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Single wavelength excited double signal-enhanced Hg<2+> fluorescent ratio method

A dual-signal, single-wavelength technology, applied in the field of Hg2+ fluorescence ratio method, can solve the problems of ratiometric fluorescence analysis method, such as difficulty in realization and signal reduction, and achieve the effect of good selectivity, high accuracy and high sensitivity

Active Publication Date: 2018-02-09
SHANGQIU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, during the detection process, the signal of DAPI is enhanced while the signal of NMM is decreased. Compared with the fluorescence quenching, people are more willing to develop fluorescence-enhanced analysis methods.
However, due to technical limitations, the ratiometric fluorescence analysis method in which the fluorescence at two wavelengths is simultaneously enhanced is difficult to achieve

Method used

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  • Single wavelength excited double signal-enhanced Hg&lt;2+&gt; fluorescent ratio method
  • Single wavelength excited double signal-enhanced Hg&lt;2+&gt; fluorescent ratio method
  • Single wavelength excited double signal-enhanced Hg&lt;2+&gt; fluorescent ratio method

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Experimental program
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Effect test

Embodiment 1

[0048] Embodiment 1 Sensing system of the present invention is to Hg 2+ Fluorescent response of

[0049] containing Mg 2+ Tris-HNO 3 Add single-stranded DNA molecule A, single-stranded DNA molecule B, NMM and DAPI to the buffer solution to obtain mixed solution I, so that the concentration of DNA molecule A in mixed solution I is 0.5 μmol / L, and the concentration of DNA molecule B is 0.5 μmol / L L, the concentration of NMM is 0.5 μmol / L, the concentration of DAPI is 0.2 μmol / L; among them, the base sequence of single-stranded DNA molecule A is 5'-TTTTTTTTTT GTGGGT-3', the base sequence of single-stranded DNA molecule B is 5'- GGGTGGGTGG TTATTTATT-3', wherein, single-stranded DNA molecule A and single-stranded DNA molecule B can form dsDNA with 9 T-T mismatched bases. Add different concentration of Hg to the mixture Ⅰ 2+ samples, 14 kinds of mixed solutions II were obtained, and the Hg in the 14 mixed solutions II 2+ The concentrations are 0, 0.05, 0.1, 0.3, 0.5, 0.7, 1.0, ...

Embodiment 2

[0053] Embodiment 2 Sensing system of the present invention is to the selectivity of other metal ions

[0054] Similar to Example 1, the sensing system of the present invention has been investigated under optimal conditions for some metal ions (Na + 、K + , Ca 2+ , Fe 3+ , Zn 2+ 、Co 2+ 、Cu 2+ 、Ag + 、Cr 3+ , Mn 2+ 、Cd 2+ 、Ni 2+ , Pb 2+ )the response to. like Figure 10 , adding the same concentration of Hg to the system 2+ Or interfering metal ions (2μmol / L), with Hg 2+ In contrast, the fluorescent signal of interfering ions is very low and the interference is very small. To further understand the selectivity of the method, competition experiments (such as Figure 11 shown), add the same concentration of Hg to the system together 2+ and interfering ions (2μmol / L), with only Hg 2+ Compared with the system, the detection system containing interfering metal ions is more effective for Hg 2+ There is no significant change in the response, indicating that the sensin...

Embodiment 3

[0055] Hg in embodiment 3 fetal bovine serum 2+ Content detection

[0056] Determination of Hg in Fetal Bovine Serum by Adding Standard Recovery Method 2+ First, pre-treat the fetal bovine serum. The pretreatment method is: mix and shake the fetal bovine serum and ethanol at a ratio of 1:1, put it in the refrigerator (0-4°C) overnight, take it out the next day, 15,000 Centrifuge at r / min for 10 min, take the supernatant in an ultrafiltration centrifuge tube, molecular cut-off (Amicon Ultra-0.5mL, Millipore) 3kDa, centrifuge at 13,000r / min 4°C for 20min, take the filtrate, and freeze it in the refrigerator until use. A certain amount of Hg2+ was added to the pretreated fetal bovine serum to obtain 4 samples, and the Hg in the 4 samples 2+ The concentrations were 0, 0.5, 1.0 and 2.0 μmol / L, respectively, and then the optimal sensing system described in Example 1 was used to detect the Hg in fetal bovine serum samples. 2+ The test was carried out, and the results are shown in...

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Abstract

The invention relates to a single wavelength excited double signal-enhanced Hg<2+> fluorescent ratio method and belongs to the technical field of chemical / biological sensing and analysis detection. The method comprises adding a single-stranded DNA molecule A, a single-stranded DNA molecule B, NMM and DAPI into a buffer solution containing Mg<2+> to obtain a mixed solution I, wherein the molecule Acomprises a part I and a part II, the molecule B comprises a part III and a part IV, the part I in the molecule A and the part III in the molecule B are partially complementary sequences with T-T basic group mismatching and the part II in the molecule A and the part IV in the molecule B form a four-stranded G-rich sequence, adding Hg<2+> into the mixed solution I, and carrying out vibration and measuring fluorescence intensity in a measuring range of 400-700nm and excitation wavelength of 380nm. The method is a double signal-enhanced novel Hg<2+> ratio-type fluorescence analysis method, is used for detection of Hg<2+> in a sample and has high selectivity, accuracy and reliability.

Description

technical field [0001] The invention belongs to the technical field of chemical / biological sensing and analysis and detection, and in particular relates to a single-wavelength excited double-signal enhanced Hg 2+ Fluorescence ratio method. Background technique [0002] Hg 2+ is a heavy metal ion, Hg 2+ Pollution not only affects the environment, but also seriously threatens human health. Therefore, Hg 2+ The quantitative analysis is of great significance. Fluorescence sensor has the characteristics of high sensitivity, good selectivity, simple method, fast response, no damage, real-time detection, etc. Compared with chemiluminescence, colorimetry and electrochemical analysis methods, fluorescence analysis method can 2+ It has obvious advantages in quantitative analysis. [0003] Over the past few decades, many Hg 2+ Fluorescence sensing system. Most of these sensors are excited by a single wavelength, and the maximum emission wavelength does not move. The change of fl...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 王永祥耿凤华姜香宇瞿鹏徐茂田
Owner SHANGQIU NORMAL UNIVERSITY
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