| Food safty has already become the global focus of attention. In recent years, a lot of news about food poisoning resulting from clenbuterol (CBL) residues in food has been reported. Currently, there is appreciable concern about the potential consumer health hazards associated with the illegal use of clenbuterol as a growth promoting agent.Time-resolved fluoroimmunoassay (TRFIA) developed in recent ten years is one kind of infant nonradiolabeled immunoassay, which is rapid, selectivity and well-reproducibility to screen large samples.In order to provide a rapid and selectivity TRFIA kit with own intellectual property rights for the determination of clenbuterol, clenbuterol was diazotized firstly and then coupled to Keyholelimpet hemocyanin (KLH) and Ovalbumin (OVA) to synthesis immunogen and coating antigen respectively. Anti-CBL antibody was raised by immunization against CBL-KLH in New Zealand white rabbits. The assay results showed that the rabbit's antiserum titre is as high as 1∶204800. Based on this antibody, a competitive indirect TRFIA was designed. CBL-OVA was coated by physical adsorption onto the microtitre plate, CBL or sample with CBL as a competitor. Both them were incubated with limited anti-CBL antibody, and a goat antirabbit IgG-Eu3+ conjugate was used as a tracer. A competitive indirect CBL-TRFIA kit was developed. It provided a limit of detection of 0.0076μg/L, recoveris (body fluid and tissues) between 80 and 120 percent. Preliminary evaluation of assay performance through following items: reproducibility, sensitivity, precision, and accuracy showed that this TRFIA kit can applied to practical detection for CBL. Moreover, it was compared with the Enzyme-linked immunosorbent assay (ELISA) kit.
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