The misuse of androgenic anabolic steroids in sports was banned in 1976 by the International Olympic Committee and global sports community. The illegal use of anabolic steroids has reached disturbing levels worldwide. This worldwide problem is fueled partially by an ever-increasing demand for better athletic performance. The World Anti-Doping Agency has formulated strict guidelines for minimum allowable concentrations of exogenous anabolic steroids and their metabolites. The standard test methods for doping control are analyzed in urine samples with trimethyl-silyl derivatization. Urine is a complex and difficult biological matrix. This research shows the advantages of using comprehensive two-dimensional gas chromatography–time-of-flight-mass spectrometry (GCÃ-GC–TOF-MS) and illustrates the capability of GCÃ-GC-TOF-MS to be an effective instrumental option for antidoping control screening.