Method and sensor for rapid visualization analysis of total amount of various phthalate pollutants

A technology of phthalic acid and pollutants, which is applied in the direction of analysis by chemical reaction of materials, material analysis by observing the impact on chemical indicators, color/spectral property measurement, etc. Detection, equipment are expensive, complex processing and other problems, to achieve the effect of more intuitive and visual detection results, easy operation and high sensitivity

Inactive Publication Date: 2019-09-20
TONGJI UNIV
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AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a broad-spectrum nucleic acid aptamer-based PAEs in order to overcome the problems of complex pretreatment process, expensive instrument, time-consuming analysis, and inability to realize one-step total detection in the existing PAEs detection technology. Total colorimetric sensing method

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  • Method and sensor for rapid visualization analysis of total amount of various phthalate pollutants

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Experimental program
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Embodiment 1

[0022] A colorimetric sensor based on a broad-spectrum nucleic acid aptamer to detect the total amount of various PAEs, using a broad-spectrum nucleic acid aptamer that can recognize a class of PAEs as a recognition element, and the absorbance is generated according to the change of the aggregation state of gold nanoparticles Ratio A 646nm / A 520nm The change of the PAEs concentration is quickly visualized and analyzed, and the sensor is constructed through the following steps:

[0023] (1) Use Mfold software to simulate the secondary structure of the long-chain nucleic acid aptamer sequence of PAEs, and use the Loop ring as the main body to cut and design its molecules by intercepting different branches on both sides to obtain nucleic acid aptamers of different lengths fragment. AutoDock molecular docking simulation experiments were performed on different aptamer fragments with DEHP, DBP, and BBP, and the truncated fragments that retained the binding sites of PAEs, had spec...

Embodiment 2

[0028] The experimental parameters affecting the sensitivity of the nucleic acid aptamer colorimetric sensor were optimized, including: the concentration of sodium chloride, the concentration of the nucleic acid aptamer and the incubation time of the system. The specific process is: respectively adding nucleic acid aptamer solution and sodium chloride solution with concentration ranges of 0.06-0.47 μmol / L and 17.24-103.45 mmol / L to the gold nano-sol system. The absorbance of different colorimetric systems was measured by a UV-visible spectrophotometer, according to A 646nm / A 520nm The optimal concentration of nucleic acid aptamer was determined to be 0.28μmol / L, and the optimal concentration of sodium chloride was 43.1mM. A after 4 minutes of reaction 646nm / A 520nm Tend to a steady state, the best time for system detection was determined to be 4min.

Embodiment 3

[0030] Weigh 1 g of DEHP, DBP and BBP in turn, and dissolve them in a 10 ml volumetric flask with ethanol as a solvent to obtain three PAEs stock solutions with a concentration of 0.1 g / mL. Then mix the three equal concentrations of PAEs solutions in equal volumes to obtain a standard mixture stock solution with a ratio of DEHP, DBP, and BBP of 1:1:1 and a total concentration of PAEs of 0.1 g / mL, which can then be diluted to obtain PAEs standards of different concentrations. Mix the pollutant solution. By mixing DEHP, DBP and BBP in different volumes, different proportions of PAEs mixed pollutant solutions can be obtained.

[0031] The concentration range of the PAEs mixed pollutants added to the nucleic acid aptamer colorimetric sensor constructed in Example 1 is 0.003 μg / L to 10 μg / L. After sufficient incubation, a UV-visible spectrophotometer is used to detect the change in the absorbance of the system and calculate A of gold nanoparticles 646nm / A 520nm value. figure 1...

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Abstract

The invention belongs to the field of environmental engineering and water pollution detection and treatment, and relates to a nucleic acid aptamer colorimetric sensor and a preparation method and application thereof: a method for rapid visualization analysis of total amount of various phthalate pollutants based on the nucleic acid aptamer colorimetric sensor. A broad-spectrum nucleic acid aptamer with a specific base sequence and specific recognition ability for a class of structurally similar PAEs is combined with a highly sensitive colorimetric analysis technology to construct the nucleic acid aptamer colorimetric sensor for detecting the total amount of PAEs. According to the invention, the broad-spectrum nucleic acid aptamer capable of binding various PAEs with high affinity is used as a recognition component to successfully realizes high sensitivity, rapid and visualization detection analysis of the total amount of the substance; the linear detection range of PAEs is 0.003 mu g / L to 10 mu g / L, and the detection limit is as low as 1ng / L; and compared with the prior art, the sensor has the advantages of simple operation, fast response, easy miniaturization and detection result visualization.

Description

technical field [0001] The invention belongs to the fields of environmental engineering and water pollution detection and treatment, and in particular relates to the construction of a colorimetric sensor based on a broad-spectrum nucleic acid aptamer and its highly sensitive, simple, fast and visual detection and analysis of the total amount of various PAEs. Background technique [0002] Phthalates (PAEs), as the most commonly used plasticizers in the plastics industry, have refractory and bioaccumulation effects and interact with plastic products through non-covalent bonding. Over time, it is easily transferred to the environment, and the widespread application of globalization makes it a large number of residues in actual water bodies. In recent years, the Yangtze River, Yellow River, Huangpu River, Songhua River, Chaohu Lake and other major water basins in my country have been polluted by PAEs to varying degrees, which has seriously affected residents' water use and daily...

Claims

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Application Information

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IPC IPC(8): G01N21/33G01N21/78
CPCG01N21/33G01N21/78
Inventor 赵国华陈雨晴
Owner TONGJI UNIV
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