CN1414371A - Conventional pesticide residue quick detecting method - Google Patents
Conventional pesticide residue quick detecting method Download PDFInfo
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- CN1414371A CN1414371A CN 02148533 CN02148533A CN1414371A CN 1414371 A CN1414371 A CN 1414371A CN 02148533 CN02148533 CN 02148533 CN 02148533 A CN02148533 A CN 02148533A CN 1414371 A CN1414371 A CN 1414371A
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Abstract
A quick residual detecting method for conventional pesticide features that the acetylcholine esterase from the liver of white rats is used as the source of the enzyme. The acetylcholine is used as the substrate. Under the action of acetylcholine esterase, the substrate is hydrolyzed so as to generate the sulfocholine and DTNB. The peak of the maximum absorption in the color reaction is at 412 nm. With concentration of enzyme being fixed, comparing OD412 value of the pesticide in different concentrations establishes the standard curvature. With being dipped and extracted, the sample to be measured is tested so as to obtain the value of OD412. Base on the value of OD412 of the sample to be measured, the residual concentration and the depression rate of the pesticide in the sample to be measured in determined.
Description
Technical field:
The present invention relates to the quick residue detection technology of a kind of conventional pesticide.
Background technology:
Along with people's living standard improves day by day, people are more and more high to the demand of agricultural byproducts such as vegetables and fruits, in order to satisfy market demand, make Pesticidal products such as vegetables and fruits avoid the invasion and attack of disease and pest, promote its growth, and various agricultural chemicals, chemical fertilizer are widely used.And be trapped in objectionable impurities in the agricultural product, and directly threatening people's normal life, the security of the continuous reinforcement agricultural product that the technical trade measure (TBT) that countries in the world ginseng Pesticidal products is produced is emphasized has become the hot issue that the whole world is paid close attention to.Wherein, residues of pesticides become the critical limitation index of developed country to agricultural product in the agricultural product, the relevant control and the Fast Detection Technique of residues of pesticides have been strengthened to abovementioned technology in countries in the world, and he mainly is: with the vegetables and fruits blades with alcohol dipping after extraction or with ethanol and the extraction of bromine water dipping; In color comparison tube, can go into damping fluid and make substrate reactions; Under the 412mn wavelength, read the light absorption value rate of change,, obtain inhibiting rate by the difference of sample and control group light absorption value.Mainly there is following point in this detection mode: at first, plant is after alcohol dipping, and plant pigment is extracted simultaneously, disregards to produce color interference; Substrate and developer add simultaneously, though more convenient, can influence molecular balance, because developer is yellow a little a little, this method is not corrected.
Summary of the invention:
The object of the invention is: at the problem that exists in the remains of pesticide fast detecting in the present agricultural product, provide a kind of conventional pesticide quick method for detecting residue.
The object of the present invention is achieved like this: the quick method for detecting residue of a kind of conventional pesticide, it is characterized in that: with this enzyme of small white mouse liver acetylcholine fat as the enzyme source, is substrate with acetyl for choline, substrate is under the effect of acetyl for CHE, hydrolysis generates thiocholine and two sulphur biconjugate nitrobenzoic acids (DTNB), playing chromogenic reaction has maximum absorption band at the 412nm place, compares the OD of variable concentrations pesticide effect by the immobilized enzyme activity
412Value is formulated typical curve, records OD after testing sample is extracted through dipping
412Value is according to the OD of testing sample
412Value is judged agricultural chemicals residual concentration and inhibiting rate in the determinand.
The difference of the methods such as Zheng Yun of the present invention and Taiwan farming examination institute is: the liver of the small white mouse that enzyme of the present invention is got, and the enzyme that is contained is similar to the formation of human body agricultural chemicals target enzyme, and method enzymes such as Zheng Yun are the acetylcholinesterase of employing housefly head; The present invention disturbs by activated charcoal elimination pigment after extracting with the acetone solution dipping, and simultaneously, acetone solution is good to the extraction rate was acquired of medicament, the recovery height of medicament; The present invention adopts oxidizer and the dual processing of non-oxidizer, and methods such as Zheng Yun only adopt the processing that adds the bromine water oxidation, this method has facilitation to the degraded of thionic phosphoric acid fat pesticide, simultaneously, developer of the present invention finishes the back at substrate reactions and adds, need not consider the influence of developer, need not to use reaction terminating agent course of reaction.The present invention and the comparison of surveying the phosphorus method fast
Each agricultural chemicals (not oxidation) is to the inhibiting rate and the detection lower limit of acetylcholine esterase
Each agricultural chemicals (1% bromine water oxidation) is to the inhibiting rate and the detection lower limit of acetylcholine esterase
Each agricultural chemicals not oxidation and 1% bromine water oxidation respectively to concentration in the inhibition of acetylcholine esterase
| The present invention | Survey the phosphorus method fast | |
| Sample treatment | Clean and rub juice | Clean |
| Detected object | Organophosphorus, carbamic acid lipid | Organophosphorus |
| Sensitivity and high accuracy (recovery) repeatability (mutation coefficient) | 0.5ppm 90-110％ ≤0.5％ | 10ppm ≤70-130％ ≤5％ |
| Expense | Cheaply | Cheaply |
| Detection time | 20min | 5-10min |
| Other |
| Medicament | Concentration (mg/ml) | Inhibiting rate (%) | Detect lower limit ppm | Medicament | Concentration (mg/ml) | Inhibiting rate (%) | Detect lower limit ppm |
| Acephatemet | ?0.104 | ?91.83 | ?5-10 | Parathion-methyl | ?0.0104 | ?71.12 | ?1-1.5 |
| ?0.052 | ?74.21 | ?0.0052 | ?42.32 | ||||
| ?0.026 | ?49.45 | ?0.0026 | ?20.72 | ||||
| ?0.013 | ?25.02 | ?0.0013 | ?8.17 | ||||
| The furans pellet | ?1.11 | ?90-65 | ?100-150 | Hostathion | ?0.00237 | ?79.72 | ?0.25-0.5 |
| ?0.555 | ?72.02 | ?0.00118 | ?55.22 | ||||
| ?0.2775 | ?41.69 | ?0.00059 | ?31.43 | ||||
| ?0.13875 | ?16.13 | ?0.00025 | ?11.15 | ||||
| Azodrin | ?0.901 | ?94.47 | ?20-50 | Orthene | ?0.0468 | ?97.71 | ?0.1-0.5 |
| ?0.4505 | ?77.89 | ?0.00468 | ?81.90 | ||||
| ?0.22525 | ?46.79 | ?0.000468 | ?21.52 | ||||
| ?0.112625 | ?28.83 | ?0.000047 | ?2.68 | ||||
| Parathion | ?0.0191 | ?89.31 | ?0.002-0.02 | Flolimat | ?1.787 | ?44.17 | ?200-300 |
| ?0.00191 | ?85.16 | ?0.894 | ?36.60 | ||||
| ?0.000191 | ?77.31 | ?0.447 | ?29.96 | ||||
| ?0.000019 | ?46.18 | ?0.2235 | ?8.94 | ||||
| Thimet | ?0.01896 | ?94.34 | ?0.002-0.02 | Isocarbophos | ?0.88 | ?85.60 | ?0.5-1.0 |
| ?0.001896 | ?92.82 | ?0.088 | ?68.48 | ||||
| ?0.000190 | ?86.29 | ?0.0088 | ?35.82 | ||||
| ?0.000019 | ?49.94 | ?0.00088 | ?19.13 | ||||
| Poison upright stone tablet with poison | ?0.00595 | ?84.44 | ?0.8-1.5 | Rogor | ?0.0049 | ?88.62 | ?0.01-0.25 |
| ?0.002975 | ?56.03 | ?0.00245 | ?60.29 | ||||
| ?0.0014875 | ?25.91 | ?0.00049 | ?41.16 | ||||
| ?0.00074375 | ?6.50 | ?0.000245 | ?26.24 |
| Medicament | Concentration (mg/ml) | Inhibiting rate (%) | Detect lower limit ppm | Medicament | Concentration (mg/ml) | Inhibiting rate (%) | Detect lower limit ppm |
| Acephatemet | ?0.104 | ?62.80 | ?0.01-0.1 | Parathion-methyl | ?0.104 | ?50.38 | ?0.1-0.5 |
| ?0.0104 | ?55.35 | ?0.0104 | ?28.02 | ||||
| ?0.00104 | ?47.99 | ?0.00104 | ?17.37 | ||||
| ?0.000104 | ?0.000104 | ?14.03 | |||||
| The furans pellet | ?0.128 | ?87.06 | ?0.1-1 | Hostathion | ?0.237 | ?85.06 | ?0.05-0.2 |
| ?0.0128 | ?65.39 | ?0.0237 | ?73.53 | ||||
| ?0.00128 | ?8.85 | ?0.00237 | ?54.60 | ||||
| ?0.000128 | -9.12 | ?0.000237 | ?39.36 | ||||
| Azodrin | ?0.146 | ?82.56 | ?1-1.5 | Orthene | ?0.0468 | ?97.71 | ?0.1-0.5 |
| ?0.0146 | ?68.16 | ?0.00468 | ?81.90 | ||||
| ?0.00146 | ?17.31 | ?0.000468 | ?21.52 | ||||
| ?0.000146 | -8.32 | ?0.000047 | ?2.86 | ||||
| To sulfuric acid | ?0.0191 | ?89.31 | ?0.001-0.01 | Flolimat | ?0.3574 | ?33.97 | ?50-100 |
| ?0.00191 | ?85.16 | ?0..1787 | ?15.02 | ||||
| ?0.000191 | ?77.31 | ?0.0894 | ?6.41 | ||||
| ?0.0000191 | ?46.18 | ?0.0447 | ?4.67 | ||||
| Thimet | ?0.011896 | ?94.34 | ?0.001-0.01 | Isocarbophos | ?0.88 | ?60.67 | ?0.5-1 |
| ?0.01896 | ?92.82 | ?0.088 | ?44.33 | ||||
| ?0.000190 | ?86.29 | ?0.0088 | ?16.73 | ||||
| ?0.000019 | ?49.94 | ?0.00088 | ?7.50 | ||||
| Poison upright stone tablet with poison | ?0..019 | ?87.33 | ?1-2.5 | Rogor | ?0.049 | ?78.60 | ?0.1-0.5 |
| ?0.0095 | ?72.90 | ?0.0245 | ?70.29 | ||||
| ?0.00475 | ?42.60 | ?0.0049 | ?51.16 | ||||
| ?0.0002375 | ?18.78 | ?0.00245 | ?46.80 |
| Each agricultural chemicals (1% bromine water oxidation) is to concentration in the inhibition of acetylcholine esterase | Concentration during each agricultural chemicals (not oxidation) suppresses acetylcholine esterase | ||||||
| Medicament | Concentration in the inhibition (mg/ml) | Medicament | Concentration in the inhibition (mg/ml) | Medicament | Concentration in the inhibition (mg/ml) | Medicament | Concentration in the inhibition (mg/ml) |
| Acephatemet | 0.01 | Parathion | 0.0001 | Acephatemet | 0.03 | Parathion | 0.00002 |
| Parathion-methyl | 0.1 | Orthene | 0.001 | Parathion-methyl | 0.006 | Orthene | 0.001 |
| The furans pellet | 0.01 | Thimet | 0.00002 | The furans pellet | 0.3 | Thimet | 0.00002 |
| Hostathion | 0.002 | Isocarbophos | 0.4 | Hostathion | 0.001 | Isocarbophos | 0.04 |
| Azodrin | 0.01 | Poison upright stone tablet with poison | 0.005 | Azodrin | 0.25 | Poison upright stone tablet with poison | 0.0025 |
| Flolimat | 0.5 | Rogor | 0.0048 | Flolimat | 2.0 | Rogor | 0.0012 |
The invention has the advantages that: can fast measuring organophosphorus and the residual quantity of carbamic acid lipid agricultural chemicals in sample, the organic solvent that uses among the present invention to the influence of test result less than 5%.
Description of drawings:
Fig. 1 is acephatemet inhibiting rate a---concentration standard curve.
Fig. 2 is Hostathion inhibiting rate a---concentration standard curve.
Embodiment:
Embodiment 1: the method for making of acephatemet inhibiting rate---concentration standard curve
1, gets 96 hole microwell plates
2, the Kong Zhongyong charger adds 50ul enzyme dilution (30 times) and 50ul acephatemet serial dilutions (control group adds PB liquid) and 50ul substrate in 37 ℃-38 ℃ warm down baths about 15 minutes, and blank column only adds 50ul PB liquid and 50ul substrate.
3, temperature is bathed the every hole adding in back 50ulDTNB developer.
4, at the light absorption value of measuring on the microplate reader under the 412nm (OD value).
5, with each the agricultural chemicals inhibiting rate typical curve under the inhibiting rate formula calculating series concentration (25.02%, 49.45%, 74.21%, 91.83%).
Embodiment 2: the method for making of Hostathion inhibiting rate---concentration standard curve.
1, gets 96 hole microwell plates
2, the Kong Zhongyong charger adds 50ul enzyme dilution (30 times) and 50ul Hostathion serial dilutions (control group adds PB liquid) and 50ul substrate in 37 ℃-38 ℃ warm down baths about 15 minutes, and blank column only adds 50ul PB liquid and 50ul substrate.
3, temperature is bathed the every hole adding in back 50ulDTNB developer.
4, at the light absorption value of measuring on the microplate reader under the 412nm (OD value).
5, with each the agricultural chemicals inhibiting rate typical curve under the inhibiting rate formula calculating series concentration (11.15%, 31.43%, 55.22%, 79.72%).
Embodiment 3: the method for quick of leaf class sample.
1, sampling: from leaf vegetables 10-15 sheet sample, get 2 grams and be working sample.
2, extracted 3 minutes with 3ml acetone and 50ml bromine water (1%), in the 20ml syringe, cross the high carbon column of 2cm, and left standstill 2 minutes.
3, in 96 hole microwell plates, add 50ul acetylcholine esterase and 50ul substrate A TCI (establishing blank), 37 ℃-38 ℃ warm down baths 15 minutes.
4, temperature is bathed back every hole adding 50ul extract and 50ul developer DTNB, in the light absorption value of measuring on the microplate reader under the 412nm (OD value).
5, calculate its inhibiting rate, and compare to determine agricultural chemicals residual concentration and inhibiting rate in the leaf class sample with typical curve.
Embodiment 4: the method for quick of rhizome and melon and fruit class sample.
1, sampling: rhizome and melon and fruit class, are got 2 grams and are working sample about thick in the outer 20-25 gram sample with 1.5cm.
2, extracted 3 minutes with 3ml acetone and 50ml bromine water (1%), in the 20ml syringe, cross the high carbon column of 2cm, and left standstill 2 minutes.
3, in 96 hole microwell plates, add 50ul extract and 50ul acetylcholine esterase and 50ul substrate A TCI (establishing blank), 37 ℃-38 ℃ warm down baths 15 minutes.
4, temperature is bathed the every hole adding in back 50ul developer DTNB, in the light absorption value of measuring on the microplate reader under the 412nm (OD value).
5, calculate its inhibiting rate, and compare to determine agricultural chemicals residual concentration and inhibiting rate in rhizome and the melon and fruit class sample with typical curve.
In actual detected, usually the phosphorothionate organophosphorus pesticide needs the oxidizer oxidation, and then effect is obvious, other organophosphorus pesticides, also oxidizer oxidation processes not.
Claims (2)
1, the quick method for detecting residue of a kind of conventional pesticide, it is characterized in that: with small white mouse liver acetylcholinesterase as the enzyme source, is substrate with acetyl for choline, substrate is under the effect of acetyl for cholinesterase, hydrolysis generates thiocholine and two sulphur biconjugate nitrobenzoic acids (DTNB), playing chromogenic reaction has maximum absorption band at the 412nm place, compares the OD of variable concentrations pesticide effect by the immobilized enzyme activity
412Value is formulated typical curve, records OD after testing sample is extracted through dipping
412Value is according to the OD of testing sample
412Value is judged agricultural chemicals residual concentration and inhibiting rate in the determinand.
2, the quick method for detecting residue of a kind of conventional pesticide according to claim 1 is characterized in that: the sample extracting solution after the extraction adopts oxidizer to handle and the not dual processing of oxidizer.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02148533 CN1414371A (en) | 2002-12-13 | 2002-12-13 | Conventional pesticide residue quick detecting method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02148533 CN1414371A (en) | 2002-12-13 | 2002-12-13 | Conventional pesticide residue quick detecting method |
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| CN1414371A true CN1414371A (en) | 2003-04-30 |
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Cited By (13)
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| CN1908622B (en) * | 2005-08-05 | 2010-07-07 | 五鼎生物技术股份有限公司 | Analysis method of enzyme inhibiting agent and module thereof |
| CN101498701B (en) * | 2009-03-09 | 2011-05-25 | 南京大学 | Detection method for phthalate ester residue |
| CN102134559A (en) * | 2010-11-15 | 2011-07-27 | 广州吉家庄生物科技有限公司 | Bacillus brevis, method for extracting bacillus brevis esterase and application of same |
| CN102183396A (en) * | 2011-03-17 | 2011-09-14 | 广州绿洲生化科技有限公司 | Sample pretreatment method for detecting pesticide residue applicable to enzymatic inhibition method, and pesticide detection method and box |
| CN101694450B (en) * | 2009-10-16 | 2012-01-11 | 长安大学 | Method for detecting dose of polymers in modified asphalt based on viscosity-temperature curve |
| CN103323415A (en) * | 2013-07-01 | 2013-09-25 | 广州市酒类检测中心 | Enzyme inhibition method for detecting carbamates |
| CN106093032A (en) * | 2016-08-31 | 2016-11-09 | 孟月志 | A kind of Fast Determination of Pesticide Residue eliminates false-positive detection method |
| CN107305213A (en) * | 2016-04-25 | 2017-10-31 | 赵芳 | A kind of method and kit for detecting organophosphate and carbamate pesticide |
| CN107305212A (en) * | 2016-04-25 | 2017-10-31 | 赵芳 | The immunological detection method and kit of a kind of organophosphate and carbamate pesticide |
| 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 |
| CN109270033A (en) * | 2018-09-10 | 2019-01-25 | 南京市产品质量监督检验院 | The rapid detection method of Hazard factor in a kind of food |
| CN110487732A (en) * | 2019-08-01 | 2019-11-22 | 李孝艳 | The rapid detection method of Organophosphorus and carbamate pesticides pesticide residue in chestnut mushroom |
| US10915548B2 (en) * | 2017-04-18 | 2021-02-09 | Chinese Academy Of Inspection And Quarantine | Method of online tracing pesticide residues and visualizing warning on basis of high resolution mass spectrum, internet and geographic information |
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2002
- 2002-12-13 CN CN 02148533 patent/CN1414371A/en active Pending
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1908622B (en) * | 2005-08-05 | 2010-07-07 | 五鼎生物技术股份有限公司 | Analysis method of enzyme inhibiting agent and module thereof |
| CN101498701B (en) * | 2009-03-09 | 2011-05-25 | 南京大学 | Detection method for phthalate ester residue |
| CN101694450B (en) * | 2009-10-16 | 2012-01-11 | 长安大学 | Method for detecting dose of polymers in modified asphalt based on viscosity-temperature curve |
| CN102134559A (en) * | 2010-11-15 | 2011-07-27 | 广州吉家庄生物科技有限公司 | Bacillus brevis, method for extracting bacillus brevis esterase and application of same |
| CN102134559B (en) * | 2010-11-15 | 2012-12-19 | 广州吉家庄生物科技有限公司 | Bacillus brevis, method for extracting bacillus brevis esterase and application of same |
| CN102183396A (en) * | 2011-03-17 | 2011-09-14 | 广州绿洲生化科技有限公司 | Sample pretreatment method for detecting pesticide residue applicable to enzymatic inhibition method, and pesticide detection method and box |
| CN103323415A (en) * | 2013-07-01 | 2013-09-25 | 广州市酒类检测中心 | Enzyme inhibition method for detecting carbamates |
| CN103323415B (en) * | 2013-07-01 | 2015-08-05 | 广州市酒类检测中心 | A kind of inhibiting AChE detecting carbamates |
| CN107305212B (en) * | 2016-04-25 | 2019-08-30 | 赵芳 | A kind of immunological detection method and kit of organophosphate and carbamate pesticide |
| CN107305213A (en) * | 2016-04-25 | 2017-10-31 | 赵芳 | A kind of method and kit for detecting organophosphate and carbamate pesticide |
| CN107305212A (en) * | 2016-04-25 | 2017-10-31 | 赵芳 | The immunological detection method and kit of a kind of organophosphate and carbamate pesticide |
| CN106093032A (en) * | 2016-08-31 | 2016-11-09 | 孟月志 | A kind of Fast Determination of Pesticide Residue eliminates false-positive detection method |
| US10915548B2 (en) * | 2017-04-18 | 2021-02-09 | Chinese Academy Of Inspection And Quarantine | Method of online tracing pesticide residues and visualizing warning on basis of high resolution mass spectrum, internet and geographic information |
| 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 |
| CN108535249B (en) * | 2018-04-25 | 2021-02-26 | 郑州贝贝生物科技有限公司 | Pesticide residue fast detection point card |
| CN109270033A (en) * | 2018-09-10 | 2019-01-25 | 南京市产品质量监督检验院 | The rapid detection method of Hazard factor in a kind of food |
| CN110487732A (en) * | 2019-08-01 | 2019-11-22 | 李孝艳 | The rapid detection method of Organophosphorus and carbamate pesticides pesticide residue in chestnut mushroom |
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