CN102323262A - Phenolethanolamine A rapid detection chip, and detection method thereof - Google Patents
Phenolethanolamine A rapid detection chip, and detection method thereof Download PDFInfo
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- CN102323262A CN102323262A CN201110227826A CN201110227826A CN102323262A CN 102323262 A CN102323262 A CN 102323262A CN 201110227826 A CN201110227826 A CN 201110227826A CN 201110227826 A CN201110227826 A CN 201110227826A CN 102323262 A CN102323262 A CN 102323262A
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Abstract
The invention discloses a phenolethanolamine A rapid detection chip, and a detection method thereof. The invention belongs to the technical field of phenolethanolamine A detections. The phenolethanolamine A rapid detection chip comprises a detection strip in a housing. The detection strip comprises components orderly stuck on a supporting backing, wherein the components comprise a sample pad, a colloidal gold membrane or a latex particle membrane, a nitrocellulose filter and a water absorbing membrane. The colloidal gold membrane or the latex particle membrane is a fiberglass membrane containing phenolethanolamine A monoclonal antibody colloidal gold or latex particle markers. Two detection lines are transversally arranged on the nitrocellulose filter, wherein one line is a detection line containing phenolethanolamine A protein conjugate, and the other is a control ling containing anti-rabbit or anti-rat antibodies. The detection chip provided by the invention is advantaged in that: with the detection chip, phenolethanolamine A content in samples such as feedstuffs, tissue samples, urine or blood serum can be directly detected with an immunologic method. The phenolethanolamine A rapid detection chip provided by the invention is easy to prepare, convenient to use, and provides accurate results.
Description
Technical field
The present invention relates to a kind of phenolethanolamine A rapid detection card and detection method thereof, is a kind of appliance that detects phenolethanolamine A, belongs to phenolethanolamine A detection technique field.
Background technology
Modern animal husbandry provides extremely abundant various food for people; But there are a large amount of phenomenons of culturing adjuvant of using in the animal husbandry simultaneously; Particularly use the beta receptor activator in the Domestic Animal feed, example hydrochloric acid Clenbuterol (clenbuterol hydrochloride), Ractopamine etc., this type adjuvant have the effect that promotes lipolysis and increase muscle growth can improve lean meat percentage; The people toxicity symptom can occur after having eaten the livestock products of raising with this type adjuvant.Therefore, China has expressly provided and has forbidden to use.Phenolethanolamine A is that latest find in 2011 is added on the beta-receptor excitant in the feed, has another name called Ke Lunba amine, and molecular formula is C
19H
24N
2O
4Chemical structural formula is 2-[4-(4-nitrobenzophenone) butyl-2-hydroxy is amino]-1-methoxybenzene ethanol; Be the isomers of Formoterol, it is the accessory substance of the synthetic Ractopamine of Lilly Co., Eli., has with clenobuterol hydrochloride effect and the effect identical with Ractopamine.
At present, the data of phenolethanolamine A detection method aspect is less in research organization or the feed, mainly is that utilization high performance liquid chromatography and LC-MS etc. carry out qualitative and quantitative analysis.These methods require high to experimental instrument and equipment, complex operation is high to operating personnel's professional knowledge requirement, wastes time and energy.Therefore setting up a kind of detection method that detects phenolethanolamine A in the food accurately and reliably again quickly and easily is very important.
The present invention sends out bright a kind of based on having antigenic immunological detection method after the small-molecule substance phenolethanolamine A coupling protein matter; Take immunology competition ratio juris; A kind of quick detection reagent of association colloid gold or latex particle labelling technique and immunochromatographic method design can detect the phenolethanolamine A in the samples such as feed or tissue fast specifically.The phenolethanolamine A content in the sample is judged in the colour developing that has or not according to test card sample detection line (T) and nature controlling line (C).Reaction is to utilize phenolethanolamine A protein conjugate antigenicity and a kind of detection method of setting up, has the advantage of high specificity, the flase drop problem that can avoid the false positive of other detection method to cause.Test card is simple to operate simultaneously need not special instrument and equipment fast, is fit to execute-in-place.Because sample size needs less, easyly be applicable to the examination of great amount of samples fast, the result is accurate, and is highly sensitive, and accuracy rate is big.
Summary of the invention
The object of the present invention is to provide a kind of appliance that can detect phenolethanolamine A; Directly the method for applied immunology detects feed or tissue; Content like phenolethanolamine A in the samples such as muscle, liver or kidney; Overcome the shortcomings such as complex operation that existing chromatography detects, set up based on the specific detection method of immunology.
Technical scheme of the present invention: a kind of phenolethanolamine A rapid detection card, it is for being provided with test-strips 2 in the test card shell 1 of rectangular flat shell shape, and the test card case surface has fenestra of detection 3 and well 4; Test-strips 2 is formed by pasting sample pad 8, collaurum film or latex particle film 7, nitrocellulose filter 6 and 9 of absorbing membranes above that successively through adhesive sticker on the supporting backboard 5; The stacked stickup nitrocellulose filter 6 in supporting backboard middle part; One endlap of supporting backboard is put and is pasted absorbing membrane 9; The stacked stickup sample pad 8 of the other end; Absorbing membrane is inner to overlap with nitrocellulose filter respectively with sample pad the inner separately, at the clinch of sample pad and nitrocellulose filter, inserts and puts between the two and pastes one section collaurum film or latex particle film 7; Collaurum film or latex particle film are the glass fibre membrane that contains anti-phenolethanolamine A antibody colloidal gold or latex particle mark; There is one to detect band and a quality control band on the nitrocellulose filter; Be successively: the detection that contains phenolethanolamine A protein conjugate is with 10, contains the quality control band 11 of anti-rabbit antibody or anti-mouse antibody, and test-strips is inserted in the test card shell; Sample pad is over against well, and nitrocellulose filter is over against detecting fenestra.
Adopt above-mentioned phenolethanolamine A rapid detection card, test sample is feed, tissue sample, urine or serum, and disposal route is following:
A, urine sample or serum: get urine or serum directly as test sample, if urine sample is muddy shape, must be earlier through filter paper filtering or at 15 ℃, centrifugal 10min under the 4000r/min condition;
B, feed: take by weighing 0.5-1g feed sample, add 1-2mL sample extract, in test tube, stir 1 min, leave standstill 10 min after, get limpid upper strata liquid and make specimen;
C, tissue sample (muscle, liver or kidney) get that 0.5-1g shreds or animal tissue's sample of homogenate, place the test tube that contains 1-2mL sample extract, tighten the pipe lid; Behind jolting sample 1 min; Through 100 ℃ of water-baths 5 minutes, be cooled to room temperature, get limpid center section and make specimen.
The sample extract is: 0.01mol/L PBST solution (pH7.4) includes 0.15mol/LNaCl, mass concentration 0.5% Tween-20.
When detection be with 10 with quality control band 11 manifest the brownish red trace simultaneously, testing result is negative, representes that phenolethanolamine A content does not exceed standard in the sample to be detected.
Be with 10 Show Colors not when detection, have only quality control band 11 to manifest the brownish red trace, testing result is positive, representes phenolethanolamine A content overproof in the sample to be detected.
Be with 10 all not develop the color with quality control band 11 when detection, show that then test-strips lost efficacy.
Beneficial effect of the present invention: advantage of the present invention is to detect the phenolethanolamine A that contains in feed, the tissue sample samples such as (muscle, liver or kidneys), is easy to preparation and saves testing cost, and is easy to use, detects fast, and highly sensitive, the result is accurate.
Description of drawings
Fig. 1 phenolethanolamine A rapid detection card shell and test-strips arrangement figure in the enclosure.1, shell, 2, test-strips, 3, detect fenestra, 4, well.
Show the trace synoptic diagram on the nitrate glass tunica fibrosa of Fig. 2 test-strips.6, nitrocellulose filter, 10, detect band, 11, quality control band.
The test-strips structural representation of Fig. 3 phenolethanolamine A rapid detection card.5, supporting backboard, 6, nitrocellulose filter, 7, collaurum film or latex particle film, 8, sample pad, 9, absorbing membrane.
Embodiment
The present invention combines specific embodiment to further specify as follows referring to accompanying drawing:
A kind of phenolethanolamine A rapid detection card, it is that test-strips 2 is set in test card shell 1, the setting of the structure of test card shell 1 and test-strips 2 is referring to accompanying drawing 1.Test card shell 1 is rectangular flat shell shape shell, long 70mm, and wide 20mm, thick 5mm, shell wall thickness 1mm is processed by engineering plastics.Test card shell 1 is connected with the shell block two halves by cap and constitutes, and fenestra of detection 3 and well 4 are arranged on test card shell 1 cap.Test-strips 2 is placed in the shell block of test card shell 1.
Test-strips 2 is fillet thin slices of sandwich construction, and sheet is about 60mm, the wide about 4mm of sheet, the about 2.5mm of thickness.Test-strips 2 is sandwich constructions: bottom is a supporting backboard 5, is the igelite thin slice, thick about 0.5mm, long 60mm, wide 4mm; The stacked stickup nitrocellulose filter 6 in supporting backboard 5 middle parts, cellulose nitrate thickness 0.5mm, long 20-25mm, wide 4mm.Two isolated horizontal demonstration trace bands are arranged on the nitrocellulose filter 6, show in the trace band referring to 2, two in accompanying drawing, one is to detect to be with 10, contains phenolethanolamine A protein conjugate; Another is a quality control band 11, contains anti-rabbit antibody or anti-mouse antibody.Stacked stickup absorbing membrane 9 on supporting backboard 5 one ends; Stacked stickup sample pad 8 on supporting backboard 5 the other end; Absorbing membrane 9 the inners overlap with nitrocellulose filter 6 one ends respectively with sample pad 8 the inners separately; At the clinch of sample pad 8 with nitrocellulose filter 6, insert and put between the two and paste one section collaurum film or latex particle film 7, collaurum film or latex particle film 7 are the glass fibre membranes that contain anti-phenolethanolamine A monoclonal antibody collaurum or latex particle label.Collaurum film or latex particle film 7 thick 0.5-0.8mm, long 5mm, wide 4mm.Sample pad 8 thick 0.5-0.8mm, long 20mm, wide 4mm.Absorbing membrane 9 thick 0.5-0.8mm, long 16-20mm, wide 4mm.Collaurum film or latex particle film 7, sample pad 8 and absorbing membrane 9 are 1-2mm with the nitrocellulose filter lap of splice.Absorbing membrane 9 materials are thieving paper, and sample pad 8 materials are water adsorption glass tunica fibrosas.
Phenolethanolamine A test card of the present invention is used for detecting the detection of the phenolethanolamine A of samples such as feed, tissue sample (muscle, liver or kidney), urine sample or serum.
Embodiment 2
The disposal route of sample is following:
A, urine sample or serum: urine or serum is directly as test sample, if urine sample is muddy shape, must be earlier through filter paper filtering or under 15 ℃, 4000r/min condition centrifugal 10min;
B, feed: take by weighing 1g feed sample, add 2mL sample extract, in test tube, stir 1 min, leave standstill 10 min after, get limpid upper strata liquid and make specimen.
C, tissue sample: (like muscle, liver or kidney) got that 1g shreds or animal tissue's sample of homogenate, placed the test tube that contains 2mL sample extract, tightens the pipe lid; Behind the jolting sample 1min; Through 100 ℃ of water-baths 5 minutes, be cooled to room temperature, get limpid center section and make specimen.
Sample extract PBST is 0.01mol/L, the pH7.4 PBST solution that contains 0.15mol/LNaCl, contains 0.5% Tween-20.
When using phenolethanolamine A rapid detection card to detect; Earlier sample liquid to be detected is instilled into from well 4 on the sample pad 8 of phenolethanolamine A rapid detection card; Because syphonic effect principle; Drive contained anti-phenolethanolamine A monoclonal antibody collaurum of sample liquid to be detected and collaurum film or latex particle film 7 or latex particle labels together to nitrocellulose filter 6 diffusions, observations in the 5-10min.
The key reaction of test card is immunologic antigen and antibody response, on nitrocellulose filter, moves
The antibody of collaurum or latex particle mark, on calibration tape respectively with contain phenolethanolamine A protein conjugate, and the anti-rabbit antibody of quality control band or anti-mouse antibody reaction forms the brownish red band.If there is phenolethanolamine A to be checked to exist in the sample, after sample adds, promptly react, and can not react with contained phenolethanolamine A protein conjugate with detection with antibody, do not develop the color thereby detect band.The result of key reaction has following 3 kinds of situation:
1, when detection be with 10 and quality control band 11 manifest the brownish red trace simultaneously, the expression testing result negative, explain that phenolethanolamine A content does not exceed standard in the sample to be detected.
2, be with 10 Show Colors not when detection, have only quality control band 11 to manifest the brownish red trace, testing result is positive, and phenolethanolamine A content overproof in the sample to be detected is described.
3, when detection be with 10 and quality control band 11 all do not develop the color, show that then detecting strip lost efficacy.
Claims (2)
1. phenolethanolamine A rapid detection card, it is characterized in that: in the test card shell (1) of rectangular flat shell shape, test-strips (2) is set, the test card case surface has detection fenestra (3) and well (4); Test-strips (2) is upward pasted sample pad (8), collaurum film or latex particle film (7), nitrocellulose filter (6) and absorbing membrane (9) above that successively through adhesive sticker by supporting backboard (5) and is formed; Supporting backboard middle part stacked stickup nitrocellulose filter (6); One endlap of supporting backboard is put and is pasted absorbing membrane (9); The stacked stickup sample pad of the other end (8); Absorbing membrane is inner to overlap with nitrocellulose filter respectively with sample pad the inner separately, at the clinch of sample pad and nitrocellulose filter, inserts and puts between the two and pastes one section collaurum film or latex particle film (7); Collaurum film or latex particle film are the glass fibre membrane that contains anti-phenolethanolamine A antibody colloidal gold film or latex particle mark; There is one to detect band and a quality control band on the nitrocellulose filter; Be successively: contain the detection band (10) of phenolethanolamine A protein conjugate, contain the quality control band (11) of anti-rabbit antibody or anti-mouse antibody, test-strips is inserted in the test card shell; Sample pad is over against well, and nitrocellulose filter is over against detecting fenestra.
2. with the detection method of the described phenolethanolamine A of claim 1 rapid detection card, it is characterized in that the content of phenolethanolamine A in the direct test sample of method of applied immunology, test sample is feed, tissue sample, urine or serum, disposal route is following:
A, urine sample or serum: urine or serum is directly as test sample, if urine sample is muddy shape, must be earlier through filter paper filtering or under 15 ℃, 4000r/min condition centrifugal 10min;
B, feed: take by weighing 0.5-1g feed sample, add 1-2mL sample extract, in test tube, stir 1 min, leave standstill 10 min after, get limpid upper strata liquid and make specimen;
C, tissue sample: like muscle, liver or kidney; Get that 0.5-1g shreds or the animal tissue of homogenate: meat, liver or kidney sample, place the test tube that contains 1-2mL sample extract, tighten the pipe lid; Behind jolting sample 1 min; Through 100 ℃ of water-baths 5 minutes, be cooled to room temperature, get limpid center section and make specimen;
The sample extract is: pH7.4,0.01mol/L PBST solution include the NaCl of 0.15mol/L, mass concentration 0.5% Tween-20;
When detection be with 10 with quality control band 11 manifest the brownish red trace simultaneously, testing result is negative, representes that phenolethanolamine A content does not exceed standard in the sample to be detected;
Be with 10 Show Colors not when detection, have only quality control band 11 to manifest the brownish red trace, testing result is positive, representes phenolethanolamine A content overproof in the sample to be detected;
Be with 10 all not develop the color with quality control band 11 when detection, show that then test-strips lost efficacy.
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| CN201110227826A CN102323262A (en) | 2011-08-10 | 2011-08-10 | Phenolethanolamine A rapid detection chip, and detection method thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102527358A (en) * | 2012-02-01 | 2012-07-04 | 宁波市农业科学研究院 | Preparation method and application of phenylethanolamine A molecularly-imprinted material |
| CN102735834A (en) * | 2012-05-11 | 2012-10-17 | 中国兽医药品监察所 | Enzyme-linked immunoassay kit for detecting phenylethanolamine A |
| CN102901818A (en) * | 2012-09-27 | 2013-01-30 | 江苏维赛科技生物发展有限公司 | Five-conjugate testing card for detecting beta-stimulant drugs and preparation method thereof |
| CN103792226A (en) * | 2012-11-03 | 2014-05-14 | 江苏维赛科技生物发展有限公司 | Chemiluminescence immunodetection kit used for detecting phenylethanolamine A |
| CN104914250A (en) * | 2015-06-10 | 2015-09-16 | 长沙安迪生物科技有限公司 | Rapid mutton detection card and detection method thereof |
| CN105092849A (en) * | 2014-09-16 | 2015-11-25 | 北京勤邦生物技术有限公司 | Test paper strip and method for detecting phenylethanolamine A |
| CN105283761A (en) * | 2013-06-10 | 2016-01-27 | 旭化成纤维株式会社 | Immunochromatographic diagnosis kit |
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| CN1888906A (en) * | 2006-07-17 | 2007-01-03 | 长沙安迪生物科技有限公司 | Clenbuterol hydrochloride-Ractopamine dual union detection card and method for processing detected samples thereof |
| CN101915842A (en) * | 2010-07-26 | 2010-12-15 | 河南省科学院生物研究所有限责任公司 | Binary detection test strip of beta-exhilarant ractopamine and salbutamol and preparation method thereof |
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Patent Citations (2)
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| CN1888906A (en) * | 2006-07-17 | 2007-01-03 | 长沙安迪生物科技有限公司 | Clenbuterol hydrochloride-Ractopamine dual union detection card and method for processing detected samples thereof |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102527358A (en) * | 2012-02-01 | 2012-07-04 | 宁波市农业科学研究院 | Preparation method and application of phenylethanolamine A molecularly-imprinted material |
| CN102527358B (en) * | 2012-02-01 | 2014-02-26 | 宁波市农业科学研究院 | Preparation method of phenylethanolamine A molecularly-imprinted material |
| CN102735834A (en) * | 2012-05-11 | 2012-10-17 | 中国兽医药品监察所 | Enzyme-linked immunoassay kit for detecting phenylethanolamine A |
| CN102735834B (en) * | 2012-05-11 | 2014-09-03 | 中国兽医药品监察所 | Enzyme-linked immunoassay kit for detecting phenylethanolamine A |
| CN102901818A (en) * | 2012-09-27 | 2013-01-30 | 江苏维赛科技生物发展有限公司 | Five-conjugate testing card for detecting beta-stimulant drugs and preparation method thereof |
| CN103792226A (en) * | 2012-11-03 | 2014-05-14 | 江苏维赛科技生物发展有限公司 | Chemiluminescence immunodetection kit used for detecting phenylethanolamine A |
| CN105283761A (en) * | 2013-06-10 | 2016-01-27 | 旭化成纤维株式会社 | Immunochromatographic diagnosis kit |
| CN105283761B (en) * | 2013-06-10 | 2018-02-09 | 旭化成株式会社 | Immunochromatography diagnostic kit |
| CN105092849A (en) * | 2014-09-16 | 2015-11-25 | 北京勤邦生物技术有限公司 | Test paper strip and method for detecting phenylethanolamine A |
| CN104914250A (en) * | 2015-06-10 | 2015-09-16 | 长沙安迪生物科技有限公司 | Rapid mutton detection card and detection method thereof |
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Application publication date: 20120118 |