM. Groups 1 and 2 reacted for 30 minutes after adding TMB, then added deionized water and continued to incubate for 30 minutes; after adding TMB and reacted for 30 minutes, group 3 added glutathione solution with a concentration of 0.4 mM and continued to incubate for 30 minutes. The reaction temperature for each set of experiments was 37ºC.

[0031] Table 1 Feasibility study scheme of chitosan mimic peroxidase for colorimetric analysis of glutathione

[0032]

[0033] Experimental results such as Figure 6 Shown: In the first gro...

Embodiment 3

[0035] This example considers H 2 o 2 also exhibit some oxidative properties, the excess H 2 o 2 The impact on the detection of glutathione is more obvious, so the H used in the detection 2 o 2 The concentration was optimized, and the optimization experiment was specifically as follows:

[0036] Four groups of experiments numbered 1-4 were set up, and each group contained two experiments of background item and signal item. The background item protocol for each group was 200 μL of chitosan solution and 50 μL of HO 2 o 2 And 50 μL of TMB solution was reacted at 37 ºC for 30 min, and then 100 μL of water was added to continue incubation for 30 min. The signal item protocol for each group is 200 μL of chitosan solution and 50 μL of HO 2 o 2 And 50 μL of TMB solution was reacted at 37 ºC for 30 min, and then 100 μL of glutathione solution was added to continue incubation for 30 min. The initial concentration of chitosan solution used in the experiment was 1.0mg / mL, the in...

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Abstract

The invention discloses a rapid detection method of glutathione. The detection method specifically comprises the following steps of mixing a chitosan solution, an H2O2 solution and a 3, 3', 5, 5'-tetramethyl benzidine soluction to react, and then respectively adding the glutathione solutions with different concentration gradients into a mixed reaction system to continuously react, and observing the color of the solution added with the glutathione with different concentration gradients by naked eyes and photographing through a camera after the reaction is finished so as to semi-quantitatively detect the concentration of glutathione according to the color of the solution to be detected; or measuring the absorbance of the solution by using an ultraviolet spectrophotometer, constructing a linear relationship between the absorbance and the concentration of glutathione, and rapidly detecting the concentration of glutathione in the solution to be detected by using the linear relationship. Themethod has the advantages of being simple to operate, being able to realize the semi-quantitative and quantitative detection of glutathione rapidly and flexibly, and has the good application prospect.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a rapid detection method for glutathione. Background technique [0002] Glutathione (GSH) is a tripeptide containing a thiol group, which is formed by condensation of glutamic acid, cysteine ​​and glycine, and is distributed in the tissue fluid and body fluid of humans and animals. Glutathione plays an important role in the body's immune regulation, integrated detoxification, cell proliferation, apoptosis, etc., and the occurrence of some diseases, such as HIV, Parkinson's syndrome, liver injury, inflammation, etc. will cause glutathione in the body to Abnormal levels. Therefore, the establishment of a sensitive glutathione analysis method has important application value for biomedical research and clinical medical testing. [0003] At present, the detection methods of glutathione mainly include electrochemical method, fluorescence method, high performance liquid...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/33G01N31/10
CPCG01N21/78G01N21/33G01N31/10
Inventor 陈金阳孙梦彤王妤欣库苠月刘祥茹
Owner HUBEI NORMAL UNIV
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