Study On Enzyme-Linked Immunosorbent Assay And Colloidal Gold Immunochromatographic Rapid Test Strip For Salbutamol

In this paper, the competitive direct enzyme-linked immunosorbent assays (cd-ELISA) and the colloidal gold immunochromatographic rapid test strip for the determination of Salbutamol(SAL) residues in pig muscles had been developed. We investigated the method of coupling hapten to carrier protein. SAL was coupled with Bovine Serum Albumin (BSA) or HRP by mixed acid anhydride method and Ovalbumin (OVA) by carbodiimide method, producing the immune antigen (SAL-BSA), the enzyme trace (SAL-HRP) and the coating antigen (SAL-OVA). A sensitive polyclonal antibody against the Salbutamol(SAL) was obtained. In the cd-ELISA, the limit of detection (IC15) was0.06±0.02μg/L, and the sensitivity (IC50) was0.80±0.10μg/L. The relative standard deviations (RSD) of intra-and inter-assay were less than20%. Three concentrations (7.0μg/kg、24.0μg/kg、50.0μg/kg) of salbutamol spiked pig muscles were detected by simple extraction and dilution using PBS without any concentration or cleanup steps. The mean recoveried were between85%and100%.On the other side, the colloidal gold immunochromatographic rapid test strip was developed for the determination of salbutamol residues, the limit of detection was1.00μg/L, which After filtered by0.22μm membrane, the samples were directly used for analysis.This method was effectively applied to identify trace salbutamol in pig muscles sample and the limit of detection of salbutamol test strip was10μg/kg. The results obtained by test strip were coherence with the results obtained by cd-ELISA.