CN2869868Y - Colloid gold test-paper for quick detecting parathion, methyl parathion residue - Google Patents

Colloid gold test-paper for quick detecting parathion, methyl parathion residue Download PDF

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Publication number
CN2869868Y
CN2869868Y CN 200520142624 CN200520142624U CN2869868Y CN 2869868 Y CN2869868 Y CN 2869868Y CN 200520142624 CN200520142624 CN 200520142624 CN 200520142624 U CN200520142624 U CN 200520142624U CN 2869868 Y CN2869868 Y CN 2869868Y
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parathion
methyl
test paper
monoclonal antibody
colloid gold
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CN 200520142624
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Chinese (zh)
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万积成
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Abstract

A colloidal gold test paper of rapid detecting the residuas of parathion and methyl parathion, which comprises a base plate, a water absorption board, a pyroxylin membrane, a monoclonal antibody gold label mat and a specimen imbibition layer. The middle of the base plate is a pyroxylin membrane on which is provided with parathion synthetic immunogen test lead or methyl parathion synthetic immunogen test lead and a multi- clone antibody control line; one end tip of the base plate is provided with a water absorption board, and the other end tip is provided with a specimen imbibition layer; the two ends of pyroxylin membrane are separately overlapped for connection each other with the water absorption board and the monoclonal antibody gold label mat; the top of the monoclonal antibody gold label mat is pressed with a specimen imbibition layer; the test paper is made by means of a colloidal gold immunity competition method; the immunity colloidal gold method is used to detect if the parathion or methyl parathion level of residue in garden stuff is overproof. The method has the features of speediness, sensitivity, simplicity of operation, low cost, no necessity of detecting by professional, real attainment of integrating the complicated principle with the simplicity of operation organically, as well as convenience of preservation and long period of validity.

Description

Fast detecting parathion, the residual colloid gold test paper of parathion-methyl
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.
Parathion (O, O-diethyl-O-(p-nitrophenyl)-thiophosphate), high malicious broad spectrum activity thionic formula monothio phosphoric acid ester insecticidal/acaricidal agent, China is also referred to as 1.Very high to higher mammal toxicity, medicament is as entering in the higher mammal body also danger close by eyes, respiratory tract, skin.Fumigation action was more obvious when temperature raise.Absorption in not having has and oozes effect in certain.
Parathion-methyl (O, O-dimethyl-O-p-nitrophenyl thiophosphate) belongs to high malicious pesticide.Parathion-methyl can cause poisoning by esophagus, respiratory tract and skin, should control pulmonary edema, encephaledema and respiration inhibition.
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/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 parathion collaurum competition law and detect test paper, parathion-methyl 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 parathion, parathion-methyl 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:
At parathion, parathion-methyl 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 and are connected (the overlapping coupling part is 1~2 millimeter scope) with parathion, parathion-methyl monoclonal antibody gold mark pad with water accepting layer respectively, are pressed with the sample liquid-adsorption layer on parathion, parathion-methyl 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 of agricultural chemicals parathion or parathion-methyl residue detection test paper is formed by sheep anti mouse polyclonal antibody bag, test wire detect parathion be by parathion synthetic immunogen bag by, to detect parathion-methyl be by parathion-methyl synthetic immunogen bag quilt.The sample liquid-adsorption layer end that detects the residual colloid gold test paper of parathion or parathion-methyl 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, parathion in the sample and parathion monoclonal antibody gold mark probe, parathion-methyl and parathion-methyl monoclonal antibody gold mark probe generation specific bond, be fixed with parathion synthetic immunogen test wire when moving to, during parathion-methyl synthetic immunogen test wire, because parathion in antibody in the parathion monoclonal antibody gold mark pad and the sample, parathion-methyl preferentially is combined into compound and loses it and combine with parathion or parathion-methyl synthetic immunogen in antibody in the parathion-methyl monoclonal antibody gold mark pad and the sample, 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 parathion, parathion-methyl in the opposite sample, antibody in the monoclonal antibody gold mark pad moves on the test wire, the parathion monoclonal antibody will with parathion synthetic immunogen, parathion-methyl monoclonal antibody will with parathion-methyl 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.Parathion or parathion-methyl content just are inversely proportional in the shade of test wire and the sample.Carry out the transition to feminine gender from the positive, since the institute parathion of surveying or parathion-methyl content difference, the color of test wire also the difference, form a red color gradient difference, detect the OD value with this principle, thereby infer that test wire reacts the amount of actual parathion or parathion-methyl.
With competition law make test paper its detect parathion, parathion-methyl is set more than the detected level, 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 the fuzzy hacures of bar 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 parathion, parathion-methyl 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, a kind of fast detecting parathion provided by the present invention, the residual colloid gold test paper of parathion-methyl have 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 residual colloid gold test paper of a kind of fast detecting parathion of the present invention, parathion-methyl.
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 parathion molecular structure, infer the antigen determining area in its molecule, select for use the parathion pesticide original medicine synthetic a kind of close with the parathion 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 parathion immunizing antigen.The same parathion of parathion-methyl antigen preparation method.
2. parathion 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 parathion molecule is used for colloid gold label.The same parathion of parathion-methyl 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) the collaurum liquid of parathion labeling of monoclonal antibody, on the adsorbing fiber material, dry back is standby.
The same parathion of parathion-methyl 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 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 parathion 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.Being lower than the following boundary value of 0.1mg/kg takes the colour code control methods to judge dosage.

Claims (6)

1. a fast detecting parathion, the residual colloid gold test paper of parathion-methyl is characterized in that being made up of base plate (7), water sucting plate (1), nitrocellulose filter (2), parathion, parathion-methyl monoclonal antibody gold mark pad (5), sample liquid-adsorption layer (6), 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. a kind of fast detecting parathion according to claim 1, the residual colloid gold test paper of parathion-methyl is characterized in that control line is to be made into by sheep anti mouse polyclonal antibody bag.
3. a kind of fast detecting parathion according to claim 1, the residual colloid gold test paper of parathion-methyl is characterized in that test wire is to be made into by parathion synthetic immunogen or parathion-methyl synthetic immunogen bag.
4. according to claim 1 or 3 described a kind of fast detecting parathion, the residual colloid gold test paper of parathion-methyl, it is characterized in that this test paper can detect parathion, the former medicine of parathion-methyl or potpourri according to the composition of regulating gold mark pad and test wire encrusting substance.
5. a kind of fast detecting parathion according to claim 1, the residual colloid gold test paper of parathion-methyl is characterized in that gold mark pad is as colloid gold label by monoclonal antibody.
6. according to the described a kind of fast detecting parathion of claim 1, the residual colloid gold test paper of parathion-methyl, it is parathion 0.01~5mg/kg, parathion-methyl 0.01~5mg/kg that the detected value that it is characterized in that test wire is set at limit of identification.
CN 200520142624 2005-12-06 2005-12-06 Colloid gold test-paper for quick detecting parathion, methyl parathion residue Expired - Fee Related CN2869868Y (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103102415A (en) * 2012-07-11 2013-05-15 南京农业大学 Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl
CN105572357A (en) * 2014-10-07 2016-05-11 江苏维赛科技生物发展有限公司 Parathion-methyl colloidal gold detection card

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103102415A (en) * 2012-07-11 2013-05-15 南京农业大学 Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl
CN103102415B (en) * 2012-07-11 2014-07-30 南京农业大学 Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl
CN105572357A (en) * 2014-10-07 2016-05-11 江苏维赛科技生物发展有限公司 Parathion-methyl colloidal gold detection card

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Granted publication date: 20070214

Termination date: 20100106