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Detection method of glutathione

A detection method, glutathione technology, applied in the detection field, can solve the problems of the fluorescence method being susceptible to interference, the stability is greatly affected, and the chromatography operation is time-consuming, achieving high accuracy, rapid real-time detection, and low detection limit Effect

Inactive Publication Date: 2019-01-15
ANHUI NORMAL UNIV
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Problems solved by technology

Among them, the enzymatic method has high requirements on the experimental conditions, and the stability of the enzyme has a great influence on the experimental results; the chromatographic method is time-consuming and cumbersome to operate; the fluorescence method is easily disturbed and has large errors; and the spectrophotometric method requires cheap instruments. and easy to operate

Method used

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  • Detection method of glutathione
  • Detection method of glutathione
  • Detection method of glutathione

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preparation example Construction

[0028] Preparation of citric acid-sodium citrate buffer solution:

[0029] Get 2.1013g citric acid (C 6 h 8 o 7 ·H 2 O) and 2.9421g sodium citrate (Na 3 C 6 h 5 o 7 .2H 2 0), dissolved in deionized water, ultrasonically oscillated for 3 minutes, then transferred to a 100mL volumetric flask and constant volume, that is, prepared as 0.1M citric acid solution and 0.1M sodium citrate solution, and prepared as a series Citric acid-sodium citrate buffer solution at different pH.

Embodiment 1

[0031] A method for detecting glutathione, specifically:

[0032] At 25°C, take 270μL 0.5mM 3,3′,5,5′-tetramethylbenzidine TMB solution and 90μL 1mM ceric sulfate solution and mix them in a series of test tubes, repeat the above system for 10 parts, add 0μL, 30μL, 60μL, 90μL, 120μL, 150μL, 180μL, 210μL, 240μL and 270μL of 0.1mM glutathione solution, and then add 50μL of 0.1M citric acid-sodium citrate buffer solution to adjust to pH=6, Add deionized water to make up to 1.5mL, the final concentration of glutathione is: 0μM, 2μM, 4μM, 6μM, 8μM, 10μM, 12μM, 14μM, 16μM and 18μM, after adding glutathione solution to the above system , the blue color of the system gradually becomes lighter, and the degree of lightening is related to the amount of glutathione added. Glutathione is within a certain concentration range, and the absorbance of the solution is linearly related to the concentration of glutathione added ( Figure 4 , Figure 5 ), obtain the linear equation A=0.496-0.022C...

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Abstract

The invention provides a detection method of glutathione. The specific principle is as follows: Ce(IV) ion can enable 3,3',5,5'-tetramethyl benzidine (TBM) to generate oxidation reaction to produce blue TBM imide; when trace glutathione is added in the Ce(IV) / TMB system, the glutathione generates redox reaction with the TMB imide, so that the blue of the system is shallowed, and the shallowing degree and the concentration of the glutathione are in direct proportion. According to the above operations, the method for detecting the glutathione is established, the method is simple in operation, and can quickly detect the concentration of the glutathione in real time. Compared with the prior art, the method provided by the invention is simple in operation, can quickly detect the concentration of the glutathione in real time; and the detection limit is low, and can be lowest to 2 microns; the detection method is high in sensitivity, high in accuracy and small in error.

Description

technical field [0001] The invention belongs to the detection field, and in particular relates to a glutathione detection method. Background technique [0002] Glutathione (glutathione, r-glutamyl cysteingl+glycine, GSH) is a tripeptide containing γ-amide bond and sulfhydryl group, which is composed of glutamic acid, cysteine ​​and glycine. Present in nearly every cell of the body. Glutathione can help maintain normal immune system functions, and has antioxidant and integrated detoxification effects. It is widely used in functional foods such as anti-aging, enhancing immunity, and anti-tumor. Therefore, it is of great theoretical and practical value to develop and design a method that can detect low-concentration glutathione. [0003] At present, the main detection methods of glutathione include enzyme cycling method, colorimetric method, high performance liquid chromatography, high performance capillary electrophoresis, flow cytometry, fluorescence method and so on. Amon...

Claims

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Application Information

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IPC IPC(8): G01N21/33
CPCG01N21/33
Inventor 汪鹏敏刘金水
Owner ANHUI NORMAL UNIV
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