CN1834628A - Fast detection method and device for pesticide residual toxin - Google Patents
Fast detection method and device for pesticide residual toxin Download PDFInfo
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- CN1834628A CN1834628A CN 200510135569 CN200510135569A CN1834628A CN 1834628 A CN1834628 A CN 1834628A CN 200510135569 CN200510135569 CN 200510135569 CN 200510135569 A CN200510135569 A CN 200510135569A CN 1834628 A CN1834628 A CN 1834628A
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Abstract
This invention relates to high detecting sensitivity, fast detecting speed, high selectivity pesticide residual toxicity detecting method that fits the situation of our country. The procedures are that (1) making following reagent for using, (2) testing of contrasting solution, (3) testing of sample solution, (4) inhibition ratio computing, the ratio is equal to [(AC-AS)/AC]*100% according to formula, and AC is absorbance of the contrasting solution, AS is absorbance of the sample solution. It is used to the pesticide residual toxicity quick detecting equipment, its features are that it includes light signal creating device, electro optical signal switching device, signal amplifying circuit, testing device and output device. Light signal created by the device is switched into electrical signal through the switching device, then the electrical signal is input into testing device for detecting after it through the amplifier circuit, finally, detecting result is outputted through output device by testing device.
Description
Technical field
The invention belongs to food residual toxicity detection method field, especially a kind of residual toxicity method for quick and equipment.
Background technology
Agricultural product are carried out residual toxicity to be detected and to be one and to have very important thing.On the one hand, because the fruits and vegetables pollution problem becomes increasingly conspicuous, it is healthy to have a strong impact on people.Since the nineties in 20th century, China vegetables and fruit residues of pesticides cause that people and animals' death incident frequently takes place.Pesticide poisoning incident 247349 examples take place in 26 provinces and cities in 1992 to 1996 whole nations, dead 24612 people, and China every year, the number because of pesticide poisoning accounted for 50% of the similar accident number in the world.Along with the raising day by day of people's living standard, require agricultural product not only quality to get well, and want health pollution-free, in the international market, strict restriction is made to the persticide residue of farm imports in many developed countries and area.Because the many agricultural product of China exist residues of pesticides to a certain extent, developed country's " green barrier " is also more and more higher, causes the foreign exchange earning of agricultural product to be a greater impact.
At present, following several method is arranged in the world aspect Fast Determination of Pesticide Residue:
1, the use housefly detects the residual hazard in the vegetables
In the later stage sixties in last century, China Taiwan adopts bioassay method to carry out Detecting Pesticide, its principle be put raise hypersensitivity housefly in dish juice, after 4-5 hour the housefly mortality ratio be below 10% qualified.The advantage of this method be process simple, need not complex instrument.Shortcoming is that detection time is longer, is only applicable to the vegetables of not gathering in the field, and this method only responds to the minority medicament, can't differentiate the kind of remains of pesticide, and accuracy is lower.
2, enzyme immunodetection
The enzyme-linked immunosorbent detection method is called for short enzyme immunoassay, is the microanalysis that antigen and antibody response are combined with modern means of science and technology.The advantage of this method is to reduce sample pre-treatment and purification step, can analyze great amount of samples at short notice, is suitable for making the on-the-spot screening of kit, and its shortcoming is that precision is not high, and Antibody Preparation is difficult for and specificity arranged.
3, enzyme suppresses method
This method is a kind of Detecting Pesticide method that develops rapidly this year, has characteristics such as easy, quick, accurate, is applicable to the fast detecting of organophosphorus and carbamate pesticide residue in the fruits and vegetables.The ultimate principle that enzyme suppresses method is:
Acetylcholinesterase (AChE)+organophosphorus or carbamate chemicals for agriculture-enzymatic activity are suppressed;
Acetylcholinesterase (AChE)+sample then contains organophosphorus or carbamate chemicals for agriculture as activity inhibited, is not suppressed as activity, does not then contain this type of agricultural chemicals.
Residual toxicity screening fast is to be arranged in the first-line inspection technology of basic unit.The detection technique that meets this screening must be simple, do not need large-scale instrument and complex apparatus, and low and result check of its cost reflects soon, adopts this method can within a short period of time the fruits and vegetables of " super malicious " to be screened.In this year, China has also carried out the research that enzyme inhibition method is detected residues of pesticides, but the single-point development process of measuring the enzyme inhibitory reaction that adopts is difficult for promoting more.
Summary of the invention
A kind of residual toxicity detection method that the purpose of this invention is to provide a kind of detection sensitivity height, detection speed is fast, selectivity is high suitable China's national situation.
Another object of the present invention provides a kind of residual toxicity checkout equipment.
Technical scheme of the present invention is: the residual toxicity method for quick is characterized in that comprising the following steps:
(1), the following reagent of making is standby: the phosphate buffer of PH=7.5, the substrate of acetylthiocholine iodide, 5, the developer (DTNB) of 5 '-two thiobiss-2-dinitrobenzoic acid, the enzyme liquid acetylcholine esterase (AChE) that extracts from housefly;
(2) control solution test: in beaker, add the 3ml phosphate buffer, add 50 μ l enzymes and 50 μ l developers again, place under the temperature 25-35 ℃ condition and cultivated 12-18 minute, take out the back and add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(3) sample solution test: choose sample from the different plants blade, fruit and vegetable are taken a sample from epidermis 1-1.5cm depths, leaf vegetables is got the 2g sample, non-leaf vegetables is got the 4g sample, with the sample chopping that obtains, put into extraction flask, add the 20ml phosphate buffer, vibrated 1-2 minute, left standstill 3-5 minute, pour 3ml sample extraction liquid, 50 μ l enzymes, 50 μ l developers in vitro respectively into, place to cultivate under the temperature 25-35 ℃ condition and took out in 12-18 minute, add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(4) inhibiting rate calculates: according to formula inhibiting rate=[(A
C-A
S)/A
C] * 100%, wherein A
CThe absorbance of contrast solution, A
SAbsorbance for sample solution.
Preferably, in control solution test, the cultivation temperature of mixing material is that 30 ℃, incubation time are 15 minutes.
Preferably, in the sample solution test, 30 ℃ of the cultivation temperature of mixing material, incubation time is 15 minutes.
Be applicable to the residual toxicity device for fast detecting of residual toxicity detection method, it is characterized in that comprising optical signal generation apparatus, the photosignal conversion equipment, signal amplification circuit, measurement mechanism and output unit, the light signal that optical signal generation apparatus produces converts electric signal to by the photosignal conversion equipment, electric signal is through behind the amplifying circuit, being input to measurement mechanism detects, measurement mechanism is exported testing result by output unit, wherein optical signal generation apparatus comprises light source, optical filter, lens and cuvette are formed, the light that light source sends passes through optical filter successively, behind lens and the cuvette, shine on the photosignal conversion equipment, described photosignal conversion equipment is a photoelectric cell, and measuring pick-up unit is CPU and A/D change-over circuit.
Described output unit comprises display, printer and/or network connection device.
Effect of the present invention is: the residual toxicity method for quick comprises the following steps: (1), to make following reagent standby; (2) control solution test; (3) sample solution test; (4) inhibiting rate calculates: according to formula inhibiting rate=[(A
C-A
S)/A
C] * 100%, wherein A
CThe absorbance of contrast solution, A
SAbsorbance for sample solution.
Method of the present invention is that this toxicology principle that the insect poisoning is caused death is carried out " fast detecting " to residues of pesticides according to the activity of organophosphorus and carbamate chemicals for agriculture energy acetylcholine esterase inhibition.As not containing agricultural chemicals in the extract of fruit and vegetable or residual quantity is very low, the activity of enzyme is not suppressed, and the substrate of adding is just by water decomposition, and the developer generation color reaction or the hydrolysate itself of hydrolysate and adding have color.Contain agricultural chemicals and residual quantity in the extract as fruit vegetable when higher, the activity of enzyme is suppressed, and substrate just can not be hydrolyzed, when adding developer, just do not develop the color or change color very little.With being applicable to that the residual toxicity device for fast detecting of residual toxicity detection method measures the variation of absorbance, just can judge organophosphorus or carbamate pesticide residue situation in the vegetables.
Method of the present invention adapts to the characteristics of China fruit, vegetables production and sales, can carry out scalping to fruit, the vegetables that contain residues of pesticides, and the fruit, the vegetables that a part are contained residual toxicity are controlled at outside the market, avoids the pesticide poisoning accident to take place.
Be applicable to the residual toxicity device for fast detecting of residual toxicity detection method, comprise optical signal generation apparatus, photosignal conversion equipment, signal amplification circuit, measurement mechanism and output unit, the light signal that optical signal generation apparatus produces converts electric signal to by the photosignal conversion equipment, electric signal is through behind the amplifying circuit, be input to measurement mechanism and detect, measurement mechanism is exported testing result by output unit.
The residual toxicity device for fast detecting not only is useful in big wholesale vegetable market, the fast detecting station is set up in important vegetables producing region, also is applicable to village, villages (towns), large-scale canteen, even can set up mobile detection station at the sub-transaction of greengrocer scene.Can really be implemented to field speed and survey, can prevent people and animals to eat by mistake to contain the vegetable and fruit of high poison, persistent pesticide, guarantee people's safe diet that secures good health.
The present invention is described further below in conjunction with drawings and Examples.
Description of drawings
Fig. 1 is the index path of residual toxicity pick-up unit of the present invention;
Fig. 2 is a residual toxicity pick-up unit circuit block diagram of the present invention;
Fig. 3 is the circuit theory diagrams of residual toxicity pick-up unit of the present invention.
Embodiment
Embodiment 1: the residual toxicity method for quick is characterized in that comprising the following steps:
(1), the following reagent of making is standby: the phosphate buffer of PH=7.5, the substrate of acetylthiocholine iodide, 5, the developer (DTNB) of 5 '-two thiobiss-2-dinitrobenzoic acid, the enzyme liquid acetylcholine esterase (AChE) that extracts from housefly;
(2) control solution test: in beaker, add the 3ml phosphate buffer, add 50 μ l enzymes and 50 μ l developers again, place under the temperature 25-35 ℃ condition and cultivated 12-18 minute, take out the back and add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(3) sample solution test: choose sample from the different plants blade, fruit and vegetable are taken a sample from epidermis 1-1.5cm depths, leaf vegetables is got the 2g sample, non-leaf vegetables is got the 4g sample, with the sample chopping that obtains, put into extraction flask, add the 20ml phosphate buffer, vibrated 1-2 minute, left standstill 3-5 minute, pour 3ml sample extraction liquid, 50 μ l enzymes, 50 μ l developers in vitro respectively into, place to cultivate under the temperature 25-35 ℃ condition and took out in 12-18 minute, add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(4) inhibiting rate calculates: according to formula, and inhibiting rate=[(A
C-A
S)/A
C] * 100%, wherein A
CThe absorbance of contrast solution, A
SAbsorbance for sample solution.
The residual toxicity device for fast detecting, comprise optical signal generation apparatus, the photosignal conversion equipment, signal amplification circuit, measurement mechanism and output unit, the light signal that optical signal generation apparatus produces converts electric signal to by the photosignal conversion equipment, electric signal is through behind the amplifying circuit, being input to measurement mechanism detects, measurement mechanism is exported testing result by output unit, wherein optical signal generation apparatus comprises light source 1, optical filter 2, lens 3 and cuvette 4 are formed, the light that light source 1 sends is successively by optical filter 2, behind lens 3 and the cuvette 4, shine on the photosignal conversion equipment, described photosignal conversion equipment is photoelectric cell 5 (referring to Fig. 1), after photoelectric cell 5 switching signals, be input to the measurement pick-up unit by signal amplification circuit 6, measuring pick-up unit is CPU and A/D change-over circuit 7, output unit comprises display, printer or network connection device 9, LCD and the keyboard 8 (referring to Fig. 2) in addition that link to each other with CPU.
Among Fig. 3, the 5th, photoelectric cell, the 6th, the signal amplification circuit of forming by integrated circuit CA3140, the 7th, CPU and A/D change-over circuit, the 8th, keyboard, the 9th, printer interface and network interface, the 10th, code translator, the 11st, program storage, the 12nd, data-carrier store, the 13rd, LCD interface, the 14th, auditory tone cues device, the 15th, power module, the 16th, source of stable pressure.
The principle of work of residual toxicity device for fast detecting is: this instrument is the activity change degree according to acetylcholinesterase, utilize organophosphorus and carbamate chemicals for agriculture to the inhibition of acetylcholinesterase for principle design, it can measure the shade of reactant in the enzyme reaction process, convert corresponding inhibition ratio then to, show residual toxicity content.
Inhibiting rate I=[(A
C-A
S)/A
C] * 100%, wherein A
CBe the absorbance of contrast solution, A
SAbsorbance for sample solution.
Under the catalysis of acetylcholinesterase, do not having to be blank under the situation of residues of pesticides, liquid to be measured is yellow, and inhibiting rate I=0 is when containing residual agricultural chemicals in the test sample, liquid to be measured is light yellow, when the residues of pesticides severe overweight, and liquid to be measured variable color hardly, when the activity of acetylcholinesterase is suppressed fully, liquid nondiscolouring to be measured, I=100%, when inhibiting rate I＜50, illustrate that sampling is defective, when inhibiting rate I＞50%, illustrate that the dish sample is contaminated, not edible.
Embodiment 2: the residual toxicity method for quick is characterized in that comprising the following steps:
(1), the following reagent of making is standby: the phosphate buffer of PH=7.5, the substrate of acetylthiocholine iodide, 5, the developer (DTNB) of 5 '-two thiobiss-2-dinitrobenzoic acid, the enzyme liquid acetylcholine esterase (AChE) that extracts from housefly;
(2) control solution test: in beaker, add the 3ml phosphate buffer, add 50 μ l enzymes and 50 μ l developers again, place under 30 ℃ of conditions of temperature and cultivated 15 minutes, take out the back and add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(3) sample solution test: choose sample from the different plants blade, fruit and vegetable are taken a sample from epidermis 1-1.5cm depths, leaf vegetables is got the 2g sample, non-leaf vegetables is got the 4g sample, with the sample chopping that obtains, put into extraction flask, add the 20ml phosphate buffer, vibrated 1-2 minute, everywhere extract, left standstill 3-5 minute, pour 3ml sample extraction liquid, 50 μ l enzymes, 50 μ l developers respectively in vitro, place to cultivate under 30 ℃ of conditions of temperature and took out in 15 minutes, add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(4) inhibiting rate calculates: according to formula, and inhibiting rate=[(A
C-A
S)/A
C] * 100%, wherein A
CThe absorbance of contrast solution, A
SAbsorbance for sample solution.
Claims (5)
1, residual toxicity method for quick is characterized in that comprising the following steps:
(1), the following reagent of making is standby: the phosphate buffer of PH=7.5, the substrate of acetylthiocholine iodide, 5, the developer (DTNB) of 5 '-two thiobiss-2-dinitrobenzoic acid, the enzyme liquid acetylcholine esterase (AChE) that extracts from housefly;
(2) control solution test: in beaker, add the 3ml phosphate buffer, add 50 μ l enzymes and 50 μ l developers again, place under the temperature 25-35 ℃ condition and cultivated 12-18 minute, take out the back and add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(3) sample solution test: choose sample from the different plants blade, fruit and vegetable are taken a sample from epidermis 1-1.5cm depths, leaf vegetables is got the 2g sample, non-leaf vegetables is got the 4g sample, with the sample chopping that obtains, put into extraction flask, add the 20ml phosphate buffer, vibrated 1-2 minute, left standstill 3-5 minute, pour 3ml sample extraction liquid, 50 μ l enzymes, 50 μ l developers in vitro respectively into, place to cultivate under the temperature 25-35 ℃ condition and took out in 12-18 minute, add 50 μ l substrates, pour in the cuvette, carry out the absorbance test;
(4) inhibiting rate calculates: according to formula inhibiting rate=[(A
C-A
S)/A
C] * 100%, wherein A
CThe absorbance of contrast solution, A
SAbsorbance for sample solution.
2, residual toxicity method for quick according to claim 1 is characterized in that in control solution test, and the cultivation temperature of mixing material is that 30 ℃, incubation time are 15 minutes.
3, residual toxicity method for quick according to claim 1 and 2 is characterized in that in the sample solution test, 30 ℃ of the cultivation temperature of mixing material, and incubation time is 15 minutes.
4, a kind of residual toxicity device for fast detecting that is applicable to the described residual toxicity detection method of above-mentioned each claim, it is characterized in that comprising optical signal generation apparatus, the photosignal conversion equipment, signal amplification circuit, measurement mechanism and output unit, the light signal that optical signal generation apparatus produces converts electric signal to by the photosignal conversion equipment, electric signal is through behind the amplifying circuit, being input to measurement mechanism detects, measurement mechanism is exported testing result by output unit, wherein optical signal generation apparatus comprises light source, optical filter, lens and cuvette are formed, the light that light source sends passes through optical filter successively, behind lens and the cuvette, shine on the photosignal conversion equipment, described photosignal conversion equipment is a photoelectric cell, and measuring pick-up unit is CPU and A/D change-over circuit.
5, residual toxicity device for fast detecting according to claim 4 is characterized in that described output unit comprises display, printer and/or network connection device.
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|---|---|---|---|
| CN 200510135569 CN1834628A (en) | 2005-12-30 | 2005-12-30 | Fast detection method and device for pesticide residual toxin |
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|---|---|---|---|
| CN 200510135569 CN1834628A (en) | 2005-12-30 | 2005-12-30 | Fast detection method and device for pesticide residual toxin |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102002367A (en) * | 2010-11-17 | 2011-04-06 | 中国科学院南京土壤研究所 | Eluent for soil polluted by organochlorine pesticides and repair method by using same |
| CN102095772A (en) * | 2011-01-27 | 2011-06-15 | 上海交通大学 | Pesticide residue detecting instrument based on three-electrode enzyme sensor |
| CN102460151A (en) * | 2009-06-12 | 2012-05-16 | 阿迪森真空产品公司 | Device and method for analysing gas and associated measurement station |
| CN103472019A (en) * | 2013-09-30 | 2013-12-25 | 重庆大学 | Spectrographic method for fast detecting pesticide residues |
| CN105021604A (en) * | 2015-07-15 | 2015-11-04 | 江苏大学 | Fast imaging detection method of heavy metal element content and device thereof |
| CN105181681A (en) * | 2013-09-03 | 2015-12-23 | 驷丞生物科技股份有限公司 | Pesticide detection device |
| CN106093032A (en) * | 2016-08-31 | 2016-11-09 | 孟月志 | A kind of Fast Determination of Pesticide Residue eliminates false-positive detection method |
| CN108693126A (en) * | 2017-04-05 | 2018-10-23 | 中国科学院大连化学物理研究所 | A kind of rapid detection method of carbamate chemicals for agriculture |
-
2005
- 2005-12-30 CN CN 200510135569 patent/CN1834628A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102460151A (en) * | 2009-06-12 | 2012-05-16 | 阿迪森真空产品公司 | Device and method for analysing gas and associated measurement station |
| CN102002367A (en) * | 2010-11-17 | 2011-04-06 | 中国科学院南京土壤研究所 | Eluent for soil polluted by organochlorine pesticides and repair method by using same |
| CN102095772A (en) * | 2011-01-27 | 2011-06-15 | 上海交通大学 | Pesticide residue detecting instrument based on three-electrode enzyme sensor |
| CN102095772B (en) * | 2011-01-27 | 2013-05-01 | 上海交通大学 | Pesticide residue detecting instrument based on three-electrode enzyme sensor |
| CN105181681A (en) * | 2013-09-03 | 2015-12-23 | 驷丞生物科技股份有限公司 | Pesticide detection device |
| CN103472019A (en) * | 2013-09-30 | 2013-12-25 | 重庆大学 | Spectrographic method for fast detecting pesticide residues |
| CN103472019B (en) * | 2013-09-30 | 2015-12-23 | 重庆大学 | The residual spectrum method for quick of agriculture |
| CN105021604A (en) * | 2015-07-15 | 2015-11-04 | 江苏大学 | Fast imaging detection method of heavy metal element content and device thereof |
| CN106093032A (en) * | 2016-08-31 | 2016-11-09 | 孟月志 | A kind of Fast Determination of Pesticide Residue eliminates false-positive detection method |
| CN108693126A (en) * | 2017-04-05 | 2018-10-23 | 中国科学院大连化学物理研究所 | A kind of rapid detection method of carbamate chemicals for agriculture |
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