CN116203092A - Preparation method and detection method of electrochemical sensor for detecting di (2-ethylhexyl) phthalate - Google Patents

Preparation method and detection method of electrochemical sensor for detecting di (2-ethylhexyl) phthalate Download PDF

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CN116203092A
CN116203092A CN202310208965.9A CN202310208965A CN116203092A CN 116203092 A CN116203092 A CN 116203092A CN 202310208965 A CN202310208965 A CN 202310208965A CN 116203092 A CN116203092 A CN 116203092A
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ethylhexyl
aptamer
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陈智栋
董美华
蒋鼎
王文昌
贾树勇
徐仿敏
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Changzhou University
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Abstract

The invention relates to the field of electrochemiluminescence detection, in particular to a preparation method and a detection method of an electrochemiluminescence sensor for detecting di (2-ethylhexyl) phthalate. The technical key points are as follows: the electrochemical luminescence aptamer sensor is formed by loading carboxylated ligand on the surface of a composite material Zr-MOF/GDY modified glassy carbon electrode; wherein the Zr-MOF/GDY composite material is formed by electrostatic interaction between Zr-MOF and GDY; carboxylated aptamers are: an aptamer comprising a base sequence of 5'-GGGTAGGGCGGGAAGTTACTGTCTTACTGTCGTA-3'. The detection method realizes the detection of the DEHP through the recovery effect of the DEHP on the ECL signal intensity of the electrochemical luminescence aptamer sensor of the GDY and Zr-MOF composite material, and has the advantages of simple operation, good selectivity and high sensitivity.

Description

Preparation method and detection method of electrochemical sensor for detecting di (2-ethylhexyl) phthalate
Technical Field
The invention relates to the field of electrochemiluminescence detection, in particular to a preparation method and a detection method of an electrochemiluminescence sensor for detecting di (2-ethylhexyl) phthalate.
Background
Bis (2-ethylhexyl) phthalate (DEHP) is the most widely used plasticizer in the production of plastics articles, producing about 100 to 400 tens of thousands of tons per year. Since DEHP molecules are linked to plastic polymers by purely physical bonds rather than chemical bonds, they are released from the microplastic into the water and soil environment after undergoing weathering and photolysis, further into the human body through the food chain. DEHP, an endocrine disrupter, can cause endocrine disorders in humans, induce cancer or cause reproductive toxicity. Has been currently classified as an environmentally preferred contaminant by the U.S. Environmental Protection Agency (EPA) and the ecological environment of the people's republic of China. For public health safety, there is an urgent need to establish a rapid and convenient detection method to monitor DEHP in an aqueous environment.
The DEHP detection methods currently used include gas chromatography and high performance liquid chromatography (GC-HPLC), gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and gas chromatography flame ionization detection (GC-FID), requiring complex pretreatment steps and specialized technicians. The detection method is tedious and can not obtain the detection result rapidly.
In view of the above-mentioned drawbacks of the existing detection methods, the present inventors have studied and innovated based on the years of experience and expertise in such materials, in combination with theoretical analysis, and developed a method for preparing and detecting an electrochemical sensor for detecting di (2-ethylhexyl) phthalate.
Disclosure of Invention
The invention aims to develop a preparation method of an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate (DEHP), which utilizes Zr-MOF/GDY nanocomposite material to modify the surface of a glassy carbon electrode to obtain a Zr-MOF/GDY/GCE modified electrode, and improves detection sensitivity through high signal to noise ratio, so that the aim of trace detection of a target object can be achieved, the sensitivity and stability of electrochemical luminescence of the di (2-ethylhexyl) phthalate (DEHP) detection are obviously improved, and the preparation method has the advantages of high sensitivity, low background, easiness in control, strong specificity, short detection time and the like.
The technical aim of the invention is realized by the following technical scheme:
in the preparation method of the electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate, the electrochemical luminescence aptamer sensor is formed by loading carboxylated ligand on the surface of a composite material Zr-MOF/GDY modified glassy carbon electrode;
wherein the Zr-MOF/GDY composite material is formed by electrostatic interaction between Zr-MOF and GDY;
carboxylated aptamers are:
an aptamer comprising a base sequence of 5'-GGGTAGGGCGGGAAGTTACTGTCTTACTGTCGTA-3'.
Further, the preparation method provided by the invention specifically comprises the following operation steps:
dispersing GDY in water to obtain solution 1; dispersing Zr-MOF in DMF to obtain solution 2; dropping the solution 1 on the surface of the glassy carbon electrode; dropping the solution 2 on the surface of a GDY/glassy carbon electrode; naturally airing to obtain a Zr-MOF/GDY modified glassy carbon electrode; by NH 2 And (3) carrying out dehydration condensation reaction with COOH to load carboxylated aptamer on the surface of a glassy carbon electrode modified by a composite material Zr-MOF/GDY, and naturally airing to prepare the electrochemiluminescence aptamer sensor.
In the present invention, NH is used 2 Besides dehydration condensation reaction with COOH, electrostatic adsorption effect exists between carboxylated aptamer and composite material, the carboxylated aptamer is loaded on the surface of a glassy carbon electrode modified by Zr-MOF/GDY composite material, and after incubation for 6 hours at room temperature, an electrochemiluminescence aptamer sensor (COOH-apt/Zr-MOF/GDY/GCE)。
Further, the Zr-MOF preparation method comprises the following steps: zrCl is added to 4 And lauric acid is dissolved in an organic solvent, 2-amino terephthalic acid is added after ultrasonic mixing is uniform, and solvent thermal synthesis reaction is carried out after ultrasonic mixing to obtain Zr-MOF.
Further, the preparation method of GDY specifically comprises the following steps: mixing and stirring hexaalkyl- [ (trimethylsilyl) ethyl ] benzene, tetrahydrofuran (THF) solution and tetrabutylammonium fluoride to synthesize hexaethylbenzene monomer, and adopting hexaethylbenzene monomer cross coupling reaction to grow GDY film on the surface of the copper foil; the GDY film was washed and dried to give GDY powder.
Further, the concentrations of the solution 1 and the solution 2 were 1mg/L, and the amounts of the dispersions were 5. Mu.L.
Further, NH 2 The dehydration condensation reaction with COOH is specifically: first to contain KCl, naCl, mgCl 2 And adding carboxylated aptamer into Tris-HCl buffer solution of ethylenediamine tetraacetic acid to prepare an aptamer solution with the concentration of the aptamer of 1-10 mu M, and then transferring the aptamer solution and dripping the aptamer solution on the surface of the glassy carbon electrode modified by the composite material Zr-MOF/GDY.
Further, the concentration of the aptamer in the aptamer solution is 3. Mu.M, which is convenient to ensure better electrochemiluminescence intensity.
Further, the carboxylated aptamer is a carboxylated aptamer comprising
An aptamer of a 5'-COOH-ACGCATAGGGTGCGACCACATACGCCCCATGTATGTCCCTTGGTTGTGCCCTATGCGT-3' base sequence.
The second object of the invention is to provide a detection method of an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate, which successfully realizes the sensitive detection of DEHP through the recovery effect of DEHP on ECL signal intensity of an electrochemical luminescence aptamer sensor of a composite material of graphite alkyne GDY and a zirconium-based metal-organic framework Zr-MOF, and has the advantages of simple operation, good selectivity, high sensitivity and wide detection range.
The technical aim of the invention is realized by the following technical scheme:
the invention provides a detection method of an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate, which is characterized in that the electrochemical luminescence sensor is used as a working electrode, ag/AgCl is used as a reference electrode, a platinum wire electrode is used as a counter electrode to form a three-electrode system, and detection is completed through generated electrochemical luminescence signals.
Specifically, the detection method comprises the following operation steps:
the COOH-apt/Zr-MOF/GDY/GCE electrochemical luminescence aptamer sensor is used as a working electrode, ag/AgCl is used as a reference electrode, a platinum wire electrode is used as a counter electrode to form a three-electrode system, and di (2-ethylhexyl) phthalate in a sample to be detected is quantitatively captured to the surface of the sensor, and detection is realized through a generated luminescence signal.
Further, the detection method provided by the invention comprises the following specific operations:
s1, contain K 2 S 2 O 8 Preparing PBS nano bubble buffer solution;
s2, preparing standard solutions of di (2-ethylhexyl) phthalate with different concentrations;
s3, drawing a standard curve, respectively soaking the electrochemiluminescence aptamer sensor in the standard solution prepared in the same amount of step S2 and reacting for the same time, so that the electrochemiluminescence aptamer sensor is combined with the di (2-ethylhexyl) phthalate to obtain DEHP/COOH-apt/Zr-MOF/GDY/GCE, then taking the DEHP/COOH-apt/Zr-MOF/GDY/GCE as a working electrode, ag/AgCl as a reference electrode, and a platinum electrode as a counter electrode to form a three-electrode system, wherein K-containing in the step S1 is used for preparing the three-electrode system 2 S 2 O 8 Performing cyclic voltammetry scanning by using the PBS nano bubble buffer solution as electrolyte, recording a luminous intensity-time curve, and establishing a linear relation between luminous intensity before and after the electrochemical luminous aptamer sensor is combined with the di (2-ethylhexyl) phthalate and the concentration logarithmic value of the di (2-ethylhexyl) phthalate in the di (2-ethylhexyl) phthalate standard solution to obtain a corresponding linear regression equation;
s4, detecting the di (2-ethylhexyl) phthalate in the sample to be detected, directly taking out a proper amount of bottled beverage from the container under the condition of no pretreatment, reacting the bottled beverage with the surface of the electrochemiluminescence aptamer sensor for the same time according to the step S3, enabling the electrochemiluminescence aptamer sensor to be combined with the di (2-ethylhexyl) phthalate, then taking the electrochemiluminescence aptamer sensor as a working electrode, detecting the luminous intensity by adopting the method of the step S3, and then calculating the concentration of the di (2-ethylhexyl) phthalate in the sample to be detected according to a linear regression equation.
Furthermore, the condition of cyclic voltammetry scanning is that the photomultiplier is carried out at high voltage of 800V within the electrochemical window range of-2.0-0V, and the scanning speed is 0.1V/s.
Further, in step S3, the electrochemical luminescence aptamer sensor is immersed in the standard solution prepared in the same amount of step S2 for a reaction time of 20min.
In summary, the invention has the following beneficial effects:
the invention designs an electrochemiluminescence aptamer sensor based on a graphite alkyne GDY and zirconium-based metal organic framework Zr-MOF composite material, and the two materials are combined through electrostatic interaction, so that high-efficiency and stable electrochemiluminescence performance can be obtained. The invention fully utilizes the unique advantages of the aptamer and the electrochemical luminescence sensor, successfully realizes the sensitive detection of the DEHP through the recovery effect of the DEHP on the ECL signal intensity of the system, and experiments prove that the sensing platform can specifically identify the detection object DEHP.
The detection range of the invention is 1.0X10 -12 ～1.0×10 -4 g/L, the lowest detection limit is 2.53 multiplied by 10 -13 The method for detecting DEHP is simple to operate, good in selectivity, high in sensitivity and wide in detection range, and has important significance for popularization of application of the aptamer sensor in actual detection.
Drawings
FIG. 1 is a graph showing the ECL response of the electrochemiluminescence aptamer sensor constructed in example 1 after binding to different concentrations of DEHP;
FIG. 2 is a plot of the difference in luminescence intensity (ΔECL) versus the DEHP concentration vs. value for example 1 before and after DEHP addition;
FIG. 3 is a scanning electron microscope image of the Zr-MOF/GDY composite material prepared in example 1;
FIG. 4 is a graph of ECL response over time for Zr-MOF/GDY modified glassy carbon electrodes;
FIG. 5 is a graph of ECL response versus time for GDY modified glassy carbon electrodes;
FIG. 6 is a graph of ECL response over time for Zr-MOF modified glassy carbon electrodes;
FIG. 7 is a graph showing the ECL response of COOH-apt/Zr-MOF/GDY modified glassy carbon electrodes over time.
Detailed Description
In order to further illustrate the technical means and effects adopted by the invention to achieve the preset aim, the invention provides a preparation method of an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate and a detection method thereof, and specific embodiments, characteristics and effects thereof are described in detail below.
In the present embodiment, it comprises
The aptamer of the 5'-COOH-ACGCATAGGGTGCGACCACATACGCCCCATGTATGTCCCTTGGTTGTGCCCTATGCGT-3' base sequence was purchased from the division of biological engineering (Shanghai);
the concentration of DEHP in the DEHP standard solution was (a) 1.0X10, respectively -4 g/L；(b) 1.0×10 -5 g/L；(c) 1.0×10 -6 g/L；(d) 1.0×10 -7 g/L；(e) 1.0×10 -8 g/L；(f) 1.0×10 -9 g/L；(g) 1.0×10 -10 g/L；(h) 1.0×10 -11 g/L；(i) 1.0×10 -12 g/L。
Example 1: preparation method and detection method of electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate
Preparing an electrochemical luminescence sensor:
s1, preparation of Zr-MOF:
zirconium tetrachloride (80 mg,0.34 mmol) and lauric acid (2.4 g,12 mmol) were dissolved in 20 mL of DMF, the mixture was vigorously sonicated at room temperature for 20min, 2-amino terephthalic acid (31 mg,0.17 mmmol) was added to the above solution, ultrasonic mixing was continued for 5 min, and the resulting homogeneous mixture was transferred to an autoclave for reaction at 120 ℃ for 12h, the yellow precipitate was washed 3 times with DMF and ethanol, respectively, and centrifuged at 12000rpm, washed, and finally, vacuum dried at room temperature;
s2, GDY preparation of materials:
hexaethylbenzene is used as a precursor, copper foil is used as a catalyst and a substrate, the hexaethylbenzene is synthesized through cross coupling reaction, hexaalkyl- [ (trimethylsilyl) ethyl ] benzene and Tetrahydrofuran (THF) solution are mixed and stirred with tetrabutylammonium fluoride (TBAF) for 10min at 8 ℃, hexaethylbenzene monomer is synthesized, the hexaethylbenzene monomer is adopted to carry out cross coupling reaction under the nitrogen atmosphere condition at 60 ℃, a reaction time of 72h, GDY film grows on the surface of the copper foil, the film is subjected to ultrasonic treatment and is washed by acetone, then hot dimethylformamide (DMF; 80 ℃) is used for ultrasonic treatment for 1h, black solid is peeled off, the solid is sequentially refluxed in NaOH (4M), HCl (6M) and NaOH (4M) at 100 ℃ for 2h, residual copper and other impurities are removed, the product is washed and centrifugally collected through hot DMF (80 ℃) and hot ethanol (70 ℃), and dried in a vacuum oven for overnight to obtain GDY crystalline black powder;
s3, dispersing the Zr-MOF obtained in the step S1 in DMF, carrying out ultrasonic treatment to uniformly disperse the Zr-MOF to obtain a solution 1, dispersing GDY in ultrapure water, carrying out ultrasonic treatment to uniformly disperse the GDY to obtain a solution 2, wherein the concentration of the dispersion liquid of the Zr-MOF and the ultrapure water is 1mg/mL;
s4, preparing an electrochemical luminescence aptamer sensor for detecting DEHP, and polishing powder (Al 2 O 3 ) Polishing the chamois leather into a mirror surface, sequentially ultrasonically cleaning the mirror surface by using a nitric acid solution, an ethanol solution and ultrapure water, drying the mirror surface at room temperature to obtain a pretreated glassy carbon electrode, transferring 5 mu L of the solution 2 prepared in the step S3 by using a microinjector, dripping 5 mu L of the solution 1 prepared in the step S3 on the surface of the glassy carbon electrode after naturally drying the mirror surface, modifying 5 mu L of the prepared Tris-HCl buffer solution containing carboxylated aptamer, and incubating the mirror surface at room temperature for 6 hours to obtain the COOH-apt/Zr-MOF/GDY/GCE sensor serving as a sensing element for an electrochemiluminescence test.
Wherein the concentration of carboxylated aptamer in the aptamer solution is 3 μm.
Method for detecting DEHP based on COOH-apt/Zr-MOF/GDY/GCE sensor
A1, drawing a standard curve
Immersing the prepared electrochemiluminescence aptamer sensor in equal amounts of DEHP standard solutions with different concentrations and reacting for the same time to enable the electrochemiluminescence aptamer sensor to be combined with the DEHP, then taking the DEHP/COOH-apt/Zr-MOF/GDY/GCE as a working electrode, ag/AgCl as a reference electrode and a platinum electrode as a counter electrode to form a three-electrode system, wherein the pH value of the three-electrode system is 7.4 and the three-electrode system contains 0.1mol/LK 2 S 2 O 8 In the method, PBS nano bubble buffer solution is taken as electrolyte, in the electrochemical window range of-2.0-0V, a photomultiplier tube is subjected to high pressure 800V and scanning speed 0.1V/s, cyclic voltammetry scanning is carried out, a luminous intensity-time curve is recorded, and a linear relation between a luminous intensity difference value (delta ECL) before and after the electrochemical luminous aptamer sensor is combined with DEHP and a DEHP concentration logarithmic value in a DEHP standard solution is established, so that a corresponding linear regression equation is obtained; ECL=12945.1099+1002.9124 lgC (mg/mL), detection range is 1.0X10 -12 ～1.0×10 -4 g/L, detection limit of 2.53×10 -13 mg/mL；
A2, detection of sample
Adding 0.1 ng.L into river water sample -1 Immersing the prepared electrochemical luminescence aptamer sensor in the solution, and calculating the concentration of DEHP in the sample to be detected according to the linear regression equation obtained in the step A1, wherein the result is shown in Table 1;
in the embodiment, zr-MOF/GDY is used as a base material (the morphology is shown in figure 4), electrostatic interaction between the Zr-MOF and GDY is utilized to stably combine, the electrochemiluminescence intensity of the independent material can be greatly improved, the conductivity is good, the stability is good, and the sensor selectivity is good.
Comparative example 1
Preparing an electrochemical luminescence sensor:
transferring 5 mu L of the solution 1 prepared in the embodiment 1 by using a micro-sample injector, dripping the solution onto the surface of a pretreated glassy carbon electrode (the pretreatment method is the same as that of the embodiment 1), naturally airing, dripping 5 mu L of 3 mu M carboxylation aptamer on the surface of the Zr-MOF/GCE chemically modified electrode, and incubating at room temperature for 6 hours to obtain the COOH-apt/Zr-MOF/GCE sensor serving as a sensing element for electrochemiluminescence testing.
Detection of DEHP based on COOH-apt/Zr-MOF/GCE sensor
A1, drawing a standard curve
The prepared COOH-apt/Zr-MOF/GCE sensor is used as a sensing element, equivalent DEHP standard solutions with different concentrations are modified on the surface of the sensor and react for 30 min, then the sensor is used as a working electrode, ag/AgCl is used as a reference electrode, a platinum electrode is used as a counter electrode, a three-electrode system is formed, and the sensor contains 0.05mol/L K 2 S 2 O 8 And (3) taking 0.1mol/L PBS buffer solution with the pH value of 7.4 as electrolyte to measure the luminous intensity, carrying out cyclic voltammetry scanning on the luminous intensity-time curve in the electrochemical window range of-2.0-0V at the high pressure of a photomultiplier tube of 800V and the scanning speed of 0.1V/s, and establishing the linear relation between the luminous intensity difference value before and after the electrochemical luminous aptamer sensor is combined with DEHP and the DEHP concentration logarithmic value in the DEHP standard solution to obtain a corresponding linear regression equation.
A2, detection of sample
Adding 0.1 ng.L into river water sample -1 DEHP the prepared electrochemiluminescence aptamer sensor was immersed in the above solution, and the concentration of DEHP in the sample to be tested was calculated according to the linear regression equation obtained in step A1, and the results are shown in table 1.
Comparative example 2:
preparation of electrochemical sensor
Transferring 5 mu L of the solution 2 prepared in the embodiment 1 by using a microinjector, dripping the solution onto the surface of a pretreated glassy carbon electrode (the pretreatment method is the same as that of the embodiment 1), dripping 5 mu L of 3 mu M carboxylated aptamer on the surface of a GDY/GCE chemically modified electrode, and naturally airing to obtain a COOH-apt/GDY/GCE sensor serving as a sensing element for electrochemiluminescence test. (monomer test modification and sample concentration and examples maintain a single variable).
Detection of DEHP based on COOH-apt/GDY/GCE sensor
A1, drawing a standard curve
The prepared COOH-apt/GDY/GCE sensor is used as a sensing element, and equal amounts of DEHP standard solutions with different concentrations are modified on a sensor tableThe reaction is carried out for 30 min, then the reaction is carried out by taking the reaction product as a working electrode, ag/AgCl as a reference electrode and a platinum electrode as a counter electrode to form a three-electrode system, and the reaction product contains 0.1mol/L K 2 S 2 O 8 The luminous intensity is measured by taking 0.1mol/L PBS buffer solution with the pH value of 7.4 as electrolyte, the high pressure of a photomultiplier tube is 800 and V within the electrochemical window range of-2.0-0V, the scanning speed is 0.1V/s, cyclic voltammetry scanning is carried out, a luminous intensity-time curve is recorded, and a linear relation between the luminous intensity difference value before and after the electrochemical luminous sensor is combined with DEHP and the DEHP concentration logarithmic value in the DEHP standard solution is established, so that a corresponding linear regression equation is obtained.
A2, detection of sample
Adding 0.1 ng.L into river water sample -1 DEHP. The prepared electrochemical luminescence aptamer sensor was immersed in the above solution, and the concentration of DEHP in the sample to be detected was calculated according to the linear regression equation obtained in step A1, and the results are shown in table 1.
Comparative example 3:
preparation of COOH-apt/Zr-MOF@GDY/GCE sensor
And 5 mu L of the solution 1 and the solution 2 prepared in the embodiment 1 are respectively removed by a microinjector and dripped on the surface of a pretreated glassy carbon electrode (the pretreatment method is the same as that of the embodiment 1), so as to obtain a Zr-MOF@GDY/GCE chemically modified electrode, after the electrode is naturally dried, 5 mu L of 3 mu M carboxylated aptamer is dripped on the surface of the Zr-MOF@GDY/GCE chemically modified electrode, and after the electrode is incubated for 6 hours at room temperature, a COOH-apt/Zr-MOF@GDY/GCE sensor is obtained and is used as a sensing element for an electrochemiluminescence test. (monomer test modification and sample concentration and examples maintain a single variable).
Detection of DEHP based on COOH-apt/Zr-MOF@GDY/GCE sensor
A1, drawing a standard curve
Soaking the COOH-apt/Zr-MOF@GDY/GCE sensor prepared in the step A1 serving as a sensing element in equal amounts of DEHP standard solutions with different concentrations and reacting for 30 min, taking the sensor as a working electrode, taking Ag/AgCl as a reference electrode and taking a platinum electrode as a counter electrode to form a three-electrode system, wherein the three-electrode system contains 0.1mol/L K 2 S 2 O 8 Is electrically charged to a PBS buffer of pH 7.4 at 0.1mol/LAnd (3) determining the luminous intensity by solution, carrying out cyclic voltammetry scanning at a high pressure of 800V and a scanning speed of 0.1V/s in an electrochemical window range of-2.0-0V, recording a luminous intensity-time curve, and establishing a linear relation between the luminous intensity difference value of the electrochemical luminous aptamer sensor before and after the electrochemical luminous aptamer sensor is combined with DEHP and the DEHP concentration logarithmic value in the DEHP standard solution to obtain a corresponding linear regression equation.
A2, detection of sample
Adding 0.1 ng.L to river water sample -1 DEHP the prepared electrochemiluminescence aptamer sensor was immersed in the above solution, and the concentration of DEHP in the sample to be tested was calculated according to the linear regression equation obtained in step A1, and the results are shown in table 1.
TABLE 1 test for detecting DEHP in certain river samples
Figure SMS_1
Remarks:
Figure SMS_2
is the average value of three determinations
As shown in Table 1, the samples were measured in parallel for 3 times, the standard recovery rate was 97% -103%, and the relative standard deviation was less than 5%, indicating that the recovery effect was good. The above experimental results show that the sensor provided by the invention can not detect DEHP by further assembling the sensing element after the glassy carbon electrode is modified by Zr-MOF, GDY or Zr-MOF@GDY without modifying the Zr-MOF/GDY composite material, so that the sensor provided by the invention can be used for detecting DEHP of bottled beverage.
Based on the verification, the invention constructs a novel method capable of rapidly and sensitively detecting the DEHP based on the electrochemiluminescence recovery effect of the DEHP on the COOH-apt/Zr-MOF/GDY/GCE system. As Zr-MOF and GDY have good electrostatic interaction, the Zr-MOF/GDY composite material with high electrochemiluminescence intensity and good stability can be formed. In the presence of trace amounts of DEHP, the Zr-MOF in the COOH-apt/Zr-MOF/GDY/GCE system enriches the specific binding of the detector DEHP to the aptamer, alters the surface material properties of the electrode, and breaks down the DEHP and COOH-apt together from the electrode surface, resulting in an increase in ECL signal of COOH-apt/Zr-MOF/GDY/GCE. It was found by investigation that the enhancement value (ΔECL) of the ECL signal of the COOH-apt/Zr-MOF/GDY/GCE sensor system exhibited a good linear relationship with the concentration of DEHP. The electrochemical luminescence method used by the invention has the advantages of high sensitivity, high detection speed, good selectivity, wide linear range and the like, and has great application potential in quantitative analysis of DEHP in water environment.
The present invention is not limited to the above-mentioned embodiments, but is not limited to the above-mentioned embodiments, and any simple modification, equivalent changes and modification made to the above-mentioned embodiments according to the technical matters of the present invention can be made by those skilled in the art without departing from the scope of the present invention.

Claims (10)

1. The preparation method of the electrochemical luminescence sensor for detecting the di (2-ethylhexyl) phthalate is characterized in that the electrochemical luminescence aptamer sensor is formed by loading carboxylated ligands on the surface of a composite material Zr-MOF/GDY modified glassy carbon electrode; wherein the Zr-MOF/GDY composite material is formed by electrostatic interaction between Zr-MOF and GDY; the carboxylated aptamer is:
an aptamer comprising a base sequence of 5'-GGGTAGGGCGGGAAGTTACTGTCTTACTGTCGTA-3'.
2. The method for preparing an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate according to claim 1, comprising the following steps:
dispersing GDY in water to obtain solution 1; dispersing Zr-MOF in DMF to obtain solution 2; dropping the solution 1 on the surface of the glassy carbon electrode; dropping the solution 2 on the surface of a GDY/glassy carbon electrode; naturally airing to obtain Zr-MOF/GDY modified glassy carbon electricityA pole; by-NH 2 And (3) carrying out dehydration condensation reaction with-COOH to load carboxylated aptamer on the surface of a glassy carbon electrode modified by a composite material Zr-MOF/GDY, and naturally airing to prepare the electrochemiluminescence aptamer sensor.
3. The method for preparing an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate according to claim 2, wherein the method for preparing Zr-MOF comprises the following steps: zrCl is added to 4 And lauric acid is dissolved in an organic solvent, 2-amino terephthalic acid is added after ultrasonic mixing is uniform, and solvent thermal synthesis reaction is carried out after ultrasonic mixing to obtain Zr-MOF.
4. The method for preparing an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate according to claim 2, wherein the preparation method of GDY specifically comprises the following steps: mixing and stirring hexaalkyl- [ (trimethylsilyl) ethyl ] benzene, tetrahydrofuran (THF) solution and tetrabutylammonium fluoride to synthesize hexaethylbenzene monomer, and adopting hexaethylbenzene monomer cross coupling reaction to grow GDY film on the surface of the copper foil; the GDY film was washed and dried to give GDY powder.
5. The method for preparing an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate according to claim 2, wherein the concentration of the solution 1 and the solution 2 is 1mg/L, and the dispensing amount of the dispersion is 5 μl.
6. The method for producing an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate according to claim 2, wherein, -NH 2 The dehydration condensation reaction with-COOH is specifically: first to contain KCl, naCl, mgCl 2 And adding carboxylated aptamer into Tris-HCl buffer solution of ethylenediamine tetraacetic acid to prepare an aptamer solution with the aptamer concentration of 1-10 mu M, then transferring the aptamer solution and dripping the aptamer solution on the composite material Zr-MOF/GDY modified glassy carbonElectrode surfaces.
7. The method for preparing an electrochemical luminescence sensor for detecting di (2-ethylhexyl) phthalate according to claim 2, wherein said carboxylated aptamer comprises
An aptamer of a 5'-COOH-ACGCATAGGGTGCGACCACATACGCCCCATGTATGTCCCTTGGTTGTGCCCTATGCGT-3' base sequence.
8. The method for detecting the electrochemical luminescence sensor of the di (2-ethylhexyl) phthalate prepared by the preparation method according to any one of claims 1 to 7 is characterized in that the electrochemical luminescence aptamer sensor is used as a working electrode, ag/AgCl is used as a reference electrode, a platinum wire electrode is used as a counter electrode to form a three-electrode system, the di (2-ethylhexyl) phthalate in a sample to be detected is quantitatively captured on the surface of the sensor, and detection is realized through a generated luminescence signal.
9. The method for detecting the electrochemical luminescence sensor for di (2-ethylhexyl) phthalate according to claim 8, wherein the method comprises the following specific operations:
s1, contain K 2 S 2 O 8 Preparing PBS nano bubble buffer solution;
s2, preparing standard solutions of di (2-ethylhexyl) phthalate with different concentrations;
s3, drawing a standard curve, respectively soaking the electrochemiluminescence aptamer sensor in the standard solution prepared in the same amount of step S2 and reacting for the same time, combining the electrochemiluminescence aptamer sensor with the di (2-ethylhexyl) phthalate to obtain DEHP/COOH-apt/Zr-MOF/GDY/GCE, then taking the DEHP/COOH-apt/Zr-MOF/GDY/GCE as a working electrode, taking Ag/AgCl as a reference electrode, taking a platinum electrode as a counter electrode, forming a three-electrode system, and taking the K-containing in the step S1 2 S 2 O 8 PBS nanobubble buffer solution is taken as electrolyte, cyclic voltammetry scanning is carried out, and luminous intensity-time is recordedThe curve is established that the electrochemiluminescence aptamer sensor combines the linear relation between the luminous intensity before and after the di (2-ethylhexyl) phthalate and the logarithmic value of the concentration of the di (2-ethylhexyl) phthalate in the di (2-ethylhexyl) phthalate standard solution to obtain a corresponding linear regression equation;
s4, detecting the di (2-ethylhexyl) phthalate in the sample to be detected, reacting the sample to be detected with the surface of the electrochemiluminescence aptamer sensor for the same time according to the step S3, enabling the electrochemiluminescence aptamer sensor to be combined with the di (2-ethylhexyl) phthalate, then taking the electrochemiluminescence aptamer sensor as a working electrode, detecting the luminous intensity by adopting the method of the step S3, and then calculating the concentration of the di (2-ethylhexyl) phthalate in the sample to be detected according to a linear regression equation.
10. The method for detecting the electrochemical luminescence sensor for detecting the di (2-ethylhexyl) phthalate according to claim 9, wherein the cyclic voltammetry scanning is performed under the condition of a high voltage of 800V of a photomultiplier within an electrochemical window range of-2.0-0V at a scanning speed of 0.1V/s.
CN202310208965.9A 2023-03-07 2023-03-07 Preparation method and detection method of electrochemical sensor for detecting di (2-ethylhexyl) phthalate Pending CN116203092A (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116891852A (en) * 2023-07-14 2023-10-17 四川大学 Specific nucleic acid aptamer, targeted antibacterial drug-loaded gelatin microsphere modified by specific nucleic acid aptamer and application of targeted antibacterial drug-loaded gelatin microsphere

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116891852A (en) * 2023-07-14 2023-10-17 四川大学 Specific nucleic acid aptamer, targeted antibacterial drug-loaded gelatin microsphere modified by specific nucleic acid aptamer and application of targeted antibacterial drug-loaded gelatin microsphere

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