CN104820003A - Paper-based micro-fluidic system and method for detection of pesticide residues - Google Patents

Paper-based micro-fluidic system and method for detection of pesticide residues Download PDF

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Publication number
CN104820003A
CN104820003A CN201510241333.8A CN201510241333A CN104820003A CN 104820003 A CN104820003 A CN 104820003A CN 201510241333 A CN201510241333 A CN 201510241333A CN 104820003 A CN104820003 A CN 104820003A
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electrode
papery
micro
sample
detection
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CN104820003B (en
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毛罕平
施杰
严玉婷
杨宁
左志强
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Jiangsu University
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Jiangsu University
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Abstract

The invention discloses a paper-based micro-fluidic system and method for detection of pesticide residues. The paper-based micro-fluidic system is composed of a paper-based micro-fluidic chip, a three-electrode system and a data collection device, wherein a substrate and a pesticide extract liquid are injected into the paper-based micro-fluidic chip; a detection signal is transmitted to the data collection device through the three-electrode system; and the pesticide residue detection result is rapidly obtained on the basis of an enzyme inhibition principle. The paper-based micro-fluidic system is simple and convenient to operate; the automatic degree of an enzyme inhibition method detection process is improved; meanwhile, the cost is greatly reduced by use of a paper-based material; and the paper-based micro-fluidic system and the method can be applied to rapid and automatic field detection of the pesticide residues in samples such as fruits and vegetables, soil and water bodies.

Description

A kind of papery microfluidic system for Detecting Pesticide and method
Technical field
The present invention relates to the technical field of papery microfluidic chip analysis technology and the Detection Technologies of Pesticide Residues combination, be specifically related to a kind of papery microfluidic system for Detecting Pesticide and method.
Background technology
Agricultural chemicals plays an important role in agricultural production, and in China's agricultural production, the pesticide species of use is various, comprises several large class kinds more than totally 600 such as Insecticides (tech) & Herbicides (tech), germifuge, rat poison.Organophosphorus and carbamate pesticides class agricultural chemicals is the maximum agricultural chemicals of the current use amount of China, and more be used on fruit trees and vegetables.Therefore, the residues of pesticides at present in the agricultural product such as fruit, vegetables, aquatic products, soil exceed standard the most outstanding with organophosphorus pesticide and carbamate pesticide residue.For monitoring and the management of organophosphate and carbamate pesticide, stop the most important thing that agricultural product list marketing that residues of pesticides exceed standard becomes agricultural product quality and safety work.
Constantly strengthen the testing of residues of pesticides both at home and abroad, the method for detection also gets more and more.The detection method of residues of pesticides is varied, have following a few class by Cleaning Principle: 1) chromatography, comprise vapor-phase chromatography, liquid phase chromatography and capillary electrophoresis 2) spectroscopic methodology 3) immunoassay 4) inhibiting AChE 5) chemical fast determining method 6) living body biological method.Inhibiting AChE is most study and a kind of Fast Determination of Pesticide Residue technology of relative maturity, this method measures Organophosphorus and carbamate pesticides pesticide residue and has quick, easy, sensitive and low cost and other advantages, the dark welcome by vast grass-roots supervision department, production base and market.
Enzyme sheet method (quick measuring card method) is roughly divided into based on the Fast Determination of Pesticide Residue means of inhibiting AChE, PH method, spectrophotometric method (colourimetry) and hexavalent chrome bio-removal etc. in prior art.Make a general survey of these Detecting Pesticide means present situations based on inhibiting AChE principle can find, enzyme level principle itself requires priority hybrid reaction and the blank of plurality of reagents, bring complex operation step, problem that automaticity is low thus, and the detection means that Current Domestic has been reported all fails effectively to address this problem outward.
Therefore develop a kind of fast and automatically, the Detection Technologies of Pesticide Residues that is portable, low cost becomes current study hotspot.Adopt papery micro-fluidic chip, make channel architecture on its surface and the analytical technology that itself and biosensor technology merge can effectively be solved the problem, greatly reduce cost, but this technology not yet there is substantial breakthrough in Detecting Pesticide field at present simultaneously.
Summary of the invention
The object of the invention is to for the domestic and international Detecting Pesticide means ubiquity complex operation step based on inhibiting AChE at present, problem that automaticity is low, provide a kind of fast and automatically, the Detection Technologies of Pesticide Residues of portable, low cost.
In order to solve above technical matters, the present invention is based on the analytical technology of papery microfluidic chip technology and electrochemica biological sensor technological incorporation.Concrete technical scheme is as follows:
For a papery microfluidic system for Detecting Pesticide, it is characterized in that: described system is made up of papery micro-fluidic chip, three-electrode system A, three-electrode system B and data collector; Three-electrode system A and three-electrode system B is integrated on papery micro-fluidic chip, and data collector connects three-electrode system A and three-electrode system B by data line.
The channel architecture of described papery micro-fluidic chip is Y type; Described Y type channel architecture is made up of sample inlet pool A and sample inlet pool B, sample intake passage A and sample intake passage B, detection cell A and detection cell B, hybrid channel, passive mixed structure; Be connected with hybrid channel after sample intake passage A and sample intake passage B converge, sample inlet pool A and sample inlet pool B lays respectively at the upper end of sample intake passage A and sample intake passage B, detection cell A is positioned at the middle part of sample intake passage A, detection cell B is positioned at hybrid channel end, and passive mixed structure is positioned at the centre position of hybrid channel.
The making material of described papery micro-fluidic chip is any one in filter paper, chromatographic paper or nitrocellulose filter.
Described three-electrode system A and three-electrode system B is all integrated on papery micro-fluidic chip; Described three-electrode system A and three-electrode system B include working electrode, contrast electrode, to electrode, contrast electrode and electrode is laid respectively to the both sides of working electrode; Three-electrode system A and three-electrode system B lays respectively at the center of detection cell A and detection cell B.
Two working electrodes of described three-electrode system A and three-electrode system B all secure enzyme.
Described fixing fixing means is any one in absorption method, investment, cross-linking method or covalent coupling method; Described enzyme is any one in acetylcholinesterase, butyrylcholine esterase, plant esterase.
Described working electrode and be any one in platinum, gold, carbon to the material of electrode; Described contrast electrode material be the argent being covered with silver chloride layer be immersed in formed in potassium chloride or hydrochloric acid solution compound substance, silver, in carbon any one.
Described data collector is that the data acquisition circuit of core and the program of microcontroller are formed by microcontroller; Described microcontroller is any one in single-chip microcomputer, DSP, ARM; Described data acquisition circuit is made up of potentiostat circuit, amplifying circuit, filtering circuit, A/D conversion circuit, interface circuit, microcontroller peripheral circuit.
A kind of pesticide residue detection method, is characterized in that comprising following characteristics step:
Step one, pre-treatment: to the pre-treatment of testing sample, comprises and gets that testing sample is put into beaker, added extractant, soak extraction goes out agricultural chemicals extract;
Step 2, injects: by the mode of syringe pump or hand injection, substrate and agricultural chemicals extract are injected papery micro-fluidic chip from sample inlet pool A and sample inlet pool B simultaneously;
Step 3, hybrid reaction: after substrate and agricultural chemicals extract inject from sample inlet pool A and sample inlet pool B respectively, first substrate flows through the detection cell A in the middle part of sample intake passage A, when carrying out catalytic reaction without the immobilised enzymes A on three-electrode system A working electrode when agricultural chemicals extract to substrate, produce electroactive material, meanwhile agricultural chemicals extract flows through sample intake passage B, then substrate and agricultural chemicals extract converge and jointly pass through hybrid channel afterwards, mix under the passive mixed structure effect in centre position, hybrid channel, the last detection cell B simultaneously flowing into hybrid channel end, immobilised enzymes B when there being agricultural chemicals extract to suppress on three-electrode system B working electrode carries out catalytic reaction to substrate, produce electroactive material,
Step 4, Electrochemical Detection: the micro-current signal produced by three-electrode system A and three-electrode system B detection electroactive material redox reaction, thus form blank;
Step 5, signals collecting: gather micro-current signal by the data collector connecting three-electrode system A and three-electrode system B, transfer to host computer and carry out data processing, contrast calculates the residues of pesticides concentration of testing sample.
Described testing sample is any one in fruits and vegetables, soil, water body; Described substrate is any one in acetylcholine, acetylthiocholine, acetonaphthone ester; Described agricultural chemicals extract is organophosphorus pesticide or carbamate chemicals for agriculture.
Present invention employs cheap papery micro-fluidic chip, channel architecture is made and the three-electrode system of integrated two immobilized enzymes on its surface, again by data collector collection signal, automatically priority hybrid reaction and the blank of plurality of reagents in Detecting Pesticide process can be completed under paper capillary force drives, easy and simple to handle, improve the automaticity of inhibiting AChE testing process, the simultaneously use of paper base material greatly reduces cost, be suitable for fruits and vegetables, soil, water body equal samples Pesticide Residues fast and automatically, Site Detection.
Accompanying drawing explanation
Fig. 1 is the structural representation of papery microfluidic system in the present invention.
Fig. 2 is the hardware circuit composition schematic diagram of data collector in the present invention.
In figure: 1. sample inlet pool A, 2. sample intake passage A, 3. detection cell A, 4. sample inlet pool B, 5. sample intake passage B, 6. passive mixed structure, 7. hybrid channel, 8. detection cell B, 9. immobilised enzymes B, 10. three-electrode system B, 11. three-electrode system A, 12, immobilised enzymes A.
Embodiment
Below in conjunction with accompanying drawing, be detected as example with the organophosphorus pesticide of vegetables, the specific embodiment of the present invention is described in detail.
In the present embodiment, the papery microfluidic system for Detecting Pesticide comprises micro-fluidic chip and a data collector of a papery, and wherein the structure of papery microfluidic system as shown in Figure 1, and the hardware circuit composition of data collector as shown in Figure 2.
Concrete testing process is:
Step one, is first placed in the containers such as beaker by the vegetables sample of 5g, adds for 2000 μ L extractant immersion treatment a period of times, or drips extractant at vegetable surface, obtains the agricultural chemicals extract extracted.
2000 μ L substrate acetyl choline solutions are injected into chip from 1 sample inlet pool A by step 2,2000 μ L agricultural chemicals extracts are injected chip from 4 sample inlet pool B simultaneously.
Step 3, substrate acetyl choline solution flows through 2 sample intake passage A to 3 detection cell A, and with the 12 immobilised enzymes A be fixed on the working electrode of 11 three-electrode system A, catalytic reaction occurs, the electroactive material of generation forms redox current signal on the working electrode (s.
Step 4, agricultural chemicals extract flows through 5 sample intake passage B, converges in 7 hybrid channels with substrate acetyl choline solution, then fully mixes under the effect of 6 passive mixed structures.
Step 5, the mixed solution of agricultural chemicals extract and substrate flow to 8 detection cell B, and with the 9 immobilised enzymes B be fixed on the working electrode of 10 three-electrode system B, catalytic reaction occurs, the electroactive material of generation forms redox current signal on the working electrode (s.
Step 6, micro-current signal that the redox effect of the electroactive material on three-electrode system A and three-electrode system B testing electrode produces.
Step 7, two current signals that data collector collection detects, form blank, transfer to host computer, carry out data processing, obtain surveyed residue of organic phosphorus pesticides in vegetable amount by the means such as wireless telecommunications or bluetooth.
Above-mentioned step 2 ~ seven are system automation operation, the papery microfluidic system using this enforcement to provide, and can obtain the sxemiquantitative of Organophosphorus and carbamate pesticides pesticide residue or quantitative testing result in 10 ~ 15min.

Claims (9)

1. for a papery microfluidic system for Detecting Pesticide, it is characterized in that: described system is made up of papery micro-fluidic chip, three-electrode system A, three-electrode system B and data collector; Three-electrode system A and three-electrode system B is integrated on papery micro-fluidic chip, and data collector connects three-electrode system A and three-electrode system B by data line.
2. a kind of papery microfluidic system for Detecting Pesticide according to claim 1, is characterized in that: the channel architecture of described papery micro-fluidic chip is Y type; Described Y type channel architecture is made up of sample inlet pool A and sample inlet pool B, sample intake passage A and sample intake passage B, detection cell A and detection cell B, hybrid channel, passive mixed structure; Be connected with hybrid channel after sample intake passage A and sample intake passage B converge, sample inlet pool A and sample inlet pool B lays respectively at the upper end of sample intake passage A and sample intake passage B, detection cell A is positioned at the middle part of sample intake passage A, detection cell B is positioned at hybrid channel end, and passive mixed structure is positioned at the centre position of hybrid channel.
3. a kind of papery microfluidic system for Detecting Pesticide according to claim 2, is characterized in that: the making material of described papery micro-fluidic chip is any one in filter paper, chromatographic paper or nitrocellulose filter.
4. a kind of papery microfluidic system for Detecting Pesticide according to claim 1, is characterized in that: described three-electrode system A and three-electrode system B is all integrated on papery micro-fluidic chip; Described three-electrode system A and three-electrode system B include working electrode, contrast electrode, to electrode, contrast electrode and electrode is laid respectively to the both sides of working electrode; Three-electrode system A and three-electrode system B lays respectively at the center of detection cell A and detection cell B.
5. a kind of papery microfluidic system for Detecting Pesticide according to claim 4, is characterized in that: two working electrodes of described three-electrode system A and three-electrode system B all secure enzyme.
6. a kind of papery microfluidic system for Detecting Pesticide according to claim 5, is characterized in that: described fixing fixing means is any one in absorption method, investment, cross-linking method or covalent coupling method; Described enzyme is any one in acetylcholinesterase, butyrylcholine esterase, plant esterase.
7. a kind of papery microfluidic system for Detecting Pesticide according to claim 1, is characterized in that: described data collector is that the data acquisition circuit of core and the program of microcontroller are formed by microcontroller; Described microcontroller is any one in single-chip microcomputer, DSP, ARM; Described data acquisition circuit is made up of potentiostat circuit, amplifying circuit, filtering circuit, A/D conversion circuit, interface circuit, microcontroller peripheral circuit.
8. the pesticide residue detection method of a kind of papery microfluidic system for Detecting Pesticide according to claim 1, is characterized in that comprising following characteristics step:
Step one, pre-treatment: to the pre-treatment of testing sample, comprises and gets that testing sample is put into beaker, added extractant, soak extraction goes out agricultural chemicals extract;
Step 2, injects: by the mode of syringe pump or hand injection, substrate and agricultural chemicals extract are injected papery micro-fluidic chip from sample inlet pool A and sample inlet pool B simultaneously;
Step 3, hybrid reaction: after substrate and agricultural chemicals extract inject from sample inlet pool A and sample inlet pool B respectively, first substrate flows through the detection cell A in the middle part of sample intake passage A, when carrying out catalytic reaction without the immobilised enzymes A on three-electrode system A working electrode when agricultural chemicals extract to substrate, produce electroactive material, meanwhile agricultural chemicals extract flows through sample intake passage B, then substrate and agricultural chemicals extract converge and jointly pass through hybrid channel afterwards, mix under the passive mixed structure effect in centre position, hybrid channel, the last detection cell B simultaneously flowing into hybrid channel end, immobilised enzymes B when there being agricultural chemicals extract to suppress on three-electrode system B working electrode carries out catalytic reaction to substrate, produce electroactive material,
Step 4, Electrochemical Detection: the micro-current signal produced by three-electrode system A and three-electrode system B detection electroactive material redox reaction, thus form blank;
Step 5, signals collecting: gather micro-current signal by the data collector connecting three-electrode system A and three-electrode system B, transfer to host computer and carry out data processing, contrast calculates the residues of pesticides concentration of testing sample.
9. a kind of pesticide residue detection method according to claim 8, is characterized in that: described testing sample is any one in fruits and vegetables, soil, water body; Described substrate is any one in acetylcholine, acetylthiocholine, acetonaphthone ester; Described agricultural chemicals extract is organophosphorus pesticide or carbamate chemicals for agriculture.
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CN105717166A (en) * 2016-02-15 2016-06-29 江苏大学 Tree growth state real-time detecting device and method based on micro-fluidic chip
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CN106199013A (en) * 2016-08-16 2016-12-07 北京大学第医院 A kind of low abundance hormone test method and device
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CN105403612A (en) * 2015-12-18 2016-03-16 重庆大学 Method for rapidly detecting pesticide residue based on plant esterase
CN105717166A (en) * 2016-02-15 2016-06-29 江苏大学 Tree growth state real-time detecting device and method based on micro-fluidic chip
CN105717166B (en) * 2016-02-15 2019-08-27 江苏大学 A kind of arboreal growth real-time detection device for state and method based on micro-fluidic chip
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CN108212232A (en) * 2018-01-09 2018-06-29 昆明理工大学 One kind is based on the double supplementary structure assisted extraction devices of microflow control technique and method
CN108548908A (en) * 2018-04-24 2018-09-18 苏州格目软件技术有限公司 A kind of residual prompt system of environmentally friendly agriculture based on processing water
CN108535249A (en) * 2018-04-25 2018-09-14 郑州贝贝生物科技有限公司 Fast Determination of Pesticide Residue point card
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CN109358043A (en) * 2018-11-01 2019-02-19 中国农业科学院茶叶研究所 A method of sample fast detecting pesticide residue is extracted with organic solvent
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