A three-fluorescence-emitting molecularly imprinted sensor and its preparation and application

A technology of molecular imprinting and fluorescence emission, which is applied in the field of analytical chemistry and rapid detection, can solve the problems of inability to accurately visualize the detection target, complex optimization process of emission peak intensity ratio, narrow range of fluorescence color change, etc., to achieve rich fluorescence color change , reduce experiment cost and shorten experiment period

Active Publication Date: 2019-06-11
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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Problems solved by technology

At present, there are two main deficiencies in the ratiometric fluorescent molecular imprinting sensor: (1) The traditional ratiometric fluorescent molecular imprinting sensor only has two emission peaks, and the fluorescence color range provided when detecting the target is narrow, and cannot accurately visualize the detection target. (2) The preparation of ratiometric fluorescent molecularly imprinted sensors often needs to optimize the fluorescence intensities of different emission peaks, and select the appropriate emission peak intensity ratio to achieve the best visual detection effect, but the emission peaks of traditional ratiometric fluorescent molecularly imprinted sensors The strength ratio optimization process is complex and tedious

Method used

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  • A three-fluorescence-emitting molecularly imprinted sensor and its preparation and application
  • A three-fluorescence-emitting molecularly imprinted sensor and its preparation and application
  • A three-fluorescence-emitting molecularly imprinted sensor and its preparation and application

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Effect test

Embodiment 1

[0036] Preparation of three fluorescence-emitting molecularly imprinted sensors:

[0037] (1) Preparation of bovine hemoglobin-imprinted microspheres, taking blue fluorescent bovine hemoglobin-imprinted microspheres as an example, see figure 1 :

[0038] To 20 mL of phosphate buffer (0.01 M, pH 7.0), add 10 mg of silica microspheres with a particle size of approximately 79 nm, 10 mg of bovine hemoglobin, and 35 μL of 3-aminopropyltriethoxysilane (APTES), and stir for 1 Hours later, 20 mg of blue fluorescent 7-hydroxycoumarin was added, and after mixing, 56 μL of ethyl orthosilicate (TEOS) and 56 μL of ammonia (NH) were added. 3 ·H 2 O), the sol-gel polymerization was carried out in the dark for 10-12 h. After the reaction was completed, the product was precipitated by centrifugation (8000 rpm, 5 minutes), the supernatant was discarded, and the bovine hemoglobin in the precipitate was eluted with 0.5% Triton X-100 to obtain a blue fluorescent bovine hemoglobin blot with a co...

Embodiment 2

[0045] The red fluorescent bovine hemoglobin-imprinted microspheres (r-MIPs) dispersion liquid, blue fluorescent bovine hemoglobin-imprinted microspheres (b-MIPs) dispersion liquid and green fluorescent bovine hemoglobin-imprinted microspheres (g-MIPs) dispersion liquid obtained above were determined according to certain conditions. The sensor was obtained by mixing the ratio in phosphate buffer (0.01 M, pH 7.0); the total volume of phosphate buffer (0.01 M, pH 7.0) was controlled at 1 mL, and the amount of red bovine hemoglobin blotting microspheres (r-MIPs) was fixed. is 5% (volume fraction), the dosage of blue bovine hemoglobin-imprinted microspheres (b-MIPs) is fixed at 6% (volume fraction), and the dosage of green bovine hemoglobin-imprinted microspheres (g-MIPs) is 0%, 3%, 5%, 7%, 9%, 11%, 13% and 15%; then different amounts of bovine hemoglobin were added to the above sensor to achieve final concentrations of 0, 0.025, 0.05, 0.075, 0.1, 0.25, 0.5, 0.75 , 1, 2 and 3 μM; ...

Embodiment 3

[0048] Different sensors are prepared according to the sensor preparation method described above. Specifically, the different microspheres obtained above are mixed in a buffer solution according to a certain ratio, that is, different sensors with different selections and different ratios are obtained; wherein the red-green-blue three fluorescence emission molecular imprinting r-MIPs in the sensor: g-MIPs: b-MIPs=5%:11%:6% (volume percentage), red-green-blue three fluorescence emission r-NIPs: g-NIPs: b-NIPs in the non-imprinted sensor =5%:11%:6% (volume percentage), r-MIPs: g-MIPs in the red-green dual fluorescence emission molecular imprinting sensor =5%:11% (volume percentage), red-blue dual fluorescence emission molecular imprinting r-MIPs in the sensor: b-MIPs=5%:6% (volume percentage), g-MIPs in the green-blue dual fluorescence emission molecular imprint sensor: b-MIPs=11%:6% (volume percentage); and, The total volume of phosphate buffer (0.01 M, pH 7.0) in each of the ab...

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Abstract

The invention belongs to the field of analytical chemistry and quick detection, and particularly relates to a red-green-blue tri-fluorescent emission molecularly imprinted sensor based on a post-imprinting mixing method and a preparation method and application thereof in accurately and visually detecting hemoglobin. The red-green-blue tri-fluorescent emission molecularly imprinted sensor is obtained by mixing red fluorescent bovine hemoglobin imprinted microspheres, green fluorescent bovine hemoglobin imprinted microspheres and blue fluorescent bovine hemoglobin imprinted microspheres throughthe post-imprinting mixing method. The sensor prepared by the method can detect bovine hemoglobin in a high-sensitivity, high-selectivity and self-correcting manner, and the sensor is wider and richerthan conventional dual-fluorescent emission molecularly imprinted sensors in fluorescent color changing range and can visually detect a target more accurately.

Description

technical field [0001] The invention belongs to the fields of analytical chemistry and rapid detection, in particular to a red-green-blue three fluorescence emission molecular imprint sensor based on a post-imprint hybrid method and its preparation and application in accurate visual detection of bovine hemoglobin. Background technique [0002] Nowadays, rapid visual inspection plays an important role in environmental inspection, food safety, clinical diagnosis and other fields. The rapid visual detection of a large number of samples to be tested, combined with the accurate detection of suspicious samples by large-scale instruments (such as high performance liquid chromatography), often constitute a complete detection process, which not only saves the money and time of detection, but also ensures the reliability of the detection results. Therefore, rapid visual detection methods with high sensitivity and high selectivity are often required. [0003] The ratiometric fluoresce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 李金花杨倩陈令新于祖海
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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