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Isolation, Identification, and Determination of Designer Anabolic Steroids Commonly Found in Dietary Supplements

The marketing of “pro-hormone” dietary supplements for athletes seeking to
increase muscle mass, strength, endurance, and recovery time has increased dramatically in recent years. Stricter drug testing regulations have prompted a few corrupt supplement manufacturers to use chemically-modified structures of existing anabolic steroids, apparently in an attempt to evade detection. These “designer” steroids are expected to convert to active hormones in the body, producing the desired effect. Although little is known about the pharmacological effects of these compounds, it is likely that they, like their banned counterparts, may cause serious long-term adverse health consequences. Several steroid-like compounds in various dietary supplements have recently been detected in our laboratory. These compounds could not be readily identified due to the lack of library reference spectra or commercially available standards. The general analytical approach to these emerging compounds will be presented, including analysis by GC-MS, LCMS, and/or HPLC-UV. Analytical scale high performance liquid chromatography with fraction collection was used to isolate and collect portions of several new designer steroids observed in samples received. The isolated compounds were then characterized by Nuclear Magnetic Resonance (NMR) spectroscopy and high resolution accurate mass-mass spectrometry (HRAM-MS) to elucidate their structure. Quantitative analysis of these emerging substances in some representative dietary supplements was achieved using HPLC-UV comparison to structurally related reference standards.