Determination of Salbutamol, Clenbuterol, and Brombuterol in Urine by a Highly Sensitive Chemiluminescence Enzyme Immunoassay

Abstract

A salbutamol-bovine serum albumin and a salbutamol-ovalbumin coating antigen were synthesized, and six New Zealand white rabbits were treated with the immunogen to obtain polyclonal antibodies to develop a rapid, sensitive, and indirect chemiluminescent enzyme immunoassay (ciCLEIA) for the analysis of swine and bovine urine. The prepared antibodies showed high cross-reactivities with clenbuterol (139.6%) and brombuterol (225%). Under the optimized conditions, the linear dynamic range and the limit of detection (LOD) for salbutamol were from 0.007 to 0.17 lg L À1 and 0.003 lg L À1 , with a correlation coefficient of 0.9965 and a half maximum inhibition concentration of 0.028 lg L À1 . Recoveries for salbutamol, clenbuterol, and brombuterol were from 78.8% to 119.0% with intra-assay and inter-assay coefficients of variation less than 13.9% and 19.7%, respectively. The reported ciCLEIA was about 10-fold more sensitive for salbutamol and 20-fold more sensitive for clenbuterol compared to conventional methods. This study showed that ciCLEIA was a reliable, convenient, and sensitive method for the simultaneous determination of salbutamol, clenbuterol, and brombuterol in swine and bovine urine.

Study on detection methods for salbutamol in food and biological samples

“…However, the illegal use of salbutamol as growth-promoting agents in animals is still a public concern due to its potential risk to the health of individuals consuming animal products contaminated with the residue of β2-agonists [28][29][30]. Thus, it is necessary to establish quick and accurate methods to detect salbutamol residues.…”

Docking guided phase display to develop fusion protein with novel scFv and alkaline phosphatase for one-step ELISA salbutamol detection

“…Immunoassay methods also have some problems. For example, most of the currently available anti-SAL antibodies are polyclonal antibodies from sheep and rabbits (Degand et al, 1993;Lei et al, 2008;Wu et al, 2014), and their specificity is usually poor. For polyclonal antibodies, the heterogeneity of antibody preparations usually leads to cross-reactions with highly similar antigens.…”

“…For polyclonal antibodies, the heterogeneity of antibody preparations usually leads to cross-reactions with highly similar antigens. For example, polyclonal anti-SAL antibody shows significant cross-reactivity to clenbuterol (Degand et al, 1993;Lei et al, 2008;Wu et al, 2014). Compared with polyclonal antibodies, monoclonal antibodies have the advantages of high specificity, and a relatively simple production process has been widely used in immunoassays.…”

Development of an immunochromatographic strip for the rapid detection of 10 β-agonists based on an ultrasensitive monoclonal antibody

“…Immunoassays, including enzyme-linked immunosorbent assay (ELISA) (Bui et al, 2016;Du et al, 2016;Wu et al, 2014), fluorescence immunoassay (FLA) (Cao et al, 2015;Zou et al, 2008) and colloidal gold immunoassay (Zhang et al, 2016), are based on antigen-antibody reactions. The ELISA is a simple and sensitive method (Ren, Zhang, Chen, & Yang, 2009), but not conducive to large-scale sample testing.…”