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
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[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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