High performance liquid chromatography to determine animal drug Clenbuterol in pork, beef and hog liver

Abstract

A semi-micro high performance liquid chromatography (HPLC) was developed to determine clenbuterol in pork, beef and hog liver. The procedure included extraction with 0.4 N perchloric acid, partitionally separation with diethyl ether, and then back-extraction with 0.2 M sulphuric acid. Analytical procedure was performed by HPLC with a Waters Cosmosil column 5C18-MS (2.0 × 150 mm), a mobile phase of 0.05 M NaH2PO4 (pH3.0)/acetonitrile solution (80/20, v/v) and absorbance at 212 nm. The recoveries of clenbuterol in pork, beef and hog liver spiked with standards of 0.0005-0.01 ppm were 80.9-90.6 %. The detection limit of clenbuterol in tested samples was 0.0001 ppm, which is lower than the officially allowed level. The equation was linear for detecting 0.0002-0.001 ppm clenbuterol. The method was validated to be usable for market samples. Three out of 150 samples were detected to contain clenbuterol (0.0001-0.00015 ppm), which were less than both JECFA and Department of Health regulation levels (hog liver 0.0006 ppm and beef 0.0002 ppm).