title: High-resolution mass spectrometry as an alternative detection method to tandem mass spectrometry for the analysis of endogenous steroids in serum.
abstract: Recently, steroid hormones quantification in blood showed a promising ability to detect testosterone doping and interesting complementarities with the urinary module of the Athlete Biological Passport (ABP). In this work, an ultra-high pressure liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) method was developed for the quantification of eleven endogenous steroids in serum. The performance of the full scan and targeted SIM acquisition modes was evaluated and compared to the performance of tandem mass spectrometry (MS/MS). Passing-Bablok regressions and Bland-Altman plots were assessed for each analyte of interest, and concentration values measured by HRMS showed high correlation with the ones obtained by MS/MS for all target hormones, with low absolute differences in the majority of cases. A slight decrease in terms of sensitivity was observed with HRMS in both acquisition modes, but performing an analysis of variance multiblock orthogonal partial least squares (AMOPLS) on the dataset obtained with all three methods revealed that only 0.8% of the total variance was related to instrumentation and acquisition methods. Moreover, the evaluation of the testosterone administration effect over time highlighted testosterone itself and dihydrotestosterone as the most promising biomarkers of exogenous testosterone administration. This conclusion suggests that HRMS could provide suitable performance for blood steroid analysis in the anti-doping field.
authors.affiliation: Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Geneva and Lausanne, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Switzerland.
authors.forename: Federico
authors.initials: F
authors.lastname: Ponzetto
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Switzerland; Swiss Center of Applied Human Toxicology (SCAHT), University of Basel, 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, Switzerland.
authors.forename: Norbert
authors.initials: N
authors.lastname: Baume
authors.affiliation: Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Geneva and Lausanne, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Switzerland.
authors.forename: Tiia
authors.initials: T
authors.lastname: Kuuranne
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Switzerland; Swiss Center of Applied Human Toxicology (SCAHT), University of Basel, Switzerland.
authors.forename: Serge
authors.initials: S
authors.lastname: Rudaz
authors.affiliation: Center of Research & Expertise in anti-doping Sciences, University of Lausanne, 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, Switzerland. Electronic address: raul.nicoli@chuv.ch.
authors.forename: Raul
authors.initials: R
authors.lastname: Nicoli
mesh_terms: D045930:Anabolic Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D013196:Dihydrotestosterone / Q000097:blood; D004300:Doping in Sports; D006801:Humans; D057230:Limit of Detection; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D013256:Steroids / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013739:Testosterone / Q000097:blood*
chemical_list: D045930:Anabolic Agents; D013256:Steroids; D013196:Dihydrotestosterone; D013739:Testosterone
keywords: AMOPLS; Doping; Serum; Testosterone; UHPLC-HRMS