Method for detecting various environmental estrogens and application thereof

An environmental estrogen and hydrothermal reaction technology is applied in the field of detecting a variety of environmental estrogens, can solve the problems of inability to detect a variety of environmental estrogens, low detection sensitivity, low detection specificity, etc., and achieves that the synthesis method is simple and feasible, High sensitivity, sensitive and accurate detection results

Pending Publication Date: 2021-11-02
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the inventors have found that this method can only detect two kinds of environmental estrogens, bisphenol A and estradiol, and cannot detect multiple environmental estrogens. When trying to detect multiple environmental estrogens, the detection specificity is relatively low. low, poor stability
At the same time, the detection limit of this method is still high, and the detection sensitivity is low

Method used

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  • Method for detecting various environmental estrogens and application thereof
  • Method for detecting various environmental estrogens and application thereof
  • Method for detecting various environmental estrogens and application thereof

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preparation example Construction

[0030] In one embodiment of the present disclosure, the preparation method of the upconverting nanoparticles coupled with oligonucleotide chains comprises: modifying the sulfhydryl group at the 5' end of the aptamer, dissolving the aptamer after centrifugation to obtain an aptamer solution ; Dissolving the up-converting nanoparticles modified with gold nanoparticles in a buffer solution, and incubating with the aptamer solution.

[0031] The 5' end of the aptamer is modified with a sulfhydryl group, which can form a strong coordination bond with the gold nanoparticles on the upconversion surface, so that the two can be stably combined. By in situ reduction method, the Au in chloroauric acid 3+ Reduction to Au, gold nanoparticles were successfully modified on the surface of upconversion nanomaterials. The excitation of external light causes the electron cloud on the gold surface to fluctuate and oscillate to form electromagnetic waves, which resonate with the external excitati...

Embodiment 1

[0043] Reagents and Instruments

[0044] Sodium citrate (C 6 h 5 Na 3 o 7 2H 2 O, 99.0%), purchased from Tianjin Guangcheng Chemical Reagent Co., Ltd.; Erbium chloride (Ⅲ) hexahydrate (ErCl 3 ·6H 2 O, 99.9%), ytterbium(III) chloride hexahydrate (YbCl 3 ·6H 2 O, 99.9%), yttrium(III) chloride hexahydrate (YCl 3 ·6H 2 O, 99.9%), bisphenol A (C 15 h 16 o 2 , 99.0%), hydrofluoric acid (HF, 40%), N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES, C 18 h 18 N 2 o 4 S, 99.5%) was purchased from Shanghai Aladdin Reagent Company; sodium hydroxide (NaOH, 96.0%), sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 O, 99.0%), sodium chloride (NaCl), ethanol (C 2 h 6 O, 99.7%), purchased from Sinopharm Chemical Reagent Co., Ltd.; ethylene glycol (C 2 h 6 o 2 , 99.7%), purchased from Shanghai Macklin Biochemical Technology Co., Ltd.; Tris(C 4 h 11 NO 3 , 99%) were purchased from Kamaishu (U.S.) company; unless otherwise specified, all reagents were analytically pu...

Embodiment 2

[0053] Reagent: chloroauric acid (HAuCl 4 ·3H 2 O, 99.9%) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; monolayer molybdenum disulfide was purchased from Nanjing Jicang Nano Technology Co., Ltd.; the aptamer sequence of bisphenol A 5'-SH-GGCGATGGGGTAGGGGGTGTGGAGGGGCCGGACGGAGGGG was provided by Beijing Qingke Biotechnology Co., Ltd. Technology Co., Ltd. Qingdao Branch Synthesis.

[0054] Modification of oligonucleotide aptamers on the surface of upconverting nanoparticles

[0055] Centrifuge the aptamer at 8000r / s for 45s, then open the lid and add 22uL ultrapure water to dissolve it. 6mgNaYF 4 : Yb 3+ , Er 3+ @Au solid was redissolved in 4mL buffer solution (0.01mol L -1 Tris+0.01mol L -1 NaCl, pH7.2), incubated at 37°C for 12h to obtain 1.5mg·mL -1 UCNPs@Au-apt solution.

[0056] Molybdenum disulfide quenches light emission from upconversion nanomaterials

[0057] Take the final concentration as 0.05mg·mL -1 UCNPs@Au-apt, add the final conce...

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Abstract

The invention belongs to the technical field of environmental estrogen detection, and particularly relates to a method for detecting various environmental estrogens and application thereof. The method comprises the following steps: adding oligonucleotide chain coupled up-conversion nanoparticles into a buffer solution, then adding environmental estrogens for incubation, after the reaction is completed, adding a molybdenum disulfide solution for incubation, and after the incubation is completed, carrying out detection. Aptamer-modified upconversion nanoparticles are used as fluorescence donors, molybdenum disulfide is used as an energy receptor, and the stable Van der Waals force between molybdenum disulfide and an oligonucleotide chain is utilized to realize fluorescence quenching of the upconversion nanoparticles through a fluorescence resonance energy transfer (FRET) principle. According to the present invention, by using the high affinity of the aptamer to the EEs and the specific recognition of the aptamer, detection of the total amount of the environmental estrogen with the phenolic hydroxyl group in the food can be achieved, the great application prospect is provided, and a new way is provided for the rapid detection of the food safety.

Description

technical field [0001] The disclosure belongs to the technical field of environmental estrogen detection, and in particular relates to a method for detecting multiple environmental estrogens and an application thereof. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Environmental estrogens (EEs) are a group of endocrine-disrupting chemicals that have serious harm to the human endocrine system. EEs are widely used in the manufacture of epoxy / polycarbonate resin products such as water bottles, baby bottles, plastic food containers, and pharmaceuticals, which can be released into surface water through industrial wastewater. Therefore, it is urgent to propose a high-throughp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6402G01N21/648G01N2021/6432G01N2021/6417
Inventor 路丽霞高建文邵欣宇宋冰婵丁林鹤
Owner SHANDONG NORMAL UNIV
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