Method for detecting reduced glutathione and/or oxidized glutathione

A detection method and technology for detecting samples, applied in the field of analytical chemistry, can solve the problems of low sensitivity, difficult to be widely used, low molecular polarizability, etc., and achieve the effects of simple operation, fast detection speed and high sensitivity

Pending Publication Date: 2019-06-14
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are SERS detection methods for glutathione in cells, including direct method and indirect method. The direct detection method is affected by the low polarizability of GSH and GSSG molecules, and the sensitivity is very low; the operation of the indirect detection method is complicated. Time-consuming, difficult to apply widely

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  • Method for detecting reduced glutathione and/or oxidized glutathione
  • Method for detecting reduced glutathione and/or oxidized glutathione
  • Method for detecting reduced glutathione and/or oxidized glutathione

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0139] The preparation of embodiment 1 solid-state silver nano-substrate

[0140] 1.1 Preparation of solid silver nano-substrates by in situ reduction method

[0141] Cut the size of the monocrystalline silicon wafer to 0.5×0.5cm 2 , placed in a mixture of concentrated sulfuric acid and hydrogen peroxide (volume ratio 7:3), heated in a constant temperature reaction box at 70°C for 15 minutes, took out the silicon chip and rinsed it with ultrapure water, N 2 Blow dry and set aside. Soak the cleaned silicon chip in 5wt% HF aqueous solution for 20min, then quickly put it into 3mM AgNO containing 10wt% HF 3 solution, react at room temperature for 10min, take out and rinse with ultrapure water, N 2 Blow dry, fix on a clean glass slide, use immediately or store in vacuum for later use. The scanning electron microscope (SEM, Japan Hitachi, model S4800) photo of the solid silver nano-substrate prepared by in situ reduction method is shown in figure 2 .

[0142] 1.2 Preparation ...

Embodiment 2

[0152] The influence of embodiment 2 different substrates on the SERS spectrogram of GSH

[0153] 2.1 Preparation of solution

[0154] Use 10mmol L -1 The PB buffer solution preparation concentration is 10mmol L -1 DTNB solution,

[0155] Prepare a concentration of 10mmol L with ultrapure water -1 GSH solution.

[0156] 2.2 Raman spectrum test

[0157] The concentration was 10mmol L -1 DTNB solution with a concentration of 10mmol L -1 Diluted with PB buffer solution to obtain a concentration of 2 μmol L -1 DTNB solution. Take 5 μL concentration as 2 μmol L -1 DTNB solution, which is added dropwise to the solid silver nano-substrate (prepared by 1.1 in Example 1) prepared by the in-situ reduction method, and another 5 μL DTNB solution is added dropwise to the solid-state gold nano-substrate (prepared by 1.3 in Example 1), until After drying, SERS spectroscopy was performed.

[0158] Take a certain volume of 10mmol L -1 DTNB solution and 10mmol L -1 GSH solution, mi...

Embodiment 3

[0160] The influence of embodiment 3 different conditions on the SERS spectrum of GSSG

[0161] 3.1 Preparation of solution

[0162] Use 10mmol L -1 The PB buffer solution was prepared with a concentration of 1mmol L -1 DTNB solution,

[0163] Prepare a concentration of 0.5 μmol L with ultrapure water -1 GSSG solution,

[0164] Prepare a mixed solution of GSH and GSSG with ultrapure water, wherein the concentration of GSH is 1000 μmol L -1 , the concentration of GSSG was 10 μmol L -1 .

[0165] 3.2 Raman spectrum test

[0166] (a) Take 200μL concentration as 0.5μmol L -1 GSSG solution, add 1mmol L -1 DTNB solution so that the final concentration of DTNB in ​​the mixture is 2 μmol L -1 , and the mixture was incubated at 37° C. for 10 min for SERS spectrum detection.

[0167] (b) Take 200μL concentration as 0.5μmol L -1 GSSG solution, add NaBH 4 Aqueous solution makes NaBH in the mixture 4 The final concentration is 5mmol L -1 , incubate the mixture at 50°C for 10...

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Abstract

The invention relates to a method for detecting reduced glutathione (GSH) and/or oxidized glutathione (GSSG) based on a surface enhanced Raman spectroscopy, and also relates to a kit for detecting GSHand/or GSSG. The detection method and kit are simple in operation, high in detection speed and high in sensitivity, and can be well used for determining the content of GSH and GSSG in an alkylating agent infected cell sample, thereby rapidly screening and detecting an alkylating agent.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, relates to a method for detecting reduced glutathione (GSH) and / or oxidized glutathione (GSSG) based on surface-enhanced Raman spectroscopy, and also relates to a method for detecting GSH and / or kits from GSSG. Background technique [0002] Glutathione is divided into reduced glutathione (GSH) and oxidized glutathione (GSSG), where the chemical name of GSH is N-(N-L-γ-glutamyl-L-cysteinyl) Glycine, the non-protein sulfhydryl compound with the highest content in cells, has important physiological functions such as anti-oxidation, regulating immune response, and resisting exogenous poisons. GSH and GSSG constitute one of the most important redox systems in cells, and their ratio GSH / GSSG is an important indicator of oxidative stress. This ratio is usually maintained in a relatively stable range, but it may change under abnormal physiological conditions. changes happened. In addition to being...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
Inventor 吴剑峰朱颖洁高中才徐华郭磊谢剑炜
Owner ACADEMY OF MILITARY MEDICAL SCI
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