CN115248237B - Copper-based MOF material modified microelectrode and application thereof in superoxide anion detection - Google Patents
Copper-based MOF material modified microelectrode and application thereof in superoxide anion detection Download PDFInfo
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- 239000013084 copper-based metal-organic framework Substances 0.000 title claims abstract description 121
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
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Abstract
The application provides a microelectrode modified by a copper-based MOF material, which is prepared by the following steps: penetrating carbon fibers into the glass capillary holes, and connecting the carbon fibers and copper wires through silver conductive adhesive; sealing the tips of the microelectrodes with nail polish; removing pollutants on the surface of the tip of the microelectrode; and placing the microelectrode in a suspension prepared by the copper-based MOF material to prepare the carbon fiber microelectrode wrapped by the copper-based MOF material. The MOF material prepared by taking metallic copper as a metal center is selected, and the MOF material imitates the active center of SOD and has copper element which is easy to change in price, so that the MOF material has good capability of catalyzing O 2 ·‑ to be disproportionated and decomposed into O 2 and H 2O2.
Description
Technical Field
The application relates to the technical field of new materials, in particular to a copper-based MOF material and application thereof in superoxide anion removal, a microelectrode modified by the copper-based MOF material and application thereof in superoxide anion detection.
Background
O 2 ·- is a Reactive Oxygen Species (ROS), a product of a common chemical biological reaction, such as Xanthine Oxidase (XOD), which catalyzes the redox reaction of xanthine with O 2 and is critical to the physiological function of an organism. O 2 ·- is involved in regulating a variety of normal cellular and in vivo function-related signaling mechanisms. To maintain redox balance in the organism, it is important to measure the concentration of O 2 ·- and to clear excess O 2 ·-. Therefore, it is necessary to design electrocatalytic and catalytic materials for the detection and removal of O 2 ·- in biological systems. The research is widely carried out by constructing a sensor by using biological enzyme (superoxide dismutase SOD), but the enzyme is expensive, the enzyme can lose activity under severe conditions or long-time storage, and the enzyme activity center is easy to be covered by protein to reduce the electron transfer rate, so the enzyme-like material with low cost, simple preparation and good stability is used for researching O 2 ·- in cells and living bodies by replacing the SOD.
MOFs are highly ordered crystalline materials formed by coordination of metal ions with organic ligands that have unique chemical and physical properties (including ultra-high porosity, large specific surface area, tunable structure, high thermal stability and chemical stability) and thus are of great interest in electrochemical sensors. In organisms, SOD active center has copper element which is easy to change valence, so the SOD active center has good capability of catalyzing O 2 ·- to be disproportionated and decomposed into O 2 and H 2O2, and therefore, we design a MOF material taking copper as metal ion to realize the detection of O 2 ·-.
Disclosure of Invention
The present application aims to solve at least one of the technical problems in the related art to some extent.
Therefore, the application aims to provide a copper-based MOF material and application thereof in superoxide anion removal, a microelectrode modified by the copper-based MOF material and application thereof in superoxide anion detection, wherein the copper-based MOF material takes copper as a metal center, a conductive ligand thiophene derivative 2, 5-dicarboxylic acid-3, 4-ethylenedioxythiophene (H 2 L) is used as an organic ligand to synthesize a copper-based metal organic framework nanomaterial (Cu-MOF), the Cu-MOF material is modified on a Carbon Fiber Electrode (CFE) and can be used as an electrocatalyst, electrochemical reduction detection of O 2 ·- can be realized at-0.05V, and on the other hand, the Cu-MOF has chemical catalytic property similar to SOD, namely disproportionation O 2 ·-. The Cu-MOF catalyst prepared by the application has the advantages of simple preparation method, easy operation, excellent electrocatalytic and SOD mimic enzyme performances, and has the function of removing O 2 ·- in living cells in cell and living body layers.
In order to achieve the above purpose, the application provides a microelectrode modified by copper-based MOF material, which is prepared by the following steps:
Penetrating carbon fibers into the glass capillary holes, and connecting the carbon fibers and copper wires through silver conductive adhesive;
Sealing the tips of the microelectrodes with nail polish;
removing pollutants on the surface of the tip of the microelectrode;
And placing the microelectrode in a suspension prepared by the copper-based MOF material to prepare the carbon fiber microelectrode wrapped by the copper-based MOF material.
Further, contaminants on the tip surface of the microelectrode are removed by applying a potentiostatic potential to the microelectrode in sodium hydroxide.
Further, the concentration of the copper-based MOF material suspension is 50-800 mg/mL.
Further, the copper-based MOF material suspension is prepared by uniformly mixing copper-based MOF material with 2% Nafion solution in an ultrasonic manner.
Further, the specific preparation method of the copper-based MOF material comprises the following steps:
Preparing a solution of 2, 5-dicarboxylic acid-3, 4-ethylenedioxythiophene and N, N-dimethylacetamide according to a molar ratio of 1.5:1-3.5:1, and simultaneously dissolving copper chloride dihydrate in distilled water to obtain a solution with mass concentration of 0.01-0.1 mol.L -1;
mixing the two solutions obtained after dissolution, adding the mixed solutions into a polytetrafluoroethylene substrate high-pressure reaction kettle, heating at a high temperature to obtain Cu-MOF crystals with good crystallization, and grinding the Cu-MOF crystals to obtain the copper-based MOF material.
Further, the high-temperature heating temperature in the polytetrafluoroethylene substrate high-pressure reaction kettle is 80-150 ℃, and the heating time is 24-60 h.
The application of a microelectrode modified by a copper-based MOF material as an electrochemical sensor in the detection of the concentration of superoxide anions in cells and living bodies.
Further, the application of the copper-based MOF material modified carbon fiber microelectrode in the detection of the concentration of superoxide anions in cells and living bodies comprises the following specific detection processes: adopting a three-electrode system, wherein a Cu-MOF modified electrode is a working electrode, ag/AgCl is a reference electrode, a platinum wire is a counter electrode, a phosphate buffer solution with the pH of 0.1M containing O 2 ·- and the pH value of 7.5 is used as electrolyte, and timing current scanning is performed under the working potential of-0.05V; in a specific O 2 ·- concentration range of 20-100uM (micromoles), the response current Ip has a good linear relation with the concentration of O 2 ·-, and the concentration of O 2 ·- is obtained through the value of the response current Ip.
Compared with the prior art, the invention has the beneficial effects that:
(1) The MOF material prepared by taking metallic copper as a metal center is selected, and the MOF material imitates the active center of SOD and has copper element which is easy to change in price, so that the MOF material has good capability of catalyzing O 2 ·- to be disproportionated and decomposed into O 2 and H 2O2.
(2) The preparation method of the synthesized Cu-MOF material is simple, the yield and purity are high, and the accurate structure of the MOF can be analyzed through single crystal diffraction.
(3) The Cu-MOF material prepared by the invention can be used for detecting and removing O 2 ·- in a biological system.
Drawings
The foregoing and/or additional aspects and advantages of the application will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings, in which:
FIG. 1 (A) is a TEM image of a copper-based MOF material synthesized in example 1 of the present invention;
FIG. 1 (B) is an SEM image of a copper-based MOF material synthesized in example 1 of the present invention;
FIG. 1 (C) is an energy dispersive X-ray spectrogram (EDX) of the copper-based MOF material synthesized in example 1 of the present invention;
FIG. 1 (D) is an XRD pattern of the copper-based MOF material synthesized in example 1 of the present invention and an XRD pattern obtained by fitting with Diamond software;
FIG. 2 (A) is an SEM image of a bare CFE of example 1 of the present invention;
FIG. 2 (B) is an SEM image of the CFE modified copper-based MOF material of example 1 of the present invention;
FIG. 3 (A) is a cyclic voltammogram of Cu-MOF/CFE in 0.1M PBS for 20 cycles at a scan rate of 100mV/s in example 1 of the present invention;
FIG. 3 (B) is a cyclic voltammogram of example 1 of the present invention before and after O 2 ·- is introduced into PBS;
FIG. 3 (C) is a cyclic voltammogram of bare CFE versus O 2 ·- in example 1 of the present invention;
FIG. 4 is a graph of the amperometric response of Cu-MOF/CFE to 20 to 100 μ M O 2 ·- at a potential of-0.05V (vs. Ag/AgCl) for each addition of O 2 ·- at a concentration of 20 μM in example 1 of the present invention;
FIG. 5 is a graph showing the selectivity of Cu-MOF/CFE to O 2 ·- and the interference immunity to other substances at a potential of-0.05V (vs. Ag/AgCl) for example 1 of the present invention, wherein 20. Mu.M AA, 5. Mu.M DA, 5. Mu.M 5-HT, 5. Mu. M H 2O2, 20. Mu.M UA and 10. Mu. M O 2 ·- were added at various times during the experiment;
FIG. 6 (A) is a bright field micrograph of Cu-MOF/CFE and a microinjection tube of example 1 of the present invention taken at-0.05V (vs. Ag/AgCl) of Hela cell release O 2 ·- by amperometric recording, wherein the inset is a Cu-MOF/CFE and microinjection tube located on a Hela cell cluster;
FIG. 6 (B) is a graph of Cu-MOF/CFE of example 1 of the present invention recording Fe 2+ stimulating O 2 ·- release from the rat cerebral cortex at a potential of-0.05V (vs. Ag/AgCl) at a microinjection rate of 2.5. Mu.L/min at 1300s, 1500s and 1700s, respectively, for 20s each;
FIG. 6 (C) is the current response of Cu-MOF/CFE to release O 2 ·- under stimulation with zymosan A at-0.05V (vs. Ag/AgCl) in example 1 of the present invention;
FIG. 6 (D) is a current response of Cu-MOF/CFE test Fe 2+ to stimulate O 2 ·- release from rat cerebral cortex at-0.05V (vs. Ag/AgCl) in example 1 of the present invention;
FIG. 7 is an experiment for observing the removal of intracellular ROS by Cu-MOF under a fluorescence microscope and corresponding bright field microscope in example 1 of the present invention.
Detailed Description
Embodiments of the present invention are described in detail below. The following examples are illustrative only and are not to be construed as limiting the invention.
The scheme of the present invention will be explained below with reference to examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the present invention and should not be construed as limiting the scope of the invention. The examples are not to be construed as limiting the specific techniques or conditions described in the literature in this field or as per the specifications of the product. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1:
In this example, a copper-based MOF material was prepared as follows, and the prepared copper-based MOF material was used to modify microelectrodes, and the resulting microelectrodes were used for detection of superoxide anions in cells and living bodies, while the copper-based MOF material was used for removal of superoxide anions in cells, the detailed preparation method and application were as follows:
the preparation method of the copper-based MOF material (Cu-MOF) comprises the following specific preparation processes:
34.1mg of H 2 L (conductive ligand thiophene derivative 2, 5-dicarboxylic acid-3, 4-ethylenedioxythiophene) was dissolved in 5mL of N, N-Dimethylacetamide (DMA), while 46.04mg of CuCl 2·2H2 O was dissolved in 10mL of H 2 O; then, the two solutions were mixed and added into a polytetrafluoroethylene substrate autoclave with a specification of 20mL, heated at 90 ℃ for 48 hours, and Cu-MOF with good crystallization was obtained, which was then ground into Cu-MOF powder in an agate mortar.
The Cu-MOF material prepared by the method is subjected to structural characterization, and the specific characterization result is as follows:
Transmission Electron Microscopy (TEM) and SEM images of Cu-MOFs are shown in fig. 1 (a) and 1 (B), from which it can be seen that the morphology of the synthesized Cu-MOFs is rectangular plate-like, with dimensions between 60 and 200 nm.
FIG. 1 (C) shows the corresponding energy dispersive X-ray spectra (EDX) of the copper-based MOF material, indicating that the prepared Cu-MOF consists of C, O, S and Cu elements, without any other elements.
FIG. 1 (D) shows the XRD pattern of the copper-based MOF material and the XRD pattern obtained by Diamond software fitting, and the synthesized Cu-MOF material has a series of sharp diffraction peaks on the XRD pattern, which are identical to the simulation model (Database Identifier IHONOK, deposition Number 1040717,Cambridge Crystallographic Data Centre) of Cu-MOF, so that we successfully synthesize Cu-MOF with the same crystal structure, the average grain size is about 76.2nm calculated by the Shelle formula, the morphology data size of Cu-MOF is combined, so that perfect Cu-MOF crystals are formed during the hydrothermal reaction, and each rectangular nano-sheet can be considered as single crystal nano-material.
(II) the preparation process of the copper-based MOF material modified microelectrode is as follows:
drawing a glass capillary by using a P-97 type drawing instrument, wherein the set parameters are as follows: HEAT 560,PULL 55,VEL 95,TIME200,P 200;
Cutting the tip of a capillary tube by using a surgical knife, penetrating carbon fiber with the diameter of 30 mu m into a hole, connecting the carbon fiber with copper wires by silver conductive adhesive, and sealing the tip of an electrode by using nail polish to avoid siphoning when immersed in a solution and influencing electrochemical measurement;
immersing a microelectrode (CFE) in ethanol to remove nail polish on the surface of the carbon fiber, and applying a constant potential of 1.5V (vs. Ag/Cl) in 0.1M NaOH for 80 seconds to remove pollutants on the surface of the CFE tip after the surface of the microelectrode is dried;
uniformly mixing Cu-MOF powder (300 mg/mL) with 2% Nafion by ultrasonic, and soaking CFE in the solution to obtain the Cu-MOF modified carbon fiber microelectrode (Cu-MOF/CFE).
The morphology and electrochemical performance characterization results of the copper-based MOF material modified carbon fiber microelectrode are as follows:
In fig. 2 (a) is an SEM image of a bare CFE, while in fig. 2 (B) is an SEM image of a Cu-MOF/CFE, it is obvious that the rectangular sheet-like Cu-MOF with a dense distribution on the carbon fiber surface in fig. 2 (B) is compared with fig. 2 (a), which shows that with the aid of Nafion, the Cu-MOF can be successfully modified on the CFE, thus laying a foundation for the subsequent electrochemical experiment.
FIG. 3 (A) is a cyclic voltammogram of Cu-MOF/CFE in 0.1M PBS for 20 consecutive turns, from which it can be seen that the reduction peak at-0.2V is due to Cu II reduction to Cu 0, and the reproducible voltammogram also shows that Cu-MOF is relatively stable on CFE for further in vivo cell analysis applications.
FIG. 3 (B) is a cyclic voltammogram before and after O 2 ·- was introduced into PBS, where the PBS curve in FIG. 3 (B) is that of Cu-MOF/CFE in 0.1M PBS, the PBS+XOD curve is that of PBS containing 1 μL XOD, and PBS+XOD+400 μ M Xanthine is that of PBS containing 1 μL XOD and 400 μM xanthine, as can be seen, the Cu-MOF/CFE shows a distinct reduction peak around-0.05V when O 2 ·- was introduced into PBS;
Also as a control, we tested the cyclic voltammetric response of bare CFE to O 2 ·-, see in particular fig. 3 (C) (where PBS is the cyclic voltammogram of bare CFE, pbs+xod+400 μ M Xanthine is the cyclic voltammogram of bare CFE containing 1 μl XOD and 400 μΜ xanthine), no significant reduction peak was found at-0.05V, indicating that O 2 ·- is responsible for the reduction peak of Cu-MOF/CFE, indicating that Cu-MOF/CFE can be used in vivo and cell analysis applications to detect O 2 ·-.
We studied the electrochemical sensing performance of Cu-MOF/CFE on O 2 ·- using chronoamperometry, which shows a very good linear response (as in fig. 4) over a concentration range of 20 to 100 μm, with a correlation coefficient (R 2) of 0.962 and a minimum detection limit of 5.18 μm (signal to noise ratio=3); as shown in FIG. 5, the selectivity of Cu-MOF/CFE was characterized, and the response of the sensor to actual physiological concentrations of AA, DA, UA, 5-HT, and H 2O2 was well below 10 μ M O 2 ·- when a constant voltage of-0.05V was applied, indicating that Cu-MOF/CFE is expected to be useful in living cell and in vivo analysis applications.
(III) the Cu-MOF/CFE prepared above is used for detecting O 2 ·- in cells and living bodies, and the specific detection process is as follows:
To monitor O 2 ·- release in real time, hela cells were cultured in an incubator for more than two days to form clusters. Before electrochemical measurement, the cell culture solution was replaced with PBS without any protein, preventing contamination of the electrodes. It is reported that this living cell cluster releases large amounts of O 2 ·- under zymosan A stimulation. As shown in fig. 6 (a), 6 (B), 6 (C) and 6 (D), a phosphate buffer solution of 0.1M ph=7.5 containing O 2 ·- was used as an electrolyte, and a chronoamperometric scan was performed at an operating potential of-0.05V; in a specific O 2 ·- concentration range of 20-100 mu M (micromoles), the response current Ip and the concentration of O 2 ·- are in good linear relation, and the concentration of O 2 ·- is obtained through corresponding current measurement; microelectrodes and stimulation microinjection tubes were placed next to each other on a cluster of living cells, and a sharp current drop signal was observed when the cells were stimulated and tested for O 2 ·-, 585s release, which signal was indicative of a large release of O 2 ·- upon zymosan A stimulation. The local maximum concentration of O 2 ·- detected on the electrode surface was about 20. Mu.M, which may be due to the vigorous metabolism of cancer cells and the massive biochemical reactions inside the cells. As O 2 ·- diffuses into the solution, the current gradually returns to the basal level prior to stimulation. Cell experiment results show that the Cu-MOF/CFE has the capability of detecting O 2 ·- at the cellular level in real time.
To test the performance of the prepared Cu-MOF/CFE sensor in vivo detection of O 2 ·-, we implanted it into the cerebral cortex of SD rats for amperometric detection and stimulated with Fe 2+ to produce O 2 ·-, then tested for O 2 ·- concentration therein, under the same specific detection conditions and conditions as described above for the Cu-MOF/CFE detection of O 2 ·- in cells, and measured by the corresponding current to obtain O 2 ·- concentration, fe 2+ was injected with a microinjection pump at a rate of 2.5 μl/min in 1300s, 1500s and 1700s, respectively, indicating that O 2 ·- was produced in the brain of the rats; the current value then returns to baseline because O 2 ·- may diffuse throughout the brain region or be decomposed by AA, GSH, etc. other protective reducing agents. These results indicate that Cu-MOF/CFE can also be successfully applied in vivo experiments. The excellent performances such as diversity and adjustability of the Cu-MOF can expand the application of the MOF in biological analysis, and a new road is opened up for the design of electrode interfaces.
(IV) the Cu-MOF material prepared by the method is used for eliminating O 2 ·- in cells, and the specific process is as follows:
the Cu-MOF dispersion was incubated with Hela cells in medium for 24 hours, and experiments were performed under a fluorescence microscope to observe green fluorescence generated by the reaction of ROS and DCFH-DA. In addition, incubation of cells with β -Lap for 6 hours resulted in the production of large amounts of Reactive Oxygen Species (ROS).
Referring to FIG. 7, (wherein (E) in the figure is a control experiment, heLa cells without any ROS initiator and scavenger, (F) cells incubated with Cu-MOF, (G) cells incubated with DCFH-DA, and (H) cells incubated with DCFH-DA and Cu-MOF), it is noted that there is a light green fluorescence in untreated cells, which may be derived from the ROS inherent in HeLa cells. After incubation with Cu-MOF, the fluorescence intensity was reduced. And after beta-Lap treatment, the green fluorescence is obviously enhanced, and the generation of a large amount of intracellular ROS is shown. However, little green fluorescence was observed after the Cu-MOF incubation of cells with β -Lap treatment, i.e., little ROS in the system. These experiments with changes in intracellular ROS concentration reflected in fluorescence intensity provide strong evidence that the Cu-MOF has excellent ability to scavenge intracellular ROS.
Example 2:
in this example, the copper-based MOF material used in example 1 can be prepared as follows:
34.1mg of H 2 L was dissolved in 5mL of N, N-Dimethylacetamide (DMA), while 92.08mg of CuCl 2·2H2 O was dissolved in 10mL of H 2 O; the two solutions were mixed and added to a 20 mL-gauge Teflon substrate autoclave and heated at 90℃for 48h. Then, a well-crystallized Cu-MOF was obtained, which was then ground into a Cu-MOF powder in an agate mortar.
Example 3:
in this example, the copper-based MOF material used in example 1 can be prepared as follows:
34.1mg of H 2 L was dissolved in 10mL of N, N-Dimethylacetamide (DMA), simultaneously 46.04mg of CuCl 2·2H2 O was dissolved in 10mL of H 2 O, and the two solutions were mixed and added to a polytetrafluoroethylene substrate autoclave of 20mL gauge, and heated at 90℃for 48 hours. Then, a well-crystallized Cu-MOF was obtained, which was then ground into a Cu-MOF powder in an agate mortar.
Example 4:
in this example, the copper-based MOF material used in example 1 can be prepared as follows:
34.1mg of H 2 L was dissolved in 5mL of N, N-Dimethylacetamide (DMA), simultaneously 46.04mg of CuCl 2·2H2 O was dissolved in 10mL of H 2 O, and the two solutions were mixed and added to a 20 mL-specification polytetrafluoroethylene substrate autoclave, and heated at 120℃for 48 hours. Then, a well-crystallized Cu-MOF was obtained, which was then ground into a Cu-MOF powder in an agate mortar.
Example 5:
in this example, the copper-based MOF material modified microelectrode of example 1 can be prepared as follows:
Drawing a glass capillary by using a P-97 type drawing instrument, cutting the tip of the capillary by using a surgical knife, penetrating carbon fibers with the diameter of 30 mu m into the holes, connecting the carbon fibers and copper wires by using silver conductive adhesive, sealing the tip of an electrode by using nail polish, and removing the nail polish on the surface of the carbon fibers by using ethanol;
A potentiostatic potential of 1.5V (vs. Ag/Cl) was applied in 0.1M NaOH for 80s to remove contaminants from the CFE tip surface. Uniformly mixing Cu-MOF powder (600 mg/mL) with 2% Nafion by ultrasonic, and soaking CFE in the solution to obtain the Cu-MOF modified carbon fiber microelectrode (Cu-MOF/CFE).
It should be noted that in the description of the present application, the terms "first," "second," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance. Furthermore, in the description of the present application, unless otherwise indicated, the meaning of "a plurality" is two or more.
Any process or method descriptions in flow charts or otherwise described herein may be understood as representing modules, segments, or portions of code which include one or more executable instructions for implementing specific logical functions or steps of the process, and further implementations are included within the scope of the preferred embodiment of the present application in which functions may be executed out of order from that shown or discussed, including substantially concurrently or in reverse order, depending on the functionality involved, as would be understood by those reasonably skilled in the art of the present application.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present application. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the present application have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the application, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the application.
Claims (5)
1. The application of the microelectrode modified by the copper-based MOF material as an electrochemical sensor in the aspect of detecting the concentration of superoxide anions in cells and living bodies is characterized in that the specific preparation process of the microelectrode is as follows:
Penetrating carbon fibers into the glass capillary holes, and connecting the carbon fibers and copper wires through silver conductive adhesive;
Sealing the tips of the microelectrodes with nail polish;
removing pollutants on the surface of the tip of the microelectrode;
And placing the microelectrode in a suspension prepared by a copper-based MOF material to prepare the carbon fiber microelectrode wrapped by the copper-based MOF material, wherein the specific preparation method of the copper-based MOF material comprises the following steps: preparing a solution of 2, 5-dicarboxylic acid-3, 4-ethylenedioxythiophene and N, N-dimethylacetamide according to a molar ratio of 1.5:1-3.5:1, and dissolving copper chloride dihydrate in distilled water to obtain a solution with mass concentration of 0.01-0.1 mol.L -1; mixing the two solutions obtained after dissolution, adding the mixed solutions into a polytetrafluoroethylene substrate high-pressure reaction kettle, heating at a high temperature to obtain Cu-MOF crystals with good crystallization, and grinding the Cu-MOF crystals to obtain a copper-based MOF material;
The application of the carbon fiber microelectrode wrapped by the copper-based MOF material as an electrochemical sensor in the aspect of detecting the concentration of superoxide anions in cells and living bodies is as follows: adopting a three-electrode system, wherein a Cu-MOF modified electrode is a working electrode, ag/AgCl is a reference electrode, a platinum wire is a counter electrode, a phosphate buffer solution with the pH of 0.1M containing O 2 •- and the pH value of 7.5 is used as electrolyte, and timing current scanning is performed under the working potential of-0.05V; in a specific concentration range of O 2 •-, the response current Ip and the concentration of O 2 •- have good linear relation, and the concentration of O 2 •- is obtained through the value of the response current Ip.
2. Use of a copper-based MOF material modified microelectrode as claimed in claim 1 for detection of superoxide anion concentration in cells and living bodies as an electrochemical sensor, characterized in that the contaminants on the tip surface of the microelectrode are removed by applying a potentiostatic potential to the microelectrode in sodium hydroxide.
3. The application of the microelectrode modified by the copper-based MOF material as an electrochemical sensor in the aspect of detecting the concentration of superoxide anions in cells and living bodies, wherein the concentration of the copper-based MOF material suspension is 50-800 mg/mL.
4. The use of a copper-based MOF material modified microelectrode as claimed in claim 1 or 3 as an electrochemical sensor for detecting superoxide anion concentration in cells and living bodies, wherein the copper-based MOF material suspension is prepared by ultrasonic uniform mixing of copper-based MOF material with 2% Nafion solution.
5. The application of the microelectrode modified by the copper-based MOF material as an electrochemical sensor in the aspect of detecting the concentration of superoxide anions in cells and living bodies, which is characterized in that the high-temperature heating temperature in a polytetrafluoroethylene substrate high-pressure reaction kettle is 80-150 ℃ and the heating time is 24-60 h.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104370943A (en) * | 2014-11-03 | 2015-02-25 | 南开大学 | Preparation method and application of [Cu2(HL)2(Mu2-OH)2(H2O)5] |
| CN107478697A (en) * | 2017-07-27 | 2017-12-15 | 华中科技大学 | Rime shape metal organic frame composite micro-electrode and in-situ preparation method and application |
| CN109406600A (en) * | 2018-09-19 | 2019-03-01 | 新乡医学院 | It is a kind of can be in the production method of body real-time detection hydrogen peroxide microelectrode |
| CN110726762A (en) * | 2019-08-30 | 2020-01-24 | 南京理工大学 | Application of copper-based three-dimensional metal organic framework material in enzyme simulation |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104370943A (en) * | 2014-11-03 | 2015-02-25 | 南开大学 | Preparation method and application of [Cu2(HL)2(Mu2-OH)2(H2O)5] |
| CN107478697A (en) * | 2017-07-27 | 2017-12-15 | 华中科技大学 | Rime shape metal organic frame composite micro-electrode and in-situ preparation method and application |
| CN109406600A (en) * | 2018-09-19 | 2019-03-01 | 新乡医学院 | It is a kind of can be in the production method of body real-time detection hydrogen peroxide microelectrode |
| CN110726762A (en) * | 2019-08-30 | 2020-01-24 | 南京理工大学 | Application of copper-based three-dimensional metal organic framework material in enzyme simulation |
Non-Patent Citations (1)
| Title |
|---|
| Nanotechnology for Electroanalytical Biosensors of Reactive Oxygen and Nitrogen Species;Rajesh Seenivasan 等;《THE CHEMICAL RECORD》;第886-901页 * |
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