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Au < 3 + > adjustment-based label-free colorimetric method for autocatalytic oxidation of o-phenylenediamine and application thereof

A determination method and a detection method technology, applied in the application field of detecting glutathione, can solve the problem of seldom paying attention to catalytic performance and the like, and achieve the effect of broadening the application

Pending Publication Date: 2020-11-24
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most analytical applications of NPs focus on their fluorescent properties, and little attention is paid to their catalytic properties.

Method used

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  • Au &lt; 3 + &gt; adjustment-based label-free colorimetric method for autocatalytic oxidation of o-phenylenediamine and application thereof
  • Au &lt; 3 + &gt; adjustment-based label-free colorimetric method for autocatalytic oxidation of o-phenylenediamine and application thereof
  • Au &lt; 3 + &gt; adjustment-based label-free colorimetric method for autocatalytic oxidation of o-phenylenediamine and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Further study of GSH on Au by TEM 3+ Influence of the reaction between and OPD. The AuNPs systems with and without GSH were characterized by TEM, and the TEM results were as follows: figure 2 As shown, Au NPs produced in the presence of GSH are completely different in size and shape from those produced in the absence of GSH. showed that GSH has an effect on OPD and Au 3+ The redox reaction between them has an influence, which further affects the synthesis of Au NPs.

Embodiment 2

[0038] OPD can be detected by Au 3+ Oxidation to generate light yellow oxidation state OPD (ox OPD), the fluorescence spectrum of the detection system is shown in image 3 As shown, it can be seen that ox OPD has strong fluorescence, the excitation wavelength is at 420 nm, and the absorption wavelength is at 568 nm. respectively will add HAuCl 4 ·3H 2 O, do not add HAuCl 4 ·3H 2 The detection system of O is characterized by UV-vis, and the results are as follows Figure 4 As shown, the raw material OPD has no absorption at 447 nm and only has an absorption peak at 288 nm, while ox OPD has an obvious absorption peak at 447 nm.

Embodiment 3

[0040] This colorimetric method detects the UV-vis of different concentrations of GSH, GSH is the abbreviation of glutathione (glutathione, L-γ-glutamyl-L-cysteinylglycine, GSH), the specific steps are as follows:

[0041] 25 μL of 0.75 mM HAuCl 4 ·3H 2O was dispersed in 0.6 mL of pH 5.0 phosphate buffer (0.01 M), and then 200 μL of different concentrations of GSH (from top to bottom were 1.14, 5.71, 11.43, 57.14, 114.29, 571.43, 1142.86, 5714.29, 11428.57, 57142.86, 114285.71 nM), and finally add 50 μL 2 mM OPD. After incubation at 10 °C for 15 min by a thermostat, the reaction system was transferred for UV-vis detection. The result is as Figure 5 As shown, it can be seen that the absorbance peak is located at 447 nm, which is attributed to ox OPD (oxidation state OPD). The blank group had the highest absorbance, and the absorbance gradually decreased with the increase of GSH concentration. Thus, the ΔA value increased systematically with increasing GSH concentration an...

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Abstract

The invention discloses an Au < 3 + > adjustment-based label-free colorimetric method for autocatalytic oxidation of o-phenylenediamine and application thereof. The method comprises the following steps: dispersing HAuCl4. 3H2O in a phosphate buffer solution, then adding GSH to be detected, finally adding OPD, incubating through a thermostat, and then transferring a reaction system for UV-vis detection. Compared with the methods in the prior art, the detection method has fewer operation steps without complex separation, immobilization or functionalization processes so that the method is simple,rapid and sensitive, can be applied to determination of the concentration of GSH in actual samples such as rat myocardial ischemia reperfusion model serum, and has a relatively good application prospect in the field of biochemical determination and analysis.

Description

technical field [0001] The invention relates to the field of biochemical determination and analysis, and designs and develops a method based on Au 3+ A label-free colorimetric method for the regulated autocatalytic oxidation of o-phenylenediamine and its application in the detection of glutathione (GSH). Background technique [0002] Glutathione (glutathione, L-γ-glutamyl-L-cysteinylglycine, GSH) is a biothiol that acts as an important endogenous antioxidant in cellular defense against toxins and free radicals plays an important role. GSH can be widely involved in immune regulation, human metabolism, energy transport and other physiological processes. Therefore, GSH has been widely used to protect the liver and kidney, for detoxification and anticancer drugs. In general, abnormal levels of GSH are closely related to several diseases, such as cancer, liver damage, human immunodeficiency virus, and diabetes, among others. Therefore, a simple and sensitive detection of GSH ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 李少光姚宏林新华张晓颖黄丽英李光文刘慧林
Owner FUJIAN MEDICAL UNIV