Method for detecting chlorpromazine by colloidal gold immune chromatography test

An immunochromatography, colloidal gold technology, applied in the field of bioengineering, to achieve the effect of strong pertinence, single detection object, and high accuracy

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An immunochromatography, colloidal gold technology, applied in the field of bioengineering, to achieve the effect of strong pertinence, single detection object, and high accuracy

CN101424688AInactive Publication Date: 2009-05-06SHANGHAI JIAO TONG UNIV

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[0031] 1. Preparation of conjugates CPZ-KLH and CPZ-BSA

[0032] ①Preparation of immune antigen: Dissolve 0.350g p-aminobenzoic acid in 3ml water and 0.7ml concentrated hydrochloric acid; add 2.5mmol sodium nitrite (1ml water) dropwise, and stir for 30min; add 0.15ml 1M sulfuric acid dropwise Ammonium, and keep stirring for 10min to remove excess nitrous acid; dissolve 2.5mmol CPZ in distilled water, gradually add the diazonium salt of p-aminobenzoic acid and keep stirring; add 1ml of 2mM sodium acetate; add 2mmol of sodium acetate after 6 hours, Use pH test paper to adjust the pH of the solution to 1.5; stir overnight at 4°C; adjust the pH to 5.6 with 0.5N NaOH, then extract with 30ml of dichloromethane; wash the organic layer twice with distilled water, and vacuum-dry the organic phase to obtain pink colored dry matter.

[0033] Use the chromatographic solution methanol-chloroform-NH4OH (20:80:5) to dissolve a certain amount of light pink dry matter, use a capillary glass t...

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Abstract

The invention relates to a method using a colloidal gold immuno chromatographic assay to test chlorpromazine in the technical field of biological engineering. CPZ hapten is connected with aminobenzoic acid through diazo reaction, and the CPZ hapten and KLH and BSA amidogens form peptide linkages by carboxyl on benzene rings so as to form the CPZ conjugates of CPZ-KLH and CPZ-BSA. The antigen CPZ-KLH is used for preparing polyclonal antibodies, the antigen CPZ-BSA is used for an indirect Elisa adsorption assay for testing polyclonal antibody titers, the polyclonal antibodies are labeled by the colloidal gold used as a tracer label, the CPZ-BSA antigen is immobilized on a test line on a membrane, and anti-rabbit polyclonal antibodies are immobilized on a control line; and the gold-labeled antibody, the corresponding antigens and the antibodies are specifically bound to perform the operation of colour development through isolation, and negative and positive results are displayed by the colour development of the test and the control lines.The invention can directly test chlorpromazine micromolecules in samples and has the advantages of fast test speed and short required time.

Description

technical field [0001] The invention relates to a detection method in the technical field of bioengineering, in particular to a method for detecting chlorpromazine by colloidal gold immunochromatography test. Background technique [0002] Chlorpromazine (CPZ, N, N-dimethyl-2-chloro-10H-phenothiazine-10-propylamine hydrochloride) is a good sedative, which has a good inhibitory effect on the central nervous system. Appropriate addition in animal feed can significantly increase daily gain, reduce feed consumption, and can be used as an anti-stress drug to prevent stress syndrome and reduce mortality during transportation. CPZ is mainly metabolized in the liver, and most of it is excreted with urine, and the excretion method is slow, which is easy to produce drug residues. CPZ has an inhibitory effect on the nervous system, respiratory system and circulatory system of the human body, and can enter the fetal blood circulation through the placental barrier. People with mild pois...

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Application Information

Patent Timeline

06 May 2009

Publication

CN101424688A

IPC: G01N33/558

Inventors: 孙建和; 史一博