Quantification of endogenous steroid sulfates and glucuronides in human urine after intramuscular administration of testosterone esters

Long‐term stability study and evaluation of intact steroid conjugate ratios after the administration of endogenous steroids

“…27 Earlier studies also report a decrease in TS concentration after the injection of T esters. 7,14,27 Hence, TS might therefore be a valuable parameter in the denominator of steroid conjugate ratios for the detection of IM T. Indeed, AS/TS was proposed as a marker for the detection of IM TU 27 and IM T cypionate. 7 In our study, AS/TS exceeded the individual reference limit as well.…”

“…However, the study of Forsdahl et al 27 state that no increase in the concentration of AG and EtioG was observed after IM TU administration.…”

“…This can be explained by the fact that E is not a metabolite of T and that production of E is suppressed after AAS use. 5,27 It can be observed that the ratios with diol glucuronides in the numerator are even higher ranked than TG/EG after BBQ analysis. EpiAS/EG (ranked 19th) and EpiAS/ES (ranked 16th) were also in the top 20 of best biomarkers (Supporting Information).…”

“…Hence, the steroid sulfates are currently not included, although sulfates have been proven to be useful in the detection and confirmation of testosterone administration. 4,7,15,[27][28][29][30] Additionally, for several steroids with multiple possible moieties that can be glucuronidated, information is lost on the potential influence of administration on the isomer distribution.…”

Annual banned‐substance review: Analytical approaches in human sports drug testing 2019/2020

“…By including conventionally determined unconjugated and glucurono‐conjugated steroidal concentrations, 29 concentration ratios with steroid sulfates were created and tentative upper limits of concentrations and ratios were suggested for future consideration in the ABP steroidal module for males and females. How these ratios perform intraindividually with and without doping scenarios will necessitate administration studies, such as done in a study by Forsdahl et al, where the intact glucurono‐ and sulfo‐conjugates of T, EpiT, A, and E were determined after intramuscular injection of either 1 g of T undecanoate or a mixture of T propionate, T phenylpropionate, T isocaproate, and T decanoate (30 mg/60 mg/60 mg/100 mg) 75 . The analytical approach was based on the SPE of 2 ml of urine followed by LC‐HRMS analysis using a C‐18 analytical column (150 × 2.1 mm, 1.7 μm particle size), 0.1% formic acid (solvent A), acetonitrile (containing 0.1% formic acid, solvent B), negative ESI, and full scan analysis with a Q/Orbitrap analyzer at a resolution of 70,000.…”