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Method for detecting GST by combining magnetic single-drop microextraction fluorescent switch with PDAcoating vesicle

A fluorescent switch, single drop technology, applied in the field of bioanalytical chemistry, to achieve the effect of broad application prospects and satisfactory recovery rate

Pending Publication Date: 2022-02-08
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Technical problem to be solved: In order to overcome the shortcomings of the existing technology and avoid the inconvenience of pre-treatment, a new method for detecting GST is proposed, which can be directly used to detect actual samples

Method used

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  • Method for detecting GST by combining magnetic single-drop microextraction fluorescent switch with PDAcoating vesicle
  • Method for detecting GST by combining magnetic single-drop microextraction fluorescent switch with PDAcoating vesicle
  • Method for detecting GST by combining magnetic single-drop microextraction fluorescent switch with PDAcoating vesicle

Examples

Experimental program
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Effect test

Embodiment 1

[0044] A magnetic single-drop micro-extraction fluorescent switch combined with a method for detecting GST by PDA-coated vesicles, the method comprising the following steps:

[0045] (1) Fe 3 o 4 Preparation of:

[0046] Weigh 0.82g FeCl 3 ·6H 2 O, dissolve in 40mL ethylene glycol, stir until the solution is clear and yellow and transparent, then add 3.6g NaAC, stir vigorously for 30min, ultrasonicate for 10min, then pour the solution into a polytetrafluoroethylene reactor, 200 ℃ reaction 12h. After taking it out, cool to room temperature, wash with ethanol 4 times, and dry at 60°C;

[0047] (2) Fe 3 o 4 Preparation of @PDA @Aptamer @GST:

[0048] 0.025g Fe 3 o 4 Add 100mL Tris-HCl buffer solution and sonicate for 3 hours, then add 0.1g dopamine hydrochloride, keep away from light and mix mechanically for 6 hours (wrap the beaker with tin foil), and the prepared Fe 3 o 4 Wash the @PDA solution three times with water, then add 20 μL of 1 μM Aptamer, incubate at 30°C...

Embodiment 2

[0057] The method described in Example 1 was used to detect GST in urine: in order to detect the content of GST in urine, the GST in the urine sample was spiked. Using the experimental data of fluorescence measurement, the content of GST in urine is obtained. As shown in Table 1, good recoveries (100.0%-114.2%) were obtained.

[0058] Table 1 Determination of GST Spiked in Urine Samples

[0059]

[0060] a Relative recovery = (total concentration - blank concentration) / spiked concentration

Embodiment 3

[0062] The method described in Example 1 was used to detect the GST content in the blood, and the GST in the blood sample was spiked. Using the experimental data of fluorescence measurement, the content of GST in the blood is obtained. As shown in Table 2, good recoveries (95.8-104.0%) were obtained.

[0063] Table 2 Determination of GST incorporated in blood samples

[0064]

[0065] a Relative recovery = (total concentration - blank concentration) / spiked concentration

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Abstract

The invention discloses a method for detecting GST by combining a magnetic single-drop microextraction fluorescent switch with a PDA coating vesicle. The method provides a pH-dependent PDA coating vesicle / magnetic Fe3O4 single-drop microextraction (SDME) fluorescent switch, a magnetic tip is designed on a magnetic rod of a single drop, so that the stability of the volume of the liquid drop can be ensured, the liquid drop can be rapidly enriched, an independent reaction system is formed, signal interference (interference of matrix effect) can be eliminated, and inconvenience caused by pretreatment is avoided. Therefore, the method can be used for direct detection of actual samples. According to the method, the concentration of the GST can be accurately quantified, the linear range of the GST is 0.005-0.5 mug / mL (R2 is greater than 0.995), and the detection limit (LOD) is 3.7 ng / mL. Besides, the method has a satisfactory recovery rate in actual determination of GST in urine samples and blood, which indicates that the method has a wide application prospect in analysis of complex samples.

Description

technical field [0001] The invention discloses a method for detecting GST using a magnetic single-drop micro-extraction fluorescent switch combined with PDA coating vesicles, which is used for the GST detection of real biological samples, and belongs to the field of bioanalytical chemistry. Background technique [0002] Glutathione s-transferase (GST) is one of the most important phase metabolism enzymes in organisms. As a detoxifying enzyme, it plays a vital role in reducing toxicity. Its main function is to catalyze the attack of reduced glutathione (GSH) on the electrophilic center of target compounds such as endogenous superoxide radicals and toxic metabolites. Thus, tissues and cells are protected from damage. GST is widely distributed in various organs of the human body, mainly in the liver and kidney. GST content in blood and urine is extremely low in normal healthy state. When related diseases occur, the content of GST in urine and blood will increase, so it can b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N1/28G01N33/543G01N33/573
CPCG01N21/6486G01N1/28G01N33/573G01N33/54326G01N2333/91177
Inventor 王丽娜沈薇莫檬嘉唐盛张景慧曾学民陈天瑜
Owner JIANGSU UNIV OF SCI & TECH
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