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Mercury ion detection method based on enzymatic reaction double-emission fluorescent probe

A double emission fluorescence, enzymatic reaction technology, applied in the field of fluorescence biological detection

Inactive Publication Date: 2020-04-10
JIANGSU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Currently, laccase (LACC) is used to catalyze the oxidation of OPD to generate DAP and AuNCs as dual-emission fluorescent probes by Hg 2+ Inhibition of LACC activity to achieve detection purposes has not been reported

Method used

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  • Mercury ion detection method based on enzymatic reaction double-emission fluorescent probe
  • Mercury ion detection method based on enzymatic reaction double-emission fluorescent probe
  • Mercury ion detection method based on enzymatic reaction double-emission fluorescent probe

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Experimental program
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Embodiment 1

[0044] (1) In several 90 μL LACC PBS solutions (laccase concentration is 90mU / mL), respectively add 50 μL concentrations of 0,0.8,1,1.5,2,3.5,5,6.5,8,10,15,20, 35μM Hg 2+ Standard solution, incubate at 37°C for 60 min.

[0045] (2) Add 75 μL of OPD in PBS (the OPD concentration is 80 μM) to the mixed solution prepared in step (1), and incubate the mixture solution at the same temperature for 70 minutes, laccase catalyzes the oxidation of OPD to produce luminescent DAP;

[0046] (3) Synthesis of AuNCs solution;

[0047] HAuCl 4 (32.0mL, 3.0mM) The aqueous solution is heated to boiling at 140℃, and 8mL (10M) L-proline solution is quickly dropped into the boiling HAuCl 4 After heating and stirring for 10 minutes, cool to room temperature. Centrifuge at 10000rpm for 15min, collect the supernatant and store at 4℃.

[0048] The TEM characterization results of AuNCs are as follows image 3 As shown in A, it has good monodispersity, with an average particle size of 1.3nm; the UV absorption s...

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Abstract

The invention belongs to the technical field of fluorescent biological detection, and discloses a mercury ion detection method based on an enzymatic reaction double-emission fluorescent probe. The method comprises: laccase can catalytically oxidize 2, 3-diaminophenazine (DAP) generated by o-phenylenediamine (OPD), and an internal filtration effect existing between DAP and gold nanoclusters (AuNCs), so that the dual-emission fluorescent probe is obtained; using the characteristic that mercury ions have an inhibiting effect on laccase activity, and adding the mercury ions to reduce the laccase activity, so that the amount of DAP generated by catalysis is reduced, and sensitive detection of the mercury ions in an actual sample is realized. The response range of the ratiometric fluorescence sensing method based on enzymatic reaction to mercury ions is 8.0 * 10<-7>-35 * 10<-6> mol.L<-1>, the detection limit is 2.7 * 10<-7> mol.L<-1>, and a novel enzyme-based ratiometric fluorescence sensingmethod is provided for measuring mercury ions in an actual sample.

Description

Technical field [0001] The invention belongs to the technical field of fluorescent biological detection, and specifically relates to a mercury ion detection method based on an enzymatic reaction double-emitting fluorescent probe. Background technique [0002] Mercury (Hg 2+ ) Is classified as the most harmful heavy metal due to its toxicity, the Hg accumulated in the human body 2+ Can seriously damage the central nervous system. To Hg 2+ The sensitive analysis has become to avoid Hg 2+ Poisoning is the key to protecting human health. Therefore, Hg 2+ Analytical techniques have been developed, such as atomic absorption spectroscopy, electrochemistry, inductively coupled plasma mass spectrometry, and fluorescence detection. Among these methods, the fluorescence method has the advantages of high sensitivity, fast response, and convenient use, and can be used for field detection. In recent years, the ratio fluorescence spectrophotometry has attracted special attention because of it...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/643G01N2021/6419
Inventor 由天艳李文佳刘东李玉叶申秀丽
Owner JIANGSU UNIV
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