Designer Steroids and Prohormones in Dietary Supplements: A Strategy for Detection Using a Mammalian Cell‐Based Androgen Receptor BioAssay and GC/MS

TL;DRAbstract

Adulteration of dietary supplements is not uncommon, with some manufacturers using misleading labeling and ingredient aliases to deceive consumers. The purpose of this work was to evaluate the possible anabolic activity of selected supplements using a mammalian cell-based human androgen receptor (hAR) bioassay. A comprehensive database containing 82,560 supplement products, 2,813 ingredients, and 37,095 ingredient aliases was searched for products containing anabolic agents. Selected products were analyzed using full-length hAR plasmid and a PB-ARE-2 plasmid transfected into COS-7 cells using Lipofectamine 2000 CD™. After transfection and recovery, the cells were treated with the supplement extracts, testosterone standards, or vehicle. Luciferase activity was quantified using a VICTOR™ X4 Multilabel Plate Reader. Three independent reporter gene assays were performed per sample. Of the 2,813 ingredients listed in the database, 165 were identified as anabolic agents; 79 were associated w

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