CN102901821A - Colloidal gold detection card for detecting adrenoreceptor stimulant and sample processing method - Google Patents
Colloidal gold detection card for detecting adrenoreceptor stimulant and sample processing method Download PDFInfo
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- CN102901821A CN102901821A CN2012103732708A CN201210373270A CN102901821A CN 102901821 A CN102901821 A CN 102901821A CN 2012103732708 A CN2012103732708 A CN 2012103732708A CN 201210373270 A CN201210373270 A CN 201210373270A CN 102901821 A CN102901821 A CN 102901821A
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Abstract
The invention relates to a colloidal gold detection card for detecting an adrenoreceptor stimulant and a sample processing method, belonging to the technical field of clenbuterol hydrochloride detection. The detection card comprises a colloidal gold bonding pad coated with a monoclonal antibody colloidal gold marker, a cellulose nitrate membrane coated with clenobuterol hydrochloride, ractopamine, salbutamol, terbutaline hemisulfate, salbutamol sulphate, Cimaterol, dopamine hydrochloride, phenylethanolamine A protein conjugate and rabbit anti-rat IgG, a sample pad, absorbent cotton, a PVC rubber sheet and a plastic mold, etc., wherein one end of the PVC rubber sheet is orderly attached to the sample pad and the bonding pad, the middle of the PVC rubber sheet is attached to the cellulose nitrate membrane, and the other end of the PVC rubber sheet is attached to the absorbent cotton. According to the invention, 8 kinds of adrenoreceptor stimulants in urine, forage, animal tissues and liver can be simultaneously detected, the method is simple and rapid, and the result is accurate.
Description
Technical field
The present invention relates to colloidal-gold detecting-card and the sample treatment of 8 kinds of adrenoceptor agonists of a kind of detection, belong to clenbuterol hydrochloride detection technique field.
Background technology
The receptor,β activator can be strengthened heart contraction, expansion skeletal muscle blood vessel and bronchial smooth muscle, and veterinary science and medically being used for the treatment of is suffered a shock and bronchial spasm.Experiment confirm, adrenoceptor agonists class medicine has the effect of redistributing to nutriment in animal body, promote the animal muscle growth, promote the animal body protein precipitation, protein synthetic in the skeletal muscle particularly suppresses the synthetic and accumulation of animal tallow, thereby improves carcass quality, the speed of growth is accelerated, and lean meat increases more than 10% relatively.Effective because of it, economy return is high, thereby both at home and abroad many plants all illegally use this class medicine to make feed addictive, causes residual in the livestock and poultry muscle tissue, by food chain human body is produced toxic action, affects greatly the security of animal derived food.
The people has eaten the residual animal derived food of receptor,β activator, can cause the symptoms such as dizziness, palpitating speed, muscular tremor, palpitaition, expiratory dyspnea, and serious caused poisoning is very big to harm.The extensive poisoning that causes along with the meat products because of edible residual receptor,β activator of continuous generation in recent years, the countries and regions such as European Union, the U.S., China all successively legislation forbid using in Production of Livestock and Poultry the receptor,β activator of promotion growth of animal as feed addictive.The bulletin " the medicine kind catalogue of forbidding using in feed and drinking water for animals " (Ministry of Agriculture announces No. 176) that China Ministry of Agriculture, the Ministry of Public Health in 2002 send is clearly forbidden clenobuterol hydrochloride, Ractopamine, salbutamol, bricalin, salbutamol sulfate, Cimaterol, 7 kinds of adrenoceptor agonists uses in food animal of Dopamine hydrochloride, and in Dec, 2012, phenolethanolamine A was classified as " material of forbidding using " by No. 1519 bulletin of the Ministry of Agriculture in feed and drinking water for animals.
Adrenoceptor agonists is of a great variety, present immune colloid gold test card for adrenoceptor agonists in the animal derived food, main or individual event material or several materials of detecting, supervision department does not also know which kind of adrenoceptor agonists the raiser has used, there is certain blindness in detection, usually causes undetected.Therefore this collaurum that detects simultaneously 8 kinds of adrenoceptor agonists of forbidding using in feed and drinking water for animals of the present invention detects the connection jig very urgent market demand and market outlook widely.
Summary of the invention
The object of the present invention is to provide a kind of colloidal-gold detecting-card that can detect simultaneously 8 kinds of adrenoceptor agonists, overcome the one by one defective of individual event or several detections of prior art.
The colloidal-gold detecting-card of 8 kinds of adrenoceptor agonists of the present invention, comprise the collaurum pad that has been coated with monoclonal antibody colloid gold label thing, be coated with clenobuterol hydrochloride, Ractopamine, salbutamol, bricalin, salbutamol sulfate, Cimaterol, the nitrocellulose filter of Dopamine hydrochloride and phenolethanolamine A protein conjugate and rabbit anti-mouse igg, sample pad, absorbent wool, the compositions such as PVC offset plate and mould of plastics, end at the PVC offset plate adheres to sample pad successively, pad, nitrocellulose filter is pasted in the centre, and the other end adheres to absorbent wool.
The disposal route of the colloidal-gold detecting-card test sample of 8 kinds of the adrenoceptor agonists clenobuterol hydrochlorides of forbidding in feed and drinking water for animals, using, Ractopamine, salbutamol, bricalin, salbutamol sulfate, Cimaterol, salbutamol sulfate and phenolethanolamine A, test sample is urine sample, Feed Sample or the samples such as animal tissue, liver, for various test sample different disposal routes is arranged.
Disposal route for urine sample is: get urine sample to be checked, if when sample is relatively more muddy, and can be more than 3000 r/min, centrifugal 5 min or filtration are got supernatant and are detected.
Disposal route for Feed Sample is: accurately take by weighing Feed Sample behind 2 ± 0.05 g homogeneous in 50 mL centrifuge tubes; Add 10 mL, 0.05 M HCl, fully whirling motion 1 min; Add 10 mL, 0.05 M NaOH solution adjust pH to abundant whirling motion 30 s of 7-8, leave standstill 5 min, get 3-4 and drip supernatant and detect.
Disposal route for animal tissue, liver sample is: accurately take by weighing flesh tissue sample behind 2 ± 0.01 g homogeneous in 50 mL centrifuge tubes; Add 1 mL, 50 mM, pH7.0 Tris-HCl sample extract, whirling motion disperses to organizing fully; Through 100 ℃ of water-baths 10 minutes, be cooled to room temperature, discard emulsion layer, get limpid center section as specimen.
The collaurum of 8 kinds of adrenoceptor agonists of forbidding using in feed and drinking water for animals of the present invention detects the connection card can detect clenobuterol hydrochloride, Ractopamine, salbutamol, bricalin, salbutamol sulfate, Cimaterol, Dopamine hydrochloride and the phenolethanolamine A that contains in urine sample, Feed Sample or the samples such as animal tissue, liver simultaneously, this double tetrad test card is easy to preparation and saves testing cost, easy to use quick, the result is accurate, greatly improves the verification and measurement ratio of adrenoceptor agonists.
Description of drawings
Shell, test-strips arrangement figure and nitrate glass cellulose membrane in the enclosure that the collaurum of 8 kinds of adrenoceptor agonists of forbidding using in feed and drinking water for animals of Fig. 1 detects the connection card show the trace schematic diagram.1, well, 2, the clenobuterol hydrochloride protein conjugate detects band, 3, the Ractopamine protein conjugate detects band, 4, the salbutamol protein conjugate detects band, 5, the bricalin protein conjugate detects band, 6, quality control band, 7, detect fenestra, 8, test-strips, 9, the salbutamol sulfate protein conjugate detects band, 10, the Cimaterol protein conjugate detects band, 11, the Dopamine hydrochloride protein conjugate detects band, and 12, phenolethanolamine A protein conjugate detects band, 13, quality control band, 14, test-strips, 15, shell.
The collaurum of 8 kinds of adrenoceptor agonists of forbidding using in feed and drinking water for animals of Fig. 2 detects the test-strips structural representation of connection card.16, sample pad, 17, the collaurum film, 18, supporting back board, 19, nitrocellulose filter, 20, absorbent wool.
Embodiment
Embodiment 1: 1 and 2 further specify by reference to the accompanying drawings
The double tetrad colloidal-gold detecting-card of 8 kinds of adrenoceptor agonists, it is that two test- strips 8 and 14 are set in test card shell 15, the setting of the structure of test card shell 15 and test- strips 8 and 14 is referring to accompanying drawing 1, test card shell 15 is rectangular flat shell shape shell, long 7cm, wide 2cm, thick 0.5cm, shell wall thickness 0.1cm is made by engineering plastics.Test card shell 15 is connected with the shell block two halves by cap and consists of, and at test card shell 15 caps the fenestra 7 of detection and well 1 is arranged, and test- strips 8 and 14 is placed in the shell block of test card shell 15.
Test-strips 8 is fillet thin slices of sandwich construction: bottom is supporting back board 18, the stacked stickup nitrocellulose filter 19 in supporting back board 18 middle parts, be coated with four p-wires and a nature controlling line on the nitrocellulose filter 19, article one, detects and be with 2, be coated with the clenobuterol hydrochloride protein conjugate, second detects and is with 3, be coated with the Ractopamine protein conjugate, article three, detection is with 4, be coated with the salbutamol protein conjugate, article four, detection is with 5, is coated with the bricalin protein conjugate, and quality control band 6 is coated with rabbit anti-mouse igg antibody; Test-strips 14 is with test-strips 8, encrusting substance on its p-wire is followed successively by the detection of article one salbutamol sulfate protein conjugate and is with 9, second Cimaterol protein conjugate detects and is with 10, article three, the Dopamine hydrochloride protein conjugate detects and is with 11, article four, phenolethanolamine A protein conjugate detects and is with 12, detect the coated protein conjugate concentration of band and be approximately 1 mg/mL, quality control band 13 coated rabbit anti-mouse igg antibody concentration are approximately 0.1 mg/mL.Supporting back board 18 1 endlaps are put and are pasted sample pad 16, the stacked stickup absorbent wool 20 of the other end, nitrocellulose filter 19 two ends closely overlap with the front end face of sample pad 16 rear end faces and absorbent wool 20 respectively, at the clinch of sample pad 16 with nitrocellulose filter 19, sandwiched stickup number section collaurum film 17 between the two, collaurum film in the test-strips 8 is for containing anti-acid hydrochloride salt clenbuterol antibody, Anti-ractopamine antibody, the glass fibre element film of salbutamol antibody and bricalin antibody colloidal gold label, the collaurum film in the test-strips 14 is for containing sulfuric-resisting salbutamol antibody, west sieve's Bart antibody, the glass fibre element film of Dopamine hydrochloride antibody and phenolethanolamine A antibody colloidal gold label.The collaurum film is with behind antibody and the colloid gold label, with micro-rifle head it is uniformly coated on the glass fibre element film, and is for subsequent use after the oven dry.Collaurum film 17, sample pad 16 and absorbent wool 20 are 0.1 cm-0.2 cm with nitrocellulose filter 19 laps of splice.Absorbent wool 20 materials are thieving paper, and sample pad 16 materials are water adsorption glass cellulose membranes.
Embodiment 2: the disposal route of test sample
The collaurum of 8 kinds of adrenoceptor agonists of forbidding using in feed and drinking water for animals of the present invention detects the connection card and whether contains clenobuterol hydrochloride, Ractopamine, salbutamol, bricalin, salbutamol sulfate, Cimaterol, Dopamine hydrochloride and phenolethanolamine A for detecting animals urine sample, livestock feed sample or the samples such as animal tissue, liver simultaneously, guarantees accordingly the safety problem of animal derived food.
For various test sample different disposal routes is arranged:
Disposal route for urine sample is: get urine sample to be checked, if when sample is relatively more muddy, and can be more than 3000 r/min, centrifugal 5 min or filtration are got supernatant and are detected.
Disposal route for Feed Sample is: accurately take by weighing Feed Sample behind 2 ± 0.05 g homogeneous in 50 mL centrifuge tubes; Add 10 mL, 0.05 M HCl, fully whirling motion 1 min; Add 10 mL, 0.05 M NaOH solution adjust pH to abundant whirling motion 30 s of 7-8, leave standstill 5 min, get 3-4 and drip supernatant and detect.
Disposal route for animal tissue, liver sample is: accurately take by weighing flesh tissue sample behind 2 ± 0.01 g homogeneous in 50 mL centrifuge tubes; Add 1 mL, 50 mM, pH7.0 Tris-HCl sample extract, whirling motion disperses to organizing fully; Through 100 ℃ of water-baths 10 minutes, be cooled to room temperature, discard emulsion layer, get limpid center section as specimen.
The key reaction of immune colloid gold test card is to utilize the amplification that the immunoreactive character of high specific and antibody or the upper special marking thing (collaurum) of antigen (containing haptens) can occur between antigen and the antibody to come the ultramicron residue is carried out a kind of analytical approach of qualitative and quantitative detection.The antibody of the colloid gold label that moves at nitrocellulose filter, on calibration tape respectively with hydrochloric Clenbuterol protein conjugate, Ractopamine protein conjugate, salbutamol protein conjugate, bricalin protein conjugate, salbutamol sulfate protein conjugate, Cimaterol protein conjugate, Dopamine hydrochloride protein conjugate, phenolethanolamine A protein conjugate, and quality control band rabbit anti-mouse igg antibody response, form the aubergine band.Corresponding clenobuterol hydrochloride to be checked is arranged in sample, Ractopamine, salbutamol, bricalin, salbutamol sulfate, Cimaterol, Dopamine hydrochloride and phenolethanolamine A exist, then sample namely reacts with corresponding antibody after adding, and not can with corresponding detection with contained clenobuterol hydrochloride protein conjugate, the Ractopamine protein conjugate, the salbutamol protein conjugate, the bricalin protein conjugate, the salbutamol sulfate protein conjugate, the Cimaterol protein conjugate, the Dopamine hydrochloride protein conjugate, the reaction of phenolethanolamine A protein conjugate, thus detection line does not develop the color.
The present invention is through test, use the collaurum detection connection of 8 kinds of adrenoceptor agonists to block when detecting, draw sample supernatant to be checked with dropper, adding 3-4 drips in well 1,3-5 min behind the application of sample, observe to detect the colour developing district of fenestra 7, judged result preserves that the result can not change after 30 minutes.Through 200 times on probation, except 2 examples were indefinite, all the other 198 examples had all reached promising result, rate of accuracy reached 99%, therefore, it is simple that the present invention has method, convenient and swift, the characteristics such as testing result is stable, accurate are the innovations on adrenoceptor agonists detects in the animal derived food.
Claims (4)
1. the colloidal-gold detecting-card of adrenoceptor agonists, it is characterized in that being coated with monoclonal antibody colloid gold label thing the collaurum pad, be coated with nitrocellulose filter, sample pad, absorbent wool, PVC offset plate and the mould of plastics of clenobuterol hydrochloride, Ractopamine, salbutamol, bricalin, salbutamol sulfate, western sieve Bart, Dopamine hydrochloride and phenolethanolamine A protein conjugate and rabbit anti-mouse igg, end at the PVC offset plate adheres to sample pad, pad successively, nitrocellulose filter is pasted in the centre, and the other end adheres to absorbent wool.
2. the sample treatment of the colloidal-gold detecting-card of adrenoceptor agonists claimed in claim 1, it is characterized in that for the disposal route of urine sample being: get urine sample to be checked, if when sample is relatively more muddy, can be more than 3000 r/min, centrifugal 5 min or filtration are got supernatant and are detected.
3. the sample treatment of the colloidal-gold detecting-card of adrenoceptor agonists claimed in claim 1 is characterized in that for the disposal route of Feed Sample being: accurately take by weighing Feed Sample behind 2 ± 0.05g homogeneous in 50 mL centrifuge tubes; Add 10 mL, 0.05 M HCl, fully whirling motion 1 min; Add 10 mL, 0.05 M NaOH solution adjust pH to abundant whirling motion 30 s of 7-8, leave standstill 5min, get 3-4 and drip supernatant and detect.
4. the sample treatment of the colloidal-gold detecting-card of adrenoceptor agonists claimed in claim 1 is characterized in that for the disposal route of animal tissue, liver sample being: accurately take by weighing flesh tissue sample behind 2 ± 0.01 g homogeneous in 50 mL centrifuge tubes; Add 1 mL, 50 mM, pH7.0 Tris-HCl sample extract, whirling motion disperses to organizing fully; Through 100 ℃ of water-baths 10 minutes, be cooled to room temperature, discard emulsion layer, get limpid center section as specimen.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012103732708A CN102901821A (en) | 2012-09-27 | 2012-09-27 | Colloidal gold detection card for detecting adrenoreceptor stimulant and sample processing method |
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| CN2012103732708A CN102901821A (en) | 2012-09-27 | 2012-09-27 | Colloidal gold detection card for detecting adrenoreceptor stimulant and sample processing method |
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- 2012-09-27 CN CN2012103732708A patent/CN102901821A/en active Pending
Patent Citations (9)
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| CN1402004A (en) * | 2002-09-27 | 2003-03-12 | 江西中德生物工程有限公司 | Method for immunochromatography one-step detecting beta-adrenin agonist, medicine,and preparation of test paper therefor |
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Application publication date: 20130130 |