Abstract: 【Objective】The aim of the study is to establish a detection method for the immune colloidal gold of phenylethanolamine A in animal urine. 【Method】Phenylethanolamine A hapten was synthetized by nucleophilic substitution with phenylethanolamine A and bromo aldehyde. Phenylethanolamine A hapten was coupled with BSA and OVA, respectively, to prepare immunogen and envelope antigen. The coupling ratio of hapten-BSA/OVA was determined by TNBS. Monoclonal antibody was acquired from mice which were immunized with hapten-BSA. The indirect ELISA method has been established for sensitivity and specificity detection of monoclonal antibody. A total of 13 compounds whose structures were similar with phenylethanolamine A were used to test the specificity and calculate the cross reaction rate of monoclonal antibody. A rapid detection method for immune colloidal gold of phenylethanolamine A in animal urine was established by the parts of strip prepared and assembled. The strip’s detection limits, false positive rate and false negative rate were determined. Compared the immunochromatography assay (ICA) with ELISA whose limited ranges of standards’ concentration were 0, 0.1, 0.3, 0.9, 2.7 and 8.1 μg·L^-1, the detection limits for pig urine was 0.5 μg·L^-1 and the ranges of percent recovery were 75%-105%.【Result】Phenylethanolamine A derivative was prepared that amino terminal of phenylethanolamine A connected with the connecting arm whose aldehyde group was on the end. Phenylethanolamine A hapten connected with BSA and OVA successfully by TNBS and the coupling ratio between hapten with BSA and OVA were 11.38 and 10.66, respectively. The titer and ascites of MC-73 hybridoma cell strain were 1﹕2×10³ and 1﹕5×10⁷ which were higher than MC-33 and MC-29. IC₅₀ of monoclonal antibody against phenylethanolamine A was 0.365 μg·L^-1 which was more sensitive than MC-33 and MC-29. The cross reaction rate with 13 compounds whose structures were similar with phenylethanolamine A were less than 1%. The detection limit, false positive rate and false negative rate were 5 μg·L^-1, 0 and 0, respectively. Detection results of phenylethanolamine A in pig urine between ICA and ELISA were the same.【Conclusion】The phenylethanolamine A strip with high sensitivity, specificity, accuracy and rapidity was developed in this study which would be used to test animal urine in 5 min without pre-treatment of urine samples. It would be efficient for mass samples screening in primary laboratory and enterprises.

Key words: phenylethanolamine A, colloidal gold immunochromatographic assay, rapid detection strip, animal urine