CN114279902B - Organophosphorus pesticide detector based on intelligent response surface and organophosphorus pesticide detection method thereof - Google Patents
Organophosphorus pesticide detector based on intelligent response surface and organophosphorus pesticide detection method thereof Download PDFInfo
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Abstract
The invention discloses an organophosphorus pesticide detector based on an intelligent response surface and a method for detecting organophosphorus pesticide by using the organophosphorus pesticide detector, and belongs to the technical field of organophosphorus pesticide detection. The organophosphorus pesticide detector based on intelligent response surface of the invention is to immerse a glass substrate in CoCl 2 ·6H 2 O and (NH) 2 ) 2 And constructing a super-hydrophilic surface in the mixed aqueous solution of CO, then modifying by acetylcholinesterase or butyrylcholine esterase, and finally immersing in a perfluorooctyl triethoxysilane solution. The invention can realize the detection of different organophosphorus pesticides by observing the change of the contact angle of different organophosphorus pesticides on the surface of the detector, does not need a large instrument, and is not interfered by ambient light and color. In addition, the organophosphorus pesticide detector based on the intelligent response surface can be used for detecting organophosphorus pesticides on site, is convenient to carry and can be applied to organophosphorus pesticide residue detection on vegetables, fruits and grains.
Description
Technical Field
The invention belongs to the technical field of organophosphorus pesticide detection, and particularly relates to an organophosphorus pesticide detector based on an intelligent response surface and a method for detecting organophosphorus pesticide by using the organophosphorus pesticide detector.
Background
In recent years, with the continuous expansion of the scale of pesticide industry in China and the continuous upgrading of technology, the development of pesticides starts to develop towards high efficiency, low toxicity, low residue, high bioactivity and high selectivity. The production scale of agriculture in China is continuously expanding, but problems related to diseases and insect pests are generated along with the production scale, in order to ensure the grain safety, pesticides are used for preventing and controlling the insect pests in each stage of grain from planting, production and storage, and organophosphorus pesticides serving as efficient, broad-spectrum and easily degradable pesticides are widely used in the grain production and storage process, so that the organophosphorus pesticides are the most commonly used choices for chemical prevention and control. The use amount of pesticides is increased, and the pesticides are used to permeate the agricultural production all the time, so that the pesticide residues in the grains always affect the edible safety of consumers.
At present, the detection of organophosphorus pesticides mainly comprises a gas chromatography/mass spectrometry (GC/MS) technology and a High Performance Liquid Chromatography (HPLC) technology, and although the residual quantity of the pesticides can be accurately detected, the methods are complex in operation, samples are often preprocessed, and most of instruments required for analysis are expensive and heavy, and are required to be operated by professionals, so that the requirements of on-site rapid detection cannot be met. Therefore, the establishment of an analysis device and a method for detecting the organophosphorus pesticides rapidly, reliably, sensitively and practically has important practical significance for environmental protection, food safety and the like.
Disclosure of Invention
Aiming at the problems existing in the prior art, one of the purposes of the invention is to provide a preparation method of an organophosphorus pesticide detector based on an intelligent response surface, which comprises the following specific steps:
(1) Immersing a glass substrate into CoCl 2 ·6H 2 O and (NH) 2 ) 2 Carrying out hydrothermal reaction in a mixed aqueous solution of CO at 55-65 ℃ and drying to obtain the super-hydrophilic surface;
(2) Every 4-6cm 2 100 μl of the solution of the response molecule is dropped onto the hydrophilic surface of (a), and dried at room temperature;
(3) Immersing the modified hydrophilic surface obtained in the step (2) into a perfluorooctyl triethoxysilane solution for 30-40min, and drying to obtain a hydrophobic surface; obtaining the organophosphorus pesticide detector based on the intelligent response surface;
the response molecule solution is acetylcholinesterase phosphate buffer salt solution or butyrylcholinesterase phosphate buffer salt solution, and the concentration is 1 U.mL -1 . Butyrylcholinesterase and acetylcholinesterase have similar properties, different substituent groups can respond to organophosphorus pesticides, different response intensities of organophosphorus pesticides are different, and different detection arrays can be constructed by adjusting response molecular types.
The CoCl 2 ·6H 2 O and (NH) 2 ) 2 CoCl in aqueous CO-mixture 2 ·6H 2 The O concentration is 0.20-0.50g L -1 ，(NH 2 ) 2 The concentration of CO is 150-250g L -1 。
In the step (1), the time of the hydrothermal reaction is 22-26h; the glass substrate used may be any type of glass-like material.
In the step (3), the perfluorooctyl triethoxysilane solution is an ethanol aqueous solution of perfluorooctyl triethoxysilane (PFOTES, CAS number 51851-37-7), wherein the volume ratio of ethanol to water is 1:1, and the volume percentage of PFOTES is 1%.
The second object of the invention is to provide an organophosphorus pesticide detector based on intelligent response surface prepared by the method.
The invention further aims to provide the application of the organophosphorus pesticide detector based on the intelligent response surface in organophosphorus pesticide detection. Wherein the organophosphorus pesticide can be one or more of profenofos, dichlorvos, fenitrothion, chlorpyrifos, malathion, methyl parathion, glyphosate, chlorpyrifos, methomyl and phoxim, and is known but not limited to organophosphorus pesticide molecules.
The fourth object of the invention is to provide a method for detecting organophosphorus pesticides, which comprises the steps of dripping known organophosphorus pesticides with different concentrations or a mixture of known organophosphorus pesticides with different concentrations on the surface of the organophosphorus pesticide detector based on the intelligent response surface, detecting the change range of the organophosphorus pesticide on the surface contact angle of the organophosphorus pesticide detector based on the intelligent response surface, and constructing an intelligent response surface contact angle change range database based on the organophosphorus pesticide;
dropwise adding a sample to be detected to the surface of the organophosphorus pesticide detector based on the intelligent response surface, detecting the change range of the contact angle of the sample to be detected on the surface of the organophosphorus pesticide detector based on the intelligent response surface, and comparing the constructed intelligent response surface contact angle change range database based on the organophosphorus pesticide to obtain the type and concentration of the organophosphorus pesticide in the sample to be detected;
wherein the temperature is 20-40deg.C, and the detection time is 20-60min; the organophosphorus pesticide can be one or more of profenofos, dichlorvos, fenitrothion, chlorpyrifos, malathion, methyl parathion, glyphosate, chlorpyrifos, methomyl and phoxim, and is known but not limited to organophosphorus pesticide molecules.
Advantages of the technical proposal of the invention
Traditional organophosphorus pesticide detection methods often use large laboratory instruments such as electrochemistry, mass spectrum, chromatography and the like, require professional operators and relatively long detection processes, and cannot be used for on-site detection. Later colorimetric methods were developed, but the observation of color changes often requires the help of an optical excitation instrument, and the accurate test cannot be performed on samples with dim light environments and interference on color display, and some people with weak colors cannot use related detection methods. The invention indicates the type and the concentration of the organophosphorus pesticide by means of the change of the contact angle which can be observed by naked eyes, can accurately measure the change of the angle by means of mobile phone software, does not need a large instrument, and is not interfered by ambient light and color.
According to the invention, the super-hydrophilic surface is prepared on the glass substrate, acetylcholinesterase or butyrylcholinesterase is introduced for modification, a hydrophobic surface is obtained, and then detection of different organophosphorus pesticides can be realized through the change of a contact angle. The method comprises the steps of constructing an array based on organophosphorus pesticide detection by adjusting the types and the concentrations of response molecules (acetylcholinesterase and butyrylcholinesterase), constructing a complete intelligent response surface contact angle change range database based on organophosphorus pesticides by laboratory detection of organophosphorus pesticides with different concentrations and different types and mixed organophosphorus pesticide samples, and determining the types and the residual amounts of the organophosphorus pesticides by comparison with the database during field detection.
The organophosphorus pesticide detector based on the intelligent response surface can be used for detecting organophosphorus pesticides on site, is convenient to carry, and can be applied to organophosphorus pesticide residue detection on vegetables and fruits.
Drawings
FIG. 1 is a scanning electron microscope image and contact angle of a hydrophilic surface prepared in accordance with the present invention;
FIG. 2 is a scanning electron microscope image and contact angle of a hydrophobic surface prepared in accordance with the present invention;
FIG. 3 is a photograph of a hydrophobic surface prepared in accordance with the present invention;
fig. 4 shows the change in contact angle of dichlorvos solution at different concentration gradients;
FIG. 5 variation of contact angle of malathion solution with different concentration gradients;
FIG. 6 variation of contact angle of glyphosate solution at different concentration gradients;
FIG. 7 Chlorpyrifos solution contact angle variation for different concentration gradients;
FIG. 8 variation of contact angle of methyl parathion solution with different concentration gradients;
FIG. 9 variation of profenofos solution contact angle for different concentration gradients;
FIG. 10 variation of contact angle of methomyl solution at different concentration gradients;
FIG. 11 variation of contact angle of fenitrothion solution with different concentration gradients;
FIG. 12 is a bar graph of the change in contact angle for 9 organophosphorus pesticides of the same concentration at 20deg.C;
figure 13 bar graph of contact angle change for 9 organophosphorus pesticides of the same concentration at 30 ℃.
Detailed Description
The terms used in the present invention generally have meanings commonly understood by those of ordinary skill in the art unless otherwise indicated.
The invention will be described in further detail below in connection with specific embodiments and with reference to the data. The following examples are intended to illustrate the invention and are not intended to limit the scope of the invention in any way.
The principle of the invention is as follows:
the method comprises the steps of constructing a super-hydrophilic surface on the surface of a glass substrate, then adopting acetylcholinesterase or butyrylcholinesterase for modification, and generating detection interaction of the organic phosphorus pesticide molecules and damage to a hydrophobic surface structure to different degrees through the difference of interaction strength of the acetylcholinesterase (or butyrylcholinesterase) and different organic phosphorus pesticide molecules and weak substrate interaction between the acetylcholinesterase (or butyrylcholinesterase) and the super-hydrophobic surface, thereby generating change of a surface contact angle. The method comprises the steps of constructing an array based on organophosphorus pesticide detection by adjusting the types and the concentrations of response molecules (acetylcholinesterase and butyrylcholinesterase), constructing a complete intelligent response surface contact angle change range database based on organophosphorus pesticides by laboratory detection of organophosphorus pesticides with different concentrations and different types and mixed organophosphorus pesticide samples, and determining the types and the residual amounts of the organophosphorus pesticides by comparison with the database during field detection.
In the detection system, acetylcholinesterase (or butyrylcholinesterase) is an organophosphorus pesticide response molecule, and has different binding strength with different organophosphorus pesticides to generate different degrees of reaction; the reaction can cause a different degree of disruption to the hydrophobic surface structure, which in turn can generate a signal of the change in the contact angle of the surface. Different concentrations of organophosphorus pesticides will produce different contact angle changes over a range, while non-organophosphorus pesticide molecules that do not respond to acetylcholinesterase (or butyrylcholinesterase) will not produce a contact angle change signal. Dropping organophosphorus pesticide molecules to be detected on the hydrophobic surface of the prepared organophosphorus pesticide detector based on the intelligent response surface, and measuring the contact angle; different types and concentrations of organophosphorus pesticides can obtain different contact angle information, so as to obtain different organophosphorus pesticide molecular information.
Example 1
An organophosphorus pesticide detector based on intelligent response surface is prepared by the following method:
(1) Preparation of hydrophilic surface: coCl is to be processed 2 ·6H 2 O and (NH) 2 ) 2 Placing the aqueous solution of CO in a sealed container, wherein the CoCl 2 ·6H 2 O concentration of 0.36g L -1 ，(NH 2 ) 2 CO concentration of 200g L -1 Taking a common borosilicate glass slide as a deposition matrix, reacting for 24 hours at 60 ℃, washing the deposited surface with deionized water, and drying to prepare a hydrophilic surface;
(2) Modification of acetylcholinesterase: every 5cm 2 100 mu L of the hydrophilic surface of the substrate was dropped on the substrate to a concentration of 1 U.mL -1 Is dried at room temperature
(3) Preparation of hydrophobic surface: immersing the modified surface in PFOTES solution for 40min, washing the prepared surface with ethanol solution, and naturally drying to obtain a hydrophobic surface; and obtaining the organophosphorus pesticide detector based on the intelligent response surface.
In the step (3), the PFOTES solution is an ethanol water solution of PFOTES, wherein the volume ratio of ethanol to water is 1:1, and the volume percentage of PFOTES is 1%.
The scanning electron microscope image and the contact angle of the hydrophilic surface prepared in the step (1) are shown in fig. 1, the scanning electron microscope image and the contact angle of the hydrophobic surface prepared in the step (3) are shown in fig. 2, and the finally prepared image of the hydrophobic surface of the organophosphorus pesticide detector based on the intelligent response surface is shown in fig. 3.
Example 2
The method for detecting the organophosphorus pesticide by using the organophosphorus pesticide detector based on the intelligent response surface prepared in the embodiment 1 comprises the following steps:
(1) 3 mu L of known organophosphorus pesticides with different concentrations or known organophosphorus pesticide mixture with different concentrations is dripped on the surface of the organophosphorus pesticide detector based on the intelligent response surface, the change range of the organophosphorus pesticide on the surface contact angle of the organophosphorus pesticide detector based on the intelligent response surface is detected, and an intelligent response surface contact angle change range database based on the organophosphorus pesticide is constructed;
(2) Dropwise adding a sample to be detected to the surface of the organophosphorus pesticide detector based on the intelligent response surface, detecting the change range of the contact angle of the sample to be detected on the surface of the organophosphorus pesticide detector based on the intelligent response surface, and comparing the constructed intelligent response surface contact angle change range database based on the organophosphorus pesticide to obtain the type and concentration of the organophosphorus pesticide in the sample to be detected;
wherein the temperature is 20-40deg.C, and the detection time is 20-60min.
Example 3
Variation of organophosphorus pesticide molecules of different concentrations in surface contact angle of organophosphorus pesticide detector based on intelligent response surface
Different concentrations of organophosphorus pesticide molecules were respectively dropped on the hydrophobic surface of the organophosphorus pesticide detector based on the intelligent response surface prepared in example 1 above, and the detection temperature was 20 ℃. And respectively measuring the contact angle of the pesticide molecule drop when the pesticide molecule drop initially contacts the hydrophobic surface and the contact angle after 20min, 25min and 30min, plotting by taking the value of the contact angle as an ordinate and the response time of acetylcholinesterase and the organophosphorus pesticide molecule as an abscissa, and detecting the contact angle change ranges of the organophosphorus pesticide molecules with different types and different concentrations.
The results are shown in FIGS. 4-11, where-1 represents a concentration of 10 -1 mol/L, -2 represents a concentration of 10 -2 mol/L, -3 represents a concentration of 10 -3 mol/L, and so on; water represents the change in contact angle of water over the measurement time range. On the superhydrophobic surface, the response of organophosphorus pesticide molecules with different concentrations to acetylcholinesterase is detected by comparing the change of the contact angle with water. In the initial stage of detection, the contact angle difference of the solutions with four concentrations is smaller, and obvious difference appears between the contact angles of the solutions with different concentrations along with the time. The contact angle change of water is stabilized at about 10 degrees within 30 minutes, and compared with water, the change of glyphosate, chlorpyrifos and parathion methyl is not obvious. The result may be that the three pesticide molecules respond poorly to acetylcholinesterase. Malathion, profenofos, dichlorvos, methomyl and fenitrothion are respectively 10 percent -3 mol/L、10 -1 mol/L，10 -1 mol/L，10 -1 mol/L，10 -3 The greatest change in contact angle occurs in mol/L. By combining detection data of various organophosphorus pesticide molecules, the result shows that the higher the concentration of the pesticide molecules is, the more obvious the contact angle change is, but 10 is selected to ensure that the optimal detection effect can be found under the condition of low concentration -3 mol/L is the optimal detection condition.
Example 4
Variation of surface contact angle of organophosphorus pesticide molecules with same concentration on organophosphorus pesticide detector based on intelligent response surface
(1) Respectively the same concentration (10 -3 mol/L) of the organophosphorus pesticide molecule was dropped on the hydrophobic surface of the intelligent response surface-based organophosphorus pesticide detector prepared in example 1 above, and the detection temperature was 20 ℃. And respectively measuring the contact angle of the pesticide molecule drop when initially contacting the hydrophobic surface and the contact angle after 30min of action, and calculating the contact angle variation ranges of the organophosphorus pesticide molecules with the same concentration in different types.
As a result, as shown in FIG. 12, the same concentration gradient (30 min,10 -3 The contact angle of different organophosphorus pesticides with mol/L) is changed from large to small, namely chlorpyrifos Chl (57.82 degrees) to glyphosate Gly (47.63 degrees) to phosphorus sesquioxide Fent (34.92 degrees) to fenitrothion Fen (33.85 degrees) to dichlorvos DDVP (28.67 degrees) to malathion Mal (26.77 degrees) to profenofos Pro (25.23 degrees) to methyl parathion Par (20.13 degrees) to methomyl Thi (14.79 degrees). Under this condition, the contact angle of chlorpyrifos changed the most and the contact angle of methomyl changed the least. By acting with a hydrophobic surface, different organophosphorus pesticide molecules can be initially distinguished.
(2) Respectively the same concentration (10 -3 mol/L) of the organophosphorus pesticide molecules were dropped on the hydrophobic surface of the intelligent response surface-based organophosphorus pesticide detector prepared in example 1 above, and the detection temperature was 30 ℃. And respectively measuring the contact angle of the pesticide molecule drop when initially contacting the hydrophobic surface and the contact angle after 60 minutes of action, and calculating the contact angle variation ranges of the organophosphorus pesticide molecules with the same concentration in different types.
As a result, as shown in fig. 13, the contact angle of different organophosphorus pesticides with the same concentration gradient changes from large to small in the same time: dichlorvos DDVP (58.23 °) > glyphosate Gly (54.65 °) > profenofos Pro (38.55 °) > fenitrothion Fen (30.24 °) > methomyl Thi (27.98 °) > methyl parathion Par (27.77 °) > chlorpyrifos Chl (24.6 °) > phoxim pent (22.98 °) > malathion Mal (18.76 °). Under this condition, the change in contact angle of dichlorvos is greatest and the change in contact angle of malathion is smallest. Under the condition, the method is favorable for screening dichlorvos and glyphosate pesticide molecules.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the invention in any way, and any person skilled in the art may make modifications or alterations to the disclosed technical content to the equivalent embodiments. However, any simple modification, equivalent variation and variation of the above embodiments according to the technical substance of the present invention still fall within the protection scope of the technical solution of the present invention.
Claims (10)
1. The preparation method of the organophosphorus pesticide detector based on the intelligent response surface is characterized by comprising the following steps of:
(1) Immersing a glass substrate into CoCl 2 ·6H 2 O and (NH) 2 ) 2 Carrying out hydrothermal reaction in a mixed aqueous solution of CO at 55-65 ℃ and drying to obtain the super-hydrophilic surface;
(2) Every 4-6cm 2 100 μl of the solution of the response molecule is dropped onto the hydrophilic surface of (a), and dried at room temperature;
(3) Immersing the modified hydrophilic surface obtained in the step (2) into a perfluorooctyl triethoxysilane solution for 30-40min, and drying to obtain a hydrophobic surface; obtaining the organophosphorus pesticide detector based on the intelligent response surface;
the response molecule solution is acetylcholinesterase phosphate buffer salt solution or butyrylcholinesterase phosphate buffer salt solution, and the concentration is 1 U.mL -1 ；
The method for detecting the organophosphorus pesticide by adopting the organophosphorus pesticide detector based on the intelligent response surface comprises the following steps of:
dripping known organophosphorus pesticides with different concentrations or a mixture of known organophosphorus pesticides with different concentrations on the surface of the organophosphorus pesticide detector based on the intelligent response surface, detecting the variation range of the surface contact angle of the organophosphorus pesticide on the organophosphorus pesticide detector based on the intelligent response surface, and constructing an intelligent response surface contact angle variation range database based on the organophosphorus pesticide;
and (3) dropwise adding a sample to be detected to the surface of the organophosphorus pesticide detector based on the intelligent response surface, detecting the change range of the contact angle of the sample to be detected on the surface of the organophosphorus pesticide detector based on the intelligent response surface, and comparing the constructed intelligent response surface contact angle change range database based on the organophosphorus pesticide to obtain the type and concentration of the organophosphorus pesticide in the sample to be detected.
2. The method for preparing an organophosphorus pesticide detector based on an intelligent response surface according to claim 1, wherein the CoCl 2 ·6H 2 O and (NH) 2 ) 2 CoCl in aqueous CO-mixture 2 ·6H 2 The O concentration is 0.20-0.50g L -1 ，(NH 2 ) 2 The concentration of CO is 150-250g L -1 。
3. The method for preparing an organophosphorus pesticide detector based on an intelligent response surface according to claim 1, wherein in the step (1), the hydrothermal reaction time is 22-26h.
4. The method for preparing an organophosphorus pesticide detector based on an intelligent response surface according to claim 1, wherein in the step (3), the perfluorooctyl triethoxysilane solution is an ethanol aqueous solution of perfluorooctyl triethoxysilane, wherein the volume ratio of ethanol to water is 1:1, and the volume percentage of perfluorooctyl triethoxysilane is 1%.
5. An organophosphorus pesticide detector based on a smart responsive surface prepared by the method of any of claims 1-4.
6. The use of the intelligent response surface-based organophosphorus pesticide detector as defined in claim 5 in organophosphorus pesticide detection.
7. The use according to claim 6, wherein the organophosphorus pesticide is one or more of profenofos, dichlorvos, fenitrothion, malathion, methyl parathion, glyphosate, chlorpyrifos, methomyl, and fenthion.
8. A method for detecting organophosphorus pesticides, which is characterized in that organophosphorus pesticides with different known types and different concentrations or mixtures of organophosphorus pesticides with different known concentrations are dripped on the surface of the organophosphorus pesticide detector based on the intelligent response surface according to claim 5, the change range of the organophosphorus pesticide on the surface contact angle of the organophosphorus pesticide detector based on the intelligent response surface is detected, and an intelligent response surface contact angle change range database based on the organophosphorus pesticide is constructed;
dropwise adding a sample to be detected to the surface of the organophosphorus pesticide detector based on the intelligent response surface according to claim 5, detecting the change range of the contact angle of the sample to be detected on the surface of the organophosphorus pesticide detector based on the intelligent response surface, and comparing the constructed intelligent response surface contact angle change range database based on the organophosphorus pesticide to obtain the type and concentration of the organophosphorus pesticide in the sample to be detected.
9. The method for detecting organophosphorus pesticides according to claim 8, wherein the temperature is 20-40 ℃ and the detection time is 20-60min.
10. The method for detecting organophosphorus pesticides according to claim 8 or 9, wherein the organophosphorus pesticides are one or more of profenofos, dichlorvos, fenitrothion, malathion, methyl parathion, glyphosate, chlorpyrifos, methomyl and fenthion.
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