Application of high-performance liquid chromatography–UV detection to quantification of clenbuterol in bovine liver samples

Abstract

A method for the determination of clenbuterol (CLB) concentration in bovine liver by reversed-phase high-performance liquid chromatography (HPLC) with UV detection was developed. The sample was extracted with acetonitrile and isopropanol, followed by HPLC analysis. A reverse-phase column C18 was used, with a UV detector at 214 nm and 0.05 M NaH2PO4 (pH 3.0)/acetonitrile (85:15, v/v) as the mobile phase. The accuracy of the analytical method was estimated by spiking bovine liver samples with three different concentrations of CLB (5.24 ng/g, 20.98 ng/g, and 41.96 ng/g) and recovery of 111.7%, 82.0%, and 84.8%, respectively, was obtained. The precision of the method was estimated by the relative standard deviation, which was < 4.74%. The limit of detection and quantification of CLB were 0.20 ng/g and 0.42 ng/g of liver sample, respectively, and the retention time was 24.82 minutes. The recent discovery of CLB contamination in Mexican food led to the specific inspection of a distribution center for this β-agonist, involving the analysis of a total of 78 bovine liver samples. Of all samples screened, 62% of them had concentrations above the maximum residue limit of 0.6 ng/g set by the United Nations Food and Agricultural Organization for CLB. The analytical method was found to be rapid, sensitive, accurate, repeatable, and reproducible, and could be applied to the measurement of CLB concentration in bovine liver.