Determination of Beta 2-receptor Agonists in Bovine Urine and Liver by Gas Chromatography-tandem Mass Spectrometry

A highly specific and sensitive method for the simultaneous detection of seven beta 2-receptor agonists in bovine liver homogenates and urine was developed. A 10-g amount of liver was homogenized and treated with Subtilisin A. The resulting enzymatic digest was extracted with tert.-butanol-ethyl acetate (3:7) and the crude extract was purified on a 6-ml Bakerbond alumina neutral disposable extraction column. Subsequently, the hydrous eluate from the alumina column was buffered at pH 6 and loaded on top of a preconditioned 3-ml Bond-Elut Certify column. Urine was buffered and loaded onto a 3-ml Certify column without pretreatment. The analytes were eluted with dichloromethane-isopropanol (8:2) containing 2% ammonia. The extract obtained was trimethylsilylated and analysed by gas chromatography-tandem mass spectrometry using multiple selected reaction monitoring. The limits of detection for the beta 2-receptor agonists evaluated were between 0.5 and 5 ppb.