CN105403703A - Carbendazim detection ELISA (enzyme linked immunosorbent assay) kit and application thereof - Google Patents
Carbendazim detection ELISA (enzyme linked immunosorbent assay) kit and application thereof Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Abstract
The invention provides a carbendazim detection ELISA (enzyme linked immunosorbent assay) kit which includes an ELISA plate coated with a coating antigen, a carbendazim standard solution, a carbendazim antibody, an enzyme-labeled secondary antibody, a substrate coloring liquid, a terminating liquid, a washing liquid and a complex solution, the coating antigen is a carbendazim coupling antigen, and the enzyme-labeled secondary antibody is an enzyme labeled carbendazim antibody. The invention also discloses a carbendazim detection method using the ELISA kit, and the method includes first, pretreatment of a sample, then detection using the ELISA kit and final detection result analyzing. The ELISA kit can be used to detect the content of carbendazim in a tobacco leaf sample, and has the advantages of being simple in operation, low in cost, high in sensitivity, capable of monitoring on site and suitable for screening a large number of samples.
Description
Technical field
The present invention relates to enzyme linked immunosorbent detection technology, being specifically related to a kind of enzyme linked immunological kit for detecting carbendazim and application thereof, can the residual quantity of carbendazim medicine in qualitative and quantitative analysis tobacco leaf.
Background technology
China produces and uses the big country of chemical pesticide, and Long-Time Service agricultural chemicals has caused showing great attention to of people to the harm of ecologic environment and health and impact.Carbendazim [carbendazim, N-(2-benzopyrazoles base) methyl carbamate] be benzopyrazoles class, a kind of good wide spectrum, systemic fungicide, all effective to the most of pathogens in sac fungus, basidiomycetes and Fungi Imperfecti, be widely used in the disease control work of crops and Chinese herbal medicine.Carbendazim stable chemical nature, can be absorbed by the seed of plant, root and leaf, the longevity of residure is longer, all has certain toxicity to people, animal, can cause tic, absent-minded, n and V, the toxicity symptom such as uncomfortable in chest, dizzy.Therefore, about the analysis of Determination of carbendazim residue more and more comes into one's own.
Because carbendazim has a wide range of applications in the preventing and treating of corps diseases, but it has again certain toxic to human body, and current world many countries has all formulated the highest limit standard of carbendazim residual quantity in (class) not of the same race agricultural byproducts.In the vegetables such as Canada national Specification cucumber, cucurbita pepo, Determination of carbendazim residue per kilogram must not more than 0.5mg; In Malaysia regulation greengrocery, Determination of carbendazim residue per kilogram must not more than 1mg; In China hygienic standard GB14870294, in regulation veterinary antibiotics, Determination of carbendazim residue per kilogram must not more than 0.5mg.But there is no at present simply, detection method efficiently.
Summary of the invention
Object of the present invention provides a kind of enzyme linked immunological kit that can detect carbendazim drug residue in tobacco leaf for above-mentioned prior art situation just, and provide a kind of efficient, accurate, easy, be suitable for batch samples screening qualitative and quantitative analysis method, apply the residual quantity that euzymelinked immunosorbent assay (ELISA) of the present invention measures carbendazim medicine in tobacco leaf, there is the advantages such as detectability is low, high specificity, easy and simple to handle, detection speed is fast, testing cost is low, very easy popularization.
The object of the invention is to be achieved through the following technical solutions: kit of the present invention comprises: be coated with the ELISA Plate of coating antigen, carbendazim standard product solution, carbendazim antibody, ELIAS secondary antibody, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid, described coating antigen is carbendazim coupled antigen, described ELIAS secondary antibody is the carbendazim antiantibody of enzyme labeling, described carbendazim coupled antigen is obtained by carbendazim haptens and carrier protein couplet, and described carbendazim haptens is obtained by reacting by carbendazim and trifluoroacetic anhydride, ammonium nitrate.
Described carrier protein can be thyroprotein, bovine serum albumin(BSA), rabbit serum proteins, human albumin, ovalbumin, hemocyanin.
Described carbendazim hapten molecule structural formula is:
。
Described carbendazim antibody prepares using carbendazim coupled antigen as immunogene, and described carbendazim antibody is carbendazim monoclonal antibody or carbendazim polyclonal antibody, wherein preferred carbendazim monoclonal antibody.
The marker enzyme of described ELIAS secondary antibody is that horseradish peroxidase or bacterium extract alkaline phosphatase, wherein preferred horseradish peroxidase; ELIAS secondary antibody is obtained by enzyme and the coupling of carbendazim antiantibody.
In order to more convenient on-site supervision and great amount of samples examination, described kit also comprises carbendazim standard product solution, substrate nitrite ion, stop buffer, cleansing solution, redissolution liquid.
Described carbendazim standard product solution 6 bottles, concentration is respectively 0 μ g/L, 0.1 μ g/L, 0.3 μ g/L, 0.9 μ g/L, 2.7 μ g/L, 8.1 μ g/L.
When marker enzyme is horseradish peroxidase, described substrate nitrite ion is made up of substrate solution A liquid and substrate solution B liquid, A is hydrogen peroxide or urea peroxide, and B liquid is o-phenylenediamine or tetramethyl benzidine, and described stop buffer is sulfuric acid solution or the hydrochloride buffer of 1 ~ 2mol/L; When marker enzyme is bacterium extraction alkaline phosphatase, described substrate nitrite ion is to nitro phosphate buffer, and described stop buffer is 1 ~ 2mol/L sodium hydroxide solution.
It is 7.4 that described cleansing solution is preferably pH value, and the phosphate buffer containing 0.5% ~ 1.0% Tween-20,0.01 ‰ ~ 0.03 ‰ sodium azide preservatives, 0.1 ~ 0.3mol/L, number percent is wherein percent weight in volume, unit g/mL.
Described redissolution liquid is preferably that pH value is 7.0, the phosphate buffer of 0.02mol/L.
The carbonate buffer solution of wherein used in ELISA Plate preparation process bag is buffered liquid to be pH value be 9.6,0.05mol/L, confining liquid is pH value is 7.1 ~ 7.5, the phosphate buffer containing 1% ~ 3% (g/mL) casein, 0.1 ~ 0.3mol/L.
In the present invention, the preparation process of ELISA Plate is: be buffered liquid with bag and coating antigen is diluted to 20 μ g/mL, every hole adds 100 μ L, 25 DEG C of lucifuges are hatched 2h or 4 DEG C and are spent the night, and liquid in hole of inclining, washs 2 times with cleansing solution, each 30s, pat dry, in every hole, then add 150 ~ 200 μ L confining liquids, 25 DEG C of lucifuges hatch 1 ~ 2h, the liquid in hole that inclines pats dry, and preserves after dry with the vacuum seal of aluminium film.
Cleaning Principle of the present invention is:
This kit adopts direct competive ELISA method, pre-coated coupled antigen on ELISA Plate capillary strip, the ELIAS secondary antibody of coupled antigen competition resisting carbendazim pre-coated on carbendazim residual in sample and ELISA Plate capillary strip, develop the color with tmb substrate, sample absorbance becomes negative correlation with the content of residue carbendazim contained by it, compare with typical curve, then be multiplied by the extension rate of its correspondence, the residual quantity of carbendazim in sample can be drawn.
Present invention also offers a kind of method applying above-mentioned enzyme linked immunological kit detection carbendazim, it comprises step:
(1) sample pre-treatments;
(2) detect with kit;
(3) testing result is analyzed.
The enzyme linked immunological kit that the present invention detects carbendazim mainly adopts the content of carbendazim in the qualitative or quantitative detection sample of ELISA method; Require low to the pre-treatment of sample, sample pretreatment process is simple, can detect batch samples fast simultaneously; Main agents provides with the form of working fluid, and the method for inspection is convenient and easy, has that detectability is low, specificity is high, easy and simple to handle, highly sensitive, degree of accuracy is high, accuracy high.Enzyme linked immunological kit of the present invention, structure is simple, easy to use, low price, carrying convenience, detection method are efficient, accurate, easy, be suitable for the qualitative, quantitative of batch samples screening.
Accompanying drawing explanation
Fig. 1: carbendazim hapten synthesis route map,
Fig. 2: kit standard curve map.
Embodiment
The present invention is set forth further below in conjunction with specific embodiment.Should be understood that these embodiments are only for illustration of the present invention, and be not used for limiting the scope of the invention.
the preparation of embodiment 1 reagent constituents
1, the haptenic preparation of carbendazim
Carbendazim bulk drug, through nitration reaction, phenyl ring introduces nitro, obtains the haptens product with aromatic amine after reduction.
Trifluoroacetic acid 20mL adds trifluoroacetic anhydride 2mL, ice-water bath, is down to 0 DEG C, adds ammonium nitrate 0.5g, stirs 1h, adds the trifluoroacetic acid solution containing carbendazim 1.0g, continues to stir, reaction 2h.Stop reaction, neutralize neutrality, dichloromethane extraction with regard to diluted sodium hydroxide solution, washing, evaporate to dryness, washed with diethylether crystallization, obtains compound a 0.76g, yield 67%.1HNMR（CDCl3,300MHZ）δ:8.31(1H,dd,J=1.616,J=1.239),7.69(1H,dd,J=8.716,J=1.616),7.64(1H,dd,J=8.716,J=1.239),3.85(3H，s)。
Compound a 0.7g adds ethanol and dissolves, and adds 0.43g tin chloride aqueous solution 10mL, passes into nitrogen, add back flow reaction 3h.Stop reaction, revolve and steam removing ethanol, add extraction into ethyl acetate, concentrated upper silicagel column, petrol ether/ethyl acetate (1:1, v/v) wash-out is separated, and obtains hapten compound b product 0.54g, yield, 83%.1HNMR（CDCl3,300MHZ）δ：3.79(3H，s),6.27(2H,s),6.90(1H,dd,J=2.225,J=1.850),6.46(1H,dd,J=8.422,J=2.225),7.34(1H,dd,J=8.422,J=1.850),5.00(1H,s),9.15(1H,s)。
In collection of illustrative plates, chemical shift δ=6.27 is the resonance absorbing peak of aromatic amine on phenyl ring, and the existence of this absorption peak proves, hapten synthesis success.
2, the preparation of antigen
Prepared by immunogene---and carbendazim haptens and bovine serum albumin(BSA) (BSA) coupling obtain immunogene.
Take BSA50mg, make it the phosphate buffer PBS(PH7.2 being fully dissolved in 3.8mL0.1M) in, obtain solution A; Get 30mg carbodiimides (EDC) and N-hydroxy-succinamide (NHS) 0.2mL water fully dissolve after in adding in solution A, stirred at ambient temperature 30min.Get 15mg haptens, be dissolved in 1mLN, in dinethylformamide (DMF), then slowly join in protein dissolution, stirring at room temperature reaction 24h.3 dislysates are changed every day with 0.01mol/LPBS4 DEG C of dialysis 3d.Packing, saves backup in-20 DEG C.
Carbendazim haptens and hemocyanin coupling obtain immunogene.
Take hemocyanin 50mg, make it the phosphate buffer PBS(PH7.2 being fully dissolved in 3.8mL0.1M) in, obtain solution A; Get 30mg carbodiimides (EDC) and N-hydroxy-succinamide (NHS) 0.2mL water fully dissolve after in adding in solution A, stirred at ambient temperature 30min.Get 5mg haptens, be dissolved in 1mLN, in dinethylformamide (DMF), then slowly join in protein dissolution, stirring at room temperature reaction 24h.3 dislysates are changed every day with 0.01mol/LPBS4 DEG C of dialysis 3d.Packing, saves backup in-20 DEG C.
Carbendazim haptens and thyroprotein coupling obtain immunogene.
Take thyroprotein 50mg, make it the phosphate buffer PBS(PH7.2 being fully dissolved in 3.8mL0.1M) in, obtain solution A; Get 30mg carbodiimides (EDC) and N-hydroxy-succinamide (NHS) 0.2mL water fully dissolve after in adding in solution A, stirred at ambient temperature 30min.Get 3mg haptens, be dissolved in 1mLN, in dinethylformamide (DMF), then slowly join in protein dissolution, stirring at room temperature reaction 24h.3 dislysates are changed every day with 0.01mol/LPBS4 DEG C of dialysis 3d.Packing, saves backup in-20 DEG C.
Prepared by coating antigen---and carbendazim haptens and ovalbumin (OVA) coupling obtain coating antigen.
Take OVA50mg, make it fully to be dissolved in 3.8mL0.1MPBS(PH7.2) in, obtain solution B; Get 30mgEDC and NHS 0.2mL water fully dissolve after in adding in solution B, stirred at ambient temperature 30min.Get 13mg haptens, be dissolved in 1mLDMF, then slowly join in protein dissolution, stirring at room temperature reaction 24h.3 dislysates are changed every day with 0.01mol/LPBS4 DEG C of dialysis 3d.Packing, saves backup in-20 DEG C.
Carbendazim haptens and human serum albumins coupling obtain coating antigen.
Take human serum albumins 50mg, make it fully to be dissolved in 3.8mL0.1MPBS(PH7.2) in, obtain solution B; Get 30mgEDC and NHS 0.2mL water fully dissolve after in adding in solution B, stirred at ambient temperature 30min.Get 15mg haptens, be dissolved in 1mLDMF, then slowly join in protein dissolution, stirring at room temperature reaction 24h.3 dislysates are changed every day with 0.01mol/LPBS4 DEG C of dialysis 3d.Packing, saves backup in-20 DEG C.
Carbendazim haptens and albumin rabbit serum coupling obtain coating antigen.
Take albumin rabbit serum 50mg, make it fully to be dissolved in 3.8mL0.1MPBS(PH7.2) in, obtain solution B; Get 30mgEDC and NHS 0.2mL water fully dissolve after in adding in solution B, stirred at ambient temperature 30min.Get 15mg haptens, be dissolved in 1mLDMF, then slowly join in protein dissolution, stirring at room temperature reaction 24h.3 dislysates are changed every day with 0.01mol/LPBS4 DEG C of dialysis 3d.Packing, saves backup in-20 DEG C.
3, the preparation of carbendazim monoclonal antibody
Animal immune: immunogene above-mentioned steps obtained is injected in Balb/c Mice Body, immunizing dose is 150 μ g/, makes it produce antiserum.
Fusion of Cells and cloning: after mice serum measurement result is higher, get its splenocyte, in 8:1(quantitative proportion) ratio and SP2/0 myeloma cell fusion, adopt indirect competitive ELISA to measure cell conditioned medium liquid, screen positive hole.Limiting dilution assay is utilized to carry out cloning to positive hole, until obtain the hybridoma cell strain secreting carbendazim monoclonal antibody.
Cell cryopreservation and recovery: monoclonal hybridoma strain cryopreserving liquid is made 1 × 10
6the cell suspension of individual/mL, preserves for a long time in liquid nitrogen.Take out cryopreservation tube during recovery, put into 37 DEG C of water-bath middling speeds immediately and melt, after centrifugal segregation cryopreserving liquid, move into and cultivate culture in glassware.
The production of monoclonal antibody and purifying: only Balb/c mouse peritoneal is injected sterilizing paraffin oil 0.5mL/, within 7 days, pneumoretroperitoneum injects stable monoclonal hybridoma strain 5 × 10
5individual/only, gather ascites after 7 days.Carry out ascites by sad-saturated ammonium sulfate method to purify ,-20 DEG C of preservations.
4, the preparation of ELIAS secondary antibody
Using goat as immune animal, with carbendazim monoclonal antibody for immunogene carries out immunity to pathogen-free domestic goat, obtain carbendazim antiantibody.Carbendazim antiantibody and horseradish peroxidase (HRP) are carried out coupling and obtains ELIAS secondary antibody.
5, the preparation of ELISA Plate
Be buffered liquid with bag and coating antigen is diluted to 20 μ g/mL, every hole adds 100 μ L, 25 DEG C of lucifuges hatch 2h, and liquid in hole of inclining washs 2 times with cleansing solution, each 30s, pat dry, in every hole, then add 200 μ L confining liquids, 25 DEG C of lucifuges hatch 2h, the liquid in hole that inclines pats dry, and preserves after dry with the vacuum seal of aluminium film.
embodiment 2 detects the establishment of the enzyme linked immunological kit of carbendazim
Set up the enzyme linked immunological kit detecting carbendazim, make it comprise following component:
(1) bag is by the ELISA Plate of carbendazim coupled antigen;
(2) carbendazim standard product solution 6 bottles, concentration is respectively 0 μ g/L, 0.1 μ g/L, 0.3 μ g/L, 0.9 μ g/L, 2.7 μ g/L, 8.1 μ g/L;
(3) with the carbendazim antiantibody of horseradish peroxidase-labeled;
(4) substrate nitrite ion is made up of A liquid and B liquid, and A liquid is urea peroxide, and B liquid is tetramethyl benzidine;
(5) stop buffer is 2mol/L sulfuric acid;
(6) cleansing solution is pH value is 7.4, the phosphate buffer containing 0.5% ~ 1.0% Tween-20,0.01 ‰ ~ 0.03 ‰ sodium azide preservatives, 0.1 ~ 0.3mol/L;
(7) redissolve liquid to be pH value be 7.0, the phosphate buffer of 0.02mol/L.
the detection of carbendazim in embodiment 3 tobacco leaf
1, sample pre-treatments
Take 1.0 ± 0.05g tobacco leaf sample in 50ml polystyrene centrifuge tube, add 10mL tobacco leaf extract, with refiner, it is fully smashed; The sample filter membrane smashed is filtered; Pipette filtrate 50mL and add 700mL deionized water, fully mix; Pipette above liquid 50mL again and add 950mL redissolution working fluid, fully mix; Get 50mL for analyzing.
2, detect with kit
Add standard solution/sample 50mL in the micropore of correspondence, then add antibody working fluid 50mL/ hole, mixing of vibrating gently, react 30min with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate.Carefully open cover plate film, dried by liquid in hole, with wash operating solution 250mL/ hole, fully washing 4-5 time, every minor tick 10s, pats dry (bubble be not eliminated after patting dry can be poked gently with original rifle head) with thieving paper.Add ELIAS secondary antibody 100mL/ hole, mixing of vibrating gently, react 30min with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate, take out and repeat to wash plate step.Add substrate solution A liquid 50 μ L/ hole, then add substrate solution B liquid 50 μ L/ hole, mixing of vibrating gently, with the 15min that develops the color in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate.Add stop buffer 50 μ L/ hole, mixing of vibrating gently, setting microplate reader determined wavelength 450nm, reference wavelength 620nm, please runs through data in 5min), measure every hole OD value.
3, Analysis of test results
The percentage absorptance of standard items or sample equals the mean value of mean value (diplopore) divided by the absorbance of first standard items (0 standard) of the absorbance of standard items or sample, then is multiplied by 100%, obtains the percentage absorbance of standard items or sample.With standard items percentage absorptance for ordinate, with the logarithm of carbendazim standard product concentration (μ g/L) for horizontal ordinate, drawing standard curve map.The percentage absorptance of sample substituted in typical curve, read the concentration corresponding to sample from typical curve, the extension rate being multiplied by its correspondence is the actual concentrations of carbendazim in sample.
the determination test of embodiment 4 carbendazim technical parameter
1, kit sensitivity and detectability
Conventionally measure kit sensitivity, the scope of typical curve is 0.1 ~ 8.1 μ g/L, IC
50(50% inhibition concentration) domain of walker is 0.45 ~ 0.65 μ g/L; 20 increment product are detected, the concentration corresponding to each percentage absorbance is found from typical curve, add that 3 times of standard deviations represent detectability with the mean value of 20 parts of concentration of specimens, result shows, and the method is 300 μ g/kg to the detectability of carbendazim in tobacco leaf.
2, sample preci-sion and accuracy test
Using the recovery as accuracy estimating index, the testing result relative standard deviation (RSD%) of a certain concentration samples of replication is as precision evaluation index.Computing formula is: the recovery (%)=practical measurement value/theoretical value × 100%, and wherein theoretical value is the interpolation concentration of sample; Relative standard deviation RSD%=SD/X × 100%, wherein SD is standard deviation, and X is the mean value of determination data.
By 300 μ g/kg, 600 μ g/kg, two concentration carbendazim respectively to tobacco sample carry out interpolation reclaim measure, each sample do 4 parallel, measure with three batches of different reagent, average recovery rate and the precision of calculation sample the results are shown in following table.
Table 1 tobacco leaf sample precision and accuracy test
Add tobacco leaf respectively with the carbendazim of 300 μ g/kg, 600 μ g/kg, two concentration, average recovery rate is between 80.3% ~ 86.7%; Batch in, batch between relative standard deviation be all less than 10%.
3, stabilization of kit test
Kit preservation condition is 2 ~ 8 DEG C, and through the mensuration of 12 months, the maximum absorbance value (zero standard) of kit, 50% inhibition concentration, carbendazim added practical measurement value all within normal range.Consider in transport and use procedure, have improper preservation condition and occur, placed 7 days under 37 DEG C of preservation conditions by kit, carry out accelerated aging tests, result shows that this kit indices meets the requirements completely.Consider that the freezing situation of kit occurs, kit is put into-20 DEG C of refrigerator freezings 7 days, measurement result also shows that kit indices is completely normal.Can show that kit at least can be preserved more than 12 months at 2 ~ 8 DEG C from above result.
Claims (8)
1. one kind is detected the enzyme linked immunological kit of carbendazim, it is characterized in that: comprise the ELISA Plate, carbendazim standard product solution, carbendazim antibody, ELIAS secondary antibody, substrate nitrite ion, stop buffer, cleansing solution, the redissolution liquid that are coated with coating antigen, described coating antigen is carbendazim coupled antigen, described ELIAS secondary antibody is the carbendazim antiantibody of enzyme labeling, described carbendazim coupled antigen is obtained by carbendazim haptens and carrier protein couplet, and described carbendazim haptens is obtained by reacting by carbendazim and trifluoroacetic anhydride, ammonium nitrate.
2. kit as claimed in claim 1, is characterized in that: described carrier protein is thyroprotein, bovine serum albumin(BSA), rabbit serum proteins, human albumin, ovalbumin, hemocyanin.
3. kit as claimed in claim 1, is characterized in that: described carbendazim hapten molecule structural formula is:
。
4. kit as claimed in claim 1, it is characterized in that: described carbendazim antibody prepares using carbendazim coupled antigen as immunogene, described carbendazim antibody is carbendazim monoclonal antibody or carbendazim polyclonal antibody.
5. kit as claimed in claim 1, it is characterized in that: the marker enzyme of described ELIAS secondary antibody is that horseradish peroxidase or bacterium extract alkaline phosphatase, when marker enzyme is horseradish peroxidase, substrate nitrite ion A liquid is hydrogen peroxide or urea peroxide, substrate nitrite ion B liquid is o-phenylenediamine or tetramethyl benzidine, and stop buffer is sulfuric acid or the hydrochloride buffer of 1 ~ 2mol/L; When marker enzyme is bacterium extraction alkaline phosphatase, substrate nitrite ion is to nitro phosphate buffer, and stop buffer is 1 ~ 2mol/L NaOH.
6. kit as claimed in claim 1, is characterized in that: described cleansing solution is pH value is 7.4, the phosphate buffer containing 0.5% ~ 1.0% Tween-20,0.01 ‰ ~ 0.03 ‰ sodium azide preservatives, 0.1 ~ 0.3mol/L; To be pH value be described redissolution liquid 7.0, the phosphate buffer of 0.02mol/L, and the number percent in cleansing solution is percent weight in volume, unit g/mL.
7. kit as claimed in claim 1, is characterized in that: the concentration of described carbendazim standard product solution is respectively 0 μ g/L, 0.1 μ g/L, 0.3 μ g/L, 0.9 μ g/L, 2.7 μ g/L, 8.1 μ g/L.
8. utilize kit described in claim 1 to detect a method for carbendazim content in tobacco sample, it is characterized in that: comprise the following steps:
(1) sample pre-treatments;
(2) detect with kit;
(3) testing result is analyzed.
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Cited By (4)
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| CN106124766A (en) * | 2016-07-05 | 2016-11-16 | 天津师范大学 | Use the method for carbendazim content in carbendazim detection of specific antibody edible fungi |
| CN106932566A (en) * | 2017-04-17 | 2017-07-07 | 四川精卫食品检测科技有限公司 | A kind of carbendazim and the two-in-one detection enzyme linked immunological kit of thiophanate-methyl |
| CN109265404A (en) * | 2018-09-21 | 2019-01-25 | 中国烟草总公司郑州烟草研究院 | A kind of preparation method and application of carbendazim haptens and antigen |
| CN112461808A (en) * | 2019-09-06 | 2021-03-09 | 苏州市农产品质量安全监测中心 | Detection method and kit for detecting carbendazim in agricultural products |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106124766A (en) * | 2016-07-05 | 2016-11-16 | 天津师范大学 | Use the method for carbendazim content in carbendazim detection of specific antibody edible fungi |
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| CN109265404A (en) * | 2018-09-21 | 2019-01-25 | 中国烟草总公司郑州烟草研究院 | A kind of preparation method and application of carbendazim haptens and antigen |
| CN112461808A (en) * | 2019-09-06 | 2021-03-09 | 苏州市农产品质量安全监测中心 | Detection method and kit for detecting carbendazim in agricultural products |
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| CN105403703B (en) | 2017-06-30 |
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