Analytical procedure for steroid profiling valid for Athlete Biological Passport

Although various attempts have been made to eliminate doping in sport, hitherto they all have proved futile. Moreover, the main class of substances that jeopardises the fair play rule remains the same - anabolic androgenic steroids (AAS). To date, longitudinal monitoring of the fluctuations of the endogenous steroids content for a given athlete is regardeded as the most effective approach to the detection of AAS abuse. This is based on the fact that the activity of the steroid biosynthesis pathway may undergo significant changes in response to the AAS administration. This paper presents the entire analytical procedure for quantification of steroids crucial for the Athlete Biological Passport (ABP): testosterone, epitestosterone, dehydroepiandrosterone, androsterone, etiocholanolone, 5-α-androstandiol and 5-β-androstandiol. The procedure consists of a four-step sample preparation process followed by analysis by gas chromatography coupled with mass spectrometry. The limits of quantification for the substances listed above were; 0.44 ng mL