167 results about "Clenbuterol hydrochloride" patented technology

The invention relates to a method for visual rapid detection of clenbuterol by adopting a nanogold probe. According to the method, contents of three main components, namely clenbuterol hydrochloride, ractopamine and salbutamol, are visually detected by specifically using melamine-modified nanogold. The method is mainly characterized in that the nanogold probe is formed by modifying melamine onto the surface of nanogold. The nanogold probe can specifically react with the clenbuterol hydrochloride, the ractopamine and the salbutamol through hydrogen bonds to cause polymerization of the nanogold and color change of a solution. According to a color change scope of the solution, whether the clenbuterol hydrochloride, the ractopamine and the salbutamol exist in the solution and the contents of the clenbuterol hydrochloride, the ractopamine and the salbutamol can be judged with the naked eye. The detection method is simple and practicable, is low in cost and high in sensitivity and is suitable for spot rapid detection of batch samples.

The invention provides a clenbuterol (CLB) magnetic particle separation enzyme-linked immunosorbent assay (ELISA) method, belonging to the technical field of food safety immunodetection. The method comprises the following steps: the immunodetection principle of the competition method is adopted to connect CLB with bio-enzyme and prepare an enzyme labeled antigen reagent, antibody against fluorescein isothiocyanate (FITC) is absorbed on the surface of magnetic particles to prepare a magnetic separation reagent, FITC is connected with the antibody against CLB to prepare an antibody reagent; CLB in a sample and enzyme-labeled CLB compete to combine with a small quantity of FITC-labeled antibody against CLB and form an antigen-antibody composite; after the magnetic separation reagent is added, the antibody against FITC connected with the surface of magnetic particles captures the composite on the surface of magnetic particles; and washing, and finally adding substrate to detect. The invention has the following advantages: (1) magnetic particles replace the traditional ELISA plate to be used as solid carrier and ensure that the immunoreaction is performed more fully and fast under the near-liquidus condition; compared with the traditional ELISA method, the method is characterized by high specificity and good repeatability; and (2) the one-step competition method principle is adopted, thus the detection time is short.

The invention provides a group of oligonucleotide aptamers Apt-1, Apt-2 and Apt-3 which are capable of simultaneously identifying clenbuterol hydrochloride and salbutamol, an oligonucleotide aptamer CLB-2 which is capable of identifying the clenbuterol hydrochloride with high specificity, an oligonucleotide aptamer SAL-5 which is capable of identifying the salbutamol with high specificity and two oligonucleotide aptamers RAC-5 and RAC-6 which are capable of identifying ractopamine with high specificity. Through an SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technology based on Fe3O4 magnetic nanoparticle separation, a random oligonucleotide library is immobilized on avidin-enveloped magnetic nanoparticles by virtue of a complementary chain of a biotinylation marker, and the oligonucleotide aptamers, which are high in specific affinity, are finally obtained by conducting screening by 16 turns. The aptamers are broad in application prospect; and the aptamers, by virtue of marker function genes or fluorescent dyes, are applicable to detection of the clenbuterol hydrochloride, the salbutamol and the ractopamine in food; therefore, a new choice is provided for existing detection methods which depend on antibodies.

The invention discloses an enzyme linked immunosorbent assay kit for detecting residual clenbuterol hydrochloride and a detection method thereof. The kit comprises a micropore plate (1) for precoating staphylococcus protein A and coating CBL antibodies, CBL standard substance (2), concentrated washing liquid (3), concentrated enzyme labeled CBL (4), enzyme labeled diluents (5), a substrate solution (6) and a stopping solution (7). The kit adopts a direct competition method, i.e. CBL in standard substance or samples to be detected and enzyme labeled CBL compete CBL antibodies on the micropore plate. The method can be used for directly detecting clenbuterol hydrochloride in animal derived food, urine and serum samples, and has the advantages of convenient and rapid operation. The operation time only needs 50 minutes, and the sensibility can reach 0.02mug/kg.

The invention discloses a clenbuterol hydrochloride assay kit and its preparation method and use method. The clenbuterol hydrochloride assay kit comprises a kit body, a clenbuterol hydrochloride enzyme label plate, a clenbuterol hydrochloride standard solution, a clenbuterol hydrochloride enzyme conjugate, a diluent, a concentrated washing liquid, a first substrate and a second substrate. The preparation method provided by the invention comprises preparing the clenbuterol hydrochloride enzyme label plate, the clenbuterol hydrochloride standard solution, the clenbuterol hydrochloride enzyme conjugate, the diluent, the concentrated washing liquid, the first substrate and the second substrate. The use method provided by the invention comprises sample preparation, sample addition and detection, and detection result analysis. The clenbuterol hydrochloride assay kit has the advantages that sensitive, stable and reliable result analysis is realized; operation steps are less; sample pre-treatment is simple; adopted equipment can be popularized easily; and the clenbuterol hydrochloride assay kit can be utilized for determination of residual clenbuterol hydrochloride of pork, has sensitivity of 0.03 micrograms per kilogram, needs detection time shorter than detection time of an ELISA method by 30 minutes and satisfies requirements of rapid detection of clenbuterol hydrochloride residues.

The invention discloses a biosensor sensitive membrane and application in detection of clenbuterol hydrochloride. The sensitive membrane is prepared by the method of: 1) placing a gold membrane loaded on a glass substrate in an ethanol solution of n-Octadecyl mercaptan to perform soaking, and then conducting flushing and drying, thus obtaining a first self-assembled gold membrane; 2) putting the first self-assembled gold membrane in a coupling solution of amino-functionalized graphene or carboxyl-functionalized graphene oxide and goat anti-mouse IgG to perform soaking, then conducting flushing with a phosphate buffer solution and carrying out drying, thus obtaining a second self-assembled gold membrane; and 3) adding an antibody solution dropwise to the surface of the second self-assembled gold membrane to perform antibody assembling so as to make the antibody cover the surface of the second self-assembled gold membrane, conducting standing, then performing flushing with a phosphate buffer solution and carrying out drying. The biosensor sensitive membrane provided by the invention undergoes recognition combination with an antibody through a clenbuterol hydrochloride antigen and its conjugates, and can carry out rapid detection on clenbuterol hydrochloride.

The invention relates to the preparation of molecular imprinted polymer in the technical field of chemical engineering and a method for separating clenbuterol hydrochloride with the molecular imprinted polymer. In the preparation method, clenbuterol, methacrylic acid, ethylene dimethacrylate are mixed according to the mol ratio of 1: 2-6: 15, dissolved in a pore-foaming agent, placed in an ampoule, added with an initiator, processed by ultrasonic wave, fed in nitrogen, sealed in vacuum, and then carried out thermal-initiated polymerization so as to obtain bulk polymer; and the bulk polymer is taken out, crushed, sieved and separated; after the polymer is dried, soxhlet extraction, high-temperature processing and solvent elution are carried out to get rid of a template. In the separation method, the molecular imprinted polymer homogenate is filled in a small column of polypropylene solid phase extraction, and the upper and lower ends of the filling material are respectively added with a sieve plate of polyethylene and the filling material is pressed tightly; and the small column of solid phase extraction is activated, leached and eluted so as to collect clenbuterol hydrochloride. The molecular imprinted polymer prepared by the method has high selectivity and specificity, can resist to unfriendly environment and combine with ELISA in use.

The invention provides a method for analyzing developing result of a colloidal gold test strip, belongs to the technical field of colloidal gold quantitative analysis and aims to solve the problem in the prior art of qualitative observation result after clenbuterol hydrochloride detection by colloidal gold. The method comprises the following steps: A, the relation of the developing level of a testing line and the concentration of the clenbuterol hydrochloride is established; B, a developing region of a controlled line, the developing region of the testing line and a background referential region are identified; C, developing grayness with the same size of pixel range for the developing region of the controlled line, the developing region of the testing line and the background referential region is acquired respectively; D, the developing degrees of the testing line and the controlled line are obtained and the developing level of the testing line is calculated; E, the concentration of the clenbuterol hydrochloride is obtained through the developing level of the testing line according to the established relation. Through the adoption of the method, as the clenbuterol hydrochloride is detected by the colloidal gold, the quantitative numerical value of the concentration of the clenbuterol hydrochloride can be obtained by analyzing the developing result of the testing line of the colloidal gold test strip.

The invention relates to a method for detecting clenbuterol in pig hair. The detection method adopts a surface-enhanced Raman spectroscopy (SERS) and takes nano gold as an SERS substrate; the pig hair is subjected to pretreatment so as to carry out enrichment and separation on the residual clenbuterol on the hair; and an SERS spectrum of the clenbuterol is obtained through the portable Raman spectrograph. The method is simple and convenient to operate, and can be used for rapidly detecting the clenbuterol hydrochloride in the pig hair.

The invention discloses a Clenbuterol hydrochloride detection test paper and a detection method thereof. The Clenbuterol hydrochloride detection test paper comprises a support and fixing lining which is sequentially provided with a sample adsorption layer, a combination pad, a microporous siphon reaction layer and a water absorption layer; the combination pad is provided with a Clenbuterol hydrochloride antibody labeled by a trace particle and an antibody A labeled by the trace particle, and the antibody A is different from the Clenbuterol hydrochloride antibody; the microporous siphon reaction layer is sequentially provided with a blotting area and a detection area of Clenbuterol hydrochloride carrier protein solution, and the detection area comprises a detection solution blotting area and a control solution blotting area. Two lines occur in the detection area when the content of the Clenbuterol hydrochloride is more than 5 nanogram in a sample; and one line occurs in the detection area when the content of the Clenbuterol hydrochloride is less than 5 nanogram in the sample. The test paper has a detection mode and a reading manner equivalent to those of a sandwich method and low cost. The detection method is simple and rapid, and has simple operation.

The invention relates to the field of biotechnology, in particular to a homogeneous immunological detection reagent. The invention adopts a method utilizing the homogeneous enzyme multiplied immunoreaction to detect the residual amounts of harmful substances in the body fluid of animal and the extract liquor of tissue, thus harmful substance residues containing common organic small-molecule medicines and illegal medicines, such as clenbuterol hydrochloride, ractopamine, melamine, malachite green, benzodiazepines, tricyclic antidepressants and various antibiotic medicines can be detected and the invention has extremely wide application prospect and social meaning. The product developed by the invention has high sensitivity and can detect hundreds of compounds and metabolic products at the same time; and the detection is fast and accurate; and the detection data can be processed and printed automatically and can also be connected with the Internet so as to realize synchronous monitoring.

The invention relates to a fast test method for detecting the clenobuterol hydrochloride of animal foodstuff. Firstly syntheses marks the clenobuterol coupled material; then it fixes the horse radish perxidase and the anti-clenobuterol monoclonal antibody on the cellulose nitrate film; then dispositions the cellulose film coated the antibody and the enzyme inside the one time polyvinyl fluoride or the reaction hole which is made by glass material after sealed and dried by the skimmed milk powder and uses the polytef film to seal it; then it prepares the color development bottom material solution and assemblies the clenobuterol fast reacting plate; it mixes the tested sample and the enzyme mark clenobuterol coupled material and adds them on the antibody film of the reaction hole of the reaction plate; it washes it after reacting at indoor temperature and adds bottom material color development solution on the reacting hole to observe the result and then quotes the clenobuterol content by the color development degree.

The invention provides a method for preparing a clenbuterol hydrochloride molecularly imprinted membrane electrochemical light emitting sensor. The method is characterized in that the surface of a glassy carbon electrode is wrapped by a composite film, and the composite film consists of reduced oxidized graphene, upconversion nanoparticles and molecular imprinting polymers. The method has the advantages that electrochemical light emission of the upconversion nanoparticles with stable cathode signals and high light emission strength are combined with the molecular imprinting polymers with goodselectivity, and trace detection on clenbuterol hydrochloride in animal derived foods can be implemented.

Disclosed is a fore-anthocyanin which is extracted from pine needle and pine peels through the steps of collecting pine needle and pine peel, bobbing and sorting, charging the raw material into alcohol extraction tank, obtaining concentrated liquid through filtering after first alcohol extraction, secondary filtering of filtering deposit, obtaining filter liquor and filtering deposit respectively, charging 8 times of water into the filtering deposit, obtaining filter liquor and filtering deposit respectively through filtering, abandoning the filtering deposit, concentrating the concrete, storing the concrete at 1-5 deg. C, finally spray drying the concrete. The invention can be applied for substituting antibiotic medicines.

The invention discloses immune colloidal gold test paper and a method for quantitatively detecting a configurable logic block (CLB) by matching with a photoelectric sensor thereof. The immune colloidal gold test paper comprises absorbent paper, a nitrocellulose (NC) membrane, a colloidal gold protective pad and a glass fiber which are arranged on a support plate and overlapped partially and mutually in sequence. Two round holes are arranged on the support plate, round areas on the NC membrane corresponding to the two round holes on the support plate are coated by T zone stripe of CLB-bovine serum albumin (BAS) and C zone stripe of a goat- anti-mouse second antibody, and the colloidal gold protective pad comprises an antibody resisting clenbuterol hydrochloride and marked by colloidal gold. The method for quantitatively detecting the CLB includes the following steps: a tested immune colloidal gold test paper strip is placed in a detecting area of the photoelectric sensor to be detected; the photoelectric sensor converts light reflected by the detecting area into electric signals, and concentration of the CLB is calculated according to strength of the electric signals.

The invention relates to a sustained release preparation containing ambroxol hydrochloride and clenbuterol hydrochloride. The preparation comprises a sustained release part and a quick release part, wherein the quick release part comprises clenbuterol hydrochloride, a quick release adjuvant and another pharmaceutic adjuvant, the sustained release part comprises ambroxol hydrochloride, a sustained release adjuvant and another pharmaceutic adjuvant, and the another pharmaceutic adjuvant is one or more selected from the group consisting of a filler, a binder and a lubricant. The invention also relates to a preparation method for the sustained release preparation. The sustained release preparation provided in the invention has the following advantages: the quick release part has fast action, and the sustained release part has a long-acting drug effect.

The invention discloses a method for synthesizing a clenbuterol hydrochloride molecularly imprinted polymer. The method comprises the following step: by using clenbuterol hydrochloride as a template molecule and alpha-methacrylic acid as a functional monomer and performing interaction in the form of hydrogen bonds, adding a crosslinking agent N,N-methylene-bis acrylamide and an initiator ammonium persulfate to perform free radical copolymerization, thereby obtaining the clenbuterol hydrochloride molecularly imprinted polymer, wherein the mole ratio of the template molecule, functional monomer and crosslinking agent is (117-274):(1-300):(30-90). The clenbuterol hydrochloride molecularly imprinted polymer has the advantages of simple and quick synthesis process, low cost, high specificity and favorable stability, can be combined with a test paper method to prepare a test paper strip, and is used for trace on-line detection of clenbuterol hydrochloride.

The invention provides a hybridoma cell strain and an anti-hydrochloric acid clenbuterol monoclonal antibody prepared from the hybridoma cell strain and belongs to the technical field of immunochemistry. The hybridoma cell strain 6G11 generates the anti-hydrochloric acid clenbuterol monoclonal antibody 6G11'. The invention also provides a method for preparing the anti-hydrochloric acid clenbuterol monoclonal antibody 6G11', which comprises the following steps of: preparing an immunogen and a coating antigen; animal immunization; selecting immune rat serum; preparing hybridoma cells; and collecting and purifying the anti-hydrochloric acid clenbuterol monoclonal antibody 6G11'. The anti-hydrochloric acid clenbuterol monoclonal antibody with strong specificity through the rat hybridoma cell strain 6G11, has low crossing reaction rate with the salbutamol and ractopamine, can be massively produced, and can be used for preparing immunology detecting agents, such as an immunoassay kit for detecting clenbuterol hydrochloride, a colloidal gold test strip and the like so as to fast and sensitively detect clenbuterol hydrochloride medicament residue in urine, muscles and internal organs.

The invention provides a nucleic acid aptamer for detecting clenbuterol hydrochloride. The nucleic acid aptamer is selected from one of following nucleotide sequences shown in SEQ ID NO.1; the nucleicacid aptamers with same lengths are cut from both ends of the nucleotide sequence shown in SEQ ID NO.1, and the cutting length is 1 to 11bp; the nucleic acid aptamers with same cutting lengths of 1 to 11bp at the two ends respectively have the corresponding nucleotide sequences shown in SEQ ID NO.12 to SEQ ID NO.2. The invention also provides a screening method and application of the nucleic acidaptamer. The nucleic acid aptamer has the advantages that the specificity is strong, and the nucleic acid aptamer does not generate crossing reaction with clenbuterol hydrochloride analogues; the affinity is high, and the affinity constant is (42.17+/-8.97) nM; the sensitivity is high; the minimum detection limit of clenbuterol hydrochloride is 1.2ng/mL.

The invention discloses immune chromatography test paper for detecting clenbuterol hydrochloride and a preparation method thereof. The immune chromatography test paper for detecting the clenbuterol hydrochloride in the invention comprises a sample pad containing superparamagnetic composite particle labeled clenbuterol hydrochloride antibody, cellulose nitrate membrane connected at one end of the sample pad, and a water absorbent pad connected at the other end of the cellulose nitrate membrane; the cellulose nitrate membrane is coated with separated detection line and quality control line, the detection line contains a clenbuterol hydrochloride antigen and the quality control line contains an antiantibody capable of being specifically bonded to the clenbuterol hydrochloride antibody. Through the experiment demonstration, the test paper provided the invention has the advantages of high sensitivity and specificity, rapidness, simplicity and convenience in detecting the clenbuterol hydrochloride, and can realize objective detection.

The present invention provides a clenbuterol detection kit, which includes a kit body, a drawer provided at the lower right part of the box body, two left and right independent kit covers connected to the box body at one end, and the inner cavity of the box is separated by a partition The drawer at the lower right of the box body is set as a recovery area; the storage area includes a storage area and a storage area 2, and three test strip slots are set in the storage area, which are respectively used for storing Test strips for renbuterol, ractopamine, and salbutamol; dropper slots, reagent bottle slots, and sample tube slots are set in the second storage area to store droppers, reagent bottles, and sample tubes respectively; multiple fixed test strips are set in the detection area The detection slot; the left box cover is complete, and the right box cover is equipped with a sample injection hole and an observation window. The invention has higher partition detection efficiency, comprehensive functions, convenient portability, simple operation, and is suitable for on-site rapid detection, and the sealing, storage, detection, and recovery of the kit are independent from each other to avoid mutual interference and make the test results more accurate.