220 results about "Clenbuterol" patented technology

The invention discloses a preparation method of a magnetic sandwich nano immunosensor and application of the preparation method. The preparation method is characterized by comprising the following steps: mixing and stirring a collaurum solution and silane ferriferrous oxide into a colorless and transparent solution, and carrying out magnetic separation by externally applying a magnet to obtain FE3O4 / Au colloid nano magnetic beads; loading horse radish peroxidase (HRP) and secondary antibodies of an object to be measured to the FE3O4 / Au colloid nano magnetic beads to obtain a secondary antibody probe Fe3O4 / Au-HRP-Ab2; electrically depositing Au to a glassy carbon electrode, loading primary antibodies of the object to be measured, and then closing non-specific active sites by using protein to obtain an Abl / Au / GCE electrode; and finally, loading antigen to the Abl / Au / GCE electrode, and then obtaining the sandwich nano immunosensor by combining the magnet probe. The magnetic sandwich nano immunosensor can be used for measuring the concentration of melamine, tonyred, clenbuterol or estrogen in food and has the advantages of high detection speed, high accuracy and sensitivity and low cost.

The invention relates to a preparation method and application of an unmarked immunosensor for rapidly detecting clenbuterol. Clenbuterol means a medicine of a beta-stimulant type instead of a specific medicine, and has the functions of prompting lean meat growth and suppressing fat meat growth as well as extremely serious side effects, and when a large amount of clenbuterol is taken, the cardiovascular system of people can be damaged and severe neurological symptoms can be caused. According to the invention, a gold-silver-sulfide nano composite material modified electrode with a core-shell structure which combines the excellent characteristics of nano gold and nano silver sulfide is adopted for preparing the unmarked immunosensor, and a clenbuterol antibody is well immobilized, so that the unmarked immunosensor is low in cost, high in sensitivity, good in specificity, rapid in detection and simple to prepare.

The present invention discloses a method for enriching clenbuterol, ractopamine and salbutamol in a urine sample of breeding livestock, and relates to the field of analytical chemistry, particularly to a sample pretreatment method. The method comprises: carrying out a sample pretreatment, adding sulfonic group surface-modified polystyrene superparamagnetic nanometer beads or sub-micron beads to adsorb, carrying out elution, and collecting the eluent, wherein the elution solution can be directly used for liquid chromatography-mass spectrometry to quantitatively analyze contents of clenbuterol, ractopamine and salbutamol. The method has characteristics of simple operation process, high extraction recovery rate, less elution solvent use amount, and no requirement of rotary evaporation.

The invention discloses a ractopamine-clenbuterol fluorescent microsphere detection test strip and a preparation method and an application. The test strip comprises a sample absorption pad, a combined pad, a chromatography membrane and a water absorption pad, which are adhered onto a soleplate and sequentially and closely connected with one another; the combined pad comprises fluorescent polystyrene microsphere labeled protein; the chromatography membrane is provided with two detection lines and one quality control line, the detection lines are close to the combined pad, and the quality control line is close to the water absorption pad; one detection line is coated with ractopamine protein conjugate, the other detection line is coated with clenbuterol protein conjugate, the quality control line is coated with protein conjugate which can be combined with proteins in the fluorescent polystyrene microsphere labeled protein and is not combined with the ractopamine and clenbuterol; and the fluorescent polystyrene microsphere labeled protein is ractopamine monoclonal antibody and a clenbuterol monoclonal antibody labeled by the fluorescent polystyrene microsphere. The test strip can simultaneously detect the ractopamine and the cleubuterol and is rapid, simple, convenient, flexible and accurate in detection.

The invention relates to an enzyme immune agent box for detecting ª‰-excitant drugs, which comprises: enzyme mark plate which coats ª‰-excitant drugs, enzyme mark antibody, clenobuterol standard solution, base material color developing solution, ª‰-excitant drugs antibody working solution, compression cleaning liquid, ending solution and compression twin solution. The invention also discloses a method for applying the detecting method, which comprises: first doing sample front process, then using the agent box to detect, at last analyzing the detected result.

The invention discloses a method for detecting the Clenbuterol by applying competitive SERS (Surface-Enhanced Raman Scattering) and application thereof, which belong to the technical field of the detection of hormonal micro-molecule substances. In the method disclosed by the invention, a sample to be detected and a SERS nano-probe combined with a Clenbuterol antibody are sequentially dropped on the surface of a substrate with a fixed antigen conjugate; the antigen in the sample to be detected and the antigen in the substrate with the antigen conjugate have an antibody competition reaction with the antibody combined with the SERS nano-probe; after the reaction is finished, the SERS nano-probe which is marked on the substrate with the antigen conjugate is subjected to signal collection through a Raman spectroscope, and data are processed; and finally the quantitative detection is carried out on the Clenbuterol according to the amplitude and standard curve of detection signals. The method disclosed by the invention is convenient to operate, has the advantages of high sensitivity, wide detection interval and high specificity and also has wide application prospects in the fields of food safety, analeptic detection, environment analysis and the like.

The present invention provides a clenbuterol monoclonal antibody and applications thereof. The present invention discloses a clenbuterol monoclonal antibody preparation method, wherein clenbuterol hapten is synthesized, the synthesized clenbuterol hapten and carrier protein are coupled to obtain clenbuterol antigen, and the clenbuterol antigen is adopted to immunize animals to obtain a high specificity monoclonal antibody. The present invention further provides a method for application of the clenbuterol monoclonal antibody in a clenbuterol enzyme-linked immunosorbent assay kit to detect clenbuterol. The present invention further provides a method for application of the clenbuterol monoclonal antibody in a clenbuterol colloidal gold test paper card to detect clenbuterol. The prepared clenbuterol monoclonal antibody has characteristics of high specificity and low cost, wherein the clenbuterol drug detection clenbuterol enzyme-linked immunosorbent assay kit prepared by using the clenbuterol monoclonal antibody and the clenbuterol drug detection clenbuterol colloidal gold test paper card prepared by using the clenbuterol monoclonal antibody have characteristics of convenient operation, high specificity, high sensitivity, high accuracy, high precision, fast detection and the like.

The invention discloses an immune chromatography test paper strip for the quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and a preparation method thereof. The test paper strip comprises a supporting layer, an adsorption layer, and a protection layer, wherein the adsorption layer comprises an adsorption fibrous layer, a fluorescent antibody fibrous layer, a cellulose film and an absorbent material layer at the handle end, the cellulose film is provided with detection blots printed by using a CL coupling carrier protein solution and control blots printed by using goat anti-mouse IgG antibodies, and the fluorescent antibody is an NaYF4:Yb:Er nanoparticle labelled CL monoclonal antibody or polyclonal antibody. According to the invention, the application of immune chromatography based on the up-conversion fluorescent nanoparticle label in the quantitative determination of CL residues is realized, so that the detection of the CL residues has no background interference; and the test paper strip has the advantages of strong specificity, high sensitivity, simple, and accurate detection, low cost, wide application scope, and easiness in popularization and application.

The invention discloses a preparation method for unmarked electrogenerated chemiluminescence clenbuterol immune sensor and belongs to the novel nano functional material and biosensor field. The dendrite nano rod-shaped precious metal alloy nano material having core-shell structure is prepared on the titanium dioxide nano-particle substrate by the photoelectric chemical synthesis method, and the unmarked electrogenerated chemiluminescence immune sensor used for detecting the clenbuterol can be prepared and the unmarked electrogenerated chemiluminescence immune sensor is low in cost, good in specificity, fast in detection speed and simple in preparation.

The invention relates to a novel synthesis method for a multilayer protection hyperstable water-soluble single fluorescent quantum dot and a fluorescent microsphere. The method comprises the following steps: transferring an oil-soluble fluorescent quantum dot to a water phase by using an amphoteric oligomer; then wrapping a water-soluble single fluorescent quantum dot and a fluorescent microsphere in silica; allowing the surfaces of the single fluorescent quantum dot and the fluorescent microsphere wrapped by silica to carry hydrophobic groups; and finally preparing the multilayer protection hyperstable water-soluble single fluorescent quantum dot and the fluorescent microsphere by using the amphoteric oligomer again. With the method, the fluorescent quantum dot can be protected from a destructive effect of an external environment (e.g., a strong acid, a strong alkali, a salt, etc.) to a greatest extent. The multilayer protection hyperstable water-soluble single fluorescent quantum dot and the fluorescent microsphere can be used in a microporous immunostrip biological detection system for detection of human chorionic gonadotropin (HCG), human immunodeficiency virus (HIV), hepatitis and clenbuterol--ractopamine(Rac) residual and the like in swine urine.

The invention discloses a method for rapidly detecting clenbuterol in urine based on a surface-enhanced Raman spectrum. Dichloromethane or ethyl acetate is adopted as an extraction solvent, an acidic aqueous solution and an organic solvent are adopted as a purification extracting solvent for carrying out liquid-liquid extraction on an extracting solution, nano gold or nano silver colloidal solution is adopted as an enhancing reagent, and a 785nm laser source Raman spectrum with the laser energy of 200mw is adopted for scanning a detection solution on a machine; and displacements 1258 minus or plus 2 cm<-1>, 1470 minus or plus 2 cm<-1> and 1601 minus or plus 2 cm<-1> are taken as characteristic peaks for judging clenbuterol; and normalization is carried out on strength of a corresponding peak 1450 minus or plus 2 cm<-1>, a linearity curve and the strength of the corresponding peak 1470 minus or plus 2 cm<-1> are combined to calculate content of clenbuterol, and rapid detection on clenbuterol in the urine are carried out qualitatively and quantitatively. A preparation method of a nano gold size is simple, operation time of pre-treatment is short, repeatability is good, and flexibility is high, in-field rapid detection can be realized, and supervision requirement can be met.

The invention relates to an enzyme couple immune detecting field, especially disclosing a clenobuterol enzyme immune agent box, relative detecting method, and a method for preparing the sample of animal organism, wherein said box comprises a box body, a 96 / 40 porous enzyme mark plate, a horse peroxide enzyme mark antibody, a clenobuterol standard solution, a substrate liquid, a substrate buffer liquid and a reaction ending liquid; the holes of said mark plate packs the packing antibody that combined with the clenobuterol antibody. The invention uses direct competitive enzyme couple immune adsorption analysis technique, to simplify the operation and reaction time, to reduce error. The inventive method has simple operation, low cost, and short time, while the extracted sample can be directly used in enzyme couple immune method and golden mark immune laminated analysis, to be sample of prior analysis.

The invention discloses a particle used for detection of a veterinary drug micromolecule. The particle comprises a receptor particle and a donor particle and is prepared through the following steps: marking carrier protein with a veterinary drug molecule; coupling the veterinary drug molecule-marked veterinary drug with a homogeneous chemiluminescent receptor ball; and preparing a homogeneous chemiluminescent receptor ball coupled with a veterinary drug molecule antibody. The invention further discloses a one-step homogeneous chemiluminescent method for detection of the micromolecule with the particle. The one-step homogeneous chemiluminescent method is used for homogeneous, rapid and high-sensitivity detection of veterinary drug residue and for testing of the contents of frequently used veterinary drug components in feeds, e.g., clenbuterol, ractopamine, salbutamol and terbutaline.

The invention relates to a method for detecting clenbuterol through the combination of a MIT technology and a SPR technology, belonging to the field of clenbuterol detection. The method of the invention comprises: firstly using a thermal polymerization method to prepare a clenbuterol molecular imprinting polymer film on a gold film substrate, removing template molecules with eluent, and obtaining a clenbuterol molecular imprinting sensor chip; then calculating the displacement value Delta Theta of SPR resonance angle resulting from the post-reaction and the pre-adsorption of the clenbuterol standard solution and the sensor chip, and creating a standard curve of negative logarithm of Delta Theta and the concentration value of the clenbuterol standard solution; and finally according to the displacement value Delta Theta of the SPR resonance angle resulting from the post-reaction and the pre-adsorption of the clenbuterol standard solution and the sensor chip, and reading the corresponding clenbuterol content on the standard curve. The method of the invention has the characteristics of severe-environment-resistance, high selectivity, fast response and simple operation, not only improves the sensitivity and limit of detection of the method, but also correspondingly reduces the detection cost, and also almost has no adsorption to salbutamol having similar structure with clenbuterol.

The intranasal delivery of an effective amount of NGF inducers, such as clenbuterol, to the brain.

The invention is a colloid gold test paper for rapidly detecting clenbutero HCl residual, composed of bottom plate, water absorbing plate, pyroxylen film, clenbutero HCl monoclonal antibody gold label pad, and sample liquor absorbing layer. The establishment of paired monoclonal antibodies is implemented by regulating immunizing dose, where the high dose is 50% of lethal dose, i.e. immunizing in 10-60 mug; the low dose is below 5 mug and the low-dose immunization is easy to induce highly selective and highly affine antibodies; but on the contrary, the high-dose immunization paired- screens the antibodies produced in low and high dose to obtain the paired monoclonal antibodies in lager probability. The method for establishing colloid gold double antibody sandwich by this theory screens antibodies to make test paper for detecting if there is the clenbutero HCl remained in animal products and has the advantages of strong peculiarity, high sensitivity, operating simplicity, and being easy to store and transport, etc.

The invention discloses a method for detecting clenbuterol residue in hair. The method sequentially comprises the following steps: (1) pre-processing a hair sample: hydrolyzing the sample with alkali at a high temperature, extracting the sample with an organic solvent and reversely extracting the sample with acid water; and (2) taking deuterated clenbuterol-D9 aqueous solution with the concentration of 5-500ng / ml as an internal standard, and detecting the clenbuterol residue in hair by liquid chromatogramy-tandem mass spectrometry. The method has the following advantages: 1. hair is taken as a detection material, which provides easy sampling, convenient storage and repeatable sampling, sampling is simple and rapid and has trace amount and high efficiency; 2. the pre-processing method of the hair sample is optimized, which shortens the pre-processing time, simplifies the processing steps and obviously reduces the cost; and 3. an isotopic compound is taken as the internal standard, and the liquid chromatogramy-tandem mass spectrometry is used as a detection means, the minimum detection limit reaches 0.01ng / g, and the minimum quantification limit is 0.5ng / g. The method has the accurate and reliable capacity to qualitatively and quantitatively detect the clenbuterol, and can be taken as a tool for large sample screening or small sample confirmation.

The invention discloses a hybridoma cell line secreting a monoclonal antibody repelling Clenbuterol hydrochloric acid, and an antibody repelling Clenbuterol hydrochloric acid through the hybridoma cell line. The invention also discloses a kit for detecting Clenbuterol hydrochloric acid residual. The kit comprises: a glowing black test board coated by CL-BSA in advance, a Clenbuterol hydrochloric acid standard liquid, an ELISA Clenbuterol hydrochloric acid antibody, a chemical luminescence liquid and a cleaning solution. The kid for detecting Clenbuterol hydrochloric acid residual has qualitative improvements on specificity, sensitivity and stability compared with the prior kit. The lowest detection value of Clenbuterol (CLB) hydrochloric acid residual is 0.02 ng / L, thereby completely meeting the requirements of detecting the level CLB residual.

The invention relates to a preparation method and application of a miniature high-efficiency clenbuterol immuno-affinity chromatography column. The technical scheme is characterized by comprising the following steps of: preparing a high-quality antibody, synthesizing efficient prepared bromoacetyl chloride clenbuterol purified by a liquid chromatogram and thiolation hemocyanin into immunogen immune animals, and obtaining antiserum; reacting the purified bromoacetyl chloride clenbuterol with thiolation agarose to obtain clenbuterol-agarose padding, loading the padding into an antigen affinity chromatography column; and purifying a clenbuterol specific antibody in the antiserum by using the antigen affinity chromatography column. The preparation method of the clenbuterol immuno affinity chromatography column comprises the following steps of: coupling high-quality antibodies at high density to the agarose oxidized by periodic acid to prepare an antibody affinity padding, and placing 25 mul of padding in a small specially-made column to prepare the miniature immuno-affinity chromatography column. The immuno-affinity chromatography column provided by the invention can be used for specifically gathering clenbuterol in a sample to be tested at high efficiency and can increase the accuracy, reliability and sensitiveness of detection when being combined with an analytic instrument and colloidal gold test paper for use.

The invention discloses a method for detecting clenbuterol residues, which comprises the following steps: (1) preparing, identifying and biotinylating clenbuterol-calf serum albumin conjugate; (2) coupling anti-clenbuterol monoclonal antibody on fluorescent microspheres; (3) drawing standard curve for suspension chip detection of clenbuterol residues; (4) testing specificity of suspension chip detection of clenbuterol residues; and (5) measuring the concentration of clenbuterol residues sample. The method of the invention for detecting clenbuterol residues has the advantages of simple operation, rapidness, sensitivity and low cost; and the overall detection process only takes 1-2h.

The invention relates to a production method of a clenbuterol molecularly imprinted-upconversion luminescent material fluorescence probe. The probe is a novel fluorescence probe produced through combining an upconversion luminescent material YF3:Yb<3+>, Er<3+> as a carrier with a covalent bond technology prepared clenbuterol molecularly imprinted polymer. The probe combines the high specific identification performance of the molecularly imprinted polymer with the high sensitivity of the upconversion luminescent material. The production method comprises the following steps: preparing the upconversion luminescent material, preparing the clenbuterol molecularly imprinted-upconversion luminescent material fluorescence probe, and eluting clenbuterol molecules. The fluorescence probe produced in the invention emits visible light under the excitation of infrared light, and has the advantages of high sensitivity, strong specificity and small interference. The production method has the advantages of simplicity, good reappearance, and good application prospect in selective identification and detection of clenbuterol in a practical sample.

The invention discloses an electrochemical immunosensor capable of detecting various beta-adrenergic receptor stimulants (short for beta-stimulants). The sensor comprises a base electrode, a graphene / chitosan complex membrane and multi-determinant antigens of beta-stimulants, wherein the graphenee / chitosan complex membrane is modified on the surface of the base electrode, and the multi-determinant antigens of the beta-stimulants are fixed on the surface of the graphene / chitosan complex membrane. The multi-determinant antigens fixed on the surface of the sensor are salbutamol-ractopamine-BSA or salbutamol-ractopamine-OVA. The invention also provides an electrochemical immunodetection method for various beta-stimulant residues. The electrochemical immunosensor can be used for detecting six beta-stimulants including clenbuterol, salbutamol, ractopamine, terbutaline, mabuterol and tulobutezol through carrying out cross immune reaction on the beta-stimulants based on wide-spectrum specific antibodies of the beta-stimulants by taking K3[Fe(CN)6] as a probe.

The invention provides a chemiluminescent immunoassay kit for detecting clenbuterol, and the kit belongs to the field of immunoassay. The kit is composed of a non-transparent white enzyme-labeled plate which is coated by a clenbuterol-carrier protein cross-linked complex, a clenbuterol standard product, a clenbuterol-peroxidase-labeled antibody, luminescent substrate solution and concentrated buffer solution. The clenbuterol-carrier protein cross-linked complex is obtained by coupling the clenbuterol and a carrier protein through the mixed-anhydride method or the direct protein activation method, and the concentrated buffer solution contains Tween-20 buffer solution. The kit has the advantages of rapidness, simpleness, high sensitivity, low detection cost, good repeatability, high throughput, and the like, and the kit can be used in the detection of residual clenbuterol in animal urine, blood, tissues, visceral organs and other samples.

The invention provides a method for extracting and analyzing beta-agonists clenbuterol by adopting a DPX (dispersive pipette extraction) tip type dispersive solid-phase microextraction column and relates to the DPX tip type dispersive solid-phase microextraction column (DPX tips <TM>). The method can be matched with various manual and automatic pipetting systems, meat sample treatment and extraction can be completed rapidly and efficiently, and defects that traditional sample treatment is time-consuming and labor-consuming, instrument loss is increased and the like can be perfectly overcome. The simple and fast solid-phase extraction method is combined with HPLC-MS (high performance liquid chromatography-mass spectrometer) analysis, so that purification of 96-384 samples can be finished simultaneously within 10 min, and rapid, qualitative and quantitative high-flux determination for multiple kinds of beta-agonists clenbuterol in animal derived food can be realized.

The invention discloses a clenbuterol matrix standard substance and a preparation method thereof. The preparation method comprises the following steps of: uniformly mixing pig urine in which a solid substance is removed; then adding a clenbuterol standard substance to a quantity value level; uniformly mixing to obtain a mixed solution; and carrying out uniformity check on the mixed solution and then carrying out subpacking and freeze drying to obtain the clenbuterol matrix standard substance. The preparation method disclosed by the invention is scientific and reasonable and is strong in operability; the prepared matrix standard substance can be stored under the condition of -20DEGC and provides a quantity value tracing substance for detecting clenbuterol residue in daily pig urine; and according to the prepared matrix standard substance, the problem that different detection results are caused by matrix difference is solved and the monitoring of the clenbuterol residue in pig products is accurately ensured.

The invention discloses a method for preparing a pork powder standard substance with clenbuterol. The method comprises the following steps: performing simulated feeding so as to obtain a pork raw material polluted by clenbuterol, crushing a sample, directly mixing with water in a proper ratio, preparing freeze-dried pork by using a freeze-drying method directly without homogenation, and implementing steps of grinding, screening, uniform mixing, radiation and the like, thereby obtaining freeze-dried pork powder in good uniformity. Value confirmation, uniformity inspection and stability monitoring of standard substances are implemented by using a liquid chromatogram isotope dilution mass spectrometry (LC-ID-MS / MS) method for verifying accuracy through international comparison. The pork powder standard substance with clenbuterol, which is prepared by using the method, is good in uniformity and stability, and can be applied to analysis method evaluation for residue detection on clenbuterol in pork, quality control in analysis process and laboratory capability evaluation.

The invention discloses a method and a special chemical luminous immunoassay kit for detecting clenbuterol. The special chemical luminous immunoassay kit for detecting clenbuterol comprises a clenbuterol specific antibody, a coating antigen and standard solution; and the coating antigen is a coupling compound of a clenbuterol hapten and a carrier protein. The detection method has the characteristics of simple sample preprocessing process, simple and convenient operation, low cost, high specificity, high sensitivity, high accuracy and the like, can perform monitoring on site and is suitable for screening a large number of samples.

The invention discloses a chemiluminescent enzyme-linked immunosorbent assay (CELISA) kit for detecting salbutamol. The kit comprises a kit body, in which an ELISA plate with wells coated with coating antigen prepared from clenbuterol and ovalbumin by coupling, anti-ractopamine polyclonal antibody, horseradish peroxidase-labeled goat anti-rabbit antibody (enzyme-labeled secondary antibody), a series of salbutamol standard solutions, a concentrated phosphate buffer solution, a concentrated washing solution and a chemiluminescence solution are arranged. The CELISA kit has the advantages of high sensitivity, good repeatability, and good simplicity, rapidness and accuracy; and the sensitivity is improved by one order of magnitude compared with that of the conventional colorimetric ELISA method. The CELISA kit can be used for detecting residual salbutamol in animal-derived foods (such as milk and animal tissues) and urine samples.

The invention discloses a method for quickly detecting clenbuterol based on functionalized gold nanoparticles. In the method, clenbuterol can generate hydrogen-bond interaction with sulfhydryl compound-modified gold nanoparticles to cause the resonance change of plasmas on the surfaces of the gold nanoparticles and the change of the color as well as ultraviolet-visible absorption strength and peak value of the gold nanoparticle solution, and therefore, whether a solution contains clenbuterol can be quickly detected by directly observing the color change of the solution with naked eyes. The method disclosed by the invention has the advantages of simplicity in operation, low cost, high sensitivity, wide application range and the like. The nano material can be used for realizing real-time and quick detection of clenbuterol.