CN105572357A - Parathion-methyl colloidal gold detection card - Google Patents
Parathion-methyl colloidal gold detection card Download PDFInfo
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- CN105572357A CN105572357A CN201410519593.2A CN201410519593A CN105572357A CN 105572357 A CN105572357 A CN 105572357A CN 201410519593 A CN201410519593 A CN 201410519593A CN 105572357 A CN105572357 A CN 105572357A
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- parathion
- methyl
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- colloidal gold
- protein
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Abstract
The present invention relates to a parathion-methyl colloidal gold detection card, which is mainly used for rapidly detecting the parathion-methyl content in food. According to the method, a sample liquid absorbing part, a colloidal gold labeling part, a detection reaction part and a water absorbing part are sequentially adhered on the back lining of test paper, and 1 strip of detecting antigen is coated on the detection reaction part while 1 strip of anti-second genus animal protein IgG is coated as a reference line. According to the present invention, the rapid detection test paper strip has strong specificity, can perform semi-quantitative detection, can be used at an ambient temperature of 4-35 DEG C, and is suitable for rapid detection of the parathion-methyl residue in plant-derived foods in individual farmers, food hygiene and quality inspection departments, customs and other departments; and the parathion-methyl colloidal gold detection card has beneficial effects of strong specificity, high sensitivity, semi-quantitative detection achieving, simple and convenient operation, and the like.
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 parathion-methyl test paper detecting method.
Background technology
Parathion-methyl (English name: parathion-methyl) is commonly called as parathion-methyl, formal name used at school O, O-dimethyl-O-(4-nitrobenzophenone) thiophosphate, a kind of organophosphorus insecticide.Industrial products are the yellowish-brown oily liquids of band garlic odour, and sterling is white crystals, and fusing point 36 ~ 36.5 DEG C, is insoluble in water, is soluble in organic solvent, heating meeting isomerization, and high temperature or chance alkali easily decompose.Parathion-methyl causes poisoning by esophagus, respiratory tract and skin.Belong to high toxic pesticide.
Parathion-methyl tool is tagged and stomach poison function, can suppress the vigor of cholinesterase in insect nervous system and lethal, and insecticidal spectrum is wide, often be processed into missible oil or pulvis use, main application prevents and treats various agricultural insect, because toxicity is high, strictly by regulation dispenser, and safety prevention measure to be strengthened.Contact (oral, suctions, skin, mucous membrane) in a short time to contact in a large number and cause acute poisoning.Performance has headache, giddy, anorexia, Nausea and vomiting, stomachache, diarrhoea, salivation, myosis, respiratory secretions increase, hidrosis, fasciculation etc.There is pulmonary edema, encephaledema, stupor, respiratory paralysis in severe one.Some cases can intentionally, liver, renal damage.There is peripheral nerve disease in minority several cases several weeks or number friend after consciousness recovery.Indivedual several cases can Late-onset sudden death.
In maximum residue limits for pesticide table in the more established agricultural byproducts of China, the limitation of parathion-methyl is 0.1mg/kg.
Summary of the invention
The object of the invention is to provide and detects the residual of parathion-methyl in food fast.
Technical scheme of the present invention is the test card of parathion-methyl and the disposal route of detection sample thereof.The test card of parathion-methyl 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 parathion-methyl antibody in gold mark pad, nitrocellulose filter laterally there are 2 isolated colour developing marking bands, article one, be the detection zone containing parathion-methyl 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 parathion-methyl 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 parathion-methyl test card shell and test strip arrangement figure in the enclosure.
Fig. 2 is parathion-methyl 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 parathion-methyl and the second kind animal protein are used PBS(0.01mo1/L respectively, pH7.4) dissolved dilution is to 2mg/mL, add the parathion-methyl 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 parathion-methyl antibody.
Colloid gold label part process: after parathion-methyl 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 parathion-methyl and carrier mass are formed) accurate quantification; Schedule in chromatographic film in horizontal stripe shape point, for detection with a determining deviation (5mm).
Parathion-methyl 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 parathion-methyl 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 parathion-methyl 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 parathion-methyl test strip 2, due to rainbow action principle, the anti-parathion-methyl 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 parathion-methyl protein conjugate, and line of reference reacts containing the IgG of the second kind animal protein, formation brownish red band.If corresponding parathion-methyl 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 parathion-methyl 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 parathion-methyl 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, parathion-methyl 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. parathion-methyl 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 parathion-methyl antibody; Detection reaction part is coated with detection parathion-methyl 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. parathion-methyl colloidal-gold detecting-card according to claim 1, is characterized in that: parathion-methyl detection antigen is the conjugates that parathion-methyl and carrier mass are formed.
3. parathion-methyl 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. parathion-methyl 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 |
|---|---|---|---|
| CN201410519593.2A CN105572357A (en) | 2014-10-07 | 2014-10-07 | Parathion-methyl colloidal gold detection card |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410519593.2A CN105572357A (en) | 2014-10-07 | 2014-10-07 | Parathion-methyl colloidal gold detection card |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105572357A true CN105572357A (en) | 2016-05-11 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410519593.2A Pending CN105572357A (en) | 2014-10-07 | 2014-10-07 | Parathion-methyl colloidal gold detection card |
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| CN (1) | CN105572357A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2869868Y (en) * | 2005-12-06 | 2007-02-14 | 万积成 | Colloid gold test-paper for quick detecting parathion, methyl parathion residue |
| CN202583192U (en) * | 2012-03-03 | 2012-12-05 | 北京勤邦生物技术有限公司 | Parathion-methyl ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit |
| CN103698510A (en) * | 2012-12-21 | 2014-04-02 | 欧普图斯(苏州)光学纳米科技有限公司 | Analyzing chemical and biological substances using nano-structure based spectral sensing |
| 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 |
-
2014
- 2014-10-07 CN CN201410519593.2A patent/CN105572357A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2869868Y (en) * | 2005-12-06 | 2007-02-14 | 万积成 | Colloid gold test-paper for quick detecting parathion, methyl parathion residue |
| CN202583192U (en) * | 2012-03-03 | 2012-12-05 | 北京勤邦生物技术有限公司 | Parathion-methyl ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit |
| 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 |
| CN103698510A (en) * | 2012-12-21 | 2014-04-02 | 欧普图斯(苏州)光学纳米科技有限公司 | Analyzing chemical and biological substances using nano-structure based spectral sensing |
Non-Patent Citations (1)
| Title |
|---|
| 刑丽杰等: "免疫金法快速检测食品中的甲基对硫磷", 《石河子大学学报（自然科学版）》 * |
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Application publication date: 20160511 |
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