CN108896541A - A kind of residual speed of vegetables agriculture surveys the method for discrimination and card of false positive - Google Patents
A kind of residual speed of vegetables agriculture surveys the method for discrimination and card of false positive Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
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Abstract
The invention discloses method of discrimination and card that a kind of residual speed of vegetables agriculture surveys false positive.Method of discrimination includes mixing sample liquid and enzyme, and the chromogenic substrate of enzyme is added later, is determined in sample liquid according to colour developing situation with the presence or absence of the secondary metabolites for leading to false positive, wherein the activity of enzyme can be inhibited by secondary metabolites, without being inhibited by pesticide residue.Method of the invention, can effectively detect, which whether there is in sample, causes conventional choline base esterase pesticide quick measuring card the secondary substance of false positive results occur.Have the advantages that easy to use, quick simultaneously, effective supplement of existing pesticide residue fast detection card can be acted on, avoid loss caused by false positive results.
Description
Technical field
The present invention relates to Detecting Pesticide fields, in particular to for identify the residual false positive of vegetables agriculture method of discrimination and
Corresponding quick measuring card.
Background technique
Pest and disease damage often causes a large amount of economic loss to agricultural, forestry animal husbandry.Spraying insecticide is prevent pest and disease damage effective
Means.But thus some disease pest pests develop drug resistance, and need to increase the dosage sprayed insecticide to tackle insect pest.These do not conform to
Reason necessarily leads to the excessive pesticide residues such as agricultural product such as veterinary antibiotics using pesticide, thus the health of the majority of consumers, very
It is poisoned to death to directly resulting in.Therefore Detecting Pesticide is very necessary.Method master currently used for Detecting Pesticide
There are spectroscopic methodology, enzyme inhibition, chromatography, chemical fast determining method, immunoassay, In vivo detection method etc..
The advantages that enzyme inhibition detects pesticide residue and is simple and convenient to operate because of its pre-treatment, low in cost, is listed in country
Recommend to survey standard detecting method (GB/T 5009.199-2003) and be widely applied in the market.But leek, garlic bolt, blanched garlic leaves,
Contain a large amount of secondary substance in the victuals such as wild rice stem, when carrying out shredding sample treatment, the cell rupture of vegetables makes secondary
Substance release influences testing result into analyte sample fluid, or even can generate a large amount of false positive results.
In the quick detection process of vegetable pesticide residue, since secondary metabolites inhibits depositing for cholinesterase activity or pigment
In the correct intuitive performance for disturbing enzyme reaction process, the false positive results made happen occasionally.The positive is found to be for detection
Sample is confirmed due to lacking effective quick means, so that many vegetables are by with the residual exceeded processing of agriculture, and some exists
After very long instrument detection, vegetables are consumed in market, this food safety for being unfavorable for ensureing the people and maintenance dish
Agriculture interests reduce quality inspection efficiency and products in circulation speed.
CN106093032A, CN104142305A and mulberry field garden, Chai Lina, Wei Chaojun wait 4 kinds of enzyme inhibition detection pungent
Research [J] China's agronomy notification that Vegetable in chili pesticide residue false positive is eliminated, 2009,25 (11):60-64. etc. is instructed to sample
Solution is heated, to overcome false positive caused by secondary metabolites.This operating method is needed by certain instrument,
Detection time is partially long, it is difficult to determining on site.
It is to have technical problem to be solved if any false positive results are fast and effeciently determined.
Summary of the invention
The purpose of the present invention is to provide a kind of method of discrimination for identifying pesticide quick measuring card false positive and differentiate card.
The technical solution adopted by the present invention is that：
A kind of method of discrimination of the residual speed survey false positive of vegetables agriculture, including sample liquid and enzyme are mixed, the colour developing of enzyme is added later
Substrate determines in sample liquid according to colour developing situation with the presence or absence of the secondary metabolites for leading to false positive, wherein the activity of enzyme can be secondary
Raw metabolin inhibits, without being inhibited by pesticide residue.
As the further improvement of above-mentioned method of discrimination, beta galactosidase of the enzyme in hydrolase.
As the further improvement of above-mentioned method of discrimination, the corresponding chromogenic substrate of enzyme is one of Benzazole compounds.
As the further improvement of above-mentioned method of discrimination, the corresponding chromogenic substrate of enzyme is the chloro- 3- indoles-β-D- of the bromo- 4- of 5-
Galactopyranoside.
A kind of residual speed of vegetables agriculture surveys the differentiation card of false positive, including enzyme and chromogenic substrate piece, fixes on enzyme active
It can be inhibited by secondary metabolites, the enzyme without being inhibited by pesticide residue, what the absorption of chromogenic substrate piece can be developed the color by enzymic catalytic reaction
Substrate.
As the further improvement of above-mentioned differentiation card, beta galactosidase of the enzyme in hydrolase.
As the further improvement of above-mentioned differentiation card, the corresponding chromogenic substrate of enzyme is one of Benzazole compounds.
As the further improvement of above-mentioned differentiation card, the corresponding chromogenic substrate of enzyme is the chloro- 3- indoles-β-D- pyrrole of the bromo- 4- of 5-
It mutters galactoside.
As the further improvement of above-mentioned differentiation card, differentiate that the water-absorbent material that card uses is selected from filter paper.
As the further improvement of above-mentioned differentiation card, differentiation is arranged with epiphragma.
The beneficial effects of the invention are as follows：
Method of the invention, can effectively detect, which whether there is in sample, causes conventional choline base esterase pesticide quick measuring card to go out
The secondary substance of existing false positive results.Have the advantages that easy to use, quick simultaneously, existing pesticide residue fast detection card can be acted on
Effective supplement, avoid loss caused by false positive results.
Differentiation card of the invention after adding sample liquid to biological enzyme on piece, is contacted with tablet by folding, need to only use hand flicking
You can get it by reaction 3min as a result, being not necessarily to pre-reaction, and the detection used time is short, at low cost, is convenient for carrying；In the residual speed of vegetables agriculture is surveyed,
There is the vegetable sample of positive findings with the detection of cholinester enzyme inhibition, causes false positive results if contained in vegetable sample liquid
Secondary substance when, the color of biological enzyme would not change colour, and such as present colourless；It is on the contrary then discoloration can be reacted, blue is such as presented.
Detailed description of the invention
Fig. 1 is a kind of floor map of false positive differentiation card that the residual speed of vegetables agriculture is surveyed；
Fig. 2 is a kind of schematic cross-section of false positive differentiation card that the residual speed of vegetables agriculture is surveyed.
Specific embodiment
A kind of method of discrimination of the residual speed survey false positive of vegetables agriculture, including sample liquid and enzyme are mixed, the colour developing of enzyme is added later
Substrate determines in sample liquid according to colour developing situation with the presence or absence of the secondary metabolites for leading to false positive, wherein the activity of enzyme can be secondary
Raw metabolin inhibits, without being inhibited by pesticide residue.
The activity of enzyme is not inhibited by pesticide residue, refers to that the activity of enzyme is still there are in the case where the pesticide residue qualification upper limit
It may remain in 80% or more or 90% or more.
As the further improvement of above-mentioned method of discrimination, beta galactosidase of the enzyme in hydrolase.
As the further improvement of above-mentioned method of discrimination, the corresponding chromogenic substrate of enzyme is one of Benzazole compounds.
As the further improvement of above-mentioned method of discrimination, the corresponding chromogenic substrate of enzyme is the chloro- 3- indoles-β-D- of the bromo- 4- of 5-
Galactopyranoside.
A kind of residual speed of vegetables agriculture surveys the differentiation card of false positive, including enzyme and chromogenic substrate piece, fixes on enzyme active
It can be inhibited by secondary metabolites, the enzyme without being inhibited by pesticide residue, what the absorption of chromogenic substrate piece can be developed the color by enzymic catalytic reaction
Substrate.
As the further improvement of above-mentioned differentiation card, beta galactosidase of the enzyme in hydrolase.
As the further improvement of above-mentioned differentiation card, the corresponding chromogenic substrate of enzyme is one of Benzazole compounds.
As the further improvement of above-mentioned differentiation card, the corresponding chromogenic substrate of enzyme is the chloro- 3- indoles-β-D- pyrrole of the bromo- 4- of 5-
It mutters galactoside.
As the further improvement of above-mentioned differentiation card, differentiate that the water-absorbent material that card uses is selected from filter paper.
As the further improvement of above-mentioned differentiation card, differentiation is arranged with epiphragma.
Referring to Fig. 1, Fig. 2, the false positive that a kind of residual speed of vegetables agriculture is surveyed differentiates card, mainly by bottom plate (1), biological enzyme (2),
Chromogenic substrate piece (3), epiphragma (4) composition, bottom plate are fixed with biological enzyme and chromogenic substrate piece, and are capped film covering.
Bottom plate is made of the synthetic paper material that can be degraded, and upper epiphragma is made of the OPP film for being covered with heat-sensitive glue.
False positive differentiates that a specific preparating example of card is as follows：
The round absorbent filter that diameter is 11mm is fixed on the biological enzyme (2) of rectangle bottom plate and diameter is 13mm's
On the position of chromogenic substrate piece (3).Where β-galactosidase 1 U is added to enzyme (2) on filter paper, drying, then X-Gal is added
It on chromogenic substrate piece (3), then dries, epiphragma is covered on bottom plate, biological enzyme (2) and chromogenic substrate piece (3) is made to prevent it
It makes moist and loses activity.
Below with reference to embodiment, technical solution of the present invention is further illustrated.
Embodiment 1
Differentiate that the process of card is as follows using false positive：
1) a piece of differentiation card is taken to throw off epiphragma, taking the residual speed of the agriculture of 60 μ L to survey is positive vegetable sample liquid, is added drop-wise to life
On object enzyme, then card doubling will will be differentiated, with finger light press 3min.Biological enzyme is shown as nothing in sky blue i.e. sample liquid
Secondary substance contains the secondary pollutant for causing false positive if biological enzyme is shown as white or light blue that is, in vegetable sample liquid
Matter.
2) by GB/T 5009.199-2003 or CFDA publication《KJ201710》Standard, the vegetables for taking the surface 3g clean,
Being cut into about 1cm can be put into sample bottle after the fragment of square, add the phosphate buffer of 10mLpH=8, fully shake rear quiet
2min is set, supernatant is prepare liquid；
3) it takes 60 μ L prepare liquids to be added drop-wise to biological enzyme on piece, then card doubling will be differentiated, with finger light press 3min；
4) it is operated with commercialization rapid detection for pesticide residue reagent by its specification after taking 3mL prepare liquid to be filtered,
Reaction reads inhibiting rate on the residual tacheometer of agriculture.
It operates, the vegetables for being determined as non agricultural chemical residuum through detection is detected, test result such as 1 institute of table by above-mentioned detection
Show：
Table 1
Card colour developing, which for white, that is, is determined as that the vegetable sample for having secondary substance takes GB/T, to be differentiated to false positive
Whole strain extraction method processing in 5009.199-2003 or KJ201710 standard, takes 60 μ L treatment fluids to be added drop-wise to biological enzyme on piece, so
Card doubling will be differentiated afterwards, with finger light press 3min；Take 3mL treatment fluid commercialization rapid detection for pesticide residue reagent by its specification
Operation, reaction reads inhibiting rate on the residual tacheometer of agriculture.The results are shown in Table 2：
Table 2
By table 1, table 2 it is found that being sentenced when the residual quick testing reagent calibrating of vegetables commercialization agriculture is positive sample using false positive
Can Ka not quickly it confirm whether containing the Plant Secondary Metabolites for inhibiting enzyme reaction in extracting solution, so as to adjust vegetable sample
Processing mode, and examined and determine under conditions of ensuring sample treatment liquid without containing secondary metabolites with enzymatic reagent.
It is operated by above-mentioned detection, is determined as the vegetables of non agricultural chemical residuum by GB/T 5009.199-2003 standard to through detecting
It is detected after carrying out carbofuran mark-on, wherein to the vegetables determined above containing Plant Secondary Metabolites using at whole strain extraction method
It manages, while the inspection of commercialization enzymatic reagent and false positive differentiation card inspection is carried out to treatment fluid, test result is as shown in table 3：
Table 3
As shown in Table 2, the residual speed survey false positive of this agriculture differentiates that card can the positive that block or enzyme reagent kit detects effectively residual to agriculture
As a result false positive judgement is carried out, test is repeated several times and variety classes vegetables are tested, this false positive differentiates that card can effectively be sentenced
Cholinesterase bring false positive test results Chu not inhibited by secondary substance.This false positive differentiates card to detection as shown in Table 3
Pesticides are insensitive, and also comply with the inspection in the test of other pesticide mark-ons such as Profenofos, Methomyl, DDVP, metrifonate
Gauge rule, tests favorable reproducibility.
Claims (10)
1. a kind of residual speed of vegetables agriculture surveys the method for discrimination of false positive, including sample liquid and enzyme are mixed, the colour developing bottom of enzyme is added later
Object determines in sample liquid according to colour developing situation with the presence or absence of the secondary metabolites for leading to false positive, wherein the activity of enzyme can be secondary
Metabolin inhibits, without being inhibited by pesticide residue.
2. method of discrimination according to claim 1, it is characterised in that：Beta galactosidase of the enzyme in hydrolase.
3. method of discrimination according to claim 2, it is characterised in that：The corresponding chromogenic substrate of enzyme is in Benzazole compounds
One kind.
4. method of discrimination according to claim 3, it is characterised in that：The corresponding chromogenic substrate of enzyme is the chloro- 3- Yin of the bromo- 4- of 5-
Diindyl-β-D- galactopyranoside.
5. a kind of residual speed of vegetables agriculture surveys the differentiation card of false positive, including enzyme and chromogenic substrate piece, it is characterised in that：It is solid on enzyme
It is fixed it is active can be inhibited by secondary metabolites, the enzyme without being inhibited by pesticide residue, the absorption of chromogenic substrate piece can be anti-by enzymatic
The substrate that should be developed the color.
6. differentiation card according to claim 5, it is characterised in that：Beta galactosidase of the enzyme in hydrolase.
7. differentiation card according to claim 6, it is characterised in that：The corresponding chromogenic substrate of enzyme is in Benzazole compounds
It is a kind of.
8. differentiation card according to claim 7, it is characterised in that：The corresponding chromogenic substrate of enzyme is the chloro- 3- Yin of the bromo- 4- of 5-
Diindyl-β-D- galactopyranoside.
9. being blocked according to the described in any item differentiations of claim 5~8, it is characterised in that：The water-absorbent material for differentiating that card uses is selected from
Filter paper.
10. being blocked according to the described in any item differentiations of claim 5~8, it is characterised in that：Differentiation is arranged with epiphragma.
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| CN201810442070.0A CN108896541B (en) | 2018-05-10 | 2018-05-10 | Method and card for discriminating false positive of vegetable pesticide residue quick test |
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|---|---|---|---|
| CN201810442070.0A CN108896541B (en) | 2018-05-10 | 2018-05-10 | Method and card for discriminating false positive of vegetable pesticide residue quick test |
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| CN108896541A true CN108896541A (en) | 2018-11-27 |
| CN108896541B CN108896541B (en) | 2020-09-22 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109975280A (en) * | 2019-03-15 | 2019-07-05 | 广州聚佰生物科技有限公司 | A kind of pesticide residue detection method and card of recognizable false positive |
| CN110596083A (en) * | 2019-08-20 | 2019-12-20 | 广州聚佰生物科技有限公司 | Reagent and card for detecting organophosphorus and carbamate pesticide residues |
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| CN1496403A (en) * | 2001-02-08 | 2004-05-12 | ϣ | Plant-derived molecules and genetic sequences encoding same and uses therefor |
| CN101469315A (en) * | 2007-12-29 | 2009-07-01 | 中国科学院生态环境研究中心 | Two-hybrid yeast for detecting estrogen-like compound in environment and biological test method |
| CN101893523A (en) * | 2009-05-20 | 2010-11-24 | 纳米基因有限公司 | Analyte detecting method and composite |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109975280A (en) * | 2019-03-15 | 2019-07-05 | 广州聚佰生物科技有限公司 | A kind of pesticide residue detection method and card of recognizable false positive |
| CN110596083A (en) * | 2019-08-20 | 2019-12-20 | 广州聚佰生物科技有限公司 | Reagent and card for detecting organophosphorus and carbamate pesticide residues |
| CN110596083B (en) * | 2019-08-20 | 2021-10-15 | 广州聚佰生物科技有限公司 | Reagent and card for detecting organophosphorus and carbamate pesticide residues |
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| CN108896541B (en) | 2020-09-22 |
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Inventor after: Liang Jiaxin Inventor after: Zhang Lianyu Inventor after: Lu Jia Inventor before: Liang Jiaxin Inventor before: Zhang Lianyu Inventor before: Lu Jia |
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