CN101403756A - Fast detection reagent for melamine - Google Patents
Fast detection reagent for melamine Download PDFInfo
- Publication number
- CN101403756A CN101403756A CNA200810202602XA CN200810202602A CN101403756A CN 101403756 A CN101403756 A CN 101403756A CN A200810202602X A CNA200810202602X A CN A200810202602XA CN 200810202602 A CN200810202602 A CN 200810202602A CN 101403756 A CN101403756 A CN 101403756A
- Authority
- CN
- China
- Prior art keywords
- melamine
- polyester film
- sample
- reagent according
- require described
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Abstract
The invention provides a melamine quick detection reagent which adopts highly specific antibody and antigen reaction and immunity-chromatographic technology to achieve the aim of quickly detecting the melamine in a sample. The reagent has the advantages of simple, convenient and quick operation and the like, can be applied to the production fields such as bread cultivation, breeding, animal husbandry, food processing and the like for quickly detecting the melamine in the food and feed in-situ and also provides simple and convenient means for the safety detection of household food.
Description
Technical field
Fast detection reagent for melamine, the applied immunology principle, immune response by collaurum relends and helps immunochromatography technique to reach to detect the melamine purpose, be applied to the melamine in fast detecting sweet milk, dairy products, bean product, feed, baby food, the pet food.
Background technology
Because melamine is a kind of industrial chemicals, toxicity is slight, and the oral median lethal dose of rat is greater than 3 gram/kg body weight.An experiment according to 1945: the melamine of heavy dose is fed to not observing tangible intoxicating phenomenon behind rat, rabbit and the dog.Animal is taken in the infringement that melamine can cause reproduction, urinary system for a long time, bladder, kidney portion calculus, and can further bring out carcinoma of urinary bladder." international chemicals security manual " the 3rd volume and international chemicals safety card that IPCS in 1994 and European Commission EC compile in collaboration with also only illustrate: taking in melamine for a long time or repeatedly in a large number may exert an influence to kidney and bladder, causes producing calculus.Yet the initial investigation results of U.S.'s pet food contamination accident in 2007 is thought: mixed≤gluten powder of 6.6% melamine causes pet food to be poisoned.Zoopery is the result show, metabolism is very fast and can not retain in animal body for it, mainly influences urinary system.There is tangible dose-effect relationship between agent of melamine amount and the clinical disease.Melamine maximal tolerance dose in infants is 15 milligrams of per kilogram milk powder.The risk assessment that the expert carries out contaminated baby formula milk powder shows, is example with the baby of 7 kilograms of body weight, suppose to take in every day milk powder 150 and restrains, and its safe pre-value is that maximal tolerance dose is 15 milligrams of/kilogram milk powder.According to the standard of U.S.'s food and FAD, melamine tolerable every day intake is 0.63 milligram/kg body weight every day.
Because the defective of food and feed industry protein content method of testing, melamine also often are used as food additives by illegal businessman, with the protein content index in the lifting food inspection, so melamine also is " extract of protein " by person.Protein mainly is made up of amino acid.The average nitrogen content of protein is about 16%, and the nitrogen content of melamine is about 66%.General protein method of testing " Kjeldahl " is to estimate protein content by measuring nitrogen content, therefore, adds melamine and can make that the protein test content of food is higher, thereby make the test of poor quality food by Food Inspection mechanism.Someone estimates makes test protein content increase one percentage point in plant protein powder and feed, has only 20% of true protein raw materials with the cost of melamine.Melamine does not have any aroma and flavor as a kind of white crystalline powder, so be difficult for being found after mixing.The melamine nitrogen content is 66.6%, is 151 times of milk, is 23 times of milk powder.Add 0.1 gram melamine in every 100g milk, just can improve 0.625% protein in theory.Detection scheme: in the national standard that existing milk powder detects, mainly carry out detections such as protein, fat, bacterium.Melamine belongs to industrial chemicals, is not allow to add in the food, so existing standard can not comprise corresponding contents.That is the melamine detection there is no national standard at present.Milk and milk powder add melamine, mainly are because it can pretend to be protein.Food all will detect protein content in accordance with regulations.If protein is insufficient, illustrate milk convert water too much, the powder that too many anything else is arranged in the milk powder is described.But protein is not easy very much to detect, and biochemists are just found out an indirect way: because protein is nitrogenous, so as long as measure nitrogen content in the food, just can extrapolate protein content wherein.Therefore the milk powder that added melamine just be difficult to detect its protein defective the false albumen of Here it is melamine.
The method that detects melamine at present mainly contains 3 kinds, high performance liquid chromatography (HPLC), combined gas chromatography mass spectrometry (GC-MS) and enzyme-linked immuno assay (ELISA).The subject matter that exists is that equipment has high input, and the running time is long, and specialty requires high, is not suitable for the requirement of field quick detection, is not suitable for the requirement that batch samples detects.
Summary of the invention
Fast detection reagent for melamine, the applied immunology principle, the immune response by collaurum relends and helps immunochromatography technique to reach to detect the melamine purpose, and it is quick to have testing process, easy to operate, detects obvious advantages such as cost is low.
The inventive method is to realize with principle of work by the following technical programs.Specific antibody (melamine antibody) is marked on the help of indicator particles, and label is attached on the polyester film; With corresponding comlete antigen (melamine-BSA bond) bag by and fixedly on the detection zone of chromatographic film (T), chromatographic film sticks on the base plate, and there is the sample processing part of using for application of sample at top, and is terminal by forming strip for the filter paper that helps chromatography to use, or strip is inlaid into moulds in the card, form reagent.During detection, because immune response has taken place in the golden labelled antibody of corresponding antigens (melamine) and redissolution in the tested sample, envelope antigen can't be competed labelled antibody when this moment, the mixed liquor chromatography was to detection zone, response line does not appear in the detection zone, promptly positive, do not have tested antigen (melamine) as tested sample on the contrary, when then the antibody chromatography of mark is to detection zone and the antigen on the coated film in conjunction with forming purplish red line, promptly negative.In order to prove reagent validity and easily interpretation, be nature controlling line at Quality Control district (C) bag by goat anti-rabbit igg respectively, so how nature controlling line C occurs all the time regardless of the result.
Fast detection reagent for melamine, form by following part: chromatographic film, the polyester film that contains help of indicator particles, sample processing part, filter paper and base plate, detect the melamine in sweet milk, dairy products, bean product, feed, baby food, the pet food.
Description of drawings
Accompanying drawing 1 is the structural representation of the embodiment of the invention.
1. nitrocellulose filters among the figure, 2. filter paper, 3. sample processing part, 4. base plate, 5. contain the polyester film of colloid gold particle, 6. reagent shell.
Accompanying drawing 2 is positive findings synoptic diagram of the embodiment of the invention.
Accompanying drawing 3 is negative findings synoptic diagram of the embodiment of the invention.
Embodiment
The present invention is further described below in conjunction with drawings and Examples.
Embodiment, fast detection reagent kit for melamine.
In Fig. 1, the nitrocellulose filter (1) that is coated with melamine-BSA bond sticks in the middle of the base plate (4), and its top is pasted with filter paper (2), and sample processing part (3) is pasted in its bottom.The antibody colloidal gold particle of the anti-melamine of mark is attached to (5) on the polyester film.The base plate (4) that is pasted with nitrocellulose filter (1), thieving paper (2) and filter sample pad glass (3) is placed on reagent shell (6), and reagent shell (6) lid is tight.
With certain density melamine-BSA bond and goat anti-rabbit igg bag by the detection line on nitrocellulose filter and the position of control line.
2. the nitrocellulose filter that makes is sticked on the base plate successively with sample processing part, filter paper.
3. in this base plate being assemblied at the bottom of the reagent shell, the reagent casing cover that closes assembles.
4. anti-melamine antibody labeling Preparation of Colloidal Gold: reduce four chloraurides with trisodium citrate and prepare particle diameter 40～60nm collaurum, regulate behind the pH every milliliter of collaurum and 10-15ug anti-melamine antibody and be mixed into row labels and under 450nm, survey OD.
5. the collaurum with the 2-3OD anti-melamine antibody labeling of 10-20ul is added on the polyester film and oven dry, promptly adheres to the colloid gold particle through mark on the polyester film.
6. detection step:
A. draw a certain amount of milk 7000rpm centrifugal degreasing, promptly get sample to be checked.
B. take out kit, keep flat on the table.
C. draw the sample that mixes with suction pipe, drip 2-3 and drip in well, and pick up counting.
D. in 5 minutes, read the result.
E. the result judges: the positive result of Fig. 2 (red lines appear in control line C position, and detection line T position is judged to the positive when not showing red lines); The negative result of Fig. 3 (red lines appear in detection line T position, and red lines also appear in control line C position, are judged to feminine gender).
7. performance index
Sensitivity: use present embodiment, melamine concentration is positive more than 1000ng/ml.
According to disclosure of the present invention, those skilled in the art need not too much test and can implement the present invention's content required for protection, and produce a desired effect.Embodiment disclosed by the invention only describes the present invention, but is not to be construed as limiting the invention.Those skilled in the art are with similar substitute or transformation; or substitute reagent described here with some reagent like the chemofacies; as replacing collaurum with color latex; or related content of the present invention changed; but do not exceed spirit of the present invention, scope and thought, all fall into the scope of protection of present invention.
Claims (9)
1. fast detection reagent for melamine, it is characterized in that, adopt the antibody antigen reaction and the immunochromatography technique of high degree of specificity,, reach the melamine in the fast detecting sample by the principle of antibody competition in conjunction with the melamine that may contain in melamine-BSA conjugate and the sample.
2. fast detection reagent for melamine is characterized in that, detectable is made up of following part: chromatographic film, the polyester film that contains help of indicator particles, sample processing part, filter paper and base plate.
3. require described reagent according to right 1, it is characterized in that, chromatographic film is a kind of in nitrocellulose filter, the cellulose acetate membrane.
4. require described reagent according to right 1, it is characterized in that, help of indicator particles is a kind of in colloid gold particle, collargol particle, fluorescent grain, the magnetic-particle.
5. require described reagent according to right 1, it is characterized in that, sample processing part is that glass fibre, polyester film, glass fibre add polyester film, glass fibre and add filter paper, polyester film and add filter paper, glass fibre and add polyester film and add a kind of in the filter paper.
6. require described reagent according to right 1, it is characterized in that with the anti-bag of goat-anti rabbit and rabbit by and fixedly on the chromatographic film.
7. require described reagent according to right 1, it is characterized in that, the polyester film that contains help of indicator particles is with the polyester film after the even distribution of the colloid gold particle of anti-melamine antibody labeling.
8. require described reagent according to right 1, it is characterized in that sample processing part was handled with damping fluid and be used to adjust the sample pH value value.
9. require described reagent according to right 1, it is characterized in that, the sample of detection is a kind of in sweet milk, dairy products, bean product, feed, baby food, the pet food.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200810202602XA CN101403756A (en) | 2008-11-12 | 2008-11-12 | Fast detection reagent for melamine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200810202602XA CN101403756A (en) | 2008-11-12 | 2008-11-12 | Fast detection reagent for melamine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101403756A true CN101403756A (en) | 2009-04-08 |
Family
ID=40537832
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA200810202602XA Pending CN101403756A (en) | 2008-11-12 | 2008-11-12 | Fast detection reagent for melamine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101403756A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106771238A (en) * | 2016-12-14 | 2017-05-31 | 北京勤邦生物技术有限公司 | Detect fluorescent micro-ball immune chromatography test paper strip of melamine residual and its preparation method and application |
-
2008
- 2008-11-12 CN CNA200810202602XA patent/CN101403756A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106771238A (en) * | 2016-12-14 | 2017-05-31 | 北京勤邦生物技术有限公司 | Detect fluorescent micro-ball immune chromatography test paper strip of melamine residual and its preparation method and application |
| CN106771238B (en) * | 2016-12-14 | 2018-08-10 | 北京勤邦生物技术有限公司 | Detect the fluorescent micro-ball immune chromatography test paper strip and its preparation method and application of melamine residual |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101261274A (en) | Method for combining colloidal gold immune leaching and immunity-chromatography for rapidly detecting medicament residue | |
| CN101865924B (en) | Method for detecting fumonisins based on immuno-magnetic bead combined enzyme-linked immunosorbent assay | |
| CN105334319B (en) | A kind of test strips for detecting zearalenone in breast | |
| CN102087284A (en) | Preparation method of reagent board for rapidly detecting aflatoxin B1 | |
| CN101173925B (en) | Colloidal gold test paper card for detecting quinolones medicament residue | |
| CN101661043A (en) | Method for detecting fumonisin by colloidal gold immunochromatographic test | |
| CN101609095A (en) | A kind of colloidal gold immunochromatographimethod method and colloid gold immune test strip of new fast quantifying and detecting Listeria monocytogenes | |
| CN103575887B (en) | A kind of test card and application thereof detecting aflatoxin B1 | |
| CN104569399A (en) | Test strip for testing ochratoxin A and application of test strip | |
| Zhang et al. | Rapid and sensitive pathogen detection platform based on a lanthanide-labeled immunochromatographic strip test combined with immunomagnetic separation | |
| CN108918849B (en) | Method for rapidly detecting aflatoxin in medicinal materials and test paper card used by same | |
| CN101434655B (en) | Preparation of egg yolk antibody against citrinin | |
| CN108640850A (en) | A kind of malachite green hapten, artificial antigen and its application in fluorescent quantitation immunochromatography | |
| Aksoy et al. | Determination of Aflatoxin Levels in Raw Milk, Cheese and Dehulled Hazelnut Samples Consumed in Samsun Province, Turk | |
| CN101650368A (en) | Method for testing zearalenone toxin by using colloidal gold immunochromatography assay | |
| CN108761075A (en) | A kind of deoxynivalenol quantifies rapid detection card and its detection method | |
| CN202033368U (en) | Reagent plate for detecting aflatoxins M1 in dairy | |
| CN101776689A (en) | Immune colloidal gold strip for detecting residue of roxarsone and preparation method thereof | |
| CN202794185U (en) | Test paper strip for detecting aflatoxin M1 | |
| CN201269879Y (en) | Melamine detection test strip by color granule immunochromatography | |
| CN107589265A (en) | A kind of time-resolved fluorescence test strips for detecting aflatoxin B1 and its application | |
| CN109061144A (en) | A kind of qualitative, quantitative immuno-chromatographic test paper strip and its preparation method and application for detecting zearalenone | |
| CN103513035A (en) | Test strip and method for detecting aflatoxin M1 | |
| CN203148952U (en) | Colloidal gold reagent plate for rapid detection of aflatoxin B1 in feed | |
| CN101403756A (en) | Fast detection reagent for melamine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20090408 |