Semi-quantitative determination of designer steroids by high-performance liquid chromatography with ultraviolet...

New designer steroids are continually being encountered in dietary supplements that claim to increase muscle mass, but quantitative analysis of such ingredients is challenging due to the availability, quality, or cost of commercial reference materials. Although standard reference material typically becomes available for these emerging compounds, laboratories often face the challenge of finding properly certified materials from accredited suppliers, due to traceability requirements. Several of these designer steroids have been isolated and identified using multiple structural elucidation tools. Structural characteristics of these compounds of interest were evaluated and molar absorptivity data was collected and compared to several readily available steroid standards using ultraviolet/visible spectroscopy. This approach was used to find suitable compounds for use as surrogate reference materials in the semi-quantitative determination of two designer steroids, 1-dehydroepiandrosterone (1-androsterone) and 6β-chloro-4-androsten-17β-ol-3-one (6β-chlorotestosterone). Laboratory-fortified matrix samples and dietary supplement samples were analyzed using this method for the estimation of 1-androsterone and 6β-chlorotestosterone by HPLC-UV. Assay values obtained for the estimation of 1-androsterone in a dietary supplement sample using a prasterone or dehydroepiandrosterone (DHEA) standard curve were 100% of those obtained using a 1-androsterone reference standard, once it became commercially available. Estimations for 6β-chlorotestosterone in laboratory-fortified matrix samples using a testosterone standard curve were 92%-93% of those obtained using isolated 6β-chlorotestosterone as "reference material."