An electrochemiluminescence sensor for high-sensitivity detection of bisphenol a and a construction method thereof
By layer-by-layer modification of glassy carbon electrodes with D-UiO-66@CsPbBr3 and CoOOH@Au materials, and combining them with specific nucleic acid aptamers, an electrochemiluminescence sensor was constructed. This solved the problem of insufficient sensitivity in bisphenol A detection, achieving highly sensitive detection of trace amounts of bisphenol A, and has significant value for environmental and food safety applications.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- THE FIFTH EXPLORATION TEAM OF SHANDONG COALFIELD GEOLOGY BUREAU
- Filing Date
- 2026-06-11
- Publication Date
- 2026-07-24
AI Technical Summary
Existing electrochemiluminescence sensors lack sufficient sensitivity in bisphenol A detection, making it difficult to meet the needs of trace detection.
A glassy carbon electrode was modified layer by layer with D-UiO-66@CsPbBr3 composite material and CoOOH@Au composite material, and an electrochemiluminescence sensor was constructed by combining it with a specific nucleic acid aptamer. CsPbBr3 was used as the substrate luminescent material and CoOOH@Au was used as the ECL energy transfer acceptor to achieve specific recognition and signal recovery of bisphenol A.
It achieves highly sensitive detection of bisphenol A, with good reproducibility and low detection limit, and is suitable for environmental monitoring and food safety fields.
Abstract
Description
Technical Field
[0001] This invention relates to the field of electrochemical technology, specifically to an electrochemiluminescence sensor for highly sensitive detection of bisphenol A and its construction method. Background Technology
[0002] Bisphenol A (BPA) is a typical endocrine disruptor, widely present in environmental media such as surface water, soil, and sediments as an industrial emission. It is significantly toxic to the reproduction and development of both aquatic and terrestrial organisms and can disrupt the balance of ecosystems.
[0003] Compared with traditional detection methods such as liquid chromatography and gas chromatography-mass spectrometry, aptamer sensors based on the principle of electrochemiluminescence (ECL) have many advantages such as high selectivity, high sensitivity and low background noise. However, existing ECL sensors still have shortcomings in terms of the stability of luminescent materials and quenching efficiency, making it difficult to meet the ultrasensitive detection requirements of trace BPA.
[0004] In humans, bisphenol A (BPA) may cause reproductive dysfunction, such as reduced fertility, reproductive organ malformations, and thyroid disorders. The harm is particularly pronounced in sensitive populations such as infants and pregnant women. Given the hazards of BPA, my country has promulgated strict limits, clearly stipulating that the limit for BPA in drinking water is 0.01 mg / L. However, BPA is usually present at trace levels in environmental and biological samples, thus requiring highly sensitive detection methods. Summary of the Invention
[0005] The purpose of this invention is to provide an electrochemiluminescence sensor for highly sensitive detection of bisphenol A and its construction method, so as to solve the problem of insufficient detection sensitivity of BPA in the prior art.
[0006] To achieve the above objectives, the present invention provides the following technical solution: a method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A, comprising the following steps:
[0007] S1. Incubate the D-UiO-66@CsPbBr3 composite material with the first nucleic acid aptamer S1 to obtain the D-UiO-66@CsPbBr3-S1 complex solution.
[0008] S2. The CoOOH@Au composite material with reduced gold nanoparticles on the surface of cobalt hydroxyoxide nanosheets was incubated with the second nucleic acid aptamer S2 of bisphenol A to obtain a CoOOH@Au-S2 complex solution.
[0009] S3. The D-UiO-66@CsPbBr3-S1 composite solution was drop-coated onto the pretreated glassy carbon electrode surface and dried at room temperature to form a film.
[0010] S4. Bovine serum albumin solution was dropped onto the surface of the glassy carbon electrode for sealing treatment, and then it was cleaned and dried.
[0011] S5. Add CoOOH@Au-S2 complex solution to the electrode surface, incubate, clean and dry to obtain the sensor.
[0012] S6. At least two bisphenol A standard solutions of different concentrations are respectively added dropwise to the surface of the electrode obtained in step S5, incubated, washed and dried to obtain an electrochemiluminescence sensor for bisphenol A detection; the concentration is selected from 0.00001-100 ng·mL -1 Different values within the range, and at least two concentrations.
[0013] Further, the preparation of the D-UiO-66@CsPbBr3-S1 composite material in step S1 includes the following steps:
[0014] S11. Zirconium tetrachloride and 1,4-terephthalic acid were ultrasonically dissolved in N,N-dimethylformamide, and acetic acid was added as a defect modifier. After solvothermal reaction, washing and drying, defect-type UiO-66 was obtained.
[0015] S12. Defective UiO-66 was dispersed in anhydrous N,N-dimethylformamide, CsBr and PbBr2 were added and stirred to react, then oleylamine and dodecylamine were added and stirring continued. Toluene was then slowly added dropwise to allow CsPbBr3 quantum dots to be generated in situ within the pores of defective UiO-66. After centrifugation and washing, the mixture was dispersed in buffer solution to obtain the D-UiO-66@CsPbBr3 composite material.
[0016] The mass ratio of D-UiO-66 to CsPbBr3 is 1:1 to 1:5.
[0017] Furthermore, the preparation method of the D-UiO-66@CsPbBr3-S1 complex solution is as follows: the D-UiO-66@CsPbBr3 composite material is dispersed in ethanol, and APTES is added for amination modification to obtain D-UiO-66@CsPbBr3-NH2; then, the 5′-carboxyl-modified first nucleic acid aptamer S1 is activated by EDC / NHS and covalently linked with D-UiO-66@CsPbBr3-NH2 through an amide bond to obtain the D-UiO-66@CsPbBr3-S1 complex solution.
[0018] Further, the preparation of the CoOOH@Au-S2 complex solution in step S2 includes the following steps:
[0019] S21. Mix NaOH solution with CoCl2·6H2O solution, sonicate, add NaClO and continue sonicating, centrifuge, wash and dry to obtain CoOOH nanosheets;
[0020] S22. CoOOH nanosheets were dispersed in ultrapure water, HAuCl4 and polyvinylpyrrolidone were added and stirred, and then sodium citrate and NaBH4 were added in sequence for reduction reaction. The mixture was stirred in the dark, centrifuged, washed and dried to obtain CoOOH@Au dispersion.
[0021] The mass ratio of CoOOH to Au is 1:1 to 5:1.
[0022] S23. The second nucleic acid aptamer S2 of bisphenol A was reduced with TCEP to expose the thiol group, and then mixed with CoOOH@Au dispersion. The mixture was fixed by Au-S covalent bonds, incubated in the dark, centrifuged and washed, and redispersed to obtain the CoOOH@Au-S2 complex solution.
[0023] Furthermore, the sequence of the first nucleic acid aptamer S1 is shown in SEQ ID NO.1, and the sequence of the second nucleic acid aptamer S2 is shown in SEQ ID NO.2.
[0024] Further, the concentration of the D-UiO-66@CsPbBr3-S1 complex solution in step S3 is 2.0-3.0 mg·mL. -1 The drop volume is 5-8 μL.
[0025] Further, the concentration of the CoOOH@Au-S2 complex solution in step S5 is 1.5-3.0 mg·mL. -1 The drop volume is 5-8 μL.
[0026] An application of an electrochemiluminescence sensor prepared using the aforementioned method in the detection of bisphenol A, the detection method comprising the following steps:
[0027] S1. The bisphenol A solution to be tested or a series of bisphenol A standard solutions of different concentrations are dropped onto the surface of the sensor electrode, incubated, cleaned and dried; wherein, the standard solution is used to plot the standard curve, and the concentration range of the standard solution is 0.00001-100 ng·mL⁻¹;
[0028] S2. Using an Ag / AgCl electrode as the reference electrode and a platinum wire as the counter electrode, the constructed electrochemiluminescence sensor was used as the working electrode and connected to a chemiluminescence detector. The photomultiplier tube voltage was 600-750 V, and the test was performed in a PBS buffer solution containing 20-100 mmol·L⁻¹ potassium persulfate at pH 6.0-8.0.
[0029] S3. The bisphenol A standard solution was detected by electrochemiluminescence immunoassay, with a voltage scan range of 0-1.2 V;
[0030] S4. Record the electrochemiluminescence intensity in the presence of different concentrations of bisphenol A. Plot a standard curve with the logarithm of bisphenol A concentration on the x-axis and the electrochemiluminescence intensity on the y-axis to perform qualitative or quantitative analysis of bisphenol A.
[0031] Compared with existing technologies, this invention provides an electrochemiluminescence sensor for highly sensitive detection of bisphenol A and its construction method. Based on the specific binding of aptamers to bisphenol A, it utilizes defective UiO-66 loaded with CsPbBr3 as the substrate luminescent material, and reduces Au nanoparticles on the surface of cobalt hydroxyoxide nanosheets to act as ECL energy transfer acceptors to quench the ECL signal. When bisphenol A is present, the specific recognition of the aptamer causes the energy acceptor to separate from the electrode surface, thereby recovering the ECL signal. The electrochemiluminescence sensor constructed through layer-by-layer modification has good reproducibility, high sensitivity, and low detection limit, enabling rapid and highly sensitive detection of trace amounts of bisphenol A, and has important application value in the fields of environmental monitoring and food safety. Detailed Implementation
[0032] To enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below.
[0033] Example 1:
[0034] A method for fabricating a defect-type UiO-66@CsPbBr3 electrochemiluminescence sensor for highly sensitive detection of bisphenol A:
[0035] 1. Polish a glassy carbon electrode with a diameter of 4 mm using Al2O3 polishing powder, clean it with deionized water, and then add 6 μL of 3.0 mg·mL... -1 The D-UiO-66@CsPbBr3-S1 solution was dropped onto the electrode surface and dried at room temperature to form a film.
[0036] The preparation steps for the solution of defective UiO-66@CsPbBr3 bound to Apt are as follows:
[0037] (1) Preparation of defective UiO-66
[0038] 0.233 g of zirconium tetrachloride and 0.166 g of 1,4-terephthalic acid were ultrasonically dissolved in 50 mL of N,N-dimethylformamide; 60 mmol of acetic acid was added to the above solution to control defects; the mixture was transferred to a 100 mL reactor and reacted in an oven at 120 °C for 24 h; the precipitate was washed three times each with DMF and ethanol, and dried at 60 °C for 12 h to obtain a white precipitate - defect type UiO-66 (D-UiO-66).
[0039] (2) Preparation of D-UiO-66@CsPbBr3
[0040] 100 mg of D-UiO-66 was dispersed in 8 mL of anhydrous N,N-dimethylformamide, followed by the addition of 1.0 mmol / L CsBr and 0.5 mmol / L PbBr2, and stirred at room temperature for 6 h. Then, 0.4 mL of oleylamine and 0.6 mL of dodecylamine were added dropwise, and stirring continued for 1.5 h. Finally, 12 mL of toluene was slowly added dropwise to the mixture, generating CsPbBr3 quantum dots in the D-UiO-66 channels. The quantum dots were washed several times by centrifugation at 9000 rpm with anhydrous toluene, and then redispersed in 10 mL of Tris-HCl buffer (10 mM Tris, 300 mM NaCl, pH 7.4). The mixture was stored at room temperature and protected from light before use to obtain the D-UiO-66@CsPbBr3 composite material.
[0041] (3) Preparation of D-UiO-66@CsPbBr3-S1
[0042] 10 mg of D-UiO-66@CsPbBr3 was dispersed in 10 mL of ethanol, and 100 μL of 1% APTES was added. The mixture was stirred at room temperature for 6 h, then centrifuged and dried under vacuum to obtain D-UiO-66@CsPbBr3-NH2.
[0043] 100 μL of 10 μM 5′-carboxyl-modified S1-DNA (SEQ ID NO.1: 5′-COOHTGGTGCGAACCCGTGATGCGCTGGGCCATACGCGGAACGCTATCCCACCTGACCACCCACCGG-3′) was dissolved in 0.1 M MES buffer (pH 5.5) and brought to a final volume of 500 μL. Then, 3 mg of D-UiO-66@CsPbBr3-NH2, 10 mM EDC, and 25 mM NHS were added. The mixture was stirred at room temperature in the dark for 4 h to achieve covalent linkage via amide bonds, yielding the D-UiO-66@CsPbBr3-S1 complex, which was stored at 4°C in the dark for later use.
[0044] 2. Add 3 μL of 0.1% BSA solution to the electrode surface, incubate for 30 min, rinse with PBS solution at pH 7.4, and air dry at room temperature;
[0045] 3. Add 6 μL of 2.5 mg / mL solution dropwise. -1 CoOOH@Au-Apt solution was used to reduce Au nanoparticles (CoOOH@Au) on the surface of cobalt hydroxyoxide nanosheets and bind bisphenol A aptamers. After incubation for 1.5 h, the nanosheets were rinsed with PBS solution at pH 7.4 and air-dried at room temperature.
[0046] The preparation steps of the CoOOH@Au-Apt solution, which involves reducing Au nanoparticles (CoOOH@Au) on the surface of cobalt hydroxyoxide nanosheets and combining them with a bisphenol A aptamer, are as follows:
[0047] (1) Preparation of CoOOH@Au
[0048] First, 625 μL of 1.0 M NaOH and 2.5 mL of 10.0 mM CoCl2・6H2O were mixed in a beaker and sonicated for 1 min. Then, 125 μL of 0.9 M NaClO was added, and sonication was continued for 15 min. Finally, the mixture was washed with deionized water at 6500 rpm and vacuum dried at 60 °C for 12 h to obtain CoOOH nanosheets.
[0049] 30 mg of CoOOH nanosheets were dispersed in 40 mL of ultrapure water. Then, 1.5 mL of 2% HAuCl4 and 5 mg of polyvinylpyrrolidone (PVP, K30) were added to the above solution and stirred for 5 h. Subsequently, 2 mL of 50 mM sodium citrate and 100 μL of 0.1 M NaBH4 were added dropwise to reduce HAuCl4, and the mixture was stirred in the dark for 12 h. The mixture was centrifuged at 10,000 rpm for 10 min, washed three times with ultrapure water to remove unbound gold, and dried under vacuum at 60 °C for 12 h to obtain CoOOH@Au.
[0050] (2) Preparation of CoOOH@Au bisphenol A aptamer-bound solution
[0051] The BPA aptamer (SEQ ID NO.2: 5′-SHCCGGTGGGTGGTCAGGTGGGATAGCGTTCCGCGTATGGCCCAGCGCATCACGGGTTCGCACCA-3′) was first reduced with 10 mM TCEP at room temperature for 30 min to expose the thiol groups. 5 mg CoOOH@Au was dispersed in 2 mL PBS (pH 7.4), and 100 μL of the pretreated 10 μg·mL⁻¹ thiol aptamer was added, immobilizing it on the CoOOH@Au surface via Au-S covalent bonds. The mixture was incubated at 4 °C in the dark for 24 h. Unbound aptamers were removed by centrifugation and washing, and the precipitate was redissolved in PBS to obtain the CoOOH@Au-Apt solution.
[0052] 4. Add 0.00001-100 ng·mL dropwise. -1 A series of different concentrations of BPA were applied to the electrode surface, incubated for 1.5 h, rinsed with PBS solution at pH 7.4, and air-dried at room temperature to prepare an electrochemiluminescent biosensor.
[0053] Example 2:
[0054] A method for fabricating a defect-type UiO-66@CsPbBr3 electrochemiluminescence sensor for highly sensitive detection of bisphenol A:
[0055] 1. Polish a glassy carbon electrode with a diameter of 4 mm using Al2O3 polishing powder, clean it with deionized water, and then add 8 μL of 2.5 mg·mL... -1 The D-UiO-66@CsPbBr3-S1 solution was dropped onto the electrode surface and dried at room temperature to form a film.
[0056] 2. Add 3 μL of 0.1% BSA solution to the electrode surface, incubate for 30 min, rinse with PBS solution at pH 7.4, and air dry at room temperature;
[0057] 3. Add 7 μL of 1.5 mg / mL solution dropwise. -1 CoOOH@Au-Apt solution was used to reduce Au nanoparticles (CoOOH@Au) on the surface of cobalt hydroxyoxide nanosheets and bind bisphenol A aptamers. After incubation for 1.5 h, the mixture was rinsed with PBS solution at pH 7.4 and air-dried at room temperature.
[0058] 4. Add 0.00001-100 ng·mL dropwise. -1 A series of different concentrations of BPA were applied to the electrode surface, incubated for 1.5 h, rinsed with PBS solution at pH 7.4, and air-dried at room temperature to prepare an electrochemiluminescent biosensor.
[0059] Example 3:
[0060] A method for fabricating a defect-type UiO-66@CsPbBr3 electrochemiluminescence sensor for highly sensitive detection of bisphenol A:
[0061] 1. Polish a glassy carbon electrode with a diameter of 4 mm using Al2O3 polishing powder, clean it with deionized water, and then add 5 μL of 2.0 mg·mL... -1 The D-UiO-66@CsPbBr3-S1 solution was dropped onto the electrode surface and dried at room temperature to form a film.
[0062] 2. Add 3 μL of 0.1% BSA solution to the electrode surface, incubate for 30 min, rinse with PBS solution at pH 7.4, and air dry at room temperature;
[0063] 3. Add 8 μL of 3.0 mg / mL solution. -1 CoOOH@Au-Apt solution was used to reduce Au nanoparticles (CoOOH@Au) on the surface of cobalt hydroxyoxide nanosheets and bind bisphenol A aptamers. After incubation for 1.5 h, the mixture was rinsed with PBS solution at pH 7.4 and air-dried at room temperature.
[0064] 4. Add 0.00001-100 ng·mL dropwise. -1 A series of different concentrations of BPA were applied to the electrode surface, incubated for 1.5 h, rinsed with PBS solution at pH 7.4, and air-dried at room temperature to prepare an electrochemiluminescent biosensor.
[0065] Example 4:
[0066] Detection of Bisphenol A
[0067] 1. Using Ag / AgCl as the reference electrode and platinum wire as the counter electrode, the prepared electrochemiluminescence sensor was correctly connected as the working electrode in the dark box of the chemiluminescence detector. The electrochemical workstation and the chemiluminescence detector were connected together, and the high voltage of the photomultiplier tube was set to 600 V. The atmosphere contained 20 mmol·L⁻¹ -1 The test was conducted in potassium persulfate and PBS buffer solution at pH 6.
[0068] 2. The bisphenol A standard solution was detected using an electrochemiluminescence method with a voltage test range of 0-1.2 V;
[0069] 3. Observe the electrochemiluminescence intensity of the sensor before and after the addition of bisphenol A, then record the linear relationship between the electrochemiluminescence intensity value and the concentration of bisphenol A, and plot the working curve.
[0070] Example 5:
[0071] Detection of Bisphenol A
[0072] (1) Using Ag / AgCl as the reference electrode and platinum wire as the counter electrode, the prepared electrochemiluminescence sensor was correctly connected to the dark box of the chemiluminescence detector as the working electrode. The electrochemical workstation and the chemiluminescence detector were connected together, and the high voltage of the photomultiplier tube was set to 750 V. In a solution containing 60 mmol·L -1 The test was conducted using potassium persulfate in PBS buffer solution at pH 6.5.
[0073] (2) The bisphenol A standard solution was detected by electrochemiluminescence method, and the voltage test range was 0-1.2 V;
[0074] (3) Observe the electrochemiluminescence intensity of the sensor before and after the addition of bisphenol A, and then record the linear relationship between the electrochemiluminescence intensity value and the concentration of bisphenol A, and plot the working curve.
[0075] Example 6:
[0076] Detection of Bisphenol A
[0077] (1) Using Ag / AgCl as the reference electrode and platinum wire as the counter electrode, the prepared electrochemiluminescence sensor was correctly connected to the dark box of the chemiluminescence detector as the working electrode. The electrochemical workstation and the chemiluminescence detector were connected together, and the high voltage of the photomultiplier tube was set to 750 V. In a solution containing 80 mmol·L -1 The test was conducted in potassium persulfate and PBS buffer solution at pH 7.
[0078] (2) The bisphenol A standard solution was detected by electrochemiluminescence method, and the voltage test range was 0-1.2 V;
[0079] (3) Observe the electrochemiluminescence intensity of the sensor before and after the addition of bisphenol A, and then record the linear relationship between the electrochemiluminescence intensity value and the concentration of bisphenol A, and plot the working curve.
[0080] Example 7:
[0081] Detection of Bisphenol A
[0082] (1) Using Ag / AgCl as the reference electrode and platinum wire as the counter electrode, the prepared electrochemiluminescence sensor was correctly connected to the dark box of the chemiluminescence detector as the working electrode. The electrochemical workstation and the chemiluminescence detector were connected together, and the high voltage of the photomultiplier tube was set to 750 V. In a solution containing 80 mmol·L -1 The test was conducted in potassium persulfate and PBS buffer solution at pH 7.4.
[0083] (2) The bisphenol A standard solution was detected by electrochemiluminescence method, and the voltage test range was 0-1.2 V;
[0084] (3) Observe the electrochemiluminescence intensity of the sensor before and after the addition of bisphenol A, and then record the linear relationship between the electrochemiluminescence intensity value and the concentration of bisphenol A, and plot the working curve.
[0085] Example 8:
[0086] Detection of Bisphenol A
[0087] (1) Using Ag / AgCl as the reference electrode and platinum wire as the counter electrode, the prepared electrochemiluminescence sensor was correctly connected to the dark box of the chemiluminescence detector as the working electrode. The electrochemical workstation and the chemiluminescence detector were connected together, and the high voltage of the photomultiplier tube was set to 750 V. In a solution containing 100 mmol·L -1 The test was conducted in potassium persulfate and PBS buffer solution at pH 8.
[0088] (2) The bisphenol A standard solution was detected by electrochemiluminescence method, and the voltage test range was 0-1.2 V;
[0089] (3) Observe the electrochemiluminescence intensity of the sensor before and after the addition of bisphenol A, and then record the linear relationship between the electrochemiluminescence intensity value and the concentration of bisphenol A, and plot the working curve.
[0090] The foregoing has only described certain exemplary embodiments of the present invention by way of illustration. Undoubtedly, those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the above description is illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.
Claims
1. A method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A, characterized in that, Includes the following steps: S1. Incubate the D-UiO-66@CsPbBr3 composite material with the first nucleic acid aptamer S1 to obtain the D-UiO-66@CsPbBr3-S1 complex solution. S2. The CoOOH@Au composite material with reduced gold nanoparticles on the surface of cobalt hydroxyoxide nanosheets was incubated with the second nucleic acid aptamer S2 of bisphenol A to obtain a CoOOH@Au-S2 complex solution. S3. The D-UiO-66@CsPbBr3-S1 composite solution was drop-coated onto the pretreated glassy carbon electrode surface and dried at room temperature to form a film. S4. Bovine serum albumin solution was dropped onto the surface of the glassy carbon electrode for sealing treatment, and then it was cleaned and dried. S5. Add CoOOH@Au-S2 complex solution to the electrode surface, incubate, then wash and dry. S6. At least two bisphenol A standard solutions of different concentrations are respectively added dropwise to the surface of the electrode obtained in step S5, incubated, washed and dried to obtain an electrochemiluminescence sensor for bisphenol A detection; the concentration is selected from 0.00001-100 ng·mL -1 Different values within the range, and at least two concentrations.
2. The method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A according to claim 1, characterized in that, The preparation of the D-UiO-66@CsPbBr3-S1 composite material in step S1 includes the following steps: S11. Zirconium tetrachloride and 1,4-terephthalic acid were ultrasonically dissolved in N,N-dimethylformamide, and acetic acid was added as a defect modifier. After solvothermal reaction, washing and drying, defect-type UiO-66 was obtained. S12. Defective UiO-66 was dispersed in anhydrous N,N-dimethylformamide, CsBr and PbBr2 were added and stirred to react, then oleylamine and dodecylamine were added and stirring continued. Toluene was then slowly added dropwise to allow CsPbBr3 quantum dots to be generated in situ within the pores of defective UiO-66. After centrifugation and washing, the mixture was dispersed in buffer solution to obtain the D-UiO-66@CsPbBr3 composite material. The mass ratio of D-UiO-66 to CsPbBr3 is 1:1 to 1:
5.
3. The method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A according to claim 2, characterized in that, The preparation method of the D-UiO-66@CsPbBr3-S1 complex solution is as follows: the D-UiO-66@CsPbBr3 composite material is dispersed in ethanol, and APTES is added for amination modification to obtain D-UiO-66@CsPbBr3-NH2; then, the 5′-carboxyl-modified first nucleic acid aptamer S1 is activated by EDC / NHS and covalently linked with D-UiO-66@CsPbBr3-NH2 through an amide bond to obtain the D-UiO-66@CsPbBr3-S1 complex solution.
4. The method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A according to claim 1, characterized in that, The preparation of the CoOOH@Au-S2 complex solution in step S2 includes the following steps: S21. Mix NaOH solution with CoCl2·6H2O solution, sonicate, add NaClO and continue sonicating, centrifuge, wash and dry to obtain CoOOH nanosheets; S22. CoOOH nanosheets were dispersed in ultrapure water, HAuCl4 and polyvinylpyrrolidone were added and stirred, and then sodium citrate and NaBH4 were added in sequence for reduction reaction. The mixture was stirred in the dark, centrifuged, washed and dried to obtain CoOOH@Au dispersion. The mass ratio of CoOOH to Au is 1:1 to 5:
1. S23. The second nucleic acid aptamer S2 of bisphenol A was reduced with TCEP to expose the thiol group, and then mixed with CoOOH@Au dispersion. The mixture was fixed by Au-S covalent bonds, incubated in the dark, centrifuged and washed, and redispersed to obtain the CoOOH@Au-S2 complex solution.
5. The method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A according to claim 1, characterized in that, The sequence of the first nucleic acid aptamer S1 is shown in SEQ ID NO.1, and the sequence of the second nucleic acid aptamer S2 is shown in SEQ ID NO.
2.
6. The method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A according to claim 1, characterized in that, The concentration of the D-UiO-66@CsPbBr3-S1 complex solution in step S3 is 2.0-3.0 mg·mL. -1 The drop volume is 5-8 μL.
7. The method for constructing an electrochemiluminescence sensor for highly sensitive detection of bisphenol A according to claim 1, characterized in that, The concentration of the CoOOH@Au-S2 complex solution in step S5 is 1.5-3.0 mg·mL. -1 The drop volume is 5-8 μL.
8. The application of an electrochemiluminescence sensor prepared by the construction method according to any one of claims 1 to 7 in the detection of bisphenol A, characterized in that, The detection method includes the following steps: S1. The bisphenol A solution to be tested or a series of bisphenol A standard solutions of different concentrations are dropped onto the surface of the sensor electrode, incubated, cleaned and dried; wherein, the standard solution is used to plot the standard curve, and the concentration range of the standard solution is 0.00001-100 ng·mL⁻¹; S2. Using an Ag / AgCl electrode as the reference electrode and a platinum wire as the counter electrode, the constructed electrochemiluminescence sensor was used as the working electrode and connected to a chemiluminescence detector. The photomultiplier tube voltage was 600-750 V, and the test was performed in a PBS buffer solution containing 20-100 mmol·L⁻¹ potassium persulfate at pH 6.0-8.
0. S3. The bisphenol A standard solution was detected by electrochemiluminescence immunoassay, with a voltage scan range of 0-1.2 V; S4. Record the electrochemiluminescence intensity in the presence of different concentrations of bisphenol A. Plot a standard curve with the logarithm of bisphenol A concentration on the x-axis and the electrochemiluminescence intensity on the y-axis to perform qualitative or quantitative analysis of bisphenol A.