Abstract
The exogenous anabolic-androgenic steroid (AAS) stanozolol stays one of the most detected substances in professional sports. Its detection is a fundamental part of doping analysis, and the analysis of this steroid has been intensively investigated for a long time. This contribution to the detection of stanozolol doping describes for the first time the unambiguous proof for the existence of 17-epistanozolol-1'N-glucuronide and 17-epistanozolol-2'N-glucuronide in stanozolol-positive human urine samples due to the access to high-quality reference standards. Examination of excretion study samples shows large detection windows for the phase-II metabolites stanozolol-1'N-glucuronide and 17-epistanozolol-1'N-glucuronide up to 12 days and respectively up to almost 28 days. In addition, we present appropriate validation parameters for the analysis of these metabolites using a fully automatic method online solid-phase extraction (SPE) method already published before. Limits of identification (LOIs) as low as 100 pg/ml and other validation parameters like accuracy, precision, sensitivity, robustness, and linearity are given.
Highlights
The family of anabolic-androgenic steroids (AAS) belongs to one of the most common illicitly used substance class in the world of professional sports
In 1986, the team around Donike and Schänzer developed the first method for the analysis of the metabolite 30-OH-stanozolol applying gas chromatography–mass spectrometry (GC–MS).[4]
Due to access to high-quality reference samples, an elimination curve based on the absolute metabolite concentrations of all four stanozolol-N-glucuronides in human urine excretion samples was shown for the first time
Summary
The family of anabolic-androgenic steroids (AAS) belongs to one of the most common illicitly used substance class in the world of professional sports. The athletes gave permission to use the urine samples for research purposes, according to the International Standard for Laboratories (ISL).[31] Samples used for the excretion study were provided by the accredited anti-doping laboratory Cologne, Institute of Biochemistry—German Sport University Cologne, Germany. For these samples, a male healthy volunteer received a single oral dose of 5 mg of stanozolol (Winstrol®). Coefficient of variation (CV) of areas (normalized with IS) for intra- and inter-day precision for three concentration levels was calculated by measuring samples on three consecutive days
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