CN105572310A - Method for rapidly detecting content of clofentezine in fruit - Google Patents
Method for rapidly detecting content of clofentezine in fruit Download PDFInfo
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- CN105572310A CN105572310A CN201410534179.9A CN201410534179A CN105572310A CN 105572310 A CN105572310 A CN 105572310A CN 201410534179 A CN201410534179 A CN 201410534179A CN 105572310 A CN105572310 A CN 105572310A
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- clofentezine
- detection
- colloidal gold
- protein
- colloidal
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Abstract
The invention relates to a clofentezine colloidal gold detection card. The clofentezine colloidal gold detection card is mainly used for rapidly detecting the content of clofentezine in a food. The backing of a test paper is sequentially pasted with a liquid sample absorption part, a colloidal gold labeling part, a detection reaction part and a water absorption part. The detection reaction part is coated with a detection antigen 1 strip used as a detection line, and is also coated with an anti-second species animal protein IgG1 strip used as a reference line. The above rapid detection test strip has strong specificity, can be used to realize semi-quantitative detection, can be used at ambient temperature of 4-35DEG C, and is suitable for rapid detection of clofentezine residual of plant-derived foods of individual farms, food health quality inspection departments and customs. The clofentezine colloidal gold detection card has the advantages of strong specificity, high sensitivity, realization of semi-quantitative detection, and simple and convenient operation.
Description
Technical field
Belong to field of detection of food safety, be specifically related to the detection method of the Harmful Residue in food, particularly clofentezine test paper detecting method.
Background technology
Clofentezine (English name: clofentezine) has another name called 3,6-two (2-chlorphenyl)-1,2,4,5-tetrazine, molecular formula C
14h
8cl
2n, belongs to low toxic pesticide.Tetrazine class acaricide, embryonic development inhibitor, mainly kills mite ovum, but also has certain effect to young mite, invalid to one-tenth mite.Drug effect plays comparatively slow, and lasting period 50 ~ 60d, can reach the highest miticidal effect in after dispenser 2 ~ 3 weeks.For crops such as fruit tree, pea, oranges and tangerines, ornamental plant, cottons, control Panonychus citri belongs to and Tetranychus evil mite, effective especially to the winter egg of elm Panonychus citri (European red mite).
In maximum residue limits for pesticide table in the more established agricultural byproducts of China, the limitation of clofentezine is pomaceous fruits fruit, must not more than 0.5mg/kg in citrus fruit.
Summary of the invention
The object of the invention is to provide and detects the residual of clofentezine in food fast.
Technical scheme of the present invention is the test card of clofentezine and the disposal route of detection sample thereof.The test card of clofentezine arranges detector bar in the test card shell of rectangular flat shell shape, test card case surface has detection fenestra and well, it is characterized in that test-strips forms by four kinds of original papers pasted by supporting backboard: in the middle part of supporting backboard, paste nitrocellulose filter, thieving paper is pasted in backboard one end in supporting, the other end pastes sample pad, thieving paper is inner to be overlapped with nitrocellulose filter, then there is one section of gold mark pad between sample pad and nitrocellulose filter, by gold mark pad, sample pad and nitrocellulose filter are linked together.It is the potpourri containing the second kind albumen and clofentezine antibody in gold mark pad, nitrocellulose filter laterally there are 2 isolated colour developing marking bands, article one, be the detection zone containing clofentezine protein conjugate, another be the IgG(of the second kind animal protein conventional be anti-rabbit antibody or anti-mouse antibody); Test-strips is inserted in test card shell, and sample pad is just to well, and nitrocellulose filter is just to detection fenestra.
Sample treatment is: take sample 2 ± 0.05g in the centrifuge tube of 50mL, add 10mL ultrapure water, smash (about 1min) with high speed tissue pulverizer under 1800r/min condition, get viscous fluid and add 50% trichloroacetic acid 2mL and methylene chloride 10mL, high speed jolting 30min on Clothoid type oscillator.Then centrifugal 30min under 4 DEG C of 5000rpm conditions, gets supernatant 10mL, and add methylene chloride 2mL, on cyclotron vibration mixing 20min, then under 4 DEG C of 12000rpm conditions centrifugal 30min, supernatant is the extract of medicine.
Advantage of the present invention can detect the clofentezine contained in sample fast, and save testing cost, easy to use, and detect fast, highly sensitive, result is accurate.
Accompanying drawing explanation
Fig. 1 is clofentezine test card shell and test strip arrangement figure in the enclosure.
Fig. 2 is clofentezine test card test strip structural representation.
Fig. 3 test strip nitrocellulose filter shows trace band schematic diagram.
Embodiment
1. mark the preparation of collaurum
The preparation of colloidal gold solution: first by 1g gold chloride (HAuCl
4) powder is dissolved in the chlorauric acid solution that 100mL ultrapure water is configured to 1%; Get 1L ultrapure water again to heat on speciality electric furnace, the trisodium citrate aqueous solution of 10mL1% is added to boiling, heating is continued after boiling, the chlorauric acid solution of 10mL1% is added after 5 minutes, again be heated to boiling, wait for that color starts timing after becoming claret, after 10 minutes, stop heating, take off and naturally to cool afterwards, be settled to 1L, be colloidal gold solution.
Antibody labeling: determine the 0.1mol/lK needed for antibody
2cO
3ratio, get 100mL colloidal gold solution, add the K of this ratio 0.1mol/L
2cO
3mixing (about 5min), again clofentezine and the second kind animal protein are used PBS(0.01mo1/L respectively, pH7.4) dissolved dilution is to 2mg/mL, add the clofentezine antibody (or 2mL2mg/mL second kind animal protein) of 2mL2mg/mL, mixing (5min), finally adds 10%BSA2mL, mixing (5min).Stay precipitation centrifugal three times, the precipitation of collecting for these three times is gold mark clofentezine antibody.
Colloid gold label part process: after clofentezine antibody dilution, soaks into carrier (as glass fibre, polyester film etc.) by a certain percentage, and evenly, taking-up is put in 37 DEG C of constant temperature ovens and smokes 8-12h, namely as colloid gold label part after drying paving.
2. the special polymer compound film preparation of immunochromatography
First make transparent backing film with a kind of nitrocellulose filter, backing film covers one deck strong to protein bound power, the chromatographic film of certain pore size that what wetting property was good have.The effect of backing film stops organic solvent in bonding agent to the destruction of protein in chromatographic film.
3. the spotting methods of chromatographic film
By certain density happy antigen (conjugates that clofentezine and carrier mass are formed) accurate quantification; Schedule in chromatographic film in horizontal stripe shape point, for detection with a determining deviation (5mm).
Clofentezine test card arranges test strip 2 in test card shell 1, structure and the arranging of test strip 2 of test card shell 1 see accompanying drawing 1, test card shell 1 is rectangular flat shell shape shell, long 70mm, wide 20mm, thick 5mm, shell wall thickness 1mm, be made up of engineering plastics, test card shell 1 is formed by upper cover and lower cover two semi join, test card shell 1 has covered and has detected fenestra 3 and well 4.Test strip 2 is placed in the lower cover of test card shell 1.
Test strip 2 is fillet thin slices of sandwich construction, length of a film 60mm, and the wide about 4mm of sheet, thickness is less than 2.5mm.Test strip 2 is sandwich constructions: bottom is supporting backboard 5, and be polyethylene sheets, thickness is about 0.5mm, long 60mm, wide 4mm; Nitrocellulose filter 6 is pasted, cellulose nitrate thickness 0.5mm, long 20mm, wide 4mm in the middle part of supporting backboard 5.Nitrocellulose filter 6 has two isolated laterally display trace bands, see accompanying drawing 3, in two display traces, one is detection line 10, and containing clofentezine conjugate, another is line of reference 11, containing the IgG of the second kind animal protein.Thieving paper 9 is pasted in backboard 5 one end in supporting, the other end pastes sample pad 8, and thieving paper 9 is inner to be overlapped with nitrocellulose filter 6, then has one section of gold to mark pad 7 between sample pad 8 and nitrocellulose filter 6, sample pad 8 and nitrocellulose filter 6 are linked together by gold mark pad 7, lap width is at 1-2mm.Sample pad 8 material is water adsorption glass fiber or polyester film.
When using clofentezine colloidal-gold detecting-card, first by sample liquid to be detected (after the extract dilution of medicine, be directly used in detection) from well 4, instill the sample pad 8 of clofentezine test strip 2, due to rainbow action principle, the anti-clofentezine monoclonal antibody colloid gold label thing contained by sample liquid to be detected and colloidal gold film 7 is driven to spread to nitrocellulose filter 6 together, observations in 5-10 minute.
The key reaction of test card is immunologic antigen and antibody response, the antibody of the colloid gold label that nitrocellulose filter moves, on p-wire with containing clofentezine protein conjugate, and line of reference reacts containing the IgG of the second kind animal protein, formation brownish red band.If corresponding clofentezine to be measured is higher than permissible value in sample, sample adds rear elder generation and marks the antibody response in padding with gold, and can not with detection zone with clofentezine protein conjugate react, thus not develop the color.Mainly contain following 3 kinds of testing results:
1. detection line 10 and line of reference 11 manifest brownish red trace simultaneously, represent that testing result be negative, illustrate in sample and do not contain clofentezine or content lower than permissible value;
2. detection line 10 does not develop the color, and line of reference 11 manifests brownish red trace, represents that testing result is positive, illustrates that in sample, clofentezine content is higher than permissible value;
3. detection line 10 manifests brownish red trace, and line of reference 11 does not develop the color, or detection line 10 and line of reference 11 all do not develop the color, and represents that test strip lost efficacy.
Claims (4)
1. clofentezine colloidal-gold detecting-card, it is characterized in that: on the backing of test paper, post sample liquid absorption portion, colloid gold label part, detection reaction part and water absorbent portion successively, the material that colloid gold label part is labeled is the potpourri of the second kind animal protein and clofentezine antibody; Detection reaction part is coated with detection clofentezine antigen 1 bar as detection line, is also coated with the IgG1 bar of anti-second kind animal protein as line of reference simultaneously.
2. clofentezine colloidal-gold detecting-card according to claim 1, is characterized in that: clofentezine detection antigen is the conjugates that clofentezine and carrier mass are formed.
3. clofentezine colloidal-gold detecting-card according to claim 2, is characterized in that: described carrier mass, is protein, protein fragments, improvement on synthesis, semi-synthetic polypeptide or polysaccharide.
4. clofentezine colloidal-gold detecting-card according to claim 1, is characterized in that: the second described kind animal protein is the protein of non-antibody source animal.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410534179.9A CN105572310A (en) | 2014-10-11 | 2014-10-11 | Method for rapidly detecting content of clofentezine in fruit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410534179.9A CN105572310A (en) | 2014-10-11 | 2014-10-11 | Method for rapidly detecting content of clofentezine in fruit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105572310A true CN105572310A (en) | 2016-05-11 |
Family
ID=55882677
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410534179.9A Withdrawn CN105572310A (en) | 2014-10-11 | 2014-10-11 | Method for rapidly detecting content of clofentezine in fruit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105572310A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103808939A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Spectinomycin colloidal gold detection card |
| CN103808932A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Maduramicin colloidal gold detection card |
-
2014
- 2014-10-11 CN CN201410534179.9A patent/CN105572310A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103808939A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Spectinomycin colloidal gold detection card |
| CN103808932A (en) * | 2012-11-06 | 2014-05-21 | 江苏维赛科技生物发展有限公司 | Maduramicin colloidal gold detection card |
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|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20160511 |