CN111307903A - Electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid - Google Patents
Electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid Download PDFInfo
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Abstract
The invention relates to an electrochemical sensor modified by a platinum/molybdenum disulfide composite material, which can simultaneously and quantitatively detect dopamine and uric acid. The invention takes two-dimensional molybdenum disulfide as a support material, in-situ synthesizes nano platinum particles, and takes a platinum/molybdenum disulfide compound as an electrode modification material for electrode modification, so as to prepare a three-electrode system capable of practically distinguishing dopamine and uric acid potentials. The electrochemical sensor has the advantages of simple and quick detection method, quick response, good sensitivity and selectivity and low cost, and can be widely applied to the fields of biology, agriculture, medicine, scientific research and the like.
Description
Technical Field
The invention belongs to the field of bioelectrochemical sensors, and particularly relates to an electrochemical sensor capable of quantitatively detecting dopamine and uric acid simultaneously.
Technical Field
Dopamine as neurotransmitter has important effects on kidney function, cardiovascular system and central nervous system, and diseases such as drug addiction and Parkinson's disease are related to abnormal secretion of dopamine in human body. Uric acid is one of the metabolites of purine, and disorders of purine metabolism can lead to hyperuricemia and gout. The dopamine and the uric acid are commonly coexisted in body fluid of an organism, so that the measurement of the content of the dopamine and the uric acid in each other is influenced. The traditional detection method mainly comprises high performance liquid chromatography, needs expensive instruments and reagents, and is complex in operation and very inconvenient. Therefore, the development of a dopamine and uric acid detection sensor and detection method which are convenient to use, low in cost, high in sensitivity and high in selectivity has important practical significance.
The electrochemical method has the advantages of quick response, high sensitivity, simple operation, low cost and the like, and has been applied to the analysis and diagnosis research of dopamine and uric acid in recent decades. However, the potentials of dopamine and uric acid on conventional electrodes are very close, and the simultaneous quantitative measurement is difficult; the contradiction that the partial dopamine and the uric acid are always simultaneously present in body fluid limits the application of the electrochemical method. Finding a method for solving the contradiction has important scientific research and social significance.
Disclosure of Invention
In order to solve the above situation, the invention discloses an electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid, which takes two-dimensional molybdenum disulfide as a support material, in-situ synthesizes nano platinum particles, and takes a platinum/molybdenum disulfide compound as an electrode modification material to modify an electrode, so that the potential of dopamine and uric acid can be distinguished, and a simple, convenient and sensitive method capable of simultaneously detecting dopamine and uric acid is established on the basis.
In order to achieve the purpose of the invention, the invention discloses an electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid, which is prepared by the following steps:
I. preparing a molybdenum disulfide nanosheet by a lithium ion insertion stripping method:
a. uniformly mixing molybdenum disulfide with n-hexane solution containing n-butyllithium under the condition of argon;
b. adding deoxygenated ultrapure water into the mixed solution obtained in the step a, and centrifuging to remove the supernatant to obtain a single-layer molybdenum disulfide nanosheet;
II, preparing the platinum/molybdenum disulfide composite material in situ:
c. cleaning the single-layer molybdenum disulfide nanosheet obtained in the purification step b;
d. adding potassium iodide and polyvinylpyrrolidone into the potassium hypochlorite platinate solution, mixing and stirring uniformly;
e. adding the single-layer molybdenum disulfide nanosheets obtained in the step c into a dimethylformamide solution, mixing and uniformly stirring;
f. mixing and stirring the two solutions obtained in the steps d and e, performing microwave reaction, centrifuging and discarding supernatant to obtain a platinum/molybdenum disulfide concentrated solution;
preparing a platinum/molybdenum disulfide composite material modified electrode:
g. ultrasonically cleaning the polished glassy carbon electrode in ultrapure water and absolute ethyl alcohol respectively, and drying the electrode by nitrogen;
h. dropwise adding the platinum/molybdenum disulfide solution obtained in the step f onto the dried electrode, and airing to obtain a platinum/molybdenum disulfide modified glassy carbon electrode;
i. and (e) enabling the platinum/molybdenum disulfide modified glassy carbon electrode obtained in the step h, a counter electrode and a reference electrode to jointly form a three-electrode system, namely the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid.
Further, the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid comprises the following specific steps:
a. weighing 1.6g of molybdenum disulfide powder, uniformly mixing the molybdenum disulfide powder with 10ml of n-hexane solution containing 1M n-butyllithium in an argon atmosphere, and performing ultrasound for 3 hours at room temperature;
b. after the reaction is finished, standing for 30 minutes, and removing supernatant after the mixture is layered to obtain dispersion liquid; introducing argon into ultrapure water for 30 minutes to remove oxygen, then adding the deoxidized ultrapure water into the dispersion liquid according to the volume ratio of 1:1, centrifuging the mixed liquid to remove the supernatant, and purifying to obtain the purified single-layer molybdenum disulfide nanosheet.
Further, in the above electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid, the step II specifically includes:
c. and D, adding deoxygenated ultrapure water into the single-layer molybdenum disulfide nanosheet obtained in the step I, centrifuging, discarding the supernatant, and repeatedly treating for three times to clean the molybdenum disulfide nanosheet.
d. Weighing 0.1g of potassium iodide and 0.25g of polyvinylpyrrolidone, uniformly mixing with 240 mu L of 100mM potassium chloroplatinate, and stirring for 30 minutes;
e. taking 1-4 ml of dimethylformamide, uniformly mixing with the molybdenum disulfide nanosheet obtained in the step c, and stirring for 30 minutes;
f. and d, uniformly mixing the two solutions obtained in the steps d and e, magnetically stirring for 15 minutes, and transferring into a 10mL microwave reaction tube. Microwave reaction at 130 deg.c for 30 min. After the reaction is finished, the product is cooled at room temperature, centrifuged, supernatant is discarded, ultrapure water is added, and the product is centrifugally cleaned. And adding a certain amount of ultrapure water into the centrifugal product to ensure that the concentration of the platinum/molybdenum disulfide composite material is 4-20 mu g/mL, and storing at 4 ℃ for later use.
Further, in the above electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid, in the step e, the volume of the added dimethylformamide is 2 ml.
Furthermore, in the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid, in the step f, the concentration of the prepared platinum/molybdenum disulfide composite material is 4 mug/mL.
Further, in the above electrochemical sensor capable of quantitatively detecting dopamine and uric acid simultaneously, the step III specifically is:
g. ultrasonically cleaning the polished glassy carbon electrode in ultrapure water and absolute ethyl alcohol respectively, and drying the electrode by nitrogen;
h. after the nitrogen is dried, 10 mu L of the platinum/molybdenum disulfide solution obtained in the second step is dripped, and the platinum/molybdenum disulfide modified glassy carbon electrode is obtained after air drying;
i. and (e) enabling the platinum/molybdenum disulfide modified glassy carbon electrode obtained in the step h, a counter electrode and a reference electrode to jointly form a three-electrode system, namely the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid. The counter electrode can be a platinum counter electrode or a carbon counter electrode, and the reference electrode can be a saturated calomel electrode or a silver/silver chloride electrode.
Further, the detection method of the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid comprises the following steps IV:
j: one end of the three electrodes of the electrochemical sensor obtained in the step i is immersed into 0.1M phosphate solution containing dopamine and uric acid with certain concentration, the other end of the three electrodes is connected with an electrochemical workstation to collect signals, differential pulse voltammetry is selected for determination, and the scanning range is set to be-0.2-0.8V;
k, preparing a series of dopamine and uric acid solutions with gradient concentrations, detecting by the method in the step j, and drawing a standard curve by taking the concentrations of the dopamine and the uric acid as horizontal coordinates and the dopamine peak current and the uric acid current measured by a differential pulse voltammetry as vertical coordinates;
l, detecting the solution to be detected according to the method in the step j to obtain the peak current values of the dopamine and the uric acid in the differential pulse voltammetry, and substituting the peak current values into the standard curve drawn in the step four j to obtain the concentration of the dopamine and the concentration of the uric acid.
Furthermore, the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid and the method thereof are applied to the fields of biology, agriculture, medicine and scientific research.
The invention has the following beneficial effects:
(1) the method takes two-dimensional molybdenum disulfide as a support material, nano platinum particles are synthesized in situ to form a platinum/molybdenum disulfide compound, the platinum/molybdenum disulfide compound is used as an electrode modification material to modify glassy carbon, and the prepared electrochemical sensor of the three-electrode system can detect the potentials of dopamine and uric acid in the same solution with high selectivity;
(2) the electrochemical sensor is convenient to use, high in detection sensitivity and selectivity and low in preparation cost;
(3) through optimization, the detection limit of the electrochemical sensor disclosed by the invention on dopamine is as low as 0.17 mu M, and the detection limit on uric acid is as low as 0.36 mu M, so that the electrochemical sensor has obvious advantages compared with the traditional sensor, and can be widely applied to the fields of biology, agriculture, medicines, scientific research and the like.
Drawings
FIG. 1 is an electron micrograph of a platinum/molybdenum disulfide composite prepared in example 1;
FIG. 2 is a cyclic voltammogram of the electrochemical sensor in example 2 in a solution containing dopamine and uric acid;
FIG. 3 is a standard curve of optimized dopamine in example 2;
fig. 4 is a standard curve of uric acid optimized in example 2.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention; the experimental methods used in the following examples are all conventional methods unless otherwise specified; materials, reagents and the like used in the following examples are commercially available unless otherwise specified; the described to be measured
Example 1
Preparation example
An electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid is prepared by the following steps,
I. preparing a molybdenum disulfide nanosheet by a lithium ion insertion stripping method:
a. weighing 1.6g of molybdenum disulfide powder, uniformly mixing the molybdenum disulfide powder with 10ml of n-hexane solution containing 1M n-butyllithium in an argon atmosphere, and performing ultrasound for 3 hours at room temperature;
b. after the reaction is finished, standing for 30 minutes, and removing supernatant after the mixture is layered to obtain dispersion liquid; introducing argon into ultrapure water for 30 minutes to remove oxygen, then adding the deoxidized ultrapure water into the dispersion liquid according to the volume ratio of 1:1, centrifuging the mixed liquid to remove the supernatant, and purifying to obtain the purified single-layer molybdenum disulfide nanosheet.
II, preparing the platinum/molybdenum disulfide composite material in situ:
c. and D, adding deoxygenated ultrapure water into the single-layer molybdenum disulfide nanosheet obtained in the step I, centrifuging, discarding the supernatant, and repeatedly treating for three times to clean the molybdenum disulfide nanosheet.
d. Weighing 0.1g of potassium iodide and 0.25g of polyvinylpyrrolidone, uniformly mixing with 240 mu L of 100mM potassium chloroplatinate, and stirring for 30 minutes;
e. taking 1-4 ml of dimethylformamide, uniformly mixing with the molybdenum disulfide nanosheet obtained in the step c, and stirring for 30 minutes;
f. and d, uniformly mixing the two solutions obtained in the steps d and e, magnetically stirring for 15 minutes, and transferring into a 10mL microwave reaction tube. Microwave reaction at 130 deg.c for 30 min. After the reaction is finished, the product is cooled at room temperature, centrifuged, supernatant is discarded, ultrapure water is added, and the product is centrifugally cleaned. And adding a certain amount of ultrapure water into the centrifugal product to ensure that the concentration of the platinum/molybdenum disulfide composite material is 4-20 mu g/mL, wherein an electron microscope picture of the platinum/molybdenum disulfide composite material is shown in figure 1.
Preparing a platinum/molybdenum disulfide composite material modified electrode:
g. ultrasonically cleaning the polished glassy carbon electrode in ultrapure water and absolute ethyl alcohol respectively, and drying the electrode by nitrogen;
h. after the nitrogen is dried, 10 mu L of the platinum/molybdenum disulfide solution obtained in the second step is dripped, and the platinum/molybdenum disulfide modified glassy carbon electrode is obtained after air drying;
i. and (e) enabling the platinum/molybdenum disulfide modified glassy carbon electrode obtained in the step h, a counter electrode and a reference electrode to jointly form a three-electrode system, namely the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid. The counter electrode is a platinum counter electrode, and the reference electrode is a saturated calomel electrode.
And e, comparing the amount of the dimethylformamide in the step e, wherein the platinum/molybdenum disulfide composite material synthesized by 2mL of dimethylformamide is relatively uniform in shape and size, and the distinction degree of the dopamine and the uric acid is best.
And f, comparing the platinum/molybdenum disulfide composite materials with different concentrations in the step f, wherein the 4 mu g/mL platinum/molybdenum disulfide composite material modified electrode is optimal and sensitive to the discrimination of dopamine and uric acid.
Thus, the amount of dimethylformamide in step e was limited to 2mL and the platinum/molybdenum disulfide composite in step f was limited to 4 μ g/mL as the best mode for the test of example 2.
Example 2
Test examples, in which all reagents were commercially available; the electrochemical workstation is a multichannel SquidStat PLUS electrochemical workstation of Admiral company, and the differential pulse voltammetry is performed according to the instruction in the work,
j: and (e) immersing one end of the three electrodes of the electrochemical sensor obtained in the step (i) into a 0.1M phosphate solution containing dopamine and uric acid with certain concentration, connecting the other end of the three electrodes with an electrochemical workstation to collect signals, selecting a differential pulse voltammetry to measure, setting a scanning range to be-0.2-0.8V, and obtaining a result as shown in the attached figure 2, wherein a dopamine curve is positioned above a uric acid curve, and the separation degree is obvious. The result shows that the electrochemical sensor modified by the platinum/molybdenum disulfide can obviously distinguish the potentials of dopamine and uric acid in the same solution, and can be used for further experiments;
k, preparing a series of dopamine and uric acid solutions with gradient concentrations, detecting by the method in the step j, respectively drawing standard curves by taking the concentrations of the dopamine and the uric acid as horizontal coordinates and taking the peak current of the dopamine and the current of the uric acid measured by a differential pulse voltammetry as vertical coordinates, wherein the results are respectively shown in the attached drawings 3 and 4, and Y =1.172+0.0744X and R in the standard curves of the dopamine are respectively shown in the attached drawings2=0.992, detection limit 0.17 μ M; in the standard curve of uric acid, Y =0.7634+0.0345X, R2=0.984, detection limit of 0.36 μ M;
l, detecting the solution to be detected according to the method in the step j to obtain the peak current values of the dopamine and the uric acid in the differential pulse voltammetry, and substituting the peak current values into the standard curve drawn in the step four j to obtain the concentration of the dopamine and the concentration of the uric acid.
According to the embodiments, when the electrochemical sensor provided by the invention is used for simultaneously detecting dopamine and uric acid, the detection limit of the electrochemical sensor is as low as 0.17 mu M, and the detection limit of the electrochemical sensor is as low as 0.36 mu M.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that modifications can be made by those skilled in the art without departing from the principle of the present invention, and these modifications should also be construed as the protection scope of the present invention.
Claims (9)
1. An electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid is characterized by being prepared by the following steps:
I. preparing a molybdenum disulfide nanosheet by a lithium ion insertion stripping method:
a. uniformly mixing molybdenum disulfide with n-hexane solution containing n-butyllithium under the condition of argon;
b. adding deoxygenated ultrapure water into the mixed solution obtained in the step a, and centrifuging to remove the supernatant to obtain a single-layer molybdenum disulfide nanosheet;
II, preparing the platinum/molybdenum disulfide composite material in situ:
c. cleaning the single-layer molybdenum disulfide nanosheet obtained in the purification step b;
d. adding potassium iodide and polyvinylpyrrolidone into the potassium hypochlorite platinate solution, mixing and stirring uniformly;
e. adding the single-layer molybdenum disulfide nanosheets obtained in the step c into a dimethylformamide solution, mixing and uniformly stirring;
f. mixing and stirring the two solutions obtained in the steps d and e, performing microwave reaction, centrifuging and discarding supernatant to obtain a platinum/molybdenum disulfide concentrated solution;
preparing a platinum/molybdenum disulfide composite material modified electrode:
g. ultrasonically cleaning the polished glassy carbon electrode in ultrapure water and absolute ethyl alcohol respectively, and drying the electrode by nitrogen;
h. dropwise adding the platinum/molybdenum disulfide solution obtained in the step f onto the dried electrode, and airing to obtain a platinum/molybdenum disulfide modified glassy carbon electrode;
i. and (e) enabling the platinum/molybdenum disulfide modified glassy carbon electrode obtained in the step h, a counter electrode and a reference electrode to jointly form a three-electrode system, namely the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid.
2. The electrochemical sensor according to claim 1, wherein the step I specifically comprises:
a. weighing 1.6g of molybdenum disulfide powder, uniformly mixing the molybdenum disulfide powder with 10ml of n-hexane solution containing 1M n-butyllithium in an argon atmosphere, and performing ultrasound for 3 hours at room temperature;
b. after the reaction is finished, standing for 30 minutes, and removing supernatant after the mixture is layered to obtain dispersion liquid; introducing argon into ultrapure water for 30 minutes to remove oxygen, then adding the deoxidized ultrapure water into the dispersion liquid according to the volume ratio of 1:1, centrifuging the mixed liquid to remove the supernatant, and purifying to obtain the purified single-layer molybdenum disulfide nanosheet.
3. The electrochemical sensor according to claim 1, wherein the step II comprises:
c. adding deoxygenated ultrapure water into the single-layer molybdenum disulfide nanosheet obtained in the step I, centrifuging, discarding the supernatant, and repeatedly treating for three times to clean the molybdenum disulfide nanosheet;
d. weighing 0.1g of potassium iodide and 0.25g of polyvinylpyrrolidone, uniformly mixing with 240 mu L of 100mM potassium chloroplatinate, and stirring for 30 minutes;
e. taking 1-4 ml of dimethylformamide, uniformly mixing with the molybdenum disulfide nanosheet obtained in the step c, and stirring for 30 minutes;
f. d, uniformly mixing the two solutions obtained in the step e, magnetically stirring for 15 minutes, and transferring into a 10mL microwave reaction tube; carrying out microwave reaction for 30 minutes at 130 ℃; after the reaction is finished, cooling the product at room temperature, centrifuging, removing the supernatant, adding ultrapure water, and centrifugally cleaning; and adding a certain amount of ultrapure water into the centrifugal product to ensure that the concentration of the platinum/molybdenum disulfide composite material is 4-20 mu g/mL, and storing at 4 ℃ for later use.
4. The electrochemical sensor according to claim 3, wherein the volume of the dimethylformamide added in step e is 2 ml.
5. The electrochemical sensor according to claim 3, wherein the concentration of the platinum/molybdenum disulfide composite material obtained in step f is 4 μ g/mL.
6. The electrochemical sensor according to claim 1, wherein the step III specifically comprises:
g. ultrasonically cleaning the polished glassy carbon electrode in ultrapure water and absolute ethyl alcohol respectively, and drying the electrode by nitrogen;
h. after the nitrogen is dried, 10 mu L of the platinum/molybdenum disulfide solution obtained in the second step is dripped, and the platinum/molybdenum disulfide modified glassy carbon electrode is obtained after air drying;
i. and (e) enabling the platinum/molybdenum disulfide modified glassy carbon electrode obtained in the step h, a counter electrode and a reference electrode to jointly form a three-electrode system, namely the electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid.
7. The method for detecting an electrochemical sensor according to any one of claims 1 to 6, which can quantitatively detect dopamine and uric acid simultaneously, wherein the method comprises the following steps IV:
j: one end of the three electrodes of the electrochemical sensor obtained in the step i is immersed into 0.1M phosphate solution containing dopamine and uric acid with certain concentration, the other end of the three electrodes is connected with an electrochemical workstation to collect signals, differential pulse voltammetry is selected for determination, and the scanning range is set to be-0.2-0.8V;
k, preparing a series of dopamine and uric acid solutions with gradient concentrations, detecting by the method in the step j, and drawing a standard curve by taking the concentrations of the dopamine and the uric acid as horizontal coordinates and the dopamine peak current and the uric acid current measured by a differential pulse voltammetry as vertical coordinates;
l, detecting the solution to be detected according to the method in the step j to obtain the peak current values of the dopamine and the uric acid in the differential pulse voltammetry, and substituting the peak current values into the standard curve drawn in the step four j to obtain the concentration of the dopamine and the concentration of the uric acid.
8. The use of the electrochemical sensor according to any one of claims 1 to 6 for the simultaneous quantitative detection of dopamine and uric acid in the fields of biology, agriculture, medicine and scientific research.
9. The application of the method for detecting an electrochemical sensor capable of simultaneously and quantitatively detecting dopamine and uric acid according to claim 7 in the fields of biology, agriculture, medicine and scientific research.
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| CN115112737A (en) * | 2022-06-27 | 2022-09-27 | 天津大学 | Preparation and application of nitrogen-doped reduced graphene oxide-based composite tungsten disulfide nanosheet electrochemical sensor |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112525974A (en) * | 2020-11-19 | 2021-03-19 | 苏州健雄职业技术学院 | Method for measuring concentrations of dopamine and uric acid in presence of ascorbic acid |
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| CN113447547A (en) * | 2021-05-28 | 2021-09-28 | 天津大学 | Prostate cancer tumor marker detection method based on molybdenum disulfide/nano platinum-coated gold triangle/polydopamine |
| CN113447547B (en) * | 2021-05-28 | 2022-11-11 | 天津大学 | Prostate cancer tumor marker detection method based on molybdenum disulfide/nano platinum-coated gold triangle/polydopamine |
| CN115112737A (en) * | 2022-06-27 | 2022-09-27 | 天津大学 | Preparation and application of nitrogen-doped reduced graphene oxide-based composite tungsten disulfide nanosheet electrochemical sensor |
| CN115112737B (en) * | 2022-06-27 | 2024-05-07 | 天津大学 | Preparation and application of electrochemical sensor based on nitrogen-doped reduced graphene oxide composite tungsten disulfide nanosheet |
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