A method for detecting antibiotics based on two-dimensional metal ene electrochemical sensor

Abstract

The application discloses an antibiotic detection method based on a two-dimensional metal ene electrochemical sensor and belongs to the technical field of biosensors; the specific steps are as follows: first, ultrasonic dispersion two-dimensional metal ene ink is drop-coated on an electrochemical working electrode to complete preparation of the two-dimensional metal ene electrochemical sensor working electrode; when the electrochemical sensor works, a phosphate buffer solution is used as an electrolyte, an antibiotic to be detected reacts with the two-dimensional metal ene modified on the working electrode to generate an oxidation or reduction reaction, corresponding chemical signals are generated, an electric signal is output through an electrochemical workstation to obtain concentration information of the antibiotic, and super-sensitive, rapid and high-specificity amplification detection of the antibiotic is realized; the application first applies the two-dimensional metal ene to the detection of the antibiotic and fully utilizes the catalytic activity and signal amplification function of the two-dimensional metal ene and the super-sensitivity and rapidness of ECS in biological detection, so that high-specificity, rapid and super-sensitive detection of the antibiotic in a biological sample can be realized.

Description

Technical Field

[0001] This invention belongs to the field of biochemical sensors, specifically relating to an antibiotic detection method based on a two-dimensional metalene electrochemical sensor. Background Technology

[0002] Antibiotics, commonly known as antimicrobials, can kill or inhibit the growth of microorganisms, possessing antibacterial and antiparasitic activities. Therefore, antibiotics are widely used in the treatment of human diseases, as well as in promoting growth or preventing diseases in livestock, aquatic products, and crops. However, antibiotics are a double-edged sword; as a novel pollutant, they can exist in the environment through complex transformations and bioaccumulation cycles, posing serious challenges to human health and the ecological environment.

[0003] Methods for detecting antibiotic residues mainly include chromatography, spectrometry, and electrochemical methods. Chromatography is the most commonly used qualitative and quantitative analysis method, including gas chromatography, liquid chromatography, and high-performance liquid chromatography-mass spectrometry, offering advantages such as high sensitivity and stability. However, its limitations, such as expensive equipment, high operating costs, demanding experimental procedures, and inconvenience, restrict it to laboratory analysis. Spectrometry is convenient to operate, but suffers from high detection limits, poor accuracy, and susceptibility to interference from other components, making it unsuitable for rapid detection of trace amounts of antibiotics. Electrochemical sensors, containing electrochemical transducers, convert the response obtained from the interaction between the target analyte and the sensing element into a measurable signal corresponding to the analyte concentration. Electrochemical methods possess unique advantages in antibiotic residue detection due to their low cost, ease of operation, and suitability for miniaturization and portable design.

[0004] Currently, two-dimensional metalloalkenes are considered promising materials due to their unique structural characteristics. On the one hand, their ultrathin two-dimensional structure significantly increases the electrochemical active area of ​​nanomaterials, improving the utilization rate of metal atoms. On the other hand, their curved and wrinkled structure can induce a significant strain effect, thereby optimizing the electronic structure of metal atoms and enhancing the intrinsic activity of catalysts. Therefore, by fully utilizing the advantages of electrochemical sensors and two-dimensional metalloalkenes, it is hoped that a highly sensitive, specific, and rapid biosensor for antibiotic determination can be established. Summary of the Invention

[0005] To address the aforementioned problems, this invention discloses an antibiotic detection method based on a two-dimensional metalene electrochemical sensor.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows:

[0007] The working electrode of the biosensor ECS used in this invention has a two-dimensional metallene with strong electrocatalytic activity, which can generate a sensitive electrical signal at a specific potential to obtain the concentration of the antibiotic to be tested. When the ECS is working, the antibiotic to be tested reacts with the two-dimensional metallene modified on the working electrode to undergo oxidation or reduction reaction, generating a corresponding chemical signal. After processing by a converter and amplifier, the signal is output as an electrical signal such as current, potential or resistance to obtain the concentration information of the target analyte, thereby realizing the amplified detection of antibiotics with high sensitivity, speed and high specificity.

[0008] This application provides an antibiotic detection method based on a two-dimensional metalene electrochemical sensor, wherein the electrochemical sensor is an electrochemical sensor modified with a two-dimensional metalene.

[0009] The antibiotic detection method based on a two-dimensional metalene electrochemical sensor described in this invention includes first synthesizing a two-dimensional metalene, then preparing a two-dimensional metalene electrochemical sensor, and finally detecting antibiotics.

[0010] Furthermore, the synthesis of the two-dimensional metalene and the preparation process of the two-dimensional metalene electrochemical sensor include the following steps:

[0011] Step (1): Synthesis of two-dimensional metal olefins. Two-dimensional metal olefins were synthesized by low-temperature solvothermal method. First, carbonyl compounds and ascorbic acid were added to oleylamine and stirred and dissolved at 50-100℃. Then, after the mixture was cooled to room temperature, palladium diacetylacetone was added, ultrasonicated, and kept at 80℃ in an oven for 12 hours. Finally, the product was centrifuged and washed three times with ethanol to obtain two-dimensional metal olefins, which were then dispersed in cyclohexane for later use.

[0012] Step (2): Preparation of carbon-supported two-dimensional metal olefins: A certain amount of carbon material is dispersed in cyclohexane and sonicated for 1 h; then cyclohexane is added and sonicated for another 0.5 h to obtain a homogeneous mixture; then the metal olefin material from step (1) is added dropwise to the previous mixture and sonicated for 2 h; finally, the product is collected by centrifugation and dried overnight at 60 °C to obtain carbon-supported two-dimensional metal olefins.

[0013] Step (3): Sensor preparation. Carbon-supported two-dimensional metalene is ultrasonically dispersed in a mixed solution containing isopropanol, distilled water and perfluorinated resin solution to form an ink-like solution. The ink-like solution is drop-coated onto the surface of the bare glassy carbon electrode obtained in step (3), dried, and rinsed with ethanol and distilled water to complete the preparation of the two-dimensional metalene electrochemical sensor.

[0014] Bare glassy carbon electrodes (GCEs) were polished sequentially using alumina slurries of 1.0, 0.3 μM, and 0.05 μM. After each polishing, the electrodes were thoroughly cleaned in an ultrasonic bath with distilled water and anhydrous ethanol. The cleaned polished bare glassy carbon electrodes (GCEs) were then dried and subjected to electrochemical pretreatment in 0.25 M H2SO4 solution until stable cyclic voltammograms (CV) curves were obtained.

[0015] Step (4): The Pdene / C obtained in step (2) is ultrasonically dispersed in a mixed solution containing isopropanol, distilled water and perfluorinated resin solution to make ink. The ink is drop-coated onto the surface of the bare glassy carbon electrode (GCE) obtained in step (3), dried under an infrared lamp, and then the modified electrode is rinsed with anhydrous ethanol and distilled water respectively to remove loosely bound particles and re-dried in air to complete the preparation of the two-dimensional metalene electrochemical sensor.

[0016] Further, the carbonyl compound mentioned in step (1) is either hexacarbonylchromium or hexacarbonylmolybdenum.

[0017] Further, the mass ratio of the carbonyl compound and ascorbic acid in step (1) is 1:2.

[0018] Furthermore, the centrifugation speed in steps (1) and (2) is 8000 rpm.

[0019] Further, in step (3), the volume ratio of isopropanol, distilled water and perfluorinated resin solution in the mixed solution is 25:24:1.

[0020] Furthermore, the detection method involves combining the synthesized two-dimensional metalene with an electrochemical working electrode to detect antibiotics.

[0021] Furthermore, the method for synthesizing the two-dimensional metal olefin is characterized as follows: palladium diacetylacetonate provides the palladium source, which is the basic metal source for synthesizing two-dimensional metal olefins (Pdene); the carbonyl compound slowly releases CO gas at low temperature, which restricts the growth of the metal olefin in the vertical direction and promotes the growth and elongation of the metal olefin in the horizontal direction.

[0022] Furthermore, the detection uses a three-electrode system, with an Ag / AgCl electrode as the reference electrode, a Pt electrode as the counter electrode, a glassy carbon electrode modified with two-dimensional metal olefins as the working electrode, and a 0.1 M phosphate buffer solution as the electrolyte.

[0023] Furthermore, the detection method utilizes the reaction between the antibiotic to be tested and the two-dimensional metal olefin modified on the working electrode to undergo oxidation or reduction reactions, generating corresponding chemical signals. The concentration information of the antibiotic is obtained by outputting an electrical signal through an electrochemical workstation, thereby achieving amplified detection of the antibiotic.

[0024] Furthermore, the electrochemical detection method includes either cyclic voltammetry or constant voltage method.

[0025] Furthermore, the antibiotic detected was a tetracycline antibiotic.

[0026] Furthermore, the detection method utilizes an electrochemical workstation to monitor changes in electrical signals, enabling real-time detection of antibiotics.

[0027] The testing methods described in this application can be used for various diagnostic or non-diagnostic purposes, including environmental monitoring, medical or biological scientific research, and product quality testing.

[0028] The beneficial effects of this invention are as follows: This invention is the first to apply two-dimensional metalene to the detection of antibiotics, and makes full use of the catalytic activity and signal amplification function of two-dimensional metalene, as well as the ultrasensitive and rapid characteristics of ECS in biological detection, so as to achieve highly specific, rapid and ultrasensitive detection of antibiotics in biological samples. Attached Figure Description

[0029] Figure 1 This is a schematic diagram of the synthesis of two-dimensional metal olefins;

[0030] Figure 2 A schematic diagram of the fabrication of a two-dimensional metalene electrochemical sensor;

[0031] Figure 3 CV curves for the electrochemical detection of tetracycline;

[0032] Figure 4 The current-tetracycline concentration fitting curve for the electrochemical detection of tetracycline by cyclic voltammetry;

[0033] Figure 5 The it curve for the electrochemical detection of tetracycline;

[0034] Figure 6 This is a current-tetracycline concentration fitting curve for the electrochemical detection of tetracycline using the constant voltage method. Detailed Implementation

[0035] The present invention will be further illustrated below with reference to the accompanying drawings and specific embodiments. It should be understood that the following specific embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0036] Example 1: Synthesis of Two-Dimensional Metalloalkenes

[0037] Two-dimensional metal olefins were synthesized using a low-temperature solvothermal method, the specific process of which is as follows: Figure 1 As shown.

[0038] First, 10 mg of chromium hexacarbonyl and 20 mg of ascorbic acid were added to 5 mL of oleylamine and dissolved by stirring at 70 °C. Next, after the mixture cooled to room temperature, 10 mg of palladium diacetylacetone was added, and the mixture was sonicated to obtain a clear solution. The mixture was then sealed and transferred to an oven, sonicated, and maintained at 80 °C for 12 h. Finally, the product was centrifuged, washed three times with cyclohexane to obtain a two-dimensional metal olefin, and dispersed in cyclohexane for subsequent use.

[0039] Example 2: Synthesis of two-dimensional metalloalkenes

[0040] Two-dimensional metal olefins were synthesized using a low-temperature solvothermal method, the specific process of which is as follows: Figure 1 As shown in the figure. First, 10 mg of molybdenum hexacarbonyl and 20 mg of ascorbic acid were added to 5 mL of oleylamine and stirred at 70 °C to dissolve. Then, after the mixture cooled to room temperature, 10 mg of palladium diacetylacetone was added, and the mixture was sonicated to obtain a transparent solution. The mixture was then sealed and transferred to an oven and kept at 80 °C for 12 h. Finally, the product was centrifuged at 8000 rpm and washed three times with cyclohexane to obtain a two-dimensional metal olefin, which was then dispersed with cyclohexane for subsequent use.

[0041] Example 3: Fabrication of a two-dimensional metalene electrochemical sensor

[0042] Before electrochemical testing, the synthesized two-dimensional metallene was loaded onto Ketjen black. The specific process is as follows: The obtained metallene and a certain amount of carbon black were ultrasonically mixed in a solution for 2 hours, then the solid product was collected by centrifugation and washed twice with ethanol. After drying overnight at 60°C, 1 mg of carbon-supported nanomaterials was dispersed in a mixed solution containing 500 μL isopropanol, 480 μL ultrapure water, and 20 μL perfluorinated resin solution, and ultrasonicated for 1 hour to form a uniformly dispersed ink-like solution. Finally, 4-20 μL of the ink-like solution was dropped onto a 4 mm diameter glassy carbon electrode (GCE) for subsequent electrochemical performance testing. The preparation process is as follows: Figure 2 As shown.

[0043] Example 4: Cyclic voltammetry (CV) for the detection of tetracycline

[0044] The working electrode was prepared based on the material optimization results. Cyclic voltammetry was used to test its performance in 0.1M PBS solutions containing different concentrations of tetracycline. The oxidation curves of tetracycline are shown below. Figure 3 As shown, the oxidation peaks are clear and well-shaped, and the peak current of tetracycline electrooxidation increases with increasing tetracycline concentration. Figure 4 As shown, the oxidation peak current is linearly related to the tetracycline concentration, and the detection limit is 0.18 μM.

[0045] Example 5: Constant Voltage Method (it) for Detection of Tetracycline

[0046] Based on the CV test spectrum, the oxidation potential of tetracycline was found to be 0.70 V. Figure 5 As shown, the constant voltage method was used to test the response of tetracycline at this potential; the response current gradually increased with increasing concentration. Figure 6 As shown, the linear relationship between the current and tetracycline concentration fitting curve is good, proving that tetracycline has a good linear relationship in the concentration range of 1-60 μM, and the detection limit is 0.58 μM.

[0047] It should be noted that the above content merely illustrates the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. For those skilled in the art, various improvements and modifications can be made without departing from the principle of the present invention, and all such improvements and modifications fall within the scope of protection of the claims of the present invention.

Claims

1. A method for antibiotic detection based on a two-dimensional metalene electrochemical sensor, characterized in that, First, the synthesis of two-dimensional metalene was completed, and then a two-dimensional metalene electrochemical sensor was prepared to detect antibiotics. The method for synthesizing the two-dimensional metalene and preparing the two-dimensional metalene electrochemical sensor includes the following steps: Step (1): Synthesis of two-dimensional metal olefins. Two-dimensional metal olefins were synthesized by low-temperature solvothermal method. First, carbonyl compounds and ascorbic acid were added to the solution and stirred and dissolved at 50-100℃. Then, after the mixture was cooled to room temperature, palladium diacetylacetone was added, ultrasonicated, and kept at 80℃ in an oven for 12 hours. Finally, the product was centrifuged and washed three times with ethanol to obtain two-dimensional metal olefins, which were then dispersed in cyclohexane for later use. Step (2): Preparation of carbon-supported two-dimensional metal olefins: a certain amount of Ketjen black is dispersed in the mixture and sonicated for 1 hour; then cyclohexane is added and sonicated for another 0.5 hours to obtain a homogeneous mixture; then the metal olefin material from step (1) is added dropwise to the previous mixture and sonicated for 2 hours; finally, the product is collected by centrifugation and dried at 60°C overnight to obtain carbon-supported two-dimensional metal olefins. Step (3): Polish the bare glassy carbon electrode (GCE) sequentially with 1.0, 0.3 μM and 0.05 μM alumina slurries. After each polishing, thoroughly clean the electrode with distilled water and anhydrous ethanol in an ultrasonic bath. Then, dry the cleaned polished bare glassy carbon electrode (GCE) and perform electrochemical pretreatment in 0.25 M H2SO4 solution until a stable cyclic voltammogram (CV) curve is obtained. Step (4): Sensor preparation. Carbon-supported two-dimensional metalene is ultrasonically dispersed in a mixed solution containing isopropanol, distilled water and perfluorinated resin solution to form an ink-like solution. The ink-like solution is drop-coated onto the surface of the bare glassy carbon electrode obtained in step (3), dried, and rinsed with ethanol and distilled water to complete the preparation of the two-dimensional metalene electrochemical sensor.

2. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, In step (1), the carbonyl compound is either hexacarbonylchromium or hexacarbonylmolybdenum, and the mass ratio of the carbonyl compound to ascorbic acid is 1:

2.

3. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, The centrifugation speed mentioned in steps (1) and (2) is 8000 rpm.

4. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, The volume ratio of isopropanol, distilled water and perfluorinated resin solution in the mixed solution in step (3) is 25:24:

1.

5. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, The detection method involves combining a synthesized two-dimensional metalene with an electrochemical working electrode to detect antibiotics.

6. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, The detection uses a three-electrode system: an Ag / AgCl electrode as the reference electrode, a Pt electrode as the counter electrode, and the working electrode is the prepared two-dimensional metalene electrochemical sensor. The electrolyte is a 0.1 M phosphate buffer solution.

7. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, The detection method utilizes the reaction between the antibiotic to be tested and a two-dimensional metal olefin modified on the working electrode to undergo an oxidation or reduction reaction, generating a corresponding chemical signal. The concentration information of the antibiotic is obtained by outputting an electrical signal through an electrochemical workstation, thereby achieving amplified detection of the antibiotic.

8. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, The electrochemical detection method includes either cyclic voltammetry or constant voltage method.

9. The antibiotic detection method based on a two-dimensional metalene electrochemical sensor as described in claim 1, characterized in that, The antibiotics tested were tetracycline antibiotics.

10. A method for antibiotic detection based on a two-dimensional metallene electrochemical sensor as described in any one of claims 1-9, characterized in that, The testing methods include environmental monitoring, medical or biological scientific research, and product quality testing.

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