Synthesis of 3α,6β-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.

Automated summary

The convenient synthesis of two 3-glucuronide conjugates as long-term markers for endogenous androgenic anabolic steroid abuse in doping control studies is reported. These markers are resistant to enzymatic hydrolysis, which may result in them being missed by conventional GC-MS analysis protocols.