CN2847295Y - Colloidal gold test paper for quick detecting Methamidophos, fenthion and glyphosate residue - Google Patents
Colloidal gold test paper for quick detecting Methamidophos, fenthion and glyphosate residue Download PDFInfo
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- CN2847295Y CN2847295Y CN 200520142626 CN200520142626U CN2847295Y CN 2847295 Y CN2847295 Y CN 2847295Y CN 200520142626 CN200520142626 CN 200520142626 CN 200520142626 U CN200520142626 U CN 200520142626U CN 2847295 Y CN2847295 Y CN 2847295Y
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- glyphosate
- entex
- acephatemet
- monoclonal antibody
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Abstract
The utility model relates to a piece of colloidal gold test paper for rapidly detecting methamidophos, fenthion and glyphosate residues, which is composed of a bottom plate, a water absorbing plate, a nitrocellulose film, a monoclonal antibody gold label pad and a sample liquid absorbing layer, wherein the middle part of the bottom plate is provided with the nitrocellulose film. A methamidophos synthetic immunogen test line, or a fenthion synthetic immunogen test line, or a glyphosate synthetic immunogen test line and a polyclonal antibody control line are arranged on the nitrocellulose film. The end head at one end of the bottom plate is provided with the water absorbing plate, and the end head at the other end is provided with the sample liquid absorbing layer. Both ends of the nitrocellulose film are respectively and mutually overlapped and connected with the water absorbing plate and the monoclonal antibody gold label pad. The sample liquid absorbing layer is pressed on the monoclonal antibody gold label pad. The test paper is made through the immunization compete method of colloidal gold. The immunization colloidal gold method is used for detecting whether the residual quantity of methamidophos, or fenthion, or glyphosate in fruit vegetable is overproof or not. The method has the advantages of rapidity, sensitivity, convenient operation and low cost. The utility model does not need professionals to detect, and the organic unification of complicated theory and simple operation can be really achieved. Simultaneously, the utility model has the characteristics of convenient preservation and long effective period.
Description
Technical field
The present invention relates to a kind of vegetables and raw grain based food Detecting Pesticide test paper and detection method thereof of being used for, belong to the colloid gold immune detection range.
Background technology
Because the vegetable growth phase is short and disease and pest is comparatively serious, thereby, in the growing vegetables process, usually need the continuous several times dispenser.And many vegetables need repeatedly be gathered, gather and the time interval of dispenser short, cause when gathering the persticide residue in the vegetables often higher, be easy to surpass state specified standards.And persticide residue exceeds standard and can have a strong impact on level of human health in the vegetables, has become the food pollution source that can not be ignored.Government and each functional department extremely pay close attention to the residues of pesticides problem of food.But because the resistance to the action of a drug of the toxicity of agricultural chemicals and insect forms a vicious cycle, forbid thoroughly comprehensively that therefore the use of high highly toxic pesticide will have long process, even in using, low-toxin farm chemicals comprehensively, the pernicious poisoning of agricultural chemicals still takes place.Therefore new requirement and higher standard have all been proposed all many-sides such as the object of analyzing and testing, kind, quantity, scope, index.
Acephatemet (O, S-dimethyl thiophosphoryl amide), a kind of high toxicity organophosphorus insecticide, solubleness is very high in water.Making tea when drinking, most of remains of pesticide can leach in millet paste.Because acephatemet has acute poisoning and water solubility height, the Ministry of Agriculture just prohibites in tealeaves, fruits and vegetables in nineteen eighty-two and uses.Ministry of Health of the People's Republic of China and Standardization Administration of China issued on January 25th, 2005, implemented agricultural chemicals maximum residue limit (GB 2763-2005) in State Standard of the People's Republic of China's food on October 1st, 2005, standard code the maximum of agricultural chemicals acephatemet in cottonseed, paddy to allow residual quantity be 0.1mg/kg; Must not detect in the vegetables.
Entex (O, O-dimethyl-O-(3-methyl-4-methyl mercapto phenyl) thiophosphate), the novel organophosphorus fungicide of a kind of poisoning.Its mechanism of action is to suppress cholinesterase in the pest body, and acetylcholine is accumulated, the excited conduction that affects the nerves, thus make insect that spasm, paralysis death take place.Ministry of Health of the People's Republic of China and Standardization Administration of China issued on January 25th, 2005, implemented agricultural chemicals maximum residue limit (GB2763-2005) in State Standard of the People's Republic of China's food on October 1st, 2005, standard code the maximum of agricultural chemicals Entex in fruit, vegetables, wheat, paddy to allow residual quantity be 0.05mg/kg.
Glyphosate (N-(phosphoric acid methyl) glycocoll), a kind of low toxicity organic phosphates herbicide.Ministry of Health of the People's Republic of China and Standardization Administration of China issued on January 25th, 2005, implemented agricultural chemicals maximum residue limit (GB 2763-2005) in State Standard of the People's Republic of China's food on October 1st, 2005, standard code the maximum of pesticide glyphosate in fruit, paddy to allow residual quantity be 0.1mg/kg; It is 0.5mg/kg that maximum in the wheat flour allows residual quantity; It is 5mg/kg that the maximum of wheat, wholemeal allows residual quantity.
At present, the analytical approach of detection residues of pesticides mainly is to rely on vapor-phase chromatography (GC) or gas chromatography and mass spectrum (MS) coupling method (GC/MS)." enzyme inhibition rate method+spectrophotometric method " has been listed in national proposed standard (GB/T 5009.199-2003).This method is quick, sensitive, easy and simple to handle and with low cost, can realize quick primary dcreening operation, but the restriction of examined scope and precision.And vapor-phase chromatography (GC) can realize the quantitative test of precision but speed is slower, and it is higher to detect cost.In a word, these method ubiquities the sample pretreatment process complexity, sensitivity be subjected to sample purification, step such as concentrate influence very big, consuming time, checkout equipment costliness, and require to have technical professional and long analytical cycle.
Therefore, utilize the competition law principle, on the basis of traditional immunodetection, introduced the collaurum Fast Detection Technique, develop acephatemet collaurum competition law and detect test paper, Entex collaurum competition law detection test paper, glyphosate collaurum competition law detection test paper.This method has fast, sensitive, easy and simple to handle, cost is low, need not the professional detects, and really reaches the organic unity of complicated principle and ease of Use, have simultaneously preserve convenient, the characteristics that the term of validity is long.
Summary of the invention
The present invention is directed to some problems of above-mentioned existence, provide a kind of collaurum that is applicable to whether detection acephatemet, Entex, glyphosate exceed standard to measure test paper.
For solving the problems of the technologies described above, the present invention is achieved by the following technical solutions:
Detect test paper at methamidophos residue, Entex residue detection test paper, it is nitrocellulose filter that glyphosate residue detects test paper base plate middle part, a test wire and a sheep anti mouse polyclonal antibody control line are arranged on the nitrocellulose filter, in base plate one end termination is water accepting layer, other end termination is the sample liquid-adsorption layer, the nitrocellulose filter two ends respectively with water accepting layer and acephatemet, Entex, glyphosate monoclonal antibody gold mark pad overlaps mutually and connects (the overlapping coupling part is in 1-2 millimeter scope), at acephatemet, Entex, be pressed with the sample liquid-adsorption layer on the glyphosate monoclonal antibody gold mark pad.Combined method about base plate, nitrocellulose filter, thieving paper, collaurum mark pad is carried out by patent CN 2741052Y.
The control line that agricultural chemicals acephatemet, Entex or glyphosate residue detect test paper is formed by sheep anti mouse polyclonal antibody bag, and it is that to be detected Entex by acephatemet synthetic immunogen bag by, test wire be that to be detected glyphosate by Entex synthetic immunogen bag by, test wire be by glyphosate synthetic immunogen bag quilt that test wire detects acephatemet.Detecting acephatemet, the sample liquid-adsorption layer end of the colloid gold test paper of Entex or glyphosate residue is put into fruit juice, vegetables are soaked in the juice, because the capillarity sample will move along test strips water accepting layer end, when moving to monoclonal antibody gold mark pad, acephatemet in the sample and acephatemet monoclonal antibody gold mark probe, Entex and Entex monoclonal antibody gold mark probe, glyphosate and glyphosate monoclonal antibody gold mark probe generation specific bond, be fixed with the acephatemet synthetic immunogen when moving to, when Entex synthetic immunogen or glyphosate synthetic immunogen test wire, because acephatemet in antibody in the acephatemet monoclonal antibody gold mark pad and the sample, Entex in antibody in the Entex monoclonal antibody gold mark pad and the sample, glyphosate preferentially is combined into compound and loses itself and acephatemet in antibody in the glyphosate monoclonal antibody gold mark pad and the sample, Entex or the combination of glyphosate synthetic immunogen, therefore its collaurum can not be stranded on the test wire, the test wire place does not have red line to show, promptly has only a red control line positive; If do not have acephatemet, Entex, glyphosate in the opposite sample, antibody in the monoclonal antibody gold mark pad moves on the test wire, the acephatemet monoclonal antibody will with acephatemet synthetic immunogen, Entex monoclonal antibody will with Entex synthetic immunogen, glyphosate monoclonal antibody will with glyphosate synthetic immunogen generation specific bond, collaurum is stranded on the test wire, promptly two red line are negative, immune competition law principle that Here it is.Acephatemet, Entex or glyphosate content just are inversely proportional in the shade of test wire and the sample.Carry out the transition to feminine gender from the positive, because the acephatemet of surveying, Entex or glyphosate content difference, the color of test wire are also different, form a red color gradient difference, detect the OD value with this principle, thereby infer that test wire reacts the amount of actual acephatemet, Entex, glyphosate.
It detects acephatemet, Entex, glyphosate and sets more than the detected level to make test paper with competition law, and it is positive that a red line appears in the view window place; It is negative to set the two red lines of the following appearance of detected level; Be that fuzzy hacures are boundary value at the test wire place when setting detected level, thereby draw judgement.
When moving to sheep anti mouse polyclonal antibody control line, no matter have or not acephatemet, Entex, glyphosate in the sample, the gold mark probe of mark all can combine delay with the sheep anti mouse polyclonal antibody that has configured, and it is red that control line is shown.So control line does not have the colour band generation, and then the representative operation is wrong, the sample liquid level is expired above MAX line or test paper when detecting.
Because adopt technique scheme, the colloid gold test paper of a kind of fast detecting acephatemet provided by the present invention, Entex, glyphosate residue has such beneficial effect, i.e. high specificity, highly sensitive, easily store, need not the technical skill personnel operation, and readability as a result.
Description of drawings
Fig. 1 is the main TV structure figure of the colloid gold test paper of a kind of fast detecting acephatemet of the present invention, Entex, glyphosate residue.
Fig. 2 is the side-looking structural drawing of invention figure one.
Fig. 3 shows positive findings figure for detecting.
Fig. 4 shows negative findings figure for detecting.
Among the figure 1, water sucting plate, 2, nitrocellulose filter, 3, sheep anti mouse polyclonal antibody control line, 4, test wire, 5, monoclonal antibody gold mark pad, 6, the sample liquid-adsorption layer, 7, base plate, 8, the MAX line.
Specific embodiment
1. according to the molecular structure of acephatemet, infer the antigen determining area in its molecule, select for use the acephatemet synthetic immunogen synthetic a kind of close with the acephatemet molecular structure and have carboxyl or amino etc. and be easy to the be connected molecular structure (haptens) of group of protein macromolecule through chemical reaction.The haptens micromolecule is connected with protein macromolecule makes the acephatemet immunizing antigen.Entex immunizing antigen, the same acephatemet of glyphosate immunizing antigen preparation method.
2. MONOCLONAL ANTIBODIES SPECIFIC FOR
(1) with synthetic immunogen immune BALB/c mouse.Set up the knurl strain with the SP2/0 Fusion of Cells and screen, get oncocyte and be injected in the BALB/c mouse abdominal cavity, make it produce ascites.
(2) extract the screening of mouse ascites purifying, the monoclonal antibody that obtains respectively to combine with the acephatemet molecule is used for colloid gold label.
Entex, the same parathion of glyphosate method for preparing monoclonal antibody.
The preparation of collaurum and with the combining of monoclonal antibody
(1) get distilled water and add an amount of gold chloride magnetic agitation and be warmed to 85～95 ℃, add an amount of citrate three sodium and continue heated and stirred to seething with excitement 3～10 minutes, it is standby to keep in Dark Place after the cooling.
(2) on the collaurum liquid adsorbing fiber material of acephatemet labeling of monoclonal antibody, dry back is standby.
Entex, the same parathion of glyphosate monoclonal antibody association colloid gold method.
4. film-making machine system film: utilize computer control transmission speed, guarantee that the antibody amount of bag quilt on the per unit film equates.
5. test strips combination: see Figure of description Fig. 2, with patent CN 2741052Y.
6. using method: the sample liquid-adsorption layer end of test paper put into fruit juice, vegetables are soaked juice (liquid level must not surpass MAX line chart 8), taking out test paper after 1 minute keeps flat, because kapillary and syphonic effect sample will move observations in the time of 5 minutes along test strips water accepting layer end.
7. the result judges: detect more than the acephatemet concentration 0.1mg/kg, it is positive that a red line appears in the view window place; 0.1mg/kg the two red lines of following appearance are negative; 0.1mg/kg the time be that fuzzy hacures are boundary value at the test wire place, this boundary value is the vegetable pesticide residue examination criteria 0.1mg/kg of country's promulgation.Thereby draw judgement.
Claims (6)
1. the colloid gold test paper of a fast detecting acephatemet, Entex, glyphosate residue is characterized in that being made up of base plate (7), water sucting plate (1), nitrocellulose filter (2), acephatemet monoclonal antibody or Entex monoclonal antibody or glyphosate monoclonal antibody gold mark pad (5), sample liquid-adsorption layer (6); The base plate middle part is a nitrocellulose filter, a test wire (4) and a polyclonal antibody control line (3) are arranged on the nitrocellulose filter, base plate one end termination is a water sucting plate, other end termination is the sample liquid-adsorption layer, the nitrocellulose filter two ends overlap mutually with monoclonal antibody gold mark pad with water sucting plate respectively and are connected, and are pressed with the sample liquid-adsorption layer on monoclonal antibody gold mark pad.
2. the colloid gold test paper of a kind of fast detecting acephatemet according to claim 1, Entex, glyphosate residue is characterized in that control line is to be made into by sheep anti mouse polyclonal antibody bag.
3. the colloid gold test paper of a kind of fast detecting acephatemet according to claim 1, Entex, glyphosate residue is characterized in that test wire is to be made into by acephatemet synthetic immunogen or Entex synthetic immunogen or glyphosate synthetic immunogen bag.
4. according to the colloid gold test paper of claim 1 or 3 described a kind of fast detecting acephatemets, Entex, glyphosate residue, what it is characterized in that being detected is that acephatemet, Entex, the former cartridge bag of glyphosate are drawn together the potpourri of acephatemet, Entex, glyphosate preparation.
5. the colloid gold test paper of a kind of fast detecting acephatemet according to claim 1, Entex, glyphosate residue is characterized in that gold mark pad is as colloid gold label by monoclonal antibody.
6. according to the colloid gold test paper of the described a kind of fast detecting acephatemet of claim 1, Entex, glyphosate residue, it is acephatemet 0.01～2mg/kg, Entex 0.01～5mg/kg, glyphosate 0.05～5mg/kg that the detected value that it is characterized in that test wire is set at the limit of identification scope.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200520142626 CN2847295Y (en) | 2005-12-06 | 2005-12-06 | Colloidal gold test paper for quick detecting Methamidophos, fenthion and glyphosate residue |
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| CN 200520142626 CN2847295Y (en) | 2005-12-06 | 2005-12-06 | Colloidal gold test paper for quick detecting Methamidophos, fenthion and glyphosate residue |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101153841B (en) * | 2006-09-26 | 2011-07-13 | 明知大学产学协力团 | Method for detecting content of fenthion |
| CN105572311A (en) * | 2014-10-11 | 2016-05-11 | 江苏维赛科技生物发展有限公司 | Glyphosate colloidal gold detection card |
| CN109406786A (en) * | 2018-08-29 | 2019-03-01 | 郑州工程技术学院 | A kind of immune chromatography test paper detecting acephatemet |
-
2005
- 2005-12-06 CN CN 200520142626 patent/CN2847295Y/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101153841B (en) * | 2006-09-26 | 2011-07-13 | 明知大学产学协力团 | Method for detecting content of fenthion |
| CN105572311A (en) * | 2014-10-11 | 2016-05-11 | 江苏维赛科技生物发展有限公司 | Glyphosate colloidal gold detection card |
| CN109406786A (en) * | 2018-08-29 | 2019-03-01 | 郑州工程技术学院 | A kind of immune chromatography test paper detecting acephatemet |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Beijing Wanhua Bioengineering Co., Ltd. Assignor: Wan Jicheng Contract fulfillment period: 2009.10.22 to 2014.10.22 Contract record no.: 2009110000260 Denomination of utility model: Colloidal gold test paper for quick detecting Methamidophos, fenthion and glyphosate residue Granted publication date: 20061213 License type: Exclusive license Record date: 20091030 |
|
| LIC | Patent licence contract for exploitation submitted for record |
Free format text: EXCLUSIVE LICENSE; TIME LIMIT OF IMPLEMENTING CONTACT: 2009.10.22 TO 2014.10.22; CHANGE OF CONTRACT Name of requester: BEIJING WANHUA BIOLOGY ENGINEERING CO., LTD. Effective date: 20091030 |
|
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20061213 Termination date: 20100106 |