CN1512165A - Detection reagent kit for pesticide residue, its preparing method and its use - Google Patents
Detection reagent kit for pesticide residue, its preparing method and its use Download PDFInfo
- Publication number
- CN1512165A CN1512165A CNA021596689A CN02159668A CN1512165A CN 1512165 A CN1512165 A CN 1512165A CN A021596689 A CNA021596689 A CN A021596689A CN 02159668 A CN02159668 A CN 02159668A CN 1512165 A CN1512165 A CN 1512165A
- Authority
- CN
- China
- Prior art keywords
- kit
- acetylcholinesterase
- protective agent
- enzyme
- housefly
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The reagent kit for detecting pesticide residue includes acetylcholinesterase, protecting agent, primer and developing agent. The single or purified monopolymer acetylcholinesterase highly sensitive to pesticide is obtained from sensitive housefly line head. The reagent kit may be maintained at normal temperature and has high sensitivity and high measurement accuracy in detecting pesticide residue in vegetable and fruits.
Description
[technical field]
The present invention relates to a kind of residual toxicity detection kit, it comprise acetylcholinesterase, protective agent, substrate and/with developer, wherein, acetylcholinesterase is the single aggressiveness acetylcholinesterase that obtains single molecule-type or purifying from housefly strain head.
[technical background]
At present, generally use agricultural chemicals in grain, vegetables, fruit planting process, agricultural product have residual trace pesticide inevitably, and to cause harm be self-evident and agricultural chemicals has people's health.Along with growth in the living standard, people more and more pay attention to food security, and therefore, the residues of pesticides problem on the agricultural product is paid close attention to by people day by day.
It is impossible that each vegetables, fruit sample are all carried out instrumental analysis, and therefore, people press for and a kind ofly can guarantee that residual toxicity on the agricultural product is in country allows safe range to the means that a large amount of vegetables, fruit sample carry out quick primary dcreening operation.
The existing report of the research of residual toxicity Fast Detection Technique aspect.Aspect instrument, spectrophotometer is reequiped the requirement that makes it to be fit to the detection of enzyme inhibition method, become instrumentation (as Beijing Rayleigh, Shanghai photoelectricity institute etc.).Aspect existing enzyme of utilization and instrument cooperation practical application, The Institute for the Control of Agrochemicals of the Ministry of Agriculture,PRC etc. have formulated the standard of utilizing butyrylcholine esterase to detect residual toxicity.But the method that adopts instrument detecting still just can not satisfy the market demand of low cost, fast detecting.
Organophosphorus and carbamate insecticides are acetylcholine esterase inhibition (acetylcholinesterase by their main toxicity mechanism of pesticide variety of extensively generally using, AChE) activity, cause acetylcholine (acetylcholine, ACh) accumulation, influenced normal nerve conduction, insect is poisoned and caused death.
Having the multiple mode of mentioning above comprising within the method to detect wherein that enzyme inhibition method detects at present for above-mentioned two class agricultural chemicals is one of topmost method.The enzyme that is adopted in the enzyme assay method mainly contains two kinds, one, and plant esterase, referring to, for example, CN1366065A, CN1293363A, CN1279292A; Its two, cholinesterase, for example, acetylcholinesterase and butyrylcholine esterase, referring to, for example, CN1349094A, CN1224160A.But present enzyme process detects and exists false positive rate higher, to the defective of some agricultural chemicals insufficient sensitivity.
The means that suppress method principle detection residual toxicity according to enzyme have test paper method, colourimetry, immobilized enzyme method (enzyme electrode or enzyme chip, also the someone claims biology sensor).Which kind of method no matter, enzyme is the most critical factor of decision detection method sensitivity and reliability.Enzyme has determined the sensitivity of method to the susceptibility of organophosphorus commonly used and carbamate chemicals for agriculture, and the kind of enzyme has determined the reliability of method.
The present inventor to the relevant enzyme that is used for enzyme inhibition method is, compare as esterase (comprising animal sources and plant source), butyrylcholine esterase, acetylcholinesterase etc. and studies confirm that, acetylcholinesterase can narrow spectrumly be suppressed by organophosphorus and carbamate chemicals for agriculture, detect this two classes agricultural chemicals with it and have tangible selectivity and reliability than butyrylcholine esterase, carboxy-lesterase etc., the false positive rate of testing result also is starkly lower than other enzyme system, with regard to overall target, acetylcholinesterase is used for the residual toxicity fast detecting and is better than other enzyme system.
But now being used in the key issue that acetylcholinesterase exists in the residual toxicity quick detection kit is the normal temperature holding time of how to improve the susceptibility of enzyme and prolonging enzyme.
The present inventor discovers by intensive; the acetylcholinesterase of separate sources is to agricultural chemicals susceptibility difference; adopting indoor responsive housefly to carry out breeding work can make its susceptibility to agricultural chemicals further improve in addition; by adding protective agent; make it to cooperate with enzyme; can prolong the normal temperature holding time of enzyme, so finished the present invention.
[summary of the invention]
Purpose of the present invention provides a kind of residual toxicity detection kit, it comprise acetylcholinesterase, protective agent, substrate and and developer, wherein, acetylcholinesterase is the single aggressiveness acetylcholinesterase that obtains single molecule-type or purifying from housefly strain head.
Another object of the present invention provides the preparation method of single aggressiveness acetylcholinesterase of the single molecule-type that adopted in a kind of kit or purifying.
A further object of the present invention provides the application that described residual toxicity detection kit is used for detecting the residual toxicity of agricultural product.
Residual toxicity detection kit of the present invention by adopt from the housefly strain head to the agricultural chemicals sensitivity obtain to the extremely sensitive single type acetylcholinesterase of agricultural chemicals, overcome and mixed the acetylcholinesterase molecule-type and detect the enzyme source that exists in the residual toxicity the problem that susceptibility is low, linear relationship is bad, false positive rate is high agricultural chemicals.In addition,, make it to cooperate, solved the problem that kit can not the normal temperature preservation with single aggressiveness acetylcholinesterase of described single molecule-type or purifying by adding protective agent.
It is sugar and/or protein that the normal temperature preservation needs use protective agent, protective agent.The sugar that can select for use among the present invention is selected from for example sucrose, cyclodextrin and sweet mellow wine and composition thereof, preferred three's potpourri.The protein that can select for use is for example bovine serum albumin.The ratio of protective agent and acetylcholinesterase is 10-500: 1, be preferably 50-200: and 1, most preferably be 100: 1, can obtain the kit that can preserve at normal temperatures.But can utilize for example grid structure fixed member type acetylcholinesterase of cyclodextrin of protective agent.
Substrate can be for example conventional acetylcholine, for example halo acetylcholine such as acecoline, acetylcholine bromide, halo acetylthiocholine such as bromination acetylthiocholine or acetylthiocholine iodide, acetyl thio choline or Butyryl thiocholine, preferred acetyl thio choline or Butyryl thiocholine.
Developer can be DTNB (two sulphur two nitrobenzoic acids) for example.
In the present invention, the housefly strain of employing mainly is the agricultural chemicals sensitive strain of cultivating by housefly compatriot culture technique.Utilize this strain, by saltout, step such as differential centrifugation, gel permeation chromatography, affinity chromatography, freeze drying, from the single aggressiveness acetylcholinesterase of responsive housefly strain head separation and purification to extremely sensitive single molecule-type of agricultural chemicals or purifying, concrete purifying route can be referring to accompanying drawing 1 and embodiment.
To collect the dry powder that obtains (promptly by above-mentioned steps, acetylcholinesterase enzyme powder, call the enzyme powder in the following text) and protective agent, the potpourri of sucrose, cyclodextrin and mannose for example, be preferably sucrose: cyclodextrin: the potpourri of sweet mellow wine=1: 4: 5, mix by for example 1: 100 (w/w), be the enzyme source (A) of detection kit after the immobilization; With substrate and developer combination, promptly obtain the colour developing source (B) of kit.A, B and damping fluid had both got the residual toxicity detection kit together after packing.
In specific embodiment, responsive housefly strain can be passed through liquid nitrogen flash freezer 2 minutes, take out the back and its head and body are separated with oscillator, by sieve head is being separated with body.With the head that obtains with organizing cutting machine to shred, filter the back differential centrifugation, saltout, desalination, sucrose density gradient centrifugation, affinity chromatography, freeze-drying obtain the single aggressiveness acetylcholinesterase to the single molecule-type of agricultural chemicals sensitivity or purifying, this esterase detects for single band, referring to accompanying drawing 4 by native gel electrophoresis.According to above-mentioned composition and ratio, make optical density value more than 0.6/ minute by adjustment to enzyme concentration, concentration of substrate, chromogenic agent.
The purposes of kit of the present invention is to detect remaining agricultural chemicals on vegetables or the fruit, particularly organophosphorus and carbamate insecticides, acetylcholinesterase in the kit of the present invention is joined vegetables or the fruit sample detects, during detection, enzymatic activity reduces then and contains residual toxicity in the sample, otherwise then nontoxic.
The present invention adopts to separate from the mixed type acetylcholinesterase the single molecule-type of agricultural chemicals sensitivity or single aggressiveness acetylcholinesterase of purifying has been overcome the defective that mixed molecules type susceptibility is low, error is big, false positive rate is high; In addition,, it is cooperated with enzyme, kit can be preserved at normal temperature by adding protective agent.
The quick residual toxicity that the present invention can be widely used in various vegetable and fruits detects, its promotion and application can significantly reduce or avoid the generation of the agricultural chemicals acute poisoning that causes because of food (containing vegetables, fruit) and outlet problem the problem includes: residual toxicity exceeds standard problem.
[description of drawings]
1. Fig. 1 is the purification technique route map of single aggressiveness acetylcholinesterase of responsive single molecule-type or purifying.
2. Fig. 2 is the susceptibility and the optimum reaction conditions mensuration of acetylcholinesterase.
3. Fig. 3 is a vegetable and fruit sample assay method.
4. Fig. 4 is single aggressiveness acetylcholinesterase collection of illustrative plates of single molecule-type or purifying.
[embodiment]
Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
Responsive housefly head is separated behind liquid nitrogen frozen, and tiny (pH7.5,0.1mol/L) 1: 5 ratio is put into refiner homogenate with homogenate by family.Homogenate is got supernatant with ammonium sulfate precipitation (50～60% saturation degree) behind 1000g, 10000g and 100000g differential centrifugation, using the desalination of G25 post.After the desalination, dry powder is made in freeze drying.With dry powder be dissolved in damping fluid (pH7.5,0.1mol/L) in (w/v:1: 1) by G200 chromatographic column and affinity column, collect contain after the active eluant freeze drying dry powder, both be acetylcholinesterase enzyme powder.(sucrose: cyclodextrin: sweet mellow wine=1: 4: 5) mixing by 1: 100 (w/w), both had been the enzyme source (A) of detection kit after the immobilization with protective agent with the enzyme powder; Replace the enzyme powder with substrate and developer as stated above, both obtained the colour developing source (B) of kit.A, B and damping fluid (pH7.5,0.1mol/L contain 0.05%～0.1% Triton X-100) wherein, contain substrate and each 0.6mg of developer in the reaction system of 3mL damping fluid (pH7.5).After packing, both got the residual toxicity detection kit.
Embodiment 2
Responsive housefly head is separated behind liquid nitrogen frozen, and tiny (pH7.5,0.1mol/L) 1: 5 ratio is put into refiner homogenate with homogenate by family.Homogenate goes supernatant with ammonium sulfate precipitation (50～60 saturation degree) behind 1000g, 10000g and 100000g differential centrifugation, is using the desalination of G25 post.After the desalination, dry powder is made in freeze drying.Dry powder is dissolved in the distilled water (w/v:1: 1),, collect the composition of 10%～20% saccharose gradient section by sucrose density gradient centrifugation, through the G200 chromatographic column, collect contain after the active eluant freeze drying dry powder, both be acetylcholinesterase enzyme powder.(starch: cyclodextrin: sweet mellow wine=1: 5: 5) mixing by 1: 150 (w/w), both had been the enzyme source (A) of detection kit after the immobilization with protective agent with the enzyme powder; Replace the enzyme powder with substrate and developer as stated above, both obtained the colour developing source (B) of kit.A, B and damping fluid (pH7.5,0.1mol/L contain 0.05%～0.1% Triton X-100) wherein, contain substrate and each 0.6mg of developer in the 3mL damping fluid (pH7.5).After packing, both got the residual toxicity detection kit.
Biological Examples 1
With standard sample of pesticide with acetone be mixed with mother liquor (about 100～200mg/L), be diluted to required concentration with extract (containing 0.05%～0.1% Triton X-100) before using.The standard items mother liquor of agricultural chemicals is diluted to 5～8 concentration gradients with extract, get 3mL respectively and add acetylcholinesterase 0.1mL (addition is that unrestraint agent control activity is 0.8～1.0 a Δ OD412 value/3 minutes), after 15 minutes～30 minutes (time length depends on medicament kind and inhibiting rate required standard 35% or 45%), add substrate and developer (each 0.6mg), on spectrophotometer, read Δ OD412 value/3 minutes.Contrast replaces pesticide mother liquor with acetone.Each measures independent the repetition 3 times.
Calculate inhibiting rate by following formula:
Inhibiting rate (%)=(1-(chemicals treatment group Δ OD412 value/3 minutes)/(contrast Δ OD412 value/3 minutes)) * 100
Make regression equation according to inhibiting rate and corresponding drug concentration logarithm value, calculate the corresponding drug concentration value of inhibiting rate 35% or 45% (kind and the inhibitory reaction time of depending on medicament) and be susceptibility, the results are shown in following table 1.
The acetylcholinesterase of table 1 kit of the present invention is to the susceptibility of several representative medicaments
| Pesticide name | Susceptibility | Remarks |
| DDVP | ＜0.01mg/L | 0.01-0.05mg/L inhibiting rate 35-71% |
| Phoxim | ＜0.5mg/L | 0.5mg/L inhibiting rate 80-92% |
| Acephatemet | ＜3mg/L | 3-10mg/L inhibiting rate 23-86% |
| Methomyl | ＜0.3mg/L | 0.3-0.7mg/L inhibiting rate 91-95% |
| Carbosulfan | ＜0.3mg/L | 0.15-0.7mg/L inhibiting rate 19-65% |
By table 1 as seen, kit of the present invention has the susceptibility of height to organophosphorus and carbamate pesticide.Can detect the residual toxicity of low concentration.
Biological Examples 2
With standard sample of pesticide with acetone be mixed with mother liquor (about 100～200mg/L), be diluted to required concentration with extract (containing 0.05%～0.1% Triton X-100) before using.The standard items mother liquor of agricultural chemicals is diluted to the listed respective concentration of table 2 with extract, get 3mL respectively and add acetylcholinesterase 0.1mL (addition is that unrestraint agent control activity is 0.8～1.0 a Δ OD412 value/3 minutes), after 15 minutes～30 minutes (time length depends on medicament kind and inhibiting rate required standard 35% or 45%), add substrate and developer (each 0.6mg), on spectrophotometer, read Δ OD412 value/3 minutes.Contrast replaces pesticide mother liquor with acetone.Each measures independent the repetition 3 times.Whole test is established 18 ℃, 25 ℃ and 30 ℃ of processing respectively.The results are shown in following table 2.
Calculate inhibiting rate by following formula:
Inhibiting rate (%)=(1-(chemicals treatment group Δ OD412 value/3 minutes)/(contrast Δ OD412 value/3 minutes)) * 100
Table 2 temperature is to the influence of medicament susceptibility
(percent numerical value is the acetylcholinesterase inhibiting rate of kit of the present invention in the table)
| Pesticide name | Concentration (mg/L) | Temperature (℃) | ||
| ????18 | ????25 | ????30 | ||
| The malathion | ????3 | ????25％ | ????29％ | ????28％ |
| Phoxim | ????0.5 | ????91％ | ????92％ | ????89％ |
| Acephatemet | ????3 | ????- | ????34％ | ????31％ |
| DDVP | ????0.02 | ????41％ | ????55％ | ????70％ |
| DDVP | ????1 | ????75.5％ | ????70.0％ | ????71.0％ |
| Methomyl | ????1 | ????98％ | ????93.3％ | ????90.0％ |
| Carbosulfan | ????0.3 | ????54％ | ????59％ | ????62％ |
By last table 2 as seen, temperature is little to kit influence of the present invention, and kit of the present invention can determine the agricultural chemicals organophosphorus and the carbamate pesticide particularly commonly used of low concentration at normal temperatures.
Biological Examples 3
Kit according to embodiment 1 preparation was preserved for three weeks under 20-30 ℃ of lucifuge condition, adopted conventional method to measure the enzymatic activity of its each time point.Enzymatic activity value unit is Δ OD412 value/3 minutes.
Table 3 room temperature preservation is to the influence (being the mean value of 3 mensuration in the table) of acetylcholine esterase active
| Time (my god) | ????0 | ????1 | ????3 | ????7 | ????14 | ????21 |
| Enzymatic activity | ????1.0 | ????0.9 | ????1.1 | ????0.8 | ????1.0 | ????0.9 |
As above table 3 as seen, the acetylcholinesterase in the kit of the present invention can be preserved for three weeks at normal temperatures, and did not influence its enzymatic activity, therefore, can carry out residual toxicity at normal temperatures and detect.
Biological Examples 4
Kit of the present invention and conventional reagent comparison the results are shown in following table 4.
| Project | Kit of the present invention | Conventional kit |
| Storage temperature | Normal temperature, storage life is greater than 2 weeks | Cryopreservation (in refrigerator) |
| Serviceability temperature | Room temperature, 18-30 ℃ | Constant temperature |
| The enzyme source | Single aggressiveness of single molecule-type or purifying | Mixed type butyrylcholine esterase or acetylcholinesterase |
| Developer and substrate | One | Separate |
| The source | Responsive housefly head | It is various to originate, and causes poor reproducibility |
By last table 4 as seen, kit of the present invention has the advantages that obviously be better than conventional kit.
Though above the present invention is described in detail with a general description of the specific embodiments on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, not departing from the these modifications or improvements basically of spirit of the present invention, all belong to the scope of protection of present invention.
Claims (10)
1. a residual toxicity detection kit is characterized in that, it comprises single aggressiveness acetylcholinesterase, protective agent, substrate and the developer of single molecule-type or purifying.
2. according to the kit of claim 1, wherein, single aggressiveness acetylcholinesterase of described single molecule-type or purifying obtains from the separation and purification of responsive housefly strain head.
3. according to the kit of claim 2, wherein, described protective agent is sugar and/or protein.
4. according to the kit of claim 3, wherein, described sugar is selected from sucrose, cyclodextrin and sweet mellow wine and composition thereof.
5. according to arbitrary kit of claim 1-4, wherein, the weight ratio of described protective agent and acetylcholinesterase is 10-500: 1.
6. according to the kit of claim 5, wherein, the weight ratio of protective agent and acetylcholinesterase is 50-200: 1
7. according to arbitrary kit of claim 1-6, wherein, described substrate is acetyl thio choline or Butyryl thiocholine.
8. according to arbitrary kit of claim 1-7, wherein, described developer is the two nitrobenzoic acids of two sulphur.
9. according to the preparation method of single aggressiveness acetylcholinesterase of single molecule-type in arbitrary kit of aforementioned claim 1-8 or purifying, it comprises the steps: to saltout, differential centrifugation, gel permeation chromatography, affinity chromatography and freeze drying.
10. according to the method for claim 9, wherein, housefly is that culture technique born of the same parents is cultivated the responsive housefly strain to agricultural chemicals.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021596689A CN1321194C (en) | 2002-12-30 | 2002-12-30 | Detection reagent kit for pesticide residue, its preparing method and its use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021596689A CN1321194C (en) | 2002-12-30 | 2002-12-30 | Detection reagent kit for pesticide residue, its preparing method and its use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1512165A true CN1512165A (en) | 2004-07-14 |
| CN1321194C CN1321194C (en) | 2007-06-13 |
Family
ID=34237571
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021596689A Expired - Fee Related CN1321194C (en) | 2002-12-30 | 2002-12-30 | Detection reagent kit for pesticide residue, its preparing method and its use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1321194C (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102016063A (en) * | 2008-04-24 | 2011-04-13 | 微化学技研株式会社 | Apparatus, kit and method for detection of cholinesterase-inhibiting substance |
| CN101724649B (en) * | 2008-10-29 | 2012-05-02 | 中国科学院上海生命科学研究院 | Method for displaying heterologous proteins on surfaces of cells and product |
| CN105785021A (en) * | 2016-04-08 | 2016-07-20 | 广州万联生物科技有限公司 | Quick detecting card for immunochromatography of organophosphorus and carbamate pesticide multiresidue cholinesterase |
| CN106191212A (en) * | 2016-08-08 | 2016-12-07 | 陈英就 | Fast detecting reagent kit for toxicity of pesticide residue, the preparation method of stabilizer and the preparation method of acetylcholine esterase enzyme powder |
| CN107513526A (en) * | 2017-09-29 | 2017-12-26 | 郑州欧柯奇仪器制造有限公司 | The fast preparation method of the residual fast survey enzyme of agriculture |
| US10132726B2 (en) | 2016-06-07 | 2018-11-20 | Cubic Corporaton | Paper transit tickets as a medium for environmental sampling |
| CN109781718A (en) * | 2019-01-17 | 2019-05-21 | 广东省食品工业研究所有限公司 | A kind of preparation method of the compound enzyme solution for fast detecting pesticide residue |
| CN112881359A (en) * | 2020-05-08 | 2021-06-01 | 北京中检葆泰生物技术有限公司 | Method for detecting pesticide residue |
| CN112924440A (en) * | 2020-12-24 | 2021-06-08 | 广东省食品工业研究所有限公司 | Rapid detection kit for pesticide residues and preparation method thereof |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BE1008221A3 (en) * | 1994-03-28 | 1996-02-20 | Anda Biolog Sa | SOLID SUPPORT FIXING AT LEAST ONE BIOLOGICALLY ACTIVE COMPONENT COMPRISING A REACTIONAL, STABILIZED SITE AND METHOD FOR OBTAINING SAME. |
| US5716831A (en) * | 1995-05-24 | 1998-02-10 | Board Of Trustees Operating Michigan State University | Method and test kit for detecting insecticide resistance |
| US5686237A (en) * | 1995-06-05 | 1997-11-11 | Al-Bayati; Mohammed A. S. | Use of biomarkers in saliva to evaluate the toxicity of agents and the function of tissues in both biomedical and environmental applications |
| JP2002062265A (en) * | 2000-08-22 | 2002-02-28 | Satake Corp | Detector for pesticide residue |
| CN1349094A (en) * | 2000-10-13 | 2002-05-15 | 杜廷发 | Fast test method and test device for residual organophosphate and carbamate pesticide |
| CN1293363A (en) * | 2000-12-18 | 2001-05-02 | 范永新 | Quick test box of residual agricultural organophosphorus chemical and its preparing process |
| KR100441056B1 (en) * | 2001-04-16 | 2004-07-22 | 주식회사 소일테크 | Method for measuring the amount of residual pesticide from agricultural products using acetylcholineserase |
| CN1375697A (en) * | 2002-03-12 | 2002-10-23 | 王晋峰 | Fruit and vegetable pesticide pollution test paper |
| JP3473022B2 (en) * | 2002-04-15 | 2003-12-02 | 株式会社サタケ | Pesticide residue measurement method |
-
2002
- 2002-12-30 CN CNB021596689A patent/CN1321194C/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102016063A (en) * | 2008-04-24 | 2011-04-13 | 微化学技研株式会社 | Apparatus, kit and method for detection of cholinesterase-inhibiting substance |
| CN102016063B (en) * | 2008-04-24 | 2014-09-17 | 微化学技研株式会社 | Apparatus, kit and method for detection of cholinesterase-inhibiting substance |
| CN101724649B (en) * | 2008-10-29 | 2012-05-02 | 中国科学院上海生命科学研究院 | Method for displaying heterologous proteins on surfaces of cells and product |
| CN105785021A (en) * | 2016-04-08 | 2016-07-20 | 广州万联生物科技有限公司 | Quick detecting card for immunochromatography of organophosphorus and carbamate pesticide multiresidue cholinesterase |
| US10132726B2 (en) | 2016-06-07 | 2018-11-20 | Cubic Corporaton | Paper transit tickets as a medium for environmental sampling |
| CN106191212A (en) * | 2016-08-08 | 2016-12-07 | 陈英就 | Fast detecting reagent kit for toxicity of pesticide residue, the preparation method of stabilizer and the preparation method of acetylcholine esterase enzyme powder |
| CN106191212B (en) * | 2016-08-08 | 2019-11-19 | 陈英就 | The preparation method of fast detecting reagent kit for toxicity of pesticide residue, the preparation method of stabilizer and cholinesterase enzyme powder |
| CN107513526A (en) * | 2017-09-29 | 2017-12-26 | 郑州欧柯奇仪器制造有限公司 | The fast preparation method of the residual fast survey enzyme of agriculture |
| CN109781718A (en) * | 2019-01-17 | 2019-05-21 | 广东省食品工业研究所有限公司 | A kind of preparation method of the compound enzyme solution for fast detecting pesticide residue |
| CN112881359A (en) * | 2020-05-08 | 2021-06-01 | 北京中检葆泰生物技术有限公司 | Method for detecting pesticide residue |
| CN112924440A (en) * | 2020-12-24 | 2021-06-08 | 广东省食品工业研究所有限公司 | Rapid detection kit for pesticide residues and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1321194C (en) | 2007-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Badiane et al. | Use of soil enzyme activities to monitor soil quality in natural and improved fallows in semi-arid tropical regions | |
| Pritsch et al. | Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae | |
| Griepentrog et al. | Nitrogen deposition promotes the production of new fungal residues but retards the decomposition of old residues in forest soil fractions | |
| Ramos et al. | Cover crops under different managements vs. frequent tillage in almond orchards in semiarid conditions: Effects on soil quality | |
| Bossio et al. | Soil microbial community response to land use change in an agricultural landscape of western Kenya | |
| Fani et al. | A critical evaluation of cultural methods for the identification of atoxigenic Aspergillus flavus isolates for aflatoxin mitigation in pistachio orchards of Iran | |
| Probst et al. | Relationships between in vivo and in vitro aflatoxin production: reliable prediction of fungal ability to contaminate maize with aflatoxins | |
| Barbehenn et al. | Performance of a generalist grasshopper on a C 3 and a C 4 grass: compensation for the effects of elevated CO 2 on plant nutritional quality | |
| Neupane et al. | Long term crop rotation effect on subsequent soybean yield explained by soil and root-associated microbiomes and soil health indicators | |
| Stauffer et al. | Effect of willow short rotation coppice on soil properties after three years of growth as compared to forest, grassland and arable land uses | |
| CN1512165A (en) | Detection reagent kit for pesticide residue, its preparing method and its use | |
| Villani et al. | The detection and interaction of multiple organophosphorus and carbamate insecticide resistance genes in field populations of Culex pipiens from Italy | |
| Surya Kalyani et al. | Influence of endosulfan on microbial biomass and soil enzymatic activities of a tropical alfisol | |
| Hydbom et al. | Reduced tillage stimulated symbiotic fungi and microbial saprotrophs, but did not lead to a shift in the saprotrophic microorganism community structure | |
| Xiang et al. | Key indicators for renewal and reconstruction of perennial trees soil: Microorganisms and phloridzin | |
| Bell et al. | Impacts of management on soil biota in Vertosols supporting the broadacre grains industry in northern Australia | |
| Low et al. | Carbohydrate analysis of western Canadian honeys and their nectar sources to determine the origin of honey oligosaccharides | |
| He et al. | Land reclamation and short‐term cultivation change soil microbial communities and bacterial metabolic profiles | |
| Lin et al. | Detection of mold in processed foods by high performance liquid chromatography | |
| Zhang et al. | Soil microbial residue dynamics after 3-year elevated O 3 exposure are plant species-specific | |
| CN1731155A (en) | Solid enzyme agent for quick detection of residual pesticide toxicity and preparation method and using method thereof | |
| Zinkl et al. | Effects of cholinesterases of rainbow trout exposed to acephate and methamidophos | |
| Liang et al. | Tree species-specific effects on soil microbial residues in an upper Michigan old-growth forest system | |
| Whitmore et al. | Glycosidically-bound volatile phenols linked to smoke taint: Stability during fermentation with different yeasts and in finished wine | |
| Smemo et al. | Characteristics of DOC exported from northern hardwood forests receiving chronic experimental NO 3− deposition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |