35 results about "Leucomalachite green" patented technology

Compounds and procedures are provided for use in the preparation of reagents that can be used in immunoassays for Malachite Green, Leucomalachite Green and Carbinol. The compounds are Malachite Green Chromatic derivatives conjugated to several positions of the Malachite Green Chromatic structure, which in turn are conjugated to antigenic polypeptides for formation of antibodies for use in immunoassays. The antibodies produced, when employed in immunoassays with specified procedures, are found to be able to detect Malachite Green Chromatic as determined by positive results with fish tissue and water sample containing Malachite Green Chromatic that has been detected by HPLC or LC-MS.

This invention discloses Pantoea sp. M3 (CCTCC No.M207022) with the ability to degrade malachite green. Logarithmic phase Pantoea sp. M3 2-4 wt. % is added into sterile degradation medium containing 0.5-5 mg / L malachite green, and cultured at 20-30 deg.C for 5-10 days to degrade malachite green. The degradation medium with a pH value of 6.0-8.0 comprises: glucose 0.5-1 g / L, NaCl 0.01-0.1 g / L, MgSO4 0.01-0.1 g / L, NH4NO3 0.02-0.1 g / L, K2HPO4 0.01-0.1 g / L, KH2PO4 0.01-0.1 g / L, and yeast extractum 0.06-0.3 g / L. Pantoea sp. M3 can effectively remove residual malachite green and leucomalachite green in water body for fishery. The degradation rate of Pantoea sp. M3 for 0.5-2 mg / L malachite green is higher than 90%.

The invention discloses an immune colloidal gold test stripe for detecting leucomalachite green in aquatic products and a preparation method; the colloidal gold test stripe provided by the invention detects leucomalachite green with a direct competition method, and comprises a sample pad, a colloidal gold combined pad, a nitrocellulose membrane (NC), a water adsorption pad and a PVC back lining, one end of the PVC back lining is sequentially adhered with the sample pad and the colloidal gold combined pad, and the middle thereof is adhered with the NC layer, and the other end thereof is sequentially adhered with water adsorption pad; the immune colloidal gold test stripe has the characteristics that colloidal gold combined pad is enveloped with colloidal gold marker which is monoclonal antibody of the specificity of the leucomalachite green, and the NC is adsorbed with leucomalachite green-OVA and sheep anti-mouse IgG. The invention has high detection sensitivity and low price and is simple and rapid, the whole reaction only needs 30 minutes; the immune colloidal gold test stripe can be used for large-batch screening of samples and can rapidly detect the residual leucomalachite green in the aquatic products at site.

The invention relates to a leucomalachite green gold-labeled test paper detection kit. The test paper detection kit has advantages of fast and accurate detection and high sensitivity, therefore, the invention also provides a preparation method of the detection kit. The detection kit includes a test strip which is packaged in a box shell body, and a sample injection hole and an observation hole are arranged on the box shell body, the test paper detection kit is characterized in that: the test strip takes polyvinyl chloride or polyethylene as a back lining, a sample injection area at one end of the test strip is affixed and adsorbed with a glass fiber film containing a conjugate of colloidal gold and anti-leucomalachite green monoclonal antibody; a cellulose nitrate film is affixed to a testing area in the middle of the test strip, a small section of the glass fiber film overlaps the cellulose nitrate film, and a water absorption area at the other end of the test strip is affixed with a piece of absorbent paper; a detection line with material of recessive malachite green-egg white albumin and a quality control line with material of goat anti-mouse IgG are arranged in the direction from the sample injection area to the water absorption area sequentially, and the cellulose nitrate film is covered with a bovine serum albumin layer with the concentration of 1 percent; and the sample injection hole is corresponding to the glass fiber film in the sample injection area of the test strip, and the observation hole is corresponding to the testing area of the test strip.

The invention provides haptens, immunogens comprising such haptens coupled to an antigenicity-conferring carrier material, conjugates comprising such haptens bonded to a labelling agent as well as, antibodies raised against such immunogens and capable of binding with leucomalachite green and malachite green.

The invention discloses a method for detecting leucomalachite green in an aquatic product, which comprises the following steps: adding anions and leucomalachite green to a weakly acidic HAc-NaAc buffer solution, shaking up, adding amphoteric surface active agent, maintaining a constant volume, standing in a dark place for some time to obtain a concentration-known detected system, carrying out synchronous scanning in a range of 400-600 nm by utilizing a fluorophotometer under the condition that the excitation wavelength is equal to the emission wavelength to obtain the light scattering strength I and the reagent blank I0 of the system, calculating according to the formula of deltaI=I-I0 to obtain the deltaII value of the object to be detected, and calculating the leucomalachite green content of a certain unknown object to be detected according to working curves. The method has the advantages of low price and easy acquisition of reagent, moderate reaction condition, simple operation, simple equipment, high sensitivity and easy popularization.

The invention relates to a synthesis method for stable isotope labeled leucomalachite green. The stable isotope labeled leucomalachite green is obtained by reacting stable isotope labeled N, N-dimethylaniline with benzaldehyde. Compared with the prior art, after the stable isotope labeled leucomalachite green prepared by the method and malachite green are separated and purified, the chemical purity reaches over 99 percent and the isotope abundance is over 98 percent atom.

The invention discloses malachite green hapten, malachite green artificial antigen and preparation methods of the malachite green hapten and artificial antigen. The malachite green hapten retains the feature structure of malachite green, and has active arms which are derived from different microdomains of the structure of malachite green, have different electron cloud densities and different carbon chain lengths, are suitable to be coupled with macromolecule carriers, and are amide linkage with amino or carboxyl tail end. The malachite green artificial antigen prepared with leucomalachite green hapten has very high specificity and sensitivity when applied to detection of the residual quantity of leucomalachite green and malachite green, and is high in accuracy and recovery. Moreover, the methods are simple, the cost is low, and a fast, sensitive, economical and efficient malachite green immunodetection method is established.

The invention discloses a toxicant elimination and fish-odor removal culturing method for soft-shelled turtles. According to the method, rotational flows are generated with oxygenic small molecular group water in a culture pond capable of generating the rotational flows in an intermittent mode, so that the culturing mode that the turtles swim for 30 minutes and rest for 90 minutes is achieved. All residual medicines, including leucomalachite green, of the cultured turtles are not detected, and therefore the turtles become delicious foods or high-quality health care products.

The invention relates to a method for determining malachite green, quinolone and sulfonamide drugs in aquatic products. The method comprises the following steps: step 1, preparing samples; step 2, extracting to-be-detected samples to obtain to-be-processed samples; step 3, determining chromatographic conditions; step 4, determining mass spectrum conditions; step 5, drawing a standard working curve; step 6, determining sensitivity, accuracy and precision. With adoption of the method, malachite green, leucomalachite green, quinolone and sulfonamide drugs in an extracted and concentrated aquatic product substrate are determined simultaneously for the first time, the problems that time is consumed and detection cost is high due to traditional respective extraction are solved, ion response value is increased, sensitivity is enhanced, detection limit is low, the effects of being qualitative, quantitative, accurate, fast, efficient and sensitive are realized truly, and the method can be taken as a reliable detection means for the three drugs and is applicable to extraction and concentration of a batch of samples.

The invention relates to a monoclonal antibody of leucomalachite green, a hybridoma cell strain producing the monoclonal antibody and a preparation method and usage of the monoclonal antibody. The invention also relates to a composition containing the monoclonal antibody and a detection kit. According to the invention, the monoclonal antibody of leucomalachite green is successfully prepared, the monoclonal antibody has high sensitivity and strong specificity and only has cross reaction with malachite green while having no cross reaction with parafuchsin, methyl blue, crystal violet, ciprofloxacin and chloramphenicol, thereby providing key technology for rapid detection of residual leucomalachite green aquatic products and laying a foundation for research and development of the detection kit of the leucomalachite green.

The invention discloses stable isotope labeled leucomalachite green and a synthesis method thereof. The method comprises the following steps: (a), oxidizing stable isotope labeled methylbenzene in a mixed solvent of acetic acid, acetic anhydride and concentrated sulfuric acid through an oxidizing agent, so as to obtain stable isotope labeled a benzal diacetate intermediate, and performing hydrolysis under the action of ethanol and 1M sulfuric acid, so as to obtain stable isotope labeled benzaldehyde; (b), ensuring that the stable isotope labeled benzaldehyde is reacted with N, N dimethylaniline under the action of a catalyst, and performing treatment, separation and purification after reaction is finished, so as to obtain the stable isotope labeled leucomalachite green. The stable isotopelabeled leucomalachite green and the method have the advantages that the utilization of stable isotope atom is high, the synthesis steps are simple, products are easy to separate and purify, the requirement of serving as a standard reagent for the quantitative detection of the leucomalachite green is met, the use value is high, and the stable isotope labeled leucomalachite green has excellent economical efficiency.

The present invention relates to an indirect competitive enzyme-linked immunosorbent method for determining malachite green (MG) in an aquatic product or water sample. According to the method, malachite green molecules are modified, the modified malachite green molecules and a carrier protein are subjected to cross-linking to obtain immunogen and coating antigen, and multiple animal immunizations are performed to prepare the anti-malachite green polyclonal antibody, wherein the IC50 value (the corresponding determined substance concentration when the absorbance in the standard curve is inhibited to achieve 50% of the maximum absorbance value) is 0.9-2.6 [mu]g / L and the detection limit is 0.02-0.10 [mu]g / L under the optimized experiment conditions, the recovery rate of the malachite green in the water sample and the aquatic product is 76.2-95.0%, and the cross-reaction rate of the malachite green and the leucomalachite green is 95.25%. According to the present invention, the method comprises: coating liquid preparing, washing liquid preparing, termination liquid preparing, blocking liquid preparing, substrate solution preparing, antibody immobilization, immune reaction, detection signal obtaining, and other step; and with the application of the characteristics of the competitive enzyme-linked immunosorbent method to detect the malachite green content in the aquatic product, the advantages of high sensitivity, low interference, convenience, rapidness, safe operation, strong specificity, and low detection cost are provided.

The invention provides a pre-column electrochemical derivatization one-time total quantity measuring method for malachite green and leucomalachite green in aquatic products and belongs to one-time total quantity measuring methods for converting leucomalachite green into malachite green. The key of the method is that it is found that after a leucomalachite green solution is subjected to oxidation voltage +650 mv of an electrochemical Coulomb detector (ECD), averagely 98.86% of leucomalachite green is converted into malachite green, original malachite green is unchanged, and the total concentration of leucomalachite green and malachite green can be measured through a high performance liquid chromatograph in the one-time measurement process of leucomalachite green and malachite green in a sample. Sample pre-treatment time is greatly shortened, use of reagent consumables is reduced, and electrochemical precolumn derivatization is used for replacing post-column derivatization of lead dioxide in the national standard. The method has the advantages of being simple in sample pre-treatment, good in purification effect, high in derivatization rate and high in recovery rate.

The invention provides haptens, immunogens comprising such haptens coupled to an antigenicity-conferring carrier material, conjugates comprising such haptens bonded to a labelling agent as well as, antibodies raised against such immunogens and capable of binding with leucomalachite green and malachite green.

The invention relates to a method and a system for detecting leucomalachite green. The method for detecting the leucomalachite green comprises the following steps: irradiating a sample with terahertzlight to obtain an absorption spectrum of the sample; extracting a sample absorption coefficient according to the absorption spectrum; determining the concentration of the leucomalachite green in thesample according to the absorption coefficient. The system for detecting the leucomalachite green comprises a terahertz time domain spectrometer device and a processor, wherein the terahertz time domain spectrometer device is used for detecting terahertz time domain spectral data of the leucomalachite green sample, and the processor is used for executing the method for detecting the leucomalachitegreen. The terahertz time domain spectrometer device is used for irradiating the sample with the terahertz light, so that the absorption spectrum of the sample is obtained; the sample absorption coefficient is extracted according to the absorption spectrum; the concentration of the leucomalachite green in the sample is determined according to the absorption coefficient. The method is short in detection time, simple in pretreatment and low in detection cost, and does not need to be operated by a professional experimenter.

The invention discloses a leucomalachite green nucleic acid aptamer and application thereof, and belongs to the field of nucleic acid aptamers. The invention discloses a nucleic acid aptamer A5-b and a mutant thereof. The nucleic acid aptamer A5-b and the mutant thereof are single-stranded DNA of which the nucleotide sequences are respectively sequences 1-6. The nucleic acid aptamer A5-b and the mutation aptamer thereof have high specificity and affinity and can specifically recognize the LMG, so that a rapid detection method taking the LMG as a detection target can be established, the process of reducing the LMG into the MG in sample pretreatment can be avoided, the operation process is greatly simplified, and rapid on-site detection is facilitated.

The invention provides an immunodetection method capable of detecting Furaltadone metabolite AMOZ, MG (malachite green) and LMG (leucomalachite green) simultaneously. An AMOZ hapten and an LMG hapten are synthesized scientifically and coupled with carrier proteins respectively, and artificial antigens are prepared. Hybridoma cell strains aiming at the two objects are prepared respectively from the AMOZ artificial antigen and the LMG artificial antigen, a hybrid-hybridoma cell strain capable of secreting an anti-AMOZ and anti-LMG bispecific antibody is prepared with a hybrid-hybridoma technique, a multi-residue immunodetection method is established successfully, simultaneous detection of AMOZ, MG and LMG is realized, the limit of detection of AMOZ is 0.1 ng / mL, and the IC50 is 1.3 ng / mL; the limit of detection of LMG is 4.8 ng / mL, and the IC50 is 45.3 ng / mL. The method has high specificity and higher sensitivity, has broad application prospect in multi-residue detection of AMOZ, MG and LMG in food and is simple and convenient to operate.

The invention relates to a leucomalachite green gold-labeled test paper detection kit. The test paper detection kit has advantages of fast and accurate detection and high sensitivity, therefore, the invention also provides a preparation method of the detection kit. The detection kit includes a test strip which is packaged in a box shell body, and a sample injection hole and an observation hole are arranged on the box shell body, the test paper detection kit is characterized in that: the test strip takes polyvinyl chloride or polyethylene as a back lining, a sample injection area at one end ofthe test strip is affixed and adsorbed with a glass fiber film containing a conjugate of colloidal gold and anti-leucomalachite green monoclonal antibody; a cellulose nitrate film is affixed to a testing area in the middle of the test strip, a small section of the glass fiber film overlaps the cellulose nitrate film, and a water absorption area at the other end of the test strip is affixed with apiece of absorbent paper; a detection line with material of recessive malachite green-egg white albumin and a quality control line with material of goat anti-mouse IgG are arranged in the direction from the sample injection area to the water absorption area sequentially, and the cellulose nitrate film is covered with a bovine serum albumin layer with the concentration of 1 percent; and the sampleinjection hole is corresponding to the glass fiber film in the sample injection area of the test strip, and the observation hole is corresponding to the testing area of the test strip.

The invention discloses an immune colloidal gold test stripe for detecting leucomalachite green in aquatic products and a preparation method; the colloidal gold test stripe provided by the invention detects leucomalachite green with a direct competition method, and comprises a sample pad, a colloidal gold combined pad, a nitrocellulose membrane (NC), a water adsorption pad and a PVC back lining, one end of the PVC back lining is sequentially adhered with the sample pad and the colloidal gold combined pad, and the middle thereof is adhered with the NC layer, and the other end thereof is sequentially adhered with water adsorption pad; the immune colloidal gold test stripe has the characteristics that colloidal gold combined pad is enveloped with colloidal gold marker which is monoclonal antibody of the specificity of the leucomalachite green, and the NC is adsorbed with leucomalachite green-OVA and sheep anti-mouse IgG. The invention has high detection sensitivity and low price and is simple and rapid, the whole reaction only needs 30 minutes; the immune colloidal gold test stripe can be used for large-batch screening of samples and can rapidly detect the residual leucomalachite green in the aquatic products at site.

The invention discloses a toxicant elimination and fish-odor removal culturing method for soft-shelled turtles. According to the method, rotational flows are generated with oxygenic small molecular group water in a culture pond capable of generating the rotational flows in an intermittent mode, so that the culturing mode that the turtles swim for 30 minutes and rest for 90 minutes is achieved. All residual medicines, including leucomalachite green, of the cultured turtles are not detected, and therefore the turtles become delicious foods or high-quality health care products.

The invention relates to a synthetic method of pentadeuterium-substituted malachite green salt. According to the method disclosed by the invention, a traditional synthesis method in the prior art is completely abandoned; instead, a new synthetic route is employed, wherein the method comprises the following steps: firstly, preparing a Grignard reagent and pentadeuterium substituted tert-butyl benzoate; then the pentadeuterium-substituted leucomalachite green is synthesized in a nucleophilic addition mode; it is unexpectedly found that few by-products are synthesized through the method, the yield of the products is high, and pentadeuterium-substituted malachite green salt prepared from the synthesized pentadeuterium-substituted leucomalachite green is few in impurity and can serve as a high-quality candidate of malachite green standard substances. More importantly, in a traditional synthesis method, hydrogen-deuterium exchange is extremely easy to occur in the reaction process, and high-abundance deuterium-substituted malachite green salt is difficult to synthesize; however, unnecessary hydrogen-deuterium exchange can be avoided in the method, so that the high-abundance deuterium-substituted malachite green salt can be obtained under mild reaction conditions.

The invention discloses a preparation method of a leucomalachite green standard sample in snakehead meat powder. The preparation method comprises the steps of selecting an experimental animal, carrying out medicine feeding on fresh and live snakeheads in a malachite green medicated bath mode, controlling the medicated bath time, and carrying out animal metabolism to obtain a sample material containing leucomalachite green; adding an antioxidant and a preservative into fresh meat, uniformly stirring, freeze-drying, crushing, sieving, adjusting the concentration, uniformly mixing, sub-packaging, and irradiating to prepare the leucomalachite green standard sample in the snakehead meat powder. The standard sample obtained by the invention can be transported at normal temperature, has long shelf life, can be used for quality control of related detection analysis, is uniform and stable, is convenient to transport and reliable to use, and has remarkable economic value and market competitiveness, and the binding situation of a target object and a matrix is consistent with that of a real detection sample.

The invention relates to a detection method of Leucomalachite green, and specifically relates to a quick extraction and detection method of Leucomalachite green in freshwater fish. The method comprises the following steps: firstly, quickly extracting malachite green and Leucomalachite green in the freshwater fish, adding potassium borohydride powder to convert the malachite green into the Leucomalachite green, collecting the multi-wavelength chromatographic data through chromatography-spectrum combination, and treating to obtain a background spectrum database N not containing the Leucomalachite green; secondly, establishing a standard spectrum database V of the Leucomalachite green; thirdly, quickly extracting the freshwater fish sample on the market, adding the potassium borohydride powder, and then collecting a sample ultraviolet-visible spectrum c; and fourthly, quickly computing the total amount of the Leucomalachite green in the to-be-detected sample by adopting a vector-sub-spaceincluded criterion algorithm. The problem that the existing method for extracting the malachite green and the Leucomalachite green is tedious, complex in operation and low in extraction rate is solved, and the quick detection of the Leucomalachite green in the freshwater fish is realized.

The invention relates to a method for determining malachite green, quinolone and sulfonamide drugs in aquatic products. The method comprises the following steps: step 1, preparing samples; step 2, extracting to-be-detected samples to obtain to-be-processed samples; step 3, determining chromatographic conditions; step 4, determining mass spectrum conditions; step 5, drawing a standard working curve; step 6, determining sensitivity, accuracy and precision. With adoption of the method, malachite green, leucomalachite green, quinolone and sulfonamide drugs in an extracted and concentrated aquatic product substrate are determined simultaneously for the first time, the problems that time is consumed and detection cost is high due to traditional respective extraction are solved, ion response value is increased, sensitivity is enhanced, detection limit is low, the effects of being qualitative, quantitative, accurate, fast, efficient and sensitive are realized truly, and the method can be taken as a reliable detection means for the three drugs and is applicable to extraction and concentration of a batch of samples.

The invention discloses malachite green hapten, malachite green artificial antigen and preparation methods of the malachite green hapten and artificial antigen. The malachite green hapten retains the feature structure of malachite green, and has active arms which are derived from different microdomains of the structure of malachite green, have different electron cloud densities and different carbon chain lengths, are suitable to be coupled with macromolecule carriers, and are amide linkage with amino or carboxyl tail end. The malachite green artificial antigen prepared with leucomalachite green hapten has very high specificity and sensitivity when applied to detection of the residual quantity of leucomalachite green and malachite green, and is high in accuracy and recovery. Moreover, the methods are simple, the cost is low, and a fast, sensitive, economical and efficient malachite green immunodetection method is established.

The invention discloses a method for preparing leucomalachite green immunogen, which is characterized by comprising: adding bovine serum albumin, leucomalachite green immunogen, 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride into phosphate buffer solution one by one according to a molar ratio of 1:592:5216; reacting for 2 hours at room temperature; filling liquid obtained after reaction into a dialysis bag; filling into 500 milliliters of distilled water, dialyzing for 48 hours, and changing liquid once every 12 hours; and collecting liquid in the dialysis bag, pre-freezing at -20 DEG C for 2 hours, freezing at -50 DEG C, and drying to obtain leucomalachite green immunogen purple solid powder. In the invention, the preparation process is simplified, the time for preparing the leucomalachite green immunogen is shortened, the production efficiency is improved by about 49 percent, the production cost is reduced, and the promotion and use are easy.

The invention discloses a method for degrading the malachite green waste water. The malachite green is degraded through oxidation after reduction, the malachite green can be reduced into the leucomalachite green to be precipitated in a short five hours, and accordingly a COD (Chemical Oxygen Demand) value of the waste water is reduced, meanwhile the chemical raw material leucomalachite green is obtained, malachite green large molecules are decomposed into small molecules of pollutants, and the further degradation of the malachite green is facilitated and the malachite green waste water which is not fully degraded is oxidized by an oxidation method and accordingly the degradation time is reduced and the energy consumption of the electrolysis is reduced.