Method for detecting glutathione and application of sulfur-gold compound
A technology of glutathione and complexes, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of long synthesis time and high operator requirements, and achieve the effects of short detection time, high sensitivity, and simple operation and processing
Active Publication Date: 2018-08-24
NAT UNIV OF DEFENSE TECH
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Problems solved by technology
The newly developed fluorescence detection method requires the synthesis of nanomaterials with fluorescent properties, which takes a long time to synthesize and requires high requirements for operators
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Embodiment 1
[0032] (1) Prepare RCG as a 10mM stock solution and dilute it with ultrapure water to a 2mM solution for later use. RCG: A tripeptide composed of arginine-cysteine-glutamic acid, which is a positively charged tripeptide under the detection conditions (pH 7.0).
[0033] (2) Prepare a 10 mM stock solution of chloroauric acid and dilute it with ultrapure water to obtain a 1 mM solution for later use.
[0034] Take 200 μL of RCG solution and chloroauric acid solution respectively, and mix them with a shaker for 1 minute to obtain a clear and transparent solution, which is the sulfur-gold complex (or gold-mercapto coordination polymer).
[0035] (3) Preparation of GSH detection probe
[0036] Incubate the synthesized sulfur-gold complex with fluorophore-labeled DNA probes (there is no special requirement for the probe sequence, and generally select a nucleic acid probe with a base length of 20-40 bases and no complex secondary structure), and incubate The conditions are: 20-40°C,...
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The invention discloses a method for detecting glutathione and application of a sulfur-gold compound. The method for detecting the glutathione (GSH) comprises the following steps: mixing a chloroauricacid solution and a polypeptide solution with thiol and reacting for 1 to 3min to obtain a solution containing the sulfur-gold compound; hatching the solution containing the sulfur-gold compound anda fluorophore labeled DNA (Deoxyribonucleic Acid) probe; hatching for 1 to 3min under the hatching conditions that the temperature is 20 to 40 DEG C and the pH (Potential of Hydrogen) value is neutral, so as to obtain a sulfur-gold compound-DNA probe; adding the glutathione with different concentrations into the obtained sulfur-gold compound-DNA probe and detecting by utilizing a fluorescence spectrometer; detecting the GSH according to the intensity of a fluorescence signal. The detection method disclosed by the invention has unique performance in the aspects of constructing a sensing interface and enhancing the sensitivity of sensing response and has excellent performance including convenience, rapidness, no need of antibody recognition and the like; the method is a novel method for conveniently, rapidly and sensitively detecting the GSH.
Description
technical field [0001] The invention relates to a method for detecting glutathione and the application of the sulfur-gold complex. Background technique [0002] Glutathione (GSH) is a tripeptide that is combined with glutamic acid, cysteine and glycine and contains a sulfhydryl group, and is distributed in all organ cells. GSH is mainly involved in the transport of amino acids, the synthesis of proteins and nucleic acids, anti-oxidation, maintaining the reduction state of protein sulfhydryl groups, maintaining the active state of enzymes, protecting cells from damage by free radicals and toxins, etc. GSH has antioxidant and integrated detoxification effects. The abnormality of its content can be used as one of the diagnostic indicators of HIV, Parkinson's syndrome, liver damage, inflammation and other diseases. Therefore, fast, simple and accurate determination of glutathione content is of great significance for the study of the physiological significance of GSH and the ...
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IPC IPC(8): G01N21/64
Inventor 刘卓靓王建方陶呈安王芳柳天雄黄坚李玉姣阳绪衡
Owner NAT UNIV OF DEFENSE TECH