Synthesis of 3,6-Dihydroxyandrostan-17-one 3-Glucuronides for the Detection of Testosterone Misuse

The convenient synthesis of two 3-glucuronide conjugates of 6 beta-hydroxyandrosterone (6OH-And-3G 2) and 6 beta-hydroxyetiocholanolone (6OH-Etio-3G 3) is reported. The conjugates are identified as long-term markers of endogenous androgenic anabolic steroid abuse in doping control studies. Their resistance to enzymatic hydrolysis by E. coli beta-glucuronidase means they may be missed by conventional GC-MS analysis protocols. One promising strategy to quantify these markers is by direct methods, which requires sufficient quantities of high purity reference materials. Our convergent synthesis features concurrent stereoselective introduction of 5 alpha/5 beta hydrogen and 6 beta-hydroxy groups on the steroid skeleton, and a mild Schmidt trichloroacetimidate glycosylation strategy.