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Preparation method of thrombin photoelectrochemical sensor based on cyclometalation Ir(III) coordination compound

A photoelectrochemical and thrombin technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of insufficient sensitivity, specificity, background fluorescence interference, high instrument cost, etc., and achieve improved detection sensitivity, photoelectric High conversion efficiency and improved specificity

Active Publication Date: 2019-11-12
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, the technical methods used to detect thrombin mainly include colorimetry, fluorescence, electrochemistry, electrochemiluminescence, photoelectrochemistry, etc. Among them, the optical detection method is the most common, but the interference of background fluorescence is serious, and the detection is relatively complicated. High cost; although the electrochemical method is simple, the sensitivity and specificity are not high enough, and the reproducibility is poor

Method used

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  • Preparation method of thrombin photoelectrochemical sensor based on cyclometalation Ir(III) coordination compound
  • Preparation method of thrombin photoelectrochemical sensor based on cyclometalation Ir(III) coordination compound
  • Preparation method of thrombin photoelectrochemical sensor based on cyclometalation Ir(III) coordination compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Synthesis of Ir(III) Complex Photoelectric Materials

[0036] (1) Take 7-diethylamino-3-formyl coumarin (127.9mg, 0.68mmol), 1,10-phenanthroline-5,6-dione (104.1mg, 0.5mmol), ammonium acetate ( 805.4mg, 11.3mmol), put into a round bottom flask, add 13mL of glacial acetic acid to dissolve, heat and reflux for 6h under the protection of nitrogen, stop heating after observing that the raw material gradually turns orange, and let it cool naturally. Add concentrated ammonia water to the mixture to adjust the pH to about 7 to obtain a pink precipitate, which is filtered and washed with water. The sample is vacuum-dried overnight, and the crude product is purified by column chromatography (dichloromethane:methanol=8:1) to obtain a milky white solid product as Cyclometallic ligands, vacuum-dried at 60°C, weighed 0.105g, yield: 71%. 1 H NMR (500MHz, CDCl 3 ):δ=9.16(d,J=4.2Hz,2H), 8.99(s,1H),8.78(d,J=5.8Hz,2H),7.70-7.72(m,2H),7.51(d,J= 8.0Hz, 1H), 6.68(dd, J=2.0, 7.0Hz, 1H), 6...

Embodiment 2

[0040] Preparation of Nanogold Probes of Cyclometallic Ir(III) Complexes

[0041] (1) Preparation of nano-gold: Wash the glass instruments, magnets and containers for storing nano-gold particles required in the preparation process with secondary water, soak overnight with aqua regia, and then rinse with a large amount of ultrapure water to pH The value is neutral, dry for later use. Chlorauric acid HAuCl 4 (1.0mmol / L, 100mL) was placed in a cleaned single-necked flask while stirring and heated to boiling, then quickly added trisodium citrate (38.8mmol / L, 10mL) to the above solution, and continued to react for 10min. Slowly turn into deep wine red, continue to reflux for 15 minutes, stop heating, and naturally cool to room temperature while stirring, take 500 μL of gold colloid solution for centrifugal washing (10000 rpm, 10 minutes), and evenly disperse in 1 mL of 0.02% SDS solution for later use.

[0042] (2) Take 1.5 μL tris-HCl (0.5M, containing 0.1M NaCl, pH=7.4), 6 μL 1...

Embodiment 3

[0044] Preparation of photoelectrochemical sensor based on iridium complex gold nanoprobe prepared in example 2

[0045] (1) Preparation of the working electrode: The ITO electrode was ultrasonically cleaned with acetone, ethanol, and ultrapure water in sequence, and then dried under a nitrogen atmosphere. Immerse the cleaned ITO electrode in 30% H 2 o 2 ,NH 4 OH and H 2 O (volume ratio 1:1:5) mixed solution for 15 minutes, took out and rinsed with ultrapure water, and dried under nitrogen flow. Then immersed in 5% APTMS ethanol solution overnight. Wash thoroughly with ethanol and dry at 110°C for 15 min. Immerse in the AuNPs solution and incubate for 12 h to obtain the electrode surface modified with gold nanoparticles. After cleaning with ultrapure water and drying under nitrogen flow, use 3M invisible tape to control the ITO electrode area to 0.5 cm × 0.5 cm. The pre-activated hairpin DNA HP1 solution (1.0 μM, 20 μL) with a sulfhydryl group was added dropwise to the ...

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Abstract

The invention discloses a preparation method of a thrombin photoelectrochemical sensor based on a cyclometalation Ir(III) coordination compound. According to the sensor, a cyclometalation iridium coordination compound is used as a photoelectro active material, through an Au-S key, the cyclometalation iridium coordination compound is assembled on nanometer gold, and a nanometer probe is prepared. According to a thrombin recognizing system, combination of an aptamer and thrombin is adopted to produce proximity effects to initiate DNA replacement, when the thrombin exists, the local concentrationof single-stranded DNA S1 and single-stranded DNA S2 containing thrombin aptamer fragments is increased, S1 / T hybridized in advance and the S2 are induced to generate a strand displacement reaction,and single chain T is released. The released T is hybridized with a capture barrette H1 fixed on a working electrode, the self-assembly reaction of the barrette is initiated and catalyzed, a large quantity of H1 / H2 hybridization chains are formed on the surface of the working electrode, the single-stranded DNA exposed at the terminal is used for capturing a signal nanometer gold probe, response ofphotocurrent signals is realized, and the thrombin photoelectrochemical sensor has high sensitivity and favorable selectivity.

Description

technical field [0001] The invention relates to a method for preparing a photoelectrochemical sensor for quantitatively detecting thrombin by using a cyclometal Ir(III) complex as a photoelectric material, and belongs to the technical field of photoelectrochemical quantitative analysis. Background technique [0002] Thrombin is a multifunctional serine protease that can promote blood coagulation and regulate coagulation. Its concentration and activity are important indicators to measure the coagulation mechanism. It plays an important role in physiological and pathological processes such as inflammation, wound healing, cardiovascular diseases and tumors. important role. Therefore, the establishment of a rapid and highly sensitive detection method for thrombin is of great significance for clinical disease diagnosis, disease course development, prognosis, and efficacy monitoring and evaluation. So far, the technical methods used to detect thrombin mainly include colorimetry, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6825C12Q1/682C12Q1/56
CPCC12Q1/6825C12Q1/682C12Q1/56C12Q2525/205C12Q2525/301C12Q2537/1373C12Q2563/137C12Q2565/607
Inventor 李春香蔡月圆张迎涛李忠成
Owner QINGDAO UNIV OF SCI & TECH
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