Determination of 17α-methyltestosterone in Bovine Serum Using Liquid Chromatography Tandem Mass Spectrometry
PATRICIA REGAL, CAROLINA NEBOT, BEATRIZ I. VÁZQUEZ, ALBERTO CEPEDA
AND CRISTINA A. FENTE
Área de Nutrición y Bromatología. Departamento de Química Analítica, Nutrición y Bromatología, Facultad de Veterinaria,
Universidad de Santiago de Compostela, Lugo 27002, Spain
(Received: August 4, 2009; Accepted: December 29, 2009)
ABSTRACT
A high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) based method has been developed and validated to confirm the presence of the synthetic growth promoter 17α-methyltestosterone (MT) in bovine serum samples. After a single-step extraction with acetonitrile, the enhanced determination of methyltestosterone was achieved by detecting its oxime derivative. In accordance with the criteria defined in Commission Decision 2002/657/EC and using deuterated testosterone (T-d3) as an internal standard, the method has been validated in terms of decision limit (0.05 ng/mL), detection capability (0.09 ng/mL), repeatability CV (7.04%), within-laboratory reproducibility CV (11.00%), and accuracy (87.00%).
Key words: methyltestosterone, bovine serum, HPLC-MS/MS, validation