CN113203788A - Bimodal rapid detection method for acetamiprid in fruits and vegetables - Google Patents
Bimodal rapid detection method for acetamiprid in fruits and vegetables Download PDFInfo
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Abstract
The invention belongs to the technical field of food detection, and relates to a bimodal rapid detection method for acetamiprid in fruits and vegetables; the bimodal nanoprobe with both fluorescence characteristic and electrochemical characteristic is constructed by combining the fluorescence technology and the electrochemical technology by utilizing the dual-signal characteristic of the zirconium-based metal organic framework for the first time, so that the bimodal detection of the single nanomaterial on the acetamiprid is realized. The bimodal detection of acetamiprid provided by the invention can reduce or eliminate low detection accuracy or false positive results caused by external factors such as substrate concentration, external environment, instrument conditions and the like. The correlation coefficients of the electrochemical and fluorescence method correlation prediction models respectively reach 0.998 and 0.997, and the selectivity and accuracy of the nano probe in a complex system are greatly improved.
Description
Technical Field
The invention belongs to the technical field of food detection, and particularly relates to a method for quickly detecting acetamiprid in fruits and vegetables in an electrochemical/fluorescent bimodal manner.
Background
China is the first major producing country of fruits and vegetables in the world, and various functional pesticides are used in large quantities in the planting process of fruits and vegetables in order to reduce the influence of diseases, insects, grass and other pests on the yield of the fruits and vegetables. However, excessive pesticide applied in the agricultural production process can directly or indirectly remain on the surfaces of fruits and vegetables, and the eating quality of the fruits and vegetables and the physical health of consumers are seriously affected. Acetamiprid is a novel broad-spectrum insecticide with acaricidal activity, and is widely used for preventing and treating insect pests such as aphids, plant hoppers and the like in the process of planting fruits and vegetables. At present, acetamiprid detection methods with strong specificity and high sensitivity are all instrumental analysis methods with high detection cost, long period and complex operation. In view of the influence of the acetamiprid on the edibility of the fruits and vegetables and the defects of the existing detection method, the development of the method for quickly, conveniently and accurately detecting the content of the acetamiprid in the fruits and vegetables has important practical significance.
The acetamiprid detection method based on the fluorescence technology and the electrochemical technology is concerned about because of high detection precision, high speed, low cost and the like. However, the single fluorescence technology and the electrochemical technology have respective advantages and defects, and in order to avoid the problem of false positive or false negative in the detection result, a bimodal detection strategy combining the fluorescence technology and the electrochemical technology becomes an important research direction in the field of rapid detection at present. The fluorescent technology and the electrochemical technology are combined, the bimodal nanoprobe with the fluorescent characteristic and the electrochemical characteristic is constructed, the limitation of a single technology can be overcome, the detection result can be corrected in a targeted manner, the accuracy of the detection result is improved, and the advantage complementation effect is played. The existing method for detecting acetamiprid based on the bimodal nanoprobe has the following two defects that firstly, the nanoprobe constructed for realizing bimodal detection adopts complex nano materials, and the manufacturing and synthesizing process is complicated; secondly, interaction of multiple nano materials is needed to realize fluorescent and electrochemical bimodal detection, the nano materials are mutually influenced, and the stability of the probe is poor.
Disclosure of Invention
In order to make up for the defects of the prior art, the invention provides a method for quickly detecting acetamiprid in fruits and vegetables based on a zirconium-based metal organic framework, which can realize electrochemical/fluorescent bimodal quick and low-cost detection of acetamiprid in fruits and vegetables. The zirconium-based metal organic framework has the fluorescence characteristic and the electrochemical characteristic at the same time, and is simple to synthesize and low in cost.
In order to achieve the purpose, the specific technical scheme of the invention is as follows:
a bimodal rapid detection method for acetamiprid in fruits and vegetables comprises the following steps:
(1) preparation of zirconium-based metal organic frameworks
S1, dissolving zirconium tetrachloride and 2-amino terephthalic acid in N, N-dimethylformamide, uniformly mixing by ultrasonic oscillation, adding glacial acetic acid, continuously stirring to obtain a mixed solution, transferring the mixed solution into a reaction kettle, reacting at a certain temperature, sequentially washing a product for several times by using the N, N-dimethylformamide and methanol after reaction, and drying in a vacuum drying oven to obtain a purified zirconium-based metal organic framework;
s2, modifying a zirconium-based metal organic framework by an acetamiprid aptamer: mixing the acetamiprid aptamer solution with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), stirring and incubating at normal temperature, adding the zirconium-based metal organic framework purified in the step S1, incubating at normal temperature, and centrifuging the product to obtain solid precipitate, namely the acetamiprid aptamer-modified zirconium-based metal organic framework;
(2) acquisition of electrochemical and fluorescent signals
S1, electrode pretreatment: polishing and grinding the gold electrode on polishing flannelette by using alumina powder with different grain diameters, sequentially carrying out ultrasonic treatment in absolute ethyl alcohol and deionized water, and drying by using nitrogen; then placing the activated gold electrode in a sulfuric acid solution for cyclic potential scanning, and observing the oxidation and reduction peak potentials until the oxidation and reduction peaks are completely coincided to obtain an activated gold electrode;
s2, gold electrode acetamiprid aptamer complementary strand modification: dropwise adding the acetamiprid aptamer complementary strand solution to the surface of the activated gold electrode in the S1 in the step (2), and washing the unbound acetamiprid aptamer complementary strand solution on the surface with deionized water after incubation at normal temperature; then, adding Bovine Serum Albumin (BSA) solution dropwise, standing at room temperature for a period of time, and washing the surface of the electrode with deionized water to obtain the acetamiprid aptamer complementary chain modified gold electrode;
s3, measuring by using a pulse voltammetry method: preparing acetamiprid standard samples with different concentrations, and respectively marking as C1,C2,……,Cn-1,CnN is a positive integer; then adding the acetamiprid aptamer modified zirconium-based metal organic framework obtained in S2 in the step (1) into the acetamiprid standard sample to obtain a mixed solution; inserting the gold electrode modified by the complementary chain of the acetamiprid aptamer obtained in the step (2) into the mixed solution, stirring and enriching for a period of time, so that the zirconium-based metal organic framework modified by the acetamiprid aptamer is connected to the gold electrode modified by the complementary chain of the acetamiprid aptamer through base complementary pairing; after the enrichment is finished, stopping stirring, standing the solution for a period of time, and attaching the zirconium-based metal organic framework modified by the acetamiprid aptamer to the surface of the gold electrode modified by the complementary chain of the acetamiprid aptamer in the process to obtain a working electrode;
s4, electric signal acquisition stage: potential scanning is carried out on the working electrode in the step (2) S3, the potential is scanned from 1.2V to 0.5V, an electric signal of the working electrode is obtained, and acetamiprid standard samples (C) with different concentrations are measured and recorded by the working electrode1,C2,……,Cn-1,Cn) Intensity of electric signal of (E)1,E2,……,En-1,EnN is a positive integer;
s5, fluorescent signal acquisition stage: after measuring the electric signal intensity of each acetamiprid standard sample with one concentration, taking out the working electrode, washing the surface of the working electrode by using a phosphate buffer solution, collecting and washing the phosphate buffer solution with the concentration, and mixing the phosphate buffer solution with the acetamiprid standard sample with the concentration; after the operation, acetamiprid standard samples with different concentrations can be obtained and washedThe mixed solution of the phosphate buffer solution is subjected to constant volume to a certain volume, then the solution subjected to constant volume is respectively put into a quartz cuvette, the excitation wavelength of a fluorescence spectrometer is set, the optimal fluorescence emission intensity is measured and recorded, and the optimal fluorescence emission intensity is recorded as F1,F,……,Fn-1,FnN is a positive integer; (3) rapid prediction of acetamiprid concentration
S1, an electrochemical rapid detection prediction model: the intensity E of the electric signal in S4 according to the step (2)1,E2,……,En-1,EnIn the standard sample C of acetamiprid with different concentrations1,C2,……,Cn-1,CnEstablishing a model to obtain an electrochemical rapid detection prediction model E ═ f (X) of acetamiprid, wherein E is the electrochemical current, X is the acetamiprid concentration, and n is a positive integer;
s2, a fluorescence rapid detection prediction model: subjecting the fluorescence emission intensity F described in S5 of step (2)1,F,……,Fn-1,FnWith acetamiprid standard sample C of different concentrations1,C2,……,Cn-1,CnEstablishing a model to obtain a fluorescence rapid detection prediction model F ═ F (X) of acetamiprid, wherein F is fluorescence emission intensity, X is acetamiprid concentration, and n is a positive integer;
s3, rapidly predicting the acetamiprid concentration in the fruits and vegetables: taking a fruit and vegetable sample to be detected, shearing or stirring, adding the universal sample extracting solution, and fully oscillating and uniformly mixing to obtain a liquid to be detected; respectively obtaining the electric signal intensity and the fluorescence emission intensity according to the methods for measuring the electrochemical signal and the fluorescence signal in the S4 and S5 in the step (2), respectively substituting the electric signal intensity and the fluorescence emission intensity into an electrochemical rapid detection prediction model and a fluorescence rapid detection prediction model, and obtaining the acetamiprid rapid prediction concentration value obtained by an electrochemical method and a fluorescence method; rapidly predicting a concentration value of acetamiprid obtained by an electrochemical method and a fluorescence method, and performing group difference analysis by using SPSS; if there is a significant difference between the two sets of data (p <0.05), the data is discarded; if the two groups of data have no significant difference (p is greater than 0.05), taking the average value of the data as the acetamiprid concentration in the fruits and vegetables, and realizing the rapid detection of the acetamiprid concentration.
Preferably, tetrachloro is described in S1 of step (1)The molar ratio of zirconium oxide to 2-amino terephthalic acid is 1: 1; the concentration of the zirconium tetrachloride and the 2-amino terephthalic acid in the mixed solution is both 8 mmol.L-1(ii) a The volume ratio of the N, N-dimethylformamide to the glacial acetic acid is 4: 1; the ultrasonic vibration mixing time is 5-10 minutes; the reaction temperature in the reaction kettle is 110-150 ℃, and the reaction time is 8-15 hours; the washing is carried out for 1 to 4 times.
Preferably, the ratio of the acetamiprid aptamer solution, the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC), the N-hydroxysuccinimide (NHS) and the purified zirconium-based metal-organic framework in S2 of step (1) is 1mL:5mL: 1-2 g; the concentration of the acetamiprid aptamer solution is 100 mu mol/L; the concentration of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 0.1 mmol/L; the concentration of the N-hydroxysuccinimide is 0.1 mmol/L; the base sequence of the acetamiprid aptamer is as follows: 5 '- (COOH) - (CH2)6-ACA GGC TAC GAG GGA AAT GCG GTG GGT GTG GGC GAT-3'.
Preferably, the incubation time in the step (1) S2 is 1-2 hours; the incubation time after the purified zirconium-based metal organic framework is added is 10-12 hours.
Preferably, in the step (2), in the S1, the alumina powders with different particle sizes have particle sizes of 1.0 μm, 0.3 μm and 0.05 μm in sequence, and the polishing time is 3-5 min; the ultrasonic treatment time is 3-5 min; the concentration of the sulfuric acid solution is 0.5 mol/L.
Preferably, the base sequence of the complementary strand of the acetamiprid aptamer in S2 of step (2) is: 5 '- (SH) - (CH2)6-TGT CCG ATG CTC-3'; the concentration of the acetamiprid aptamer complementary strand solution is 100 mu mol/L, and the dosage of the acetamiprid aptamer complementary strand solution dripped on the surface of the activated gold electrode is 5-10 mu L; and the incubation time at the normal temperature is 2-4 hours.
Preferably, in the step (2), the mass concentration of the bovine serum albumin solution in the S2 is 1%, and the dosage of the bovine serum albumin solution dripped on the surface of the activated gold electrode is 10-20 mu L; the standing time at room temperature is 1-2 hours.
Preferably, the acetamiprid concentration in S3 in the step (2) is 0-10 mg/kg; the dosage ratio of the acetamiprid standard sample to the acetamiprid aptamer-modified zirconium-based metal organic framework is (10-15) mL: (1-2) g; the stirring and enriching time is 10-20 minutes; the standing time is 10-20 seconds.
Preferably, the pH of the phosphate buffer solution in the step (2) S4 is 7.2-7.5, and the volume of the washing working electrode is 1-2 ml; the volume after constant volume is 10 mL.
Wherein W is a specific excitation wavelength, is greater than 0 and is a positive integer; preferably, the excitation wavelength is 365 nm.
Preferably, the universal extracting solution in the step (3) S3 is a YRSPN sample universal extracting solution; the dosage ratio of the fruit and vegetable sample to be detected to the universal extracting solution of the sample is 2-2.5 g: 5 mL.
The invention has the beneficial effects that:
the invention firstly proposes that the double-signal characteristic of the zirconium-based metal organic framework is utilized for the electrochemical and fluorescent rapid detection of the acetamiprid in the fruits and vegetables, and the bimodal detection of the acetamiprid by the single nano material is firstly realized.
The bimodal detection of acetamiprid provided by the invention can reduce or eliminate low detection accuracy or false positive results caused by external factors such as substrate concentration, external environment, instrument conditions and the like. The correlation coefficients of the electrochemical and fluorescence method correlation prediction models respectively reach 0.998 and 0.997.
The acetamiprid aptamer is used for specifically identifying the acetamiprid, so that the selectivity and the accuracy of the nano probe in a complex system are greatly improved.
The invention simplifies the manufacture of the bimodal nanosensor sensing system and further reduces the detection error (the relative error with the detection result of the national standard method is less than 1%) caused by the stability problem of the sensing system.
Drawings
FIG. 1 is a topographical feature diagram of a zirconium-based metal organic framework.
FIG. 2 is a graph showing an electrochemical signal (a) and a fluorescence signal (b) of a zirconium-based metal organic framework.
Detailed description of the preferred embodiments
The present invention will be described in detail below with reference to embodiments. These examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. Furthermore, various changes or modifications may be made by one skilled in the art after reading the disclosure herein, and equivalents may fall within the scope of the invention as defined by the claims appended hereto.
The gold electrode used in the invention is purchased from Shanghai Lu major company, Inc., and the other reagents are purchased from the national medicine group chemical reagent company, Inc.
A bimodal rapid detection method for acetamiprid in fruits and vegetables comprises the following steps:
(1) preparation of zirconium-based metal organic frameworks
S1, dissolving zirconium tetrachloride and 2-amino terephthalic acid in 20mL of N, N-dimethylformamide, wherein the use amounts of the zirconium tetrachloride and the 2-amino terephthalic acid are both 0.2 mol; carrying out ultrasonic oscillation for 10 minutes, moving to a magnetic stirrer, adding 5mL of glacial acetic acid, continuously stirring and uniformly mixing for 10 minutes, and transferring the mixed solution to a 50mL reaction kettle; reacting for 15 hours at 130 ℃, washing the obtained zirconium-based metal organic framework with N, N-dimethylformamide for 2 times, washing with methanol for 2 times, and drying in a vacuum drying oven to obtain the purified zirconium-based metal organic framework.
S2, modifying a zirconium-based metal organic framework by using an acetamiprid aptamer, mixing 1mL of a 100 mu mol/L acetamiprid aptamer (5 '- (COOH) - (CH2)6-ACA GGC TAC GAG GGA AAT GCG GTG GGT GTG GGC GAT-3') solution with 5mL of 0.1mmol/L EDC solution and 5mL of 0.1mmol/L NHS solution, and incubating for 2 hours under stirring at normal temperature to activate carboxyl; and then adding 1.5g of the obtained zirconium-based metal organic framework, and incubating for 12 hours at normal temperature to obtain the acetamiprid aptamer-modified zirconium-based metal organic framework.
(2) Acquisition of electrochemical signals
Electrode pretreatment: polishing and grinding the gold electrode by using alumina powder with the grain diameters of 1.0 mu m, 0.3 mu m and 0.05 mu m in sequence for 5min on polishing flannelette, then respectively carrying out ultrasonic treatment in absolute ethyl alcohol and deionized water for 5min to remove residual alumina, and drying by using nitrogen; carrying out cyclic potential scanning in 0.5mol/L dilute sulfuric acid, and observing the oxidation peak potential and the reduction peak potential until the oxidation peak and the reduction peak are completely coincided, which represents that the surface of the gold electrode is clean, so as to obtain an activated gold electrode;
modification of gold electrode acetamiprid aptamer complementary strand: and dripping 5 mu L of 100 mu mol/L acetamiprid aptamer complementary chain (5 '- (SH) - (CH2)6-TGT CCG ATG CTC-3') solution on the surface of the activated gold electrode, incubating for 3 hours at normal temperature, and washing the unbound acetamiprid aptamer complementary chain on the surface by deionized water. Titrating 20 mu L of BSA with the mass concentration of 1%, standing at room temperature for 1 hour, and washing the surface of the electrode with deionized water to obtain the acetamiprid aptamer complementary chain modified gold electrode.
And (3) pulse voltammetry determination: preparing acetamiprid standard samples with different concentrations, wherein the concentrations of the acetamiprid are respectively 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10mg/kg, and adding 1.5g of the zirconium-based metal organic framework modified by the acetamiprid aptamer into the standard samples. Inserting the gold electrode modified by the acetamiprid aptamer complementary chain into the mixed solution, stirring to ensure that the acetamiprid aptamer complementary chain modified on the gold electrode is combined with the acetamiprid aptamer base complementary pairing modified on the zirconium-based metal organic framework in a complementary pairing manner, enriching the zirconium-based metal organic framework on the surface of the gold electrode, and continuing the enrichment process for 15 minutes to obtain a working electrode; after the enrichment is finished, stopping stirring, and standing the solution for 15 seconds; an electric signal acquisition stage: the potential of the working electrode is scanned from 1.2V to 0.5V, a zirconium-based metal organic framework is attached to the surface of the gold electrode in the process, the dissolution current is formed, and the electric signal intensities of 21.782, 18.987, 17.056, 15.71, 13.227, 11.057, 9.315, 7.1352, 5.2454, 3.189 and 0.987mA at different concentrations of acetamiprid are measured and recorded by an electrochemical workstation.
(3) Acquisition of fluorescence signals
Taking out the working electrode in the step (2), taking 1mL of phosphate buffer solution to wash the surface of the electrode, collecting the washing solution and the solution in the step (2), fixing the volume to 10mL, taking 2mL of the solution in a quartz cuvette, setting the excitation wavelength of a fluorescence spectrometer to be 365nm, and measuring and recording the optimal fluorescence emission intensity of 199.74, 248.56, 315.71, 361.24, 428.79, 479.56, 516.91, 597.16, 668.25, 719.53 and 787.15 of the mixed solution under the standard acetamiprid samples with different concentrations.
(4) Rapid prediction of acetamiprid concentration
Electrochemical rapid detection prediction model: and (3) modeling the electrochemical signals 21.782, 18.987, 17.056, 15.71, 13.227, 11.057, 9.315, 7.1352, 5.2454, 3.189 and 0.987mA obtained in the step (2) on acetamiprid standard samples with different concentrations of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 mg/kg. Obtaining an electrochemical rapid detection prediction model E-2.033X +21.411 (R) of acetamiprid2＝0.998)；
Fluorescence rapid detection prediction model: establishing models of the fluorescence signals 199.74, 248.56, 315.71, 361.24, 428.79, 479.56, 516.91, 597.16, 668.25, 719.53 and 787.15 obtained in the step (3) and acetamiprid standard samples 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10mg/kg with different concentrations to obtain an acetamiprid fluorescence rapid detection prediction model F (R) 58.532X +191.21 (R)2＝0.997)。
Example 1: rapid prediction of acetamiprid concentration in apples
Weighing 5 parts of the apple sample to be detected, wherein the parts are respectively (2.05g, 2.12g, 2.09g, 2.06g and 2.15g), and adding the cut or crushed sample into a 10mL centrifuge tube; adding 5mL of the universal extracting solution of the YRSPN sample, fully oscillating and uniformly mixing to obtain the solution to be detected.
Respectively taking 2mL of solution to be detected, adding the acetamiprid aptamer-modified zirconium-based metal organic framework into the solution to be detected, respectively obtaining a fluorescence signal and an electrochemical signal, repeatedly measuring the fluorescence signal and the electrochemical signal for three times, and obtaining the acetamiprid concentration result by using a rapid prediction model of the fluorescence method and the electrochemical method as shown in Table 1. The SPSS analysis result shows that the fluorescence method and the electrochemical method in the group 4 sample have significant difference, which indicates that the data has problems, and the group data is determined as invalid data and discarded; when no significant difference exists among the data of groups 1, 2, 3 and 5, the average value of the data obtained is 0.466mg/kg, 1.074mg/kg, 1.553mg/kg and 2.471mg/kg of acetamiprid in apples. In order to verify the detection accuracy of the method, the acetamiprid concentration analysis is carried out on the samples of the groups 1, 2, 3 and 5 by using a national standard method (GB/T23584-. The relative error of the obtained detection result is less than 1%, which shows that the bimodal rapid detection method for acetamiprid in fruits and vegetables further reduces the detection error caused by the stability problem of a sensing system and improves the detection accuracy.
TABLE 1 measurement of acetamiprid content in apple by fluorescence and electrochemical methods
Example 2: rapid prediction of acetamiprid concentration in cherry tomato
Weighing 5 parts of cherry tomato samples to be detected, wherein the parts are respectively (2.03g, 2.02g, 2.06g, 2.09g and 2.11g), and adding the cut or crushed samples into a 10mL centrifuge tube; adding 5mL of the universal extracting solution of the YRSPN sample, fully oscillating and uniformly mixing to obtain the solution to be detected.
And (3) respectively adding a zirconium-based metal organic framework into 2mL of solution to be detected to respectively obtain a fluorescence signal and an electrochemical signal, repeatedly measuring for three times, and obtaining the acetamiprid concentration result by using rapid prediction models of a fluorescence method and an electrochemical method as shown in Table 2. The SPSS analysis result shows that the fluorescence method and the electrochemical method measurement result in the group 1 sample have significant difference analysis, which indicates that the data have problems, and the group data is determined as invalid data and discarded; and if the data of groups 2, 3, 4 and 5 have no significant difference, the concentrations of the acetamiprid in the cherry tomatoes are respectively 2.312mg/kg, 3.476mg/kg, 4.683mg/kg and 5.366 mg/kg. In order to verify the detection accuracy of the method, the acetamiprid concentration analysis is carried out on the samples of the groups 2, 3, 4 and 5 by using a national standard method (GB/T23584-. The relative error of the obtained detection result is less than 1%, which shows that the bimodal rapid detection method for acetamiprid in fruits and vegetables further reduces the detection error caused by the stability problem of a sensing system and improves the detection accuracy.
Table 2. measuring the content of acetamiprid in cherry tomatoes by a fluorescence method and an electrochemical method
Description of the drawings: the above embodiments are only used to illustrate the present invention and do not limit the technical solutions described in the present invention; thus, while the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted; all such modifications and variations are intended to be included herein within the scope of this disclosure and the present invention and protected by the following claims.
Claims (10)
1. A bimodal rapid detection method for acetamiprid in fruits and vegetables is characterized by comprising the following specific steps:
(1) preparation of zirconium-based metal organic frameworks
S1, dissolving zirconium tetrachloride and 2-amino terephthalic acid in N, N-dimethylformamide, uniformly mixing by ultrasonic oscillation, adding glacial acetic acid, continuously stirring to obtain a mixed solution, transferring the mixed solution into a reaction kettle, reacting at a certain temperature, sequentially washing a product for several times by using the N, N-dimethylformamide and methanol after reaction, and drying in a vacuum drying oven to obtain a purified zirconium-based metal organic framework;
s2, modifying a zirconium-based metal organic framework by an acetamiprid aptamer: mixing the acetamiprid aptamer solution with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, stirring and incubating at normal temperature, adding the zirconium-based metal organic framework purified in the step S1, incubating at normal temperature, and centrifuging the product to obtain solid precipitate, namely the acetamiprid aptamer-modified zirconium-based metal organic framework;
(2) acquisition of electrochemical and fluorescent signals
S1, electrode pretreatment: polishing and grinding the gold electrode on polishing flannelette by using alumina powder with different grain diameters, sequentially carrying out ultrasonic treatment in absolute ethyl alcohol and deionized water, and drying by using nitrogen; then placing the activated gold electrode in a sulfuric acid solution for cyclic potential scanning, and observing the oxidation and reduction peak potentials until the oxidation and reduction peaks are completely coincided to obtain an activated gold electrode;
s2, gold electrode acetamiprid aptamer complementary strand modification: dropwise adding the acetamiprid aptamer complementary strand solution to the surface of the activated gold electrode in the S1 in the step (2), and washing the unbound acetamiprid aptamer complementary strand solution on the surface with deionized water after incubation at normal temperature; then, dropwise adding bovine serum albumin solution, standing at room temperature for a period of time, and washing the surface of the electrode with deionized water to obtain the acetamiprid aptamer complementary chain modified gold electrode;
s3, measuring by using a pulse voltammetry method: preparing acetamiprid standard samples with different concentrations, and respectively marking as C1,C2,……,Cn-1,CnN is a positive integer; then adding the acetamiprid aptamer modified zirconium-based metal organic framework obtained in S2 in the step (1) into the acetamiprid standard sample to obtain a mixed solution; inserting the gold electrode modified by the complementary chain of the acetamiprid aptamer obtained in the step (2) into the mixed solution, stirring and enriching for a period of time, so that the zirconium-based metal organic framework modified by the acetamiprid aptamer is connected to the gold electrode modified by the complementary chain of the acetamiprid aptamer through base complementary pairing; after the enrichment is finished, stopping stirring, standing the solution for a period of time, and attaching the zirconium-based metal organic framework modified by the acetamiprid aptamer to the surface of the gold electrode modified by the complementary chain of the acetamiprid aptamer in the process to obtain a working electrode;
s4, electric signal acquisition stage: potential scanning is carried out on the working electrode in the step (2) S3, the potential is scanned from 1.2V to 0.5V, an electric signal of the working electrode is obtained, the working electrode is used for measuring and recording the electric signal intensity of the acetamiprid standard samples with different concentrations, and the electric signal intensity is recorded as E1,E2,……,En-1,EnN is a positive integer;
s5, fluorescent signal acquisition stage: after measuring the electric signal intensity of each acetamiprid standard sample with one concentration, taking out the working electrode, washing the surface of the working electrode with phosphate buffer solution, and collecting and washing phosphate with the concentrationA buffer solution mixed with the acetamiprid standard sample with the concentration; after the operation, mixed liquor of acetamiprid standard samples with different concentrations and the washed phosphate buffer solution is obtained, the volumes are all fixed to a certain same volume, then the solutions with the fixed volumes are respectively put into a quartz cuvette, the excitation wavelength of a fluorescence spectrometer is set, the optimal fluorescence emission intensity is measured and recorded, and the optimal fluorescence emission intensity is recorded as F1,F,……,Fn-1,FnN is a positive integer;
(3) rapid detection of acetamiprid concentration
S1, an electrochemical rapid detection prediction model: the intensity E of the electric signal in S4 according to the step (2)1,E2,……,En-1,EnIn the standard sample C of acetamiprid with different concentrations1,C2,……,Cn-1,CnEstablishing a model to obtain an electrochemical rapid detection prediction model E ═ f (X) of acetamiprid, wherein E is the electrochemical current, X is the acetamiprid concentration, and n is a positive integer;
s2, a fluorescence rapid detection prediction model: subjecting the fluorescence emission intensity F described in S5 of step (2)1,F,……,Fn-1,FnWith acetamiprid standard sample C of different concentrations1,C2,……,Cn-1,CnEstablishing a model to obtain a fluorescence rapid detection prediction model F ═ F (X) of acetamiprid, wherein F is fluorescence emission intensity, X is acetamiprid concentration, and n is a positive integer;
s3, rapidly predicting the acetamiprid concentration in the fruits and vegetables: taking a fruit and vegetable sample to be detected, shearing or stirring, adding the universal sample extracting solution, and fully oscillating and uniformly mixing to obtain a liquid to be detected; respectively obtaining the electric signal intensity and the fluorescence emission intensity according to the methods for measuring the electrochemical signal and the fluorescence signal in the S4 and S5 in the step (2), respectively substituting the electric signal intensity and the fluorescence emission intensity into an electrochemical rapid detection prediction model and a fluorescence rapid detection prediction model, and obtaining the acetamiprid rapid prediction concentration value obtained by an electrochemical method and a fluorescence method; rapidly predicting a concentration value of acetamiprid obtained by an electrochemical method and a fluorescence method, and performing group difference analysis by using SPSS; if there is significant difference between the two sets of data, discarding the data; if the two groups of data have no significant difference, taking the average value of the data as the acetamiprid concentration in the fruits and vegetables, and realizing the rapid detection of the acetamiprid concentration.
2. The bimodal rapid detection method of acetamiprid in fruits and vegetables according to claim 1, characterized in that the molar ratio of zirconium tetrachloride to 2-amino terephthalic acid in S1 of step (1) is 1: 1; the concentration of the zirconium tetrachloride and the 2-amino terephthalic acid in the mixed solution is both 8 mmol.L-1(ii) a The volume ratio of the N, N-dimethylformamide to the glacial acetic acid is 4: 1; the ultrasonic vibration mixing time is 5-10 minutes; the reaction temperature in the reaction kettle is 110-150 ℃, and the reaction time is 8-15 hours; the washing is carried out for 1 to 4 times.
3. The bimodal rapid detection method of acetamiprid in fruits and vegetables according to claim 1, wherein the usage ratio of the acetamiprid aptamer solution, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide and purified zirconium-based metal organic framework in S2 in step (1) is 1mL:5mL: 1-2 g; the concentration of the acetamiprid aptamer solution is 100 mu mol/L; the concentration of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 0.1 mmol/L; the concentration of the N-hydroxysuccinimide is 0.1 mmol/L; the base sequence of the acetamiprid aptamer is as follows: 5 '- (COOH) - (CH2)6-ACA GGC TAC GAG GGA AAT GCG GTG GGT GTG GGC GAT-3'.
4. The bimodal rapid detection method for acetamiprid in fruits and vegetables according to claim 1, wherein the stirring incubation time in S2 of step (1) is 1-2 hours; the incubation time after the purified zirconium-based metal organic framework is added is 10-12 hours.
5. The bimodal rapid detection method for acetamiprid in fruits and vegetables according to claim 1, wherein in step (2) S1, the alumina powders with different particle sizes are 1.0 μm, 0.3 μm and 0.05 μm in sequence, and the grinding time is 3-5 min; the ultrasonic treatment time is 3-5 min; the concentration of the sulfuric acid solution is 0.5 mol/L.
6. The bimodal rapid detection method for acetamiprid in fruits and vegetables according to claim 1, wherein the base sequence of the complementary strand of acetamiprid aptamer in S2 of step (2) is: 5 '- (SH) - (CH2)6-TGT CCG ATG CTC-3'; the concentration of the acetamiprid aptamer complementary strand solution is 100 mu mol/L, and the dosage of the acetamiprid aptamer complementary strand solution dripped on the surface of the activated gold electrode is 5-10 mu L; and the incubation time at the normal temperature is 2-4 hours.
7. The bimodal rapid detection method of acetamiprid in fruits and vegetables according to claim 1, wherein in the step (2) S2, the mass concentration of the bovine serum albumin solution is 1%, and the dosage of the bovine serum albumin solution dripped on the surface of the activated gold electrode is 10-20 μ L; the standing time at room temperature is 1-2 hours.
8. The bimodal rapid detection method for acetamiprid in fruits and vegetables according to claim 1, wherein the acetamiprid concentration in S3 of step (2) is 0-10 mg/kg; the dosage ratio of the acetamiprid standard sample to the acetamiprid aptamer-modified zirconium-based metal organic framework is (10-15) mL: (1-2) g; the stirring and enriching time is 10-20 minutes; the standing time is 10-20 seconds.
9. The bimodal rapid detection method for acetamiprid in fruits and vegetables according to claim 1, wherein in step (2) S4, the pH of the phosphate buffer solution is 7.2-7.5, and the volume of the washing working electrode is 1-2 ml; the volume after constant volume is 10 mL.
10. The bimodal rapid detection method for acetamiprid in fruits and vegetables according to claim 1, wherein the universal extract in S3 in step (3) is a YRSPN sample universal extract; the dosage ratio of the fruit and vegetable sample to be detected to the universal extracting solution of the sample is 2-2.5 g: 5 mL.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113970585A (en) * | 2021-09-03 | 2022-01-25 | 江苏大学 | Enhanced adsorption electrochemical immunosensor and preparation method and detection method thereof |
| CN114437709A (en) * | 2021-09-15 | 2022-05-06 | 中国科学院海洋研究所 | Nucleic acid functionalized MOF material and preparation and application thereof |
| CN114858898A (en) * | 2022-05-23 | 2022-08-05 | 山西大学 | Fluorescence/electrochemistry double-signal mode biosensor and construction method and application thereof |
| CN115032254A (en) * | 2022-06-28 | 2022-09-09 | 河南工业大学 | Method for detecting lead ions by dual-mode biosensor based on magnetic separation technology mediation |
| CN115308171A (en) * | 2022-06-13 | 2022-11-08 | 江苏大学 | Preparation method and application of fluorescence/electrochemistry bimodal nanosensor based on covalent organic framework |
| CN118010697A (en) * | 2024-04-08 | 2024-05-10 | 泉州师范学院 | Vegetable leaf acetamiprid residue detection method and detection system |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107525834A (en) * | 2017-07-18 | 2017-12-29 | 济南大学 | A kind of DNA aptamer sensors of Cu MOF marks are used for the method for detecting Acetamiprid |
| CN108120704A (en) * | 2017-12-11 | 2018-06-05 | 东莞理工学院 | A kind of fluorescence detection method of Acetamiprid |
| US20190293588A1 (en) * | 2018-03-22 | 2019-09-26 | Kabushiki Kaisha Toshiba | Molecular detection apparatus and molecular detection method |
| CN110618272A (en) * | 2019-08-28 | 2019-12-27 | 江苏大学 | Preparation method and application of mercury ion fluorescence/electrochemical sensor |
| CN110763740A (en) * | 2019-09-06 | 2020-02-07 | 南京医科大学 | Based on Fe3O4@MnO2Method for detecting hydrogen peroxide by using electrochemical and fluorescence dual-signal sensor with carbon dots |
| US20200064325A1 (en) * | 2018-08-22 | 2020-02-27 | Endress+Hauser Conducta Gmbh+Co. Kg | Measuring arrangement for determining an ozone content of a measured medium |
| CN111551608A (en) * | 2020-06-08 | 2020-08-18 | 济南大学 | Method for simultaneously detecting acetamiprid and malathion |
-
2021
- 2021-05-21 CN CN202110561054.5A patent/CN113203788B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107525834A (en) * | 2017-07-18 | 2017-12-29 | 济南大学 | A kind of DNA aptamer sensors of Cu MOF marks are used for the method for detecting Acetamiprid |
| CN108120704A (en) * | 2017-12-11 | 2018-06-05 | 东莞理工学院 | A kind of fluorescence detection method of Acetamiprid |
| US20190293588A1 (en) * | 2018-03-22 | 2019-09-26 | Kabushiki Kaisha Toshiba | Molecular detection apparatus and molecular detection method |
| US20200064325A1 (en) * | 2018-08-22 | 2020-02-27 | Endress+Hauser Conducta Gmbh+Co. Kg | Measuring arrangement for determining an ozone content of a measured medium |
| CN110618272A (en) * | 2019-08-28 | 2019-12-27 | 江苏大学 | Preparation method and application of mercury ion fluorescence/electrochemical sensor |
| CN110763740A (en) * | 2019-09-06 | 2020-02-07 | 南京医科大学 | Based on Fe3O4@MnO2Method for detecting hydrogen peroxide by using electrochemical and fluorescence dual-signal sensor with carbon dots |
| CN111551608A (en) * | 2020-06-08 | 2020-08-18 | 济南大学 | Method for simultaneously detecting acetamiprid and malathion |
Non-Patent Citations (3)
| Title |
|---|
| XIN LIU等: "Aptamer contained triple-helix molecular switch for rapid fluorescent sensing of acetamiprid", 《TALANTA》 * |
| 何晓静等: "基于金属有机骨架材料固载的碲化镉量子点作为信号探针测定心肌肌钙蛋白的电致化学发光免疫传感器", 《化学传感器》 * |
| 候林丽等: "基于Lys-AuNPs@ZIF-8纳米复合材料比率荧光检测Hg~(2+)", 《江西师范大学学报(自然科学版)》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113970585A (en) * | 2021-09-03 | 2022-01-25 | 江苏大学 | Enhanced adsorption electrochemical immunosensor and preparation method and detection method thereof |
| CN114437709A (en) * | 2021-09-15 | 2022-05-06 | 中国科学院海洋研究所 | Nucleic acid functionalized MOF material and preparation and application thereof |
| CN114858898A (en) * | 2022-05-23 | 2022-08-05 | 山西大学 | Fluorescence/electrochemistry double-signal mode biosensor and construction method and application thereof |
| CN115308171A (en) * | 2022-06-13 | 2022-11-08 | 江苏大学 | Preparation method and application of fluorescence/electrochemistry bimodal nanosensor based on covalent organic framework |
| CN115032254A (en) * | 2022-06-28 | 2022-09-09 | 河南工业大学 | Method for detecting lead ions by dual-mode biosensor based on magnetic separation technology mediation |
| CN115032254B (en) * | 2022-06-28 | 2024-02-02 | 河南工业大学 | Method for detecting lead ions by using dual-mode biosensor based on magnetic separation technology mediation |
| CN118010697A (en) * | 2024-04-08 | 2024-05-10 | 泉州师范学院 | Vegetable leaf acetamiprid residue detection method and detection system |
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