CN103940866A - Rapid detector for quantitative detection on pesticide residue in fruits and vegetables - Google Patents
Rapid detector for quantitative detection on pesticide residue in fruits and vegetables Download PDFInfo
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Abstract
The invention discloses a rapid detector for quantitative detection on pesticide residue in fruits and vegetables, which belongs to the technical field of agricultural product safety detection. The rapid detector provided by the invention is composed of a three-electrode system with an acetylcholin esterase sensor, a signal detection and processing system, a display and printing storage system and a power supply system, wherein a fruit and vegetable pesticide type and national pesticide residue limit database, an organophosphorus and carbamic acid ester pesticide standard curve database and a BP (back propagation) neural network algorithm are embedded in a microprocessor of the detector; the measured value is compared with the common organophosphorus and carbamic acid ester pesticide standard curve database stored in a storage unit by a BP neural network so as to quantitatively determine information such as the types of contained pesticides and whether the pesticides exceed standards or not. The rapid detector provided by the invention is capable of performing qualitative and quantitative detection on pesticide residue, has the characteristics of being convenient to carry, rapid and accurate in detection and simple to operate, and is applicable to field rapid detection on pesticide residue in fruits and vegetables.
Description
Technical field
The present invention relates to a kind of fast detector of quantitative detection fruits and vegetables Pesticide Residues, belong to agricultural product security detection technique field.
Technical background
The pesticide variety of applying in production field now has reached kind more than 100, keeps good harvest volume increase to play a significant role to agricultural, and it has become a kind of important means of production.Be accompanied by deeply carrying out of reform and opening-up, in some Changshu Aeras, the speed of development of agricultural chemicals cause is very fast.Because mechanization of agriculture constantly advances, it is indispensable that the large area of agricultural chemicals is used.But because of irrational use of agricultural chemicals, cause many environmental problems, as destroyed in farmland ecosystem etc.A large amount of uses of agricultural chemicals, in having ensured a large amount of stable yields of the crops such as grain, have also brought many environmental problems.In plant, soil and water, residual agricultural chemicals has two kinds of remaining forms: the first keeps original chemical constitution; The product of the second after with chemical conversion or to be biodegradable the form of product remaining.Therefore, Pesticide Residue can not be ignored.
The main method of pesticide residue analysis is gas chromatograph, liquid chromatograph, gas chromatograph-mass spectrometer, LC-MS instrument etc. at present, although these method analysis precisions are high, quantitatively accurately, but the complex pretreatment of its sample, detect the operating personnel that length consuming time, cost are high, need to be skilled in technique.The fast determining method of China's residues of pesticides is that enzyme suppresses test paper method and enzyme inhibition-spectrophotometry (the residual fast detector of agriculture), can realize the field quick detection of organophosphorus pesticide and carbamate chemicals for agriculture, has good practical value.Quick measuring card is by the change color of visual inspection card, and the vegetable sample that therefore generally can only be used for severe overweight carries out observational measurement.The application of enzyme inhibition-spectrophotometry is also more extensive, and the residual tacheometer of domestic existing Multiple Pesticides is all based on this principle.Light splitting light ratio juris is that the variation based on absorbance detects, but in vegetable and fruit, a large amount of pigments can cause very large impact to spectrophotometric method, causes the inaccurate of testing result.And said method exists, and the recovery is low, false retrieval, undetected ratio are higher, poor repeatability, be difficult to meet low-residual and the shortcoming such as the requirement that quantitatively detects.
Summary of the invention
The object of the invention is to for the deficiencies in the prior art, a kind of fast detector that can quantitatively detect fruits and vegetables Pesticide Residues is provided, have easy to carry, detect quick and precisely, simple to operate, can qualitative, quantitative measure and in fruits and vegetables, whether contain residues of pesticides, concrete concentration and the information such as whether exceed standard.Take following technical scheme:
A kind of fast detector of quantitative detection fruits and vegetables Pesticide Residues, comprise acetylcholinesterase sensor three-electrode system, input and disposal system, show and print storage system, electric power system composition, in the microprocessor of the inside of detector, implant fruits and vegetables pesticide variety and national Pesticide Residue database, organophosphorus and carbamate pesticide typical curve database and BP neural network algorithm, detector utilizes BP neural network that the conventional organophosphorus of measured value and cell stores and carbamate chemicals for agriculture typical curve database are compared, the contained pesticide variety of qualitative definite sample and whether exceeding standard, and the concentration of the agricultural chemicals that quantitatively exceeds standard,
Described acetylcholinesterase sensor three-electrode system, forms electrode by being fixed with acetylcholinesterase working electrode, saturated calomel reference electrode and platinum, sends the current signal containing before and after agricultural chemicals collecting to input and disposal system;
The preparation method of described acetylcholinesterase sensor, step is as follows:
1) in 50mL acetic acid solution, add 0.1g shitosan to be prepared into 0.2% chitosan solution, in 4mL 0.2% chitosan solution, add 1mg tin ash and 3mg to hide carbon nano-tube, in 25 DEG C of water, after ultrasonic concussion 30min, obtain many walls carbon-tin ash-chitosan nano composite membrane;
2) get many walls carbon-tin ash-chitosan nano composite membrane that 2.5 μ L prepare and drip and be coated onto the gold electrode surfaces that pre-service is good, normal temperature leaves standstill 1h, rinses electrode surface by the phosphate buffered solution of pH7.5;
3) after electrode surface dries, the acetylcholinesterase of getting 2.5 μ L drips and is coated in electrode surface, leaves standstill 2h at 4 DEG C, then rinses surface by the phosphate buffered solution of pH7.5, and nitrogen dries up;
4) the Nafion diaphragm of getting 2.5 μ L 0.5% drips and is coated in electrode surface, leaves standstill 1.5h under normal temperature, then rinses surface by the phosphate buffered solution of pH7.5, and nitrogen dries up;
Described input and disposal system, by Detection of Weak Signals and treatment circuit, A/D conversion chip and single-chip microcomputer composition, the low current signal collecting is carried out I/V conversion by Detection of Weak Signals and treatment circuit, amplify, after filtering, after the conversion of A/D chip, digital quantity signal is sent into single-chip microcomputer and carry out routine processes;
Described demonstration and storage print system, by mini-printer, display screen and storage unit composition are also unified programming Control by single-chip microcomputer to it, display screen shows remains of pesticide concentration, inhibition degree, data are stored in storage unit so that user checks in time and carries out data analysis, and data are printed output by mini-printer;
Described electric power system, containing potentiostatic circuit provides the stable operating voltage of 500mV for working electrode and contrast electrode;
Described fruits and vegetables pesticide variety and limitation database, be to set up according to the Pesticide maximum residue limit specifying in GB2763-2012, and be implanted in advance in the microprocessor of detector;
Described organophosphorus and carbamate chemicals for agriculture typical curve database, be to be measured and set up by mark-on method, and be implanted in advance in the microprocessor of detector;
In the microprocessor of described detector inside, implant BP neural network algorithm, utilize BP neural network that the conventional organophosphorus of measured value and cell stores and carbamate chemicals for agriculture typical curve database are compared, the contained pesticide variety of qualitative definite sample and whether exceeding standard, and the concentration of the agricultural chemicals that quantitatively exceeds standard.
A kind of step of the detection method of fast detector described in claim 1 of utilizing is as follows:
1) selection will detect the kind of vegetables, then acetylcholinesterase biology sensor working electrode is put into containing liquid at the bottom of the test of agricultural chemicals, the electric current I while recording not containing agricultural chemicals
1, electric current before suppressing, or the default value of setting in employing system;
2) acetylcholinesterase biology sensor working electrode is taken out, with distilled water flushing, then working electrode is put into analyte sample fluid and hatch 10min, again use distilled water flushing, then working electrode is put into test end liquid, recorded and suppress after-current I
2;
3) operation trace routine, detector calculates inhibiting rate automatically according to computing formula;
4) detector is according to vegetable species selected in step 1), with the calculating of comparing of pesticide variety and limitation database and typical curve database, determines whether residues of pesticides exceed standard and concentration.
Described method step 1) described in to test end liquid be the pH7.5 phosphate buffered solution that contains 1.0 mM chlorinated thio acetylcholines.
Described method step 2) described in the preprocess method of testing sample be: choose vegetables or fruit sample, with moist rag by the earth wiped clean of vegetable surface, be chopped into the square fragment of 2cm, sample thief 2g, puts into extraction flask, adds 10ml phosphate buffer, put into ultrasonic extraction apparatus, ultrasonic extraction 3min, pours out extract and leaves standstill 2min, for subsequent use.
Described method step 3) described in computing formula be: I %
=(i
1-I
2)/I
1× 100%.
Described fast detector is for the field quick detection of fruit and vegetable residues of pesticides.
Detection principle and the testing process of the fast detector of a kind of quantitative detection fruits and vegetables Pesticide Residues of the present invention are: the three-electrode system of acetylcholinesterase sensor causes acetylcholine esterase active to be suppressed by before and after acetylcholinesterase biology sensor exposed to pesticides, and then the electric current that acetylcholinesterase catalytic substrate produces is reduced, current signal acetylcholinesterase biology sensor being produced by testing circuit gathers, the low current signal collecting is carried out I/V conversion by testing circuit, amplify, filtering, A/D conversion the most at last digital quantity signal is sent into single-chip microcomputer and is carried out routine processes.User inputs the kind of information of vegetable and fruit on detector, detector just can be determined the kind that may use agricultural chemicals, before and after the exposed to pesticides of measuring according to detector, at the bottom of acetylcholinesterase catalysis, chloro acetylcholine generation current changes, calculate the inhibiting rate of enzymatic activity, the typical curve database of corresponding agricultural chemicals is determined the relevant informations the such as whether kind of agricultural chemicals and concentration exceed standard.
Beneficial effect of the present invention:
The application, in the storage unit of detector, has added kind and the national Pesticide Residue standard database of common pesticides in fruits and vegetables, conventional organophosphorus and carbamate pesticide typical curve database in fruits and vegetables.Utilize BP neural network that the conventional organophosphorus of measured value and cell stores and carbamate chemicals for agriculture typical curve database are compared, quantitative definitely contain which kind of agricultural chemicals and the information such as whether exceed standard.The invention solves the defect that existing instrument for detecting pesticide residue through can only observational measurement.
Brief description of the drawings
Fig. 1 is the structured flowchart of the fast detector of a kind of quantitative detection fruits and vegetables Pesticide Residues of the present invention.
Fig. 2 is the present invention's three electrodes and potentiostatic circuit and A/D conversion treatment circuit in early stage.
Embodiment
A kind of three-electrode system that in fruits and vegetables, the residual fast detector of Multiple Pesticides is made up of acetylcholinesterase biology sensor that detects of the present invention, input and disposal system, show and print storage system, electric power system composition.Potentiostatic circuit in electric power system provides the stable operating voltage of 500mV before to working electrode and contrast electrode.In test end liquid, add certain density chloro acetylcholine substrate, by measuring the information such as working electrode before and after exposed to pesticides and concentration to mensuration exposed to pesticides corresponding to the variation of the electric current producing between electrode.Signal deteching circuit gathers by working electrode and to the low current signal producing between electrode; The low current signal collecting is carried out I/V conversion by input and disposal system, amplifies, and filtering, A/D conversion the most at last digital quantity signal is sent into single-chip microcomputer and is processed; Show and print storage system by parameter displays such as residual pesticide concentration, inhibition degree out, and data being stored so that user checks in time and carries out data analysis.Detector of the present invention includes mini-printer, data can be carried out to corresponding printout.The single-chip microcomputer of input and disposal system inside carries out programming Control to showing with the unification of printing storage system.
In the present invention, the potentiostatic circuit in testing circuit provides stable operating voltage, the embodiment that Fig. 2 is potentiostat circuit for three-electrode system current/voltage transforms testing circuit, by a two-way operational amplifier U10 TLC272CP and R44, R46, R4, C35, C16, R48 forms.The pin 6 of amplifier B is connected with 7 and is connected with 3 pin of A, 2 pin of A are connected and have connected C16 with the working electrode of three-electrode system, strengthen antijamming capability, R48 be as feedback resistance on amplifier A, the external potentiometer R46 of 5 pins of B, supply electric excitation by positive 5V, R44, R46, R4 supplies with 500mv voltage to forming potentiostatic circuit with 5 pins of B by voltage dividing potential drop, initial voltage be 500mv. amplifier A and connect+5V of pin 8 power supply, connect-5V of pin 4, the amplifier B pin 7 of U9 TLC272CP is directly connected as voltage follower with pin 6, ADE pin 3 is connected with the pin 5 of B, common ground, play jamproof effect.The pin 2 of A is directly connected with the contrast electrode of three-electrode system, pin 1 is directly connected with the auxiliary electrode of three-electrode system, between connect electric capacity 14, object has been the effect of buffer circuit, can improve load-carrying ability simultaneously, therefore U9 and U10 have formed the three-electrode system of biology sensor jointly, and test side used, by drawing here, gathers and the low current signal that detects electrochemical reaction generation.
For realizing above function, the preparation technology of a kind of acetylcholinesterase biology sensor that detects the residual fast detector of Multiple Pesticides in fruits and vegetables of the present invention is as follows: (1) modifies many walls of one deck carbon-tin ash-chitosan nano composite membrane at working electrode surface, and the preparation process of many walls carbon-tin ash-chitosan nano composite membrane is as follows: in 50mL acetic acid solution, add 0.1g shitosan to be prepared into 0.2% chitosan solution.In 4mL 0.2% chitosan solution, add 1mg tin ash, 3mg multi-walled carbon nano-tubes, ultrasonic concussion 30 minutes in 25 DEG C of water, many walls carbon-tin ash-chitosan nano composite membrane is successfully prepared.(2) get many walls carbon-tin ash-chitosan nano composite membrane that 2.5 μ L prepare and drip and be coated in the metal working electrode surface that pre-service is good, under normal temperature, leave standstill 1h, rinse electrode surface with the phosphate buffer of pH7.5; (3) after electrode surface dries, get 2.5 μ L acetylcholinesterases and drip and be coated in electrode surface, leave standstill 2h at 4 DEG C, then rinse surface with the phosphate buffer of pH7.5, nitrogen dries up; (4) get 2.5 μ L0.5%Nafion diaphragms and drip and be coated in electrode surface, leave standstill 1.5h under normal temperature, then rinse surface with the phosphate buffer of pH7.5, nitrogen dries up.
A kind of sample treatment that detects the residual fast detector of Multiple Pesticides in fruits and vegetables of the present invention is as follows: choose representational vegetables and fruit sample, with moist rag by the earth wiped clean of vegetable surface, be chopped into the square fragment of 2cm left and right, sample thief 2g, puts into extraction flask, adds 10ml phosphate buffer, put into ultrasonic extraction apparatus, ultrasonic extraction 3 minutes, pours out extract and leaves standstill 2min, for subsequent use.
For realizing above function, a kind of testing process that detects the residual fast detector of Multiple Pesticides in fruits and vegetables of the present invention is as follows: first, the acetylcholinesterase biology sensor preparing is put into not containing liquid at the bottom of the test of agricultural chemicals to the electric current I while recording not containing agricultural chemicals
1, electric current before suppressing (default value that before suppressing, electric current is set in also can employing system); Afterwards acetylcholinesterase biology sensor working electrode is taken out, with after distilled water flushing, the working electrode of acetylcholinesterase biology sensor is put into detected sample liquid hatches 10 minutes, again use distilled water flushing, then the working electrode of acetylcholinesterase biology sensor is put into test end liquid, recorded the electric current I after inhibition
2; Automatically calculate inhibiting rate (I%) by the trace routine of moving on detector, computing formula is as follows:
I%=(?I
1-?I
2)/?I
1?×100%
If conventionally detect in solution and contain agricultural chemicals, after acetylcholinesterase and Pesticide Exposure, the activity of enzyme is suppressed and reduces, and causes electric current I
2be significantly less than I
1.I
1with I
2difference more inhibiting rate is larger, the concentration that shows to detect solution Pesticide Residues is higher; I
1with I
2the less inhibiting rate of difference less, the concentration that shows to detect solution Pesticide Residues is lower.Due to according to the national standard of the Pesticide maximum residue limit specifying in GB2763-2012, in detector data-carrier store, implant the limitation to residues of pesticides in Common Vegetables fruit, therefore user can be in the time of pesticide residue determination, first input the kind of vegetable and fruit, according to the kind of vegetable and fruit, detector just can be determined kind and the agricultural chemicals maximum residue limit of corresponding common pesticides.In detector data storage cell, implant the typical curve database of measuring in advance the various agricultural chemicals of setting up by mark-on method simultaneously.The kind of the agricultural chemicals that may use according to different vegetable, the typical curve of corresponding corresponding agricultural chemicals, the information such as determine the concentration of agricultural chemicals and whether exceed standard.
A kind of residual fast detector testing process embodiment of Multiple Pesticides in fruits and vegetables that detects of the present invention: first, obtain sample liquid according to the disposal route of above-mentioned vegetable sample.Detector is placed in to duty, presses instrument switch key, enter homepage after system initialization, there are the buttons such as vegetable species options button, test button, reset, printing and storage on display screen side.The in the situation that of blank sample, Pen-down detection button, carries out detector self calibration continuously, shows that the inhibiting rate of twice measurement result is less than 5%, illustrates that detector is working properly, can start normal detection.In the kind of carrying out first selecting before vegetable sample detection vegetables, then the biology sensor that fixes acetylcholinesterase is put into test end liquid, crocodile clip is stepped up respectively on three electrodes that three-electrode system is corresponding.Click for the first time test button, start to detect and show reading I
1; Afterwards acetylcholinesterase biology sensor working electrode is taken out, with after distilled water flushing, the working electrode of acetylcholinesterase biology sensor is put into detected sample liquid hatches 10 minutes, again use distilled water flushing, then the working electrode of acetylcholinesterase biology sensor is put into test end liquid, click for the second time test button, show result I
2; Click and print, print result is also preserved.Automatically calculate the maximum inhibition of this sample liquid to immobilised enzymes by two current data.Determine kind and the concentration of agricultural chemicals according to the maximum limit standard of residues of pesticides of implanting and typical curve database, press printing key, the data of agricultural chemicals inhibiting rate, pesticide variety and the detectable concentration of twice detection in front and back are printed to output.
A kind of residual fast detector of Multiple Pesticides in fruits and vegetables that detects of the present invention, operating procedure is simple, detection time is shorter, can realize the qualitative, quantitative of residues of pesticides measures, have highly sensitive, good stability, high repeatability and other advantages, meet China's Fast Determination of Pesticide Residue technical development and internationalization requirement.
Claims (6)
1. one kind is quantitatively detected the fast detector of fruits and vegetables Pesticide Residues, it is characterized in that, comprise acetylcholinesterase sensor three-electrode system, input and disposal system, show and print storage system, electric power system composition, in the microprocessor of the inside of detector, implant fruits and vegetables pesticide variety and national Pesticide Residue database, organophosphorus and carbamate pesticide typical curve database and BP neural network algorithm, detector utilizes BP neural network that the conventional organophosphorus of measured value and cell stores and carbamate chemicals for agriculture typical curve database are compared, the contained pesticide variety of qualitative definite sample and whether exceeding standard, and the concentration of the agricultural chemicals that quantitatively exceeds standard,
Described acetylcholinesterase sensor three-electrode system, forms electrode by being fixed with acetylcholinesterase working electrode, saturated calomel reference electrode and platinum, sends the current signal containing before and after agricultural chemicals collecting to input and disposal system;
The preparation method of described acetylcholinesterase sensor, step is as follows:
In 50mL acetic acid solution, add 0.1g shitosan to be prepared into 0.2% chitosan solution, in 4mL 0.2% chitosan solution, add 1mg tin ash and 3mg to hide carbon nano-tube, in 25 DEG C of water, after ultrasonic concussion 30min, obtain many walls carbon-tin ash-chitosan nano composite membrane;
Get many walls carbon-tin ash-chitosan nano composite membrane that 2.5 μ L prepare and drip and be coated onto the gold electrode surfaces that pre-service is good, normal temperature leaves standstill 1h, rinses electrode surface by the phosphate buffered solution of pH7.5;
After electrode surface dries, the acetylcholinesterase of getting 2.5 μ L drips and is coated in electrode surface, leaves standstill 2h at 4 DEG C, then rinses surface by the phosphate buffered solution of pH7.5, and nitrogen dries up;
The Nafion diaphragm of getting 2.5 μ L 0.5% drips and is coated in electrode surface, leaves standstill 1.5h under normal temperature, then rinses surface by the phosphate buffered solution of pH7.5, and nitrogen dries up;
Described input and disposal system, by Detection of Weak Signals and treatment circuit, A/D conversion chip and single-chip microcomputer composition, the low current signal collecting is carried out I/V conversion by Detection of Weak Signals and treatment circuit, amplify, after filtering, after the conversion of A/D chip, digital quantity signal is sent into single-chip microcomputer and carry out routine processes;
Described demonstration and storage print system, by mini-printer, display screen and storage unit composition are also unified programming Control by single-chip microcomputer to it, display screen shows remains of pesticide concentration, inhibition degree, data are stored in storage unit so that user checks in time and carries out data analysis, and data are printed output by mini-printer;
Described electric power system, containing potentiostatic circuit provides the stable operating voltage of 500mV for working electrode and contrast electrode;
Described fruits and vegetables pesticide variety and limitation database, be to set up according to the Pesticide maximum residue limit specifying in GB2763-2012, and be implanted in advance in the microprocessor of detector;
Described organophosphorus and carbamate chemicals for agriculture typical curve database, be to be measured and set up by mark-on method, and be implanted in advance in the microprocessor of detector;
In the microprocessor of the inside of described detector, implant BP neural network algorithm, utilize BP neural network that the conventional organophosphorus of measured value and cell stores and carbamate chemicals for agriculture typical curve database are compared, the contained pesticide variety of qualitative definite sample and whether exceeding standard, and the concentration of the agricultural chemicals that quantitatively exceeds standard.
2. a detection method of utilizing fast detector described in claim 1, is characterized in that, step is as follows:
Selection will detect the kind of vegetables, then acetylcholinesterase biology sensor working electrode is put into containing liquid at the bottom of the test of agricultural chemicals, the electric current I while recording not containing agricultural chemicals
1, electric current before suppressing, or the default value of setting in employing system;
Acetylcholinesterase biology sensor working electrode is taken out, with distilled water flushing, then working electrode is put into analyte sample fluid and hatch 10min, again use distilled water flushing, then working electrode is put into test end liquid, record and suppress after-current I
2;
Operation trace routine, detector calculates inhibiting rate automatically according to computing formula;
Detector is according to vegetable species selected in step 1), with the calculating of comparing of pesticide variety and limitation database and typical curve database, determines whether residues of pesticides exceed standard and concentration.
3. method described in claim 2, is characterized in that, described in step 1), testing end liquid is the pH7.5 phosphate buffered solution that contains 1.0 mM chlorinated thio acetylcholines.
4. method described in claim 2, it is characterized in that step 2) described in the preprocess method of testing sample be: choose vegetables or fruit sample, with moist rag by the earth wiped clean of vegetable surface, be chopped into the square fragment of 2cm, sample thief 2g, puts into extraction flask, adds 10ml phosphate buffer, put into ultrasonic extraction apparatus, ultrasonic extraction 3min, pours out extract and leaves standstill 2min, for subsequent use.
5. method described in claim 2, is characterized in that, computing formula described in step 3) is:
I?%
=(?I
1-?I
2)/?I
1?×100%。
Described in claim 1 fast detector for the field quick detection of fruit and vegetable residues of pesticides.
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| CN105675807A (en) * | 2016-01-07 | 2016-06-15 | 中国农业大学 | Evaluation method of atrazine residue based on BP neural network |
| CN106404876A (en) * | 2016-12-22 | 2017-02-15 | 山东理工大学 | Handheld fast detector for pesticide residues based on internet of things |
| CN107290402A (en) * | 2016-04-01 | 2017-10-24 | 王方明 | Nano biological sensor fast detecting pesticide residue device and detection method |
| CN108195915A (en) * | 2017-12-27 | 2018-06-22 | 李文清 | One kind is used for Practice for Pesticide Residue in Agricultural Products fast detector |
| CN111497461A (en) * | 2020-05-26 | 2020-08-07 | 广东星创众谱仪器有限公司 | Intelligent machine for edible agricultural product qualification certificate and working method thereof |
| CN114113247A (en) * | 2020-08-31 | 2022-03-01 | 浙江纳智汇生物科技有限公司 | Electrochemical detection method, detection chip, detection kit and detection system based on enzyme inhibition method |
| CN115184534A (en) * | 2022-07-12 | 2022-10-14 | 江南大学 | Automatic pesticide multi-residue analysis system |
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