Simultaneous Determination of 11 β-Agonists in Human Urine Using High-Performance Liquid Chromatography/Tandem Mass Spectrometry with Isotope Dilution

The misuse of beta-agonists constitutes a potential risk to public health and has been forbidden in many countries. In this study, we describe a method for specific, sensitive and rapid detection of beta-agonists in human urine. Urine samples were extracted with ethyl acetate, without any additional purification step, and analyzed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) with Clenbuterol-D9 and Salbuterol-D3 as internal standards. The intra-and interday precision values of the method were all <5.60% and the accuracy ranged from 94.5 to 109%. Extraction recovery for 11 beta-agonists varied from 66.7 to 108%. One UPLC-MS-MS analysis could be completed within 12 min and the limits of detection for 11 beta-agonists were 0.1 ng/mL in the experiment. beta-Agonists in human urines from 24 volunteers were analyzed by our validated method and 1.70 ng/mL salbutamol was detected in one volunteer. The application of UPLC-MS-MS method in beta-agonists detection of human urine will be helpful in veterinary control of beta-agonists and for studying the effect of beta-agonists on human health.