CN202216956U - Organic phosphorus pesticide multi-residue gold-labeled rapid test card - Google Patents
Organic phosphorus pesticide multi-residue gold-labeled rapid test card Download PDFInfo
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- CN202216956U CN202216956U CN2011201055925U CN201120105592U CN202216956U CN 202216956 U CN202216956 U CN 202216956U CN 2011201055925 U CN2011201055925 U CN 2011201055925U CN 201120105592 U CN201120105592 U CN 201120105592U CN 202216956 U CN202216956 U CN 202216956U
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Abstract
The utility model discloses an organic phosphorus pesticide multi-residue gold-labeled rapid test card, which comprises a bottom plate, a cover sealing plate and at least two groups of cushion pads, sample pads, golden pads, detection films and absorbent pads, wherein the cushion pads, the sample pads, the golden pads, the detection films and the absorbent pads are stuck in sequence. The cushion pads are stuck onto the bottom plate, at least two sample pads are stuck onto the cushion pads in crossing mode, and the golden pads, the detection films and the absorbent pads are sequentially stuck onto the sample pads. The cover sealing plate is provided with a sample filling hole and visualization windows, the sample filling hole is opposite to the cushion pads, the visualization windows are respectively opposite to the detection films, and organic phosphorus pesticide antigen and goat-anti-mouse immunoglobulin G (IgG) are coated on the upper surfaces of the detection films in parallel mode to form a detection line and a quality control line respectively. The organic phosphorus pesticide multi-residue gold-labeled rapid test card is simple and rapid to operate, and can rapidly and simultaneously detect ten 2-ethoxy thiophosphate ester pesticides and eight 2-methoxy thiophosphate ester pesticides in samples. Sensitivity of the organic phosphorus pesticide multi-residue gold-labeled rapid test card can reach 0.001 to 0.693mg/kg and 0.026 to 1.0mg/kg, results can be observed in 3 to 5 minutes, and the organic phosphorus pesticide multi-residue gold-labeled rapid test card is suitable for rapid detection of organic phosphorus pesticides in vegetables, fruits and environmental water samples.
Description
Technical field
The utility model belongs to the detection technique field; Be specifically related to a kind of gold mark quick measuring card that can detect diethoxy thiophosphate agricultural chemicals and dimethoxy group thiophosphate agricultural chemicals simultaneously, can use the fast detecting of organophosphorus pesticide in fruit, vegetables and the environmental water sample.
Background technology
Organophosphorus pesticide is efficient as one type, the pesticide and the herbicide of wide spectrum are widely used in agricultural production, but its a large amount of back residual human and livestock health of serious harm and earth safety in agricultural product and environment of using.Because organophosphorus pesticide use phenomenon of a great variety and composite is general, many method for detecting residue that foundation can detect a plurality of objects simultaneously are very necessary.Instrument analytical method can be realized many residue detection, but needs expensive instrument and equipment and professional operating personnel, and sample pre-treatments loaded down with trivial details time-consuming, testing cost is high, is difficult to satisfy the needs of high flux, quick, online detection.
At present, develop the important means that quick many method for detecting residue become the deficiency that remedies instrumental method.From 80 years 20th century; Suppress the cholinesterase activity principle based on organophosphorus and carbamate chemicals for agriculture and set up the rapid screening that " enzyme inhibition method " begins to be widely used in organophosphorus and carbamate chemicals for agriculture in the agricultural product; Yet because its sensitivity and less stable are progressively withdrawn from the market, only minority developing countries such as China, Korea S are still using at present.It is extremely urgent with the method for quick of stability that development has high sensitivity more.It is consistent with conventional instrumental analysis to have sensitivity based on the immunoassay of antigen and antibody specific association reaction; Be fit to on-the-spot screening; Simply, fast, advantage such as cost is low, the sample aequum is few, pre-treatment is simple and convenient, be considered to the competitive and challenging ultramicron detection technique of tool of 21 century.
The gold-marking immunity chromatography is that collaurum directly is marked on the antibody molecule, utilizes the immunochromatography principle to carry out method for measuring.It is easy and simple to handle, and a few minutes can be seen the result fast, and cost is also very cheap.Present development trend is sensitive more for preparation, can carry out the gold test strip of many residue detection fast with the while.Seldom report both at home and abroad based on the how residual quick measuring card of organophosphorus pesticide of gold mark principle at present, on same quick measuring card, can detect two organophosphorus agricultural chemicals simultaneously and not appear in the newspapers as yet.
Summary of the invention
The purpose of the utility model is to overcome the deficiency of prior art; Provide a kind of organophosphorus pesticide how residual quick measuring card, can realize simultaneously, easy, detect diethoxy thiophosphate and dimethoxy group thiophosphate organophosphorus pesticide in food and the environment apace.
The object of the invention is achieved through following technical scheme:
The utility model provides a kind of organophosphorus pesticide how residual gold-marking immunity quick measuring card, comprises base plate and capping plate, and cushion pad, sample pad, gold that at least two groups are pasted successively fill up, detect film, adsorptive pads; Said cushion pad simultaneously sticks on the base plate, and the cushion pad another side is pasted sample pad, pastes the gold pad on the sample pad successively, detects film, adsorptive pads; Paste the above capping plate of bonnet successively and promptly get said quick measuring card.Said capping plate is provided with well and visual window, and said well is facing to cushion pad, and said visual window is respectively facing to detecting film; Said detection film upper surface is coated with organophosphorus pesticide antigen and sheep anti-mouse igg side by side, forms detection line and nature controlling line respectively.
Preferably, said cushion pad is as a whole, and across two sample pad, said well is facing to the cushion pad middle.
Said base plate adopts the plastic plate that does not absorb water, and plastic plate simultaneously scribbles adhesive sticker, and base plate plays fixing other ingredients of test paper of supporting.
Cushion pad and sample pad adopt filter paper or all-glass paper; Said cushion pad and sample pad are that filter paper or all-glass paper are immersed 2～5min in the phosphate buffer (PBS) of 0.1mol/L, pH6.4～7.2, take out the back and are drying to obtain in 80 ℃ of oven dry or vacuum drying or other modes.
Said golden pad is to adopt all-glass paper or filter paper, and all-glass paper or filter paper are immersed respectively in diethoxy thiophosphate agricultural chemicals monoclonal anti liquid solution and the dimethoxy group thiophosphate agricultural chemicals monoclonal anti liquid solution, and preferred 4 ℃ are stirred immersion 2h down.Said diethoxy thiophosphate agricultural chemicals monoclonal anti liquid solution reference technique document Xu et al. Analytical Chemistry, the description among 2010,82,9314 – 9321.
Said dimethoxy group thiophosphate agricultural chemicals monoclonal anti liquid solution reference technique document Li Yong is auspicious etc., analytical chemistry, and 2010,38 (11): the description in the 1550-1555 polyclonal antibody (PAb-H3) combines the conventional method preparation.
Said detection film sticks on the gold pad; Said adsorptive pads preferred glass fibers paper, filter paper or thieving paper.
The utility model detects principle: add sample through well, sample arrives sample pad through cushion pad, follows to move on to the gold pad.If contain organophosphorus pesticide in the sample; Move on to detection line on can combining with the organophosphorus pesticide antibody on the gold pad to continue; Because the organophosphorus pesticide antibody of colloid gold label has only a binding site; After organophosphorus pesticide in the sample combined with it, the organophosphorus pesticide antigen on the detection line just can not be again and the antibodies of colloid gold label, so detection line (T) is colourless; When not having organophosphorus pesticide in the sample; Seized organophosphorus pesticide antigen (conjugate of 6-((diethoxy D2EHDTPA base) amino) caproic acid and ovalbumin (OVA)) was caught when the antibody of colloid gold label arrived detection line (T), then formed the naked eyes red color visible.No matter whether contain organophosphorus pesticide in the sample, nature controlling line (C) all redness can occur.Therefore, if only contain the diethoxy thiophosphate agricultural chemicals in the sample, detect on the film above and have only a red line; If only contain dimethoxy group thiophosphate agricultural chemicals in the sample, detect on the film below and have only a red line; If two kinds of agricultural chemicals all sometimes, detect film and following detection film above and all have only a red line.When not containing above-mentioned two organophosphorus agricultural chemicals in the sample, detecting film and following detection film above all has two red lines.
The beneficial effect of the utility model:
(1) Broadspectrum specificity is good.The utility model quick measuring card adopts the monoclonal antibody of diethoxy thiophosphate agricultural chemicals and dimethoxy group thiophosphate agricultural chemicals with Broadspectrum specificity, respectively two types of agricultural chemicals is had good recognition performance.
(2) the utility model is simple to operate, and 10 diethoxy thiophosphate agricultural chemicals and 8 dimethoxy group thiophosphate agricultural chemicals in the while test sample reach 0.001～0.693mg/kg and 0.026～1.0mg/kg respectively fast.
(3) high flux.Detect when can realize diethoxy thiophosphate agricultural chemicals and two types of agricultural chemicals of dimethoxy group thiophosphate agricultural chemicals.
(4) simple to operation.Fruit, vegetables and Environmental Water sample are carried out can detecting after the simple process.This quick measuring card does not need any instrument and equipment, and common layman can operate, and with the naked eye gets final product interpretation.
(5) speed is fast, colour stable, but dripping sample 3～5min just observations later on, the color of detectability and nature controlling line can keep more than the 30min, is suitable for the organophosphorus pesticide in vegetables, fruit and the environmental water sample is carried out fast detecting.
Description of drawings
The structural representation of Fig. 1 the utility model.
Embodiment
Come further explain the utility model below in conjunction with accompanying drawing and specific embodiment.
(1) structure of the utility model
Shown in accompanying drawing 1, the utility model provides the how residual gold mark of a kind of organophosphorus pesticide quick measuring card, comprises that base plate 1 and 9, two groups of cushion pads 2 of capping plate, sample pad 3, gold fill up 4, detect film 5 and adsorptive pads 8; Said sample pad 3, gold pad 4, detect film 5 and adsorptive pads 8 sticks on the base plate successively;
Be the utility model optimized technical scheme shown in the accompanying drawing 1, said cushion pad 2 is across on two sample pad 3; Said detection film 3 upper surfaces are coated with organophosphorus pesticide antigen (detection line 6) and goat anti-mouse igg (nature controlling line 7) side by side.
The function and the preparation method of the each several part of the described test paper of the utility model are described below:
Base plate 1:, play fixing support other ingredients of quick measuring card and be merged into quick measuring card with capping plate 9 for simultaneously scribbling the plastic plate that does not absorb water of adhesive sticker.
Gold pad 4: play the fixedly antibody of colloid gold label.
The preparation of golden pad 4 and processing comprise the preparation of collaurum colloidal sol, colloid gold label organophosphorus pesticide antibody and three steps of gold pad processing:
Collaurum colloidal sol preparation: with gold chloride (HauCl
4) be configured to 1% mother liquor with ultrapure water, get the mother liquor of 1mL, with the ultrapure water constant volume to 100mL; Be made into 0.01% solution, be heated to boiling, add a certain amount of 1% trisodium citrate aqueous solution; Continue to be heated to occur transparent orange red till, promptly get collaurum colloidal sol;
Colloid gold label organophosphorus pesticide Monoclonal Antibody: with organophosphorus pesticide monoclonal antibody (Xu et al. Analytical Chemistry, 2010,82; 9314 – 9321) with Tris-HCl (0.01mol/L, pH8.0) dissolved dilution is to 3mg/mL, every 10mL collaurum colloidal sol adds the organophosphorus pesticide monoclonal antibody of 0.1mL 2 mg/mL; Vortex 5min adds 10% PEG-20000 2mL, the centrifugal 15min of 15000r/min; Remove supernatant, (0.01mol/L pH7.4) redissolves with Tris-HCl; With centrifugal 15 min of 10000 r/min, remove supernatant, will precipitate (0.01mol/L with Tris-HCl; PH7.4) dilution is 1000 times, and product indicates the machine phosphorus insecticide monoclonal antibody as gold;
The gold pad is handled: gold is indicated the machine phosphorus insecticide monoclonal antibody pour in the clean beaker, all-glass paper or filter paper immerse 1min, take out, and vacuum freeze drying is gold pad 4.
Detect film 5: detect on the film 5 and encapsulate 1 organophosphorus pesticide antigen (Li Yongxiang etc.; Analytical chemistry; 2010; 38 (11): the conjugate of haptens H7 among the 1550-1555 and ovalbumin (OVA)) as detection line 6, the sheep anti-mouse igg line that encapsulates 1 is as nature controlling line 7, and organophosphorus pesticide antigen can be discerned by two kinds of antibody simultaneously.
Adsorptive pads 8: all-glass paper, filter paper or thieving paper after the employing drying at room temperature are as adsorptive pads.Adsorptive pads 8 main effects are the sample solution that moves is up absorbed.
Capping plate 9: well 10 and visual window 11 are arranged above the plastic plate, can combine to form test card with base plate 1.
The assembling of the utility model; Shown in accompanying drawing 1; On base plate 1, paste two groups of sample pad 3, gold pad 4 successively, detect film 5 and adsorptive pads 8, cushion pad 1 across on two sample pad, is covered capping plate 9 then; Well 10 is facing to the cushion pad middle, and two visual windows 11 detect film 5 facing to two respectively.
(2) application of the utility model
The detection of environmental water sample: directly draw environmental water sample and be added drop-wise in the well 10, drip 4, leave standstill keep flat 3～5min after.If two are detected film and all present color, show that sample is negative, do not contain diethoxy thiophosphate or dimethoxy group thiophosphate organophosphorus pesticide, otherwise positive sample.No matter sample is negative or positive, and nature controlling line should develop the color, if nature controlling line does not develop the color, shows that test paper lost efficacy.
The detection of fruit and vegetable sample: fruit (getting skin portion) or vegetables are cut into small pieces, and take by weighing 1g and extract 10 min with the PBS solution that 1mL 0.05mol/L contains 10% methyl alcohol, and the centrifugal 5min of 5000 rpm gets supernatant and directly detects, the same environmental water sample of method and judged result.The detectability to organophosphorus pesticide of the utility model is as shown in table 1.
This quick measuring card of table 1 is to the detection limit (mg/kg) of organophosphorus pesticide
| The diethoxy thiophosphate agricultural chemicals | Detectability (mg/kg) | Dimethoxy group thiophosphate agricultural chemicals | Detectability (mg/kg) |
| Resistox | 0.001 | Di-captan | 0.231 |
| Parathion | 0.004 | Cynock | 0.026 |
| Phoxim | 0.017 | Parathion-methyl | 0.056 |
| Dichlofenthion | 0.084 | Fenifrothion | 0.113 |
| Quinalphos | 0.039 | Entex | 0.026 |
| Hostathion | 0.043 | Famphur | 0.178 |
| Triazotion | 0.076 | Iodfenphos | 0.910 |
| Phosalone | 0.098 | Chlorpyrifos-methyl | 1.000 |
| Disulfoton | 0.693 | ? | ? |
| Thimet | 0.383 | ? | ? |
Choose 5 blank samples (wherein 1-3 is the pond water sample, and 4 and 5 are the leek sample) and add an amount of parathion, adopt this quick measuring card and commercially available enzyme process quick measuring card to measure simultaneously, adopt the HPLC-MS/MS method to prove conclusively simultaneously, the result lists in table 2.Can find out that from result shown in the table 2 this quick measuring card and enzyme process quick measuring card can be measured negative sample preferably, but,, can't measure the sample of low concentration agricultural chemicals because the sensitivity of enzyme process quick measuring card is lower for positive.Explain that this quick measuring card has sensitivity preferably, be superior to traditional enzyme process quick measuring card.
This quick measuring card of table 2 and cholinesterase quick measuring card are to the sample determination comparing result
| ? Sequence number | This quick measuring card | The enzyme process quick measuring card |
HPLC-MS/ |
| 1 | Positive | Negative | Parathion 0.022mg/ |
| 2 | Negative | Negative | Negative |
| 3 | Positive | Positive | Parathion 0.12 mg/ |
| 4 | Negative | Negative | Negative |
| 5 | Positive | Negative | Parathion 0.014 mg/kg |
Claims (5)
1. the how residual gold mark of an organophosphorus pesticide quick measuring card it is characterized in that comprising base plate and capping plate, and cushion pad, sample pad, gold that at least two groups are pasted successively fills up, detects film, adsorptive pads; Said cushion pad simultaneously sticks on the base plate, and the cushion pad another side is pasted sample pad, pastes the gold pad on the sample pad successively, detects film, adsorptive pads; Said capping plate is provided with well and visual window, and said well is facing to cushion pad, and said visual window is respectively facing to detecting film; Said detection film upper surface is coated with organophosphorus pesticide antigen and sheep anti-mouse igg side by side, forms detection line and nature controlling line respectively;
Said gold pad is handled in the dimethoxy group thiophosphate organophosphorus pesticide polyspecific monoclonal anti liquid solution of the diethoxy thiophosphate agricultural chemicals polyspecific monoclonal anti liquid solution that immerses colloid gold label and colloid gold label respectively.
2. the how residual gold mark of organophosphorus pesticide according to claim 1 quick measuring card is characterized in that said cushion pad is as a whole, across two sample pad; Said capping plate has only a well, and said well is facing to the cushion pad middle.
3. according to the how residual gold mark of the said organophosphorus pesticide of claim 1 quick measuring card, it is characterized in that cushion pad and sample pad are respectively filter paper or the all-glass paper of handling in the PBS damping fluid that immerses 0.1mol/L, pH6.4～7.2.
4. according to the how residual gold mark of the said organophosphorus pesticide of claim 1 quick measuring card, said gold pad is all-glass paper.
5. according to the how residual gold mark of the said organophosphorus pesticide of claim 1 quick measuring card, said detection film upper surface is coated with the conjugate that organophosphorus pesticide antigen is 6-((diethoxy D2EHDTPA base) amino) caproic acid and ovalbumin OVA side by side.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011201055925U CN202216956U (en) | 2011-04-12 | 2011-04-12 | Organic phosphorus pesticide multi-residue gold-labeled rapid test card |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011201055925U CN202216956U (en) | 2011-04-12 | 2011-04-12 | Organic phosphorus pesticide multi-residue gold-labeled rapid test card |
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| CN202216956U true CN202216956U (en) | 2012-05-09 |
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| CN2011201055925U Expired - Lifetime CN202216956U (en) | 2011-04-12 | 2011-04-12 | Organic phosphorus pesticide multi-residue gold-labeled rapid test card |
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| CN103018435A (en) * | 2012-07-11 | 2013-04-03 | 南京农业大学 | Organophosphorus pesticide multiresidue fast immune-gold localization test strip and preparation method thereof |
| CN103207270A (en) * | 2013-04-09 | 2013-07-17 | 江西中德生物工程有限公司 | Cimaterol colloidal gold test strip as well as preparation method and application thereof |
| CN103513030A (en) * | 2012-06-28 | 2014-01-15 | 艾博生物医药(杭州)有限公司 | Test strip for detecting samples |
| CN103940990A (en) * | 2013-01-23 | 2014-07-23 | 北京怡成生物电子技术有限公司 | Sensitivity-enhanced gold-labeled immunochromatographic strip and preparation method thereof |
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| CN105572361A (en) * | 2014-10-11 | 2016-05-11 | 江苏维赛科技生物发展有限公司 | Phoxim colloidal gold detection card |
| CN105785021A (en) * | 2016-04-08 | 2016-07-20 | 广州万联生物科技有限公司 | Quick detecting card for immunochromatography of organophosphorus and carbamate pesticide multiresidue cholinesterase |
| CN106170699A (en) * | 2014-04-04 | 2016-11-30 | 田中贵金属工业株式会社 | Immunochromatographiassays assays method |
| CN106324243A (en) * | 2016-08-23 | 2017-01-11 | 广东志道医药科技有限公司 | Colloidal gold immunochromatography test strip and preparing and use method thereof |
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| CN103018435A (en) * | 2012-07-11 | 2013-04-03 | 南京农业大学 | Organophosphorus pesticide multiresidue fast immune-gold localization test strip and preparation method thereof |
| CN103940990A (en) * | 2013-01-23 | 2014-07-23 | 北京怡成生物电子技术有限公司 | Sensitivity-enhanced gold-labeled immunochromatographic strip and preparation method thereof |
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| CN105785021A (en) * | 2016-04-08 | 2016-07-20 | 广州万联生物科技有限公司 | Quick detecting card for immunochromatography of organophosphorus and carbamate pesticide multiresidue cholinesterase |
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| CN112858276A (en) * | 2021-01-25 | 2021-05-28 | 郑州欧柯奇仪器制造有限公司 | Soybean milk maturity rapid test card and detection method thereof |
| CN115308406A (en) * | 2022-07-19 | 2022-11-08 | 杭州协合医疗用品有限公司 | Double-sample pad colloidal gold immunochromatographic test strip |
| CN115308406B (en) * | 2022-07-19 | 2024-06-04 | 杭州协合医疗用品有限公司 | Double-sample pad colloidal gold immunochromatography test strip |
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