Method of detecting endocrine disrupting chemicals by adoption of nucleic acid aptamer visible-light electrode
A technology of endocrine disruptors and nucleic acid aptamers, applied in the direction of material electrochemical variables, etc., can solve the problems of loss of selective recognition ability, easy denaturation, etc., achieve good molecular recognition ability, maintain stability, and avoid inactivation effects
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Embodiment 1
[0039] Preparation of TiO 2 -Fe 2 o 3 The method of NTs electrode is as follows: (1) Ti plate (purity 99.9%) and Fe particles (purity 99.99%) are mixed in a certain ratio, and under the protection of argon atmosphere, they are melted in a high-temperature arc melting furnace to obtain Ti-6Fe titanium ferroalloy. It was cut into ferro-titanium alloy plates with a size of 10×10×1 mm by wire cutting machine. Wherein the mass percentage of iron in the ferro-titanium alloy is 6%. (2) Polish the titanium-iron alloy plate in step (1) with sandpaper, and ultrasonically clean it in distilled water and acetone for 15 min each. At room temperature, the pretreated titanium-iron alloy plate was used as the anode, and a two-electrode system was used with a distance of 1 cm between the electrodes to contain 0.25 mol / L NH 4 The ethylene glycol solution of F was used as the electrolyte solution, under magnetic stirring, anodized at a constant potential of +30V for 5h, cleaned with twice d...
Embodiment 2
[0042] A series of bisphenol A standard solutions with different concentrations were prepared by using 0.1M PBS buffer solution with a pH of 7. A 0.1M PBS buffer solution with a pH of 7 was used as an electrolyte, and a nucleic acid aptamer modified with a nucleic acid aptamer was used as a working electrode. First, assemble the solution containing bisphenol A or interfering substances (2,4-D, acetamiprid, chlorpyrifos or omethoate) with the nucleic acid aptamer visible photoelectrode at 37°C for 30 min to facilitate the detection Molecules and aptamers on the surface of the electrode were selectively identified and enriched, and washed three times with PBS buffer; then the electrodes before and after the action were connected with a three-electrode system, left to stand, and the visible light source was turned on; The photocurrent was measured under the condition of voltage (0.6V(Vs.SCE)), and recorded by computer. Bisphenol A is quantified by the size of the photocurrent, a...
Embodiment 3
[0044] First, wash the nucleic acid aptamer visible light electrode carefully and use it as a working electrode to contain a certain concentration of bisphenol A or an interfering substance 100 times higher than that of bisphenol A, such as 2,4-D, acetamiprid, chlorpyrifos or oxychlor The solution of the fruit is the analyte, and its pH value is adjusted with 0.1M PBS buffer solution of pH 7.4. Using the test conditions set in Example 2, measure the visible light current of the solution, and calculate the concentration of bisphenol A in the actual solution according to the working curve drawn in Example 2. The results showed that 100 times the bisphenol A concentration of other small molecule interferers had little effect on the photocurrent of bisphenol A, such as image 3 shown. It reflects the good selective analysis ability of the nucleic acid aptamer visible light electrode.
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