title: Longitudinal monitoring of endogenous steroids in human serum by UHPLC-MS/MS as a tool to detect testosterone abuse in sports.
abstract: The detection of testosterone abuse in sports is routinely achieved through the 'steroidal module' of the Athlete Biological Passport by GC-MS(/MS) quantification of selected endogenous anabolic androgenic steroids (EAAS) from athletes' urines. To overcome some limitations of the "urinary steroid profile" such as the presence of confounding factors (ethnicity, enzyme polymorphism, bacterial contamination, and ethanol), ultrahigh performance liquid chromatography (UHPLC) measurements of blood concentrations of testosterone, its major metabolites, and precursors could represent an interesting and complementary strategy. In this work, two UHPLC-MS/MS methods were developed for the quantification of testosterone and related compounds in human serum, including major progestogens, corticoids, and estrogens. The validated methods were then used for the analyses of serum samples collected from 19 healthy male volunteers after oral and transdermal testosterone administration. Results from unsupervised multiway analysis allowed variations of target analytes to be assessed simultaneously over a 96-h time period. Except for alteration of concentration values due to the circadian rhythm, which concerns mainly corticosteroids, DHEA, and progesterone, significant variations linked to the oral and transdermal testosterone administration were observed for testosterone, DHT, and androstenedione. As a second step of analysis, the longitudinal monitoring of these biomarkers using intra-individual thresholds showed, in comparison to urine, significant improvements in the detection of testosterone administration, especially for volunteers with del/del genotype for phase II UGT2B17 enzyme, not sensitive to the main urinary marker, T/E ratio. A substantial extension of the detection window after transdermal testosterone administration was also observed in serum matrix. The longitudinal follow-up proposed in this study represents a first example of 'blood steroid profile' in doping control analysis, which can be proposed in the future as a complement to the 'urinary module' for improving steroid abuse detection capabilities.
authors.affiliation: Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Geneva and Lausanne, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Chemin des Croisettes 22, 1066, Epalinges, Switzerland.
authors.forename: Federico
authors.initials: F
authors.lastname: Ponzetto
authors.affiliation: Laboratory of Pharmaceutical Analytical Chemistry, School of Pharmaceutical Sciences, University of Geneva, Pavillon des Isotopes, Boulevard d'Yvoy 20, 1211, Genève 4, Switzerland.
authors.forename: Florence
authors.initials: F
authors.lastname: Mehl
authors.affiliation: Laboratory of Pharmaceutical Analytical Chemistry, School of Pharmaceutical Sciences, University of Geneva, Pavillon des Isotopes, Boulevard d'Yvoy 20, 1211, Genève 4, Switzerland.
authors.forename: Julien
authors.initials: J
authors.lastname: Boccard
authors.affiliation: Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Geneva and Lausanne, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Chemin des Croisettes 22, 1066, Epalinges, Switzerland.
authors.forename: Norbert
authors.initials: N
authors.lastname: Baume
authors.affiliation: Laboratory of Pharmaceutical Analytical Chemistry, School of Pharmaceutical Sciences, University of Geneva, Pavillon des Isotopes, Boulevard d'Yvoy 20, 1211, Genève 4, Switzerland.
authors.forename: Serge
authors.initials: S
authors.lastname: Rudaz
authors.affiliation: Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Geneva and Lausanne, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Chemin des Croisettes 22, 1066, Epalinges, Switzerland.
authors.forename: Martial
authors.initials: M
authors.lastname: Saugy
authors.affiliation: Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Geneva and Lausanne, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Chemin des Croisettes 22, 1066, Epalinges, Switzerland. raul.nicoli@chuv.ch.
authors.forename: Raul
authors.initials: R
authors.lastname: Nicoli
mesh_terms: D000328:Adult; D004300:Doping in Sports; D006801:Humans; D008137:Longitudinal Studies; D008297:Male; D013256:Steroids / Q000097:blood*; D015813:Substance Abuse Detection / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013739:Testosterone / Q000008:administration & dosage*; D055815:Young Adult
chemical_list: D013256:Steroids; D013739:Testosterone
keywords: Multiway data analysis; Serum; Steroid profile; Testosterone doping; UHPLC-MS/MS