Abstract
Detection of anabolic steroids in animal urine samples is currently performed with GC–MS in our lab. However we found that the detection of 17α-trenbolone (17α-TbOH), 4-chloroandrost-4-ene-3,17-dion (CLAD), 16-β-OH-stanozolol (16OHstan) and α- and β-boldenone (α-Bol, β-Bol) was very difficult, if not impossible. Therefore a sensitive, specific and selective qualitative multi-analyte LC–MS–MS method was developed. The LC separation was achieved by using a Symmetry ® C 18 column and methanol–water–formic acid (54.7–44.7–0.6) as a mobile phase at a flow-rate of 0.3 ml/min. The mass spectrometer was operated in multiple reaction monitoring mode with positive electrospray interface. Validation of the method was done according to draft SANCO/1805/2000 Rev.1 and a CC β smaller then 1 ng/ml was obtained for each compound.
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