Anti-doping analysis in sport: new methods on biological samples to protect athlete health and safety

In this PhD project, original and innovative approaches for the qualitative-quantitative analysis of doping substances have been studied, developed and validated for applications in various fields such as forensic, clinical and pharmaceutical. All parameters involved in the developed analytical workflows have been adequately and accurately optimized, from sample collection to sample pretreatment to instrumental analysis. Advanced dry blood microsampling technologies have been developed, which can bring several advantages to the method as a whole, such as a significant reduction in the use of solvents, feasible storage and transport conditions and improvement of the stability of the analyte. At the same time, the use of capillary blood allows to increase the compliance of the subject and the applicability of the global method by exploiting such innovative technologies. The biological samples involved in this project were subjected to optimized pretreatment techniques developed ad hoc for each target analyte, also making use of advanced microextraction techniques. Finally, original and advanced instrumental analytical methods based on high-performance liquid chromatography and nano liquid chromatography (HPLC, Nano UHPLC) coupled to mass spectrometry were developed. In addition, a pharmacovigilance study coupled with statistical analysis was carried out to assess the health risk of certain specific doping substances. Each method has been designed to achieve highly selective, sensitive but sustainable systems and has been validated according to international guidelines. All methods developed here have proven to be suitable for the analysis of test compounds and may be useful tools in pharmaceutical chemistry, pharmaceutical analysis, clinical studies and forensic investigations.