A kind of graphene signal amplification microcystin-lr electrochemical detection method

A technology of microcystin and signal amplification, applied in the direction of material electrochemical variables, measuring devices, scientific instruments, etc., can solve the problems of complex process, high cost and cycle, and achieve good electrical conductivity, multiple electrochemical active areas, sensitive The effect of the electrochemical signal

Inactive Publication Date: 2018-08-28
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent 200810242879.5 discloses the preparation and application of a microcystin-LR quantitative rapid detection sensor, but this patent is an electrochemical sensor that uses biological antibodies as the selective mechanism. Compared with nucleic acid aptamers, the cost of antibody cultivation and The cycle is higher than that of nucleic acid aptamers, and the process is more complicated, so it is not very suitable for use in actual detection

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  • A kind of graphene signal amplification microcystin-lr electrochemical detection method
  • A kind of graphene signal amplification microcystin-lr electrochemical detection method
  • A kind of graphene signal amplification microcystin-lr electrochemical detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A microcystin-LR electrochemical detection method for graphene signal amplification, wherein the specific manufacturing process of the sensor involved is as follows:

[0034](1) First, the gold electrode is pretreated, and the treatment method is as follows: the surface of the gold electrode is placed on a freshly configured piranha (H 2 SO 4 / H 2 o 2 =7:3v / v) Soak in the lotion for 10-15 minutes, take it out and wash it with ultra-pure water, and then use aluminum oxide with a particle size of 1.0μm, 0.3μm, and 0.05μm in the word "8" Sand carefully to a smooth finish. Afterwards, the aluminum oxide on the surface of the electrode was rinsed with ultrapure water, and the electrode was placed in ultrapure water, ethanol, and ultrapure water for 5 to 10 minutes of ultrasonication. After the ultrasound is completed, take out the electrode immediately, use it as the working electrode, the platinum wire electrode as the counter electrode, and the SCE as the reference ele...

Embodiment 2

[0038] Before measuring the concentration of MC-LR, the prepared MCH / Au NPs / Au electrode was taken out, and 10 μL of endonuclease (DNase I) reaction buffer (pH=7.0, containing 100 mmol / L NaCl), 5 μL graphene-aptamer complex, 3 μL MC-LR of the concentration to be tested, 2 μL DNase I solution (1000 U / mL), and place the electrode in a biochemical incubator at 30 ° C for one hour take out. Gently rinse the electrode surface with ethanol and ultrapure water to remove weakly bound graphene. The electrodes were then dried under a nitrogen atmosphere and taken out for DPV detection. The electrochemical detection was carried out in 20 mmol / L phosphate buffer solution (pH=7.0) containing 5 mmol / L ferrocenecarboxylic acid and 0.1 mol / L sodium perchlorate.

[0039] The change of current and the concentration of MC-LR at 1.0×10 -12 ~1.0×10 -10 In the range of mol / L, there is a good linear relationship, and the correlation coefficient is about 0.99. The lowest detection limit is 8.0×10...

Embodiment 3

[0041] The prepared electrochemical aptasensor was mixed with 1.0×10 -11 mol / L MC-LR respectively with a concentration of 1.0×10 -9 The mol / L interfering substances were mixed in pairs for determination. The interference experiments of three common environmental pollutants, omethoate, glyphosate, paraquat, monosultap, acetamiprid and trichlorfon, on the determination of MC-LR were investigated. Using the test conditions in Example 2 to measure the current response, the research results show that the impact of other interfering substances with a concentration 100 times that of MC-LR on the current of MC-LR is less than 10.0%. It can be seen that the prepared aptasensor has high selectivity to MC-LR.

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Abstract

The invention relates to a microcystic toxin-LR electrochemical detection method with amplified graphene signals. The method comprises the following steps: (1) dropwise adding a mixed solution with a ratio of an endonuclease reaction buffer solution: a graphene-aptamer compound: a microcystic toxin-LR solution to be detected: an endonuclease solution of (9-11):(4-6):(2-4):(1-3) on the surface of an MCH / Au NPs / Au electrode; (2) culturing the MCH / Au NPs / Au electrode for 1 to 2 hours at a temperature of 25 to 35 DEG C, slightly washing the electrode surface by ethanol and ultrapure water to remove weakly combined graphene, drying the electrode in nitrogen gas, taking the electrode out, carrying out electrochemical detection to measure the concentration of the microcystic toxin-LR solution. According to the provided method, a high sensitive electrochemical analysis technology and an aptamer with a high specific recognition performance are effectively combined to detect trace microcystic toxin-LR in the environment, the detection is highly sensitive and highly specific, moreover, the instruments are cheap and portable, the method is simple and easy to perform, and the results can be obtained quickly.

Description

technical field [0001] The invention relates to a microcystin-LR detection method, in particular to a microcystin-LR electrochemical detection method for graphene signal amplification. Background technique [0002] In recent years, environmental problems have been particularly serious, and blue-green algae outbreaks have occurred in many waters. Microcystins (MCs) are biotoxins produced by cyanobacteria in eutrophic water bodies, which are biologically active cyclic heptapeptide compounds composed of seven amino acids. There are many types of microcystins, and there are more than 90 confirmed isomers. Among them, Microcystin-LR (Microcystin-LR, MC-LR) is one of the isomers. Microcystin-LR exists most commonly in water and is also the most toxic microcystin. It is of great environmental significance to control and highly sensitively monitor the content of microcystin-LR in water. [0003] At present, the method for determining microcystin-LR mainly adopts traditional instr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26
CPCG01N27/3277G01N27/48
Inventor 刘梅川王国强赵国华
Owner TONGJI UNIV
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