CN2893707Y - Collaturum test puper for fast detecting chlortoluron residue - Google Patents
Collaturum test puper for fast detecting chlortoluron residue Download PDFInfo
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- CN2893707Y CN2893707Y CN 200520142604 CN200520142604U CN2893707Y CN 2893707 Y CN2893707 Y CN 2893707Y CN 200520142604 CN200520142604 CN 200520142604 CN 200520142604 U CN200520142604 U CN 200520142604U CN 2893707 Y CN2893707 Y CN 2893707Y
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- chlortoluron
- monoclonal antibody
- test paper
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Abstract
A colloidal gold test paper for fast detection of chlortoluron residue is composed of a base plate, a water absorption plate, a cellulose nitrate film, a monoclonal antibody gold marking pad, and a sample absorption layer. The cellulose nitrate film is placed in the middle of the base plate, and a chlorotoluron synthetic immunogen test wire and a polyclonal antibody control wire are placed on the cellulose nitrate film. On one end of the base plate a water absorption plate is provided, and on the other end the sample absorption layer is positioned. The two ends of the cellulose nitrate film overlap and connect respectively with the water absorption plate and the monoclonal antibody gold marking pad. The sample absorption layer is pressed on the monoclonal antibody gold marking pad. The test paper is made using the colloidal gold immunity competition method which adopts the immunity colloidal gold method to detect whether the chlortoluron residue in fruits and vegetables exceeds specified standard. The method features in rapid, sensitive, simple and convenient operation which requires detection not necessarily by professional personnel, low cost which realizes the real unification of the complicated principle and simple operation provides such virtues as easy storage and long validity terms.
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.
Chlortoluron (N '-(3-chlorine 4-tolyl)-N, N-methyl urea), be a kind of selectivity inner sucting conduction type herbicide.Be applicable in the dry crops such as wheat, cotton, peanut, soybean, tobacco and prevent and treat annual gramineae, Cyperaceae and most of broad leaved weed.Persticide residue just is a key factor of weighing the food quality quality.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 chlortoluron in crops to allow residual quantity be 0.1mg/kg.
At present, the analytical approach of detection residues of pesticides mainly is dependence, 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/T5009.199-2003).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 chlortoluron collaurum competition law and detect 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 being applicable to detect the collaurum mensuration test paper whether chlortoluron exceeds standard.
For solving the problems of the technologies described above, the present invention is achieved by the following technical solutions:
At chlortoluron residue detection test paper base plate middle part is nitrocellulose filter, 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 overlap mutually with chlortoluron monoclonal antibody gold mark pad with water accepting layer respectively and are connected (the overlapping coupling part is in 1-2 millimeter scope), are pressed with the sample liquid-adsorption layer on chlortoluron monoclonal antibody gold mark pad.Combined method about base plate, nitrocellulose filter, thieving paper, collaurum mark pad is carried out by patent CN2741052Y.
The control line of chlortoluron residue detection test paper is formed by sheep anti mouse polyclonal antibody bag, and test wire is by chlortoluron synthetic immunogen bag quilt.The sample liquid-adsorption layer end that detects the residual colloid gold test paper of chlortoluron is put into Juice, because the capillarity sample will move along test strips water accepting layer end, when moving to chlortoluron monoclonal antibody gold mark pad, chlortoluron in the sample and chlortoluron monoclonal antibody gold mark probe generation specific bond, when moving to when being fixed with chlortoluron synthetic immunogen test wire, lose it and combine owing to chlortoluron in the antibody in the chlortoluron monoclonal antibody gold mark pad and the sample preferentially is combined into compound with chlortoluron-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 chlortoluron in the opposite sample, antibody in the chlortoluron monoclonal antibody gold mark pad moves on the test wire, the chlortoluron monoclonal antibody will with chlortoluron 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.Chlorotoluron content just is inversely proportional in the shade of test wire and the sample.Carry out the transition to feminine gender from the positive, because the chlorotoluron content difference of surveying, the color of test wire is also different, forms a red color gradient difference, detects the OD value with this principle, thereby infers the amount of actual chlortoluron that test wire reacts.
It detects more than the chlortoluron setting 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 chlortoluron 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.Therefore control line does not have colour band and produces that then the representative operation is wrong, during detection the sample liquid level surpass the MAX line or test paper expired.
Because adopt technique scheme, the residual colloid gold test paper of a kind of fast detecting chlortoluron provided by the present invention has such beneficial effect, promptly high specificity is highly sensitive, easily stores, 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 residual colloid gold test paper of a kind of fast detecting chlortoluron of the present invention.
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 chlortoluron, infer the antigen determining area in its molecule, select for use other reagent or chlortoluron pesticide original medicine synthetic a kind of close with the chlortoluron 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 chlortoluron immunizing antigen.
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 acquisition can combine with the chlortoluron molecule is used for colloid gold label.
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) the collaurum liquid of chlortoluron labeling of monoclonal antibody, on the adsorbing fiber material, dry back is standby.
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, method is with patent CN2741052Y.
6. using method: the sample liquid-adsorption layer end of test paper is put into vegetables soak juice (liquid level must not surpass MAX line chart 8), take out test paper after 1 minute and keep 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 chlortoluron 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. colloid gold test paper that the fast detecting chlortoluron is residual, it is characterized in that by base plate (7), water sucting plate (1), nitrocellulose filter (2), chlortoluron monoclonal antibody gold mark pad (5), sample liquid-adsorption layer (6) form, MAX line (8); 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 residual colloid gold test paper of a kind of fast detecting chlortoluron according to claim 1 is characterized in that control line is to be made into by sheep anti mouse polyclonal antibody bag.
3. the residual colloid gold test paper of a kind of fast detecting chlortoluron according to claim 1 is characterized in that test wire is to be made into by chlortoluron synthetic immunogen bag.
4. according to claim 1 or the residual colloid gold test paper of 3 described a kind of fast detecting chlortolurons, it is characterized in that the object that is detected is the potpourri that the former cartridge bag of chlortoluron is drawn together the chlortoluron preparation.
5. the residual colloid gold test paper of a kind of fast detecting chlortoluron according to claim 1 is characterized in that gold mark pad is as colloid gold label by the chlortoluron monoclonal antibody.
6. according to the residual colloid gold test paper of the described a kind of fast detecting chlortoluron of claim 1, it is chlortoluron 0.05～0.1mg/kg that the detected value that it is characterized in that test wire is set at limit of identification.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200520142604 CN2893707Y (en) | 2005-12-06 | 2005-12-06 | Collaturum test puper for fast detecting chlortoluron residue |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200520142604 CN2893707Y (en) | 2005-12-06 | 2005-12-06 | Collaturum test puper for fast detecting chlortoluron residue |
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| Publication Number | Publication Date |
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| CN2893707Y true CN2893707Y (en) | 2007-04-25 |
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| CN 200520142604 Expired - Fee Related CN2893707Y (en) | 2005-12-06 | 2005-12-06 | Collaturum test puper for fast detecting chlortoluron residue |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104655834A (en) * | 2013-11-20 | 2015-05-27 | 丹阳亿太生物科技发展有限公司 | Chemical-luminescent enzyme-linked immunoassay method for detecting bentazone |
-
2005
- 2005-12-06 CN CN 200520142604 patent/CN2893707Y/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104655834A (en) * | 2013-11-20 | 2015-05-27 | 丹阳亿太生物科技发展有限公司 | Chemical-luminescent enzyme-linked immunoassay method for detecting bentazone |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070425 Termination date: 20100106 |