CN105486868A - Acephate colloidal gold detection card - Google Patents
Acephate colloidal gold detection card Download PDFInfo
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- CN105486868A CN105486868A CN201410522807.1A CN201410522807A CN105486868A CN 105486868 A CN105486868 A CN 105486868A CN 201410522807 A CN201410522807 A CN 201410522807A CN 105486868 A CN105486868 A CN 105486868A
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- detection
- orthene
- acephate
- colloidal gold
- bar
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Abstract
The invention relates to an acephate colloidal gold detection card. The acephate colloidal gold detection card is mainly used for rapidly detecting acephate 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 orderly stuck on a backing of test paper. The detection reaction part is coated by an antigen 1 bar which is applied to detection and is used as a detection line, and at the same time is coated by an IgG1 bar which resists the second species of animal protein and is used as a reference line. The rapid detection test paper bar has high specificity, can carry out semi quantitative detection, can be used at the environmental temperature of 4 to 35 DEG C, and is suitable for an individual farmer, a food hygiene and quality inspection department, customs and other animal source food to carry out rapid detection on acephate residues. The acephate colloidal gold detection card provided by the invention has the advantages of high specificity, high sensitivity, simple and convenient operation and the like, and can carry out semi quantitative detection.
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 orthene test paper detecting method.
Background technology
Orthene (English name: Acephate) has another name called accephate, belongs to low toxic pesticide.Orthene is systemic insecticide, there is stomach toxicity and action of contace poison, and can ovicidal, there is certain fumigation action, it is slow-acting type pesticide, be applicable to the crops such as vegetables, tea tree, tobacco, fruit tree, cotton, paddy rice, wheat, rape, prevent and treat multiple chewing type, sucking pest and evil mite and sanitary insect pest.Keeping and improper use can cause person poultry poisoning.
Orthene belongs to organophosphorus compounds agricultural chemicals.Such agricultural chemicals suppresses cholinesterase in body, causes nervous physiology dysfunction.Organophosphorus pesticide acute poisoning system wrongly takes and causes.Poisoning manifestations has headache, giddy, anorexia, Nausea and vomiting, stomachache, diarrhoea, salivation, myosis, respiratory secretions increase, hidrosis, fasciculation etc.There is pulmonary edema, stupor, respiratory paralysis, encephaledema in severe, minority severe intoxication person occurs neuropathy several weeks after clinical symptom disappearance.Contact organophosphorus pesticide workman can have dizziness, headache, unable, insomnia, hidrosis, numb limb, muscle beat etc.Blood cholinesterase activity reduces.
In maximum residue limits for pesticide table in the more established agricultural byproducts of China, the limitation of orthene is cereal (in raw grain), must not more than 0.2mg/kg in vegetables; Must not more than 0.5mg/kg in fruit.
Summary of the invention
The object of the invention is to provide and detects the residual of orthene in food fast.
Technical scheme of the present invention is the test card of orthene and the disposal route of detection sample thereof.The test card of orthene 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 orthene antibody in gold mark pad, nitrocellulose filter laterally there are 2 isolated colour developing marking bands, article one, be the detection zone containing orthene 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 orthene 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 orthene test card shell and test strip arrangement figure in the enclosure.
Fig. 2 is orthene 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 orthene and the second kind animal protein are used PBS(0.01mo1/L respectively, pH7.4) dissolved dilution is to 2mg/mL, add the orthene 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 orthene antibody.
Colloid gold label part process: after orthene 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 orthene and carrier mass are formed) accurate quantification; Schedule in chromatographic film in horizontal stripe shape point, for detection with a determining deviation (5mm).
Orthene 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 orthene 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 orthene 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 orthene test strip 2, due to rainbow action principle, the anti-orthene 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 orthene protein conjugate, and line of reference reacts containing the IgG of the second kind animal protein, formation brownish red band.If corresponding orthene 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 orthene 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 orthene 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, orthene 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. orthene 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 orthene antibody; Detection reaction part is coated with detection orthene 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. orthene colloidal-gold detecting-card according to claim 1, is characterized in that: orthene detection antigen is the conjugates that orthene and carrier mass are formed.
3. orthene 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. orthene 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 |
|---|---|---|---|
| CN201410522807.1A CN105486868A (en) | 2014-10-08 | 2014-10-08 | Acephate colloidal gold detection card |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410522807.1A CN105486868A (en) | 2014-10-08 | 2014-10-08 | Acephate colloidal gold detection card |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105486868A true CN105486868A (en) | 2016-04-13 |
Family
ID=55673985
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410522807.1A Pending CN105486868A (en) | 2014-10-08 | 2014-10-08 | Acephate colloidal gold detection card |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105486868A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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
- 2014-10-08 CN CN201410522807.1A patent/CN105486868A/en active Pending
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 |
| 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 |
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| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
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Application publication date: 20160413 |