The role of high-resolution mass spectrometry in advancing the detection of SARMs and peptide hormones for sports antidoping

The relentless pursuit of athletic excellence is shadowed by the equally persistent evolution of performance-enhancing drugs (PEDs). In recent years, the antidoping landscape has been profoundly shaped by the rise of two sophisticated classes of substances: selective androgen receptor modulators (SARMs) and peptide hormones. These agents present formidable analytical challenges due to their structural diversity, low dosage regimens, and, in the case of peptides, their similarity to endogenous biomolecules. Mass spectrometry (MS), the cornerstone of doping control, has evolved in response, with high-resolution mass spectrometry (HRMS) emerging as the preeminent technology to counter these threats. This review provides a critical and exhaustive analysis of the role of HRMS in the detection of SARMs and peptide hormones. We dissect the entire analytical workflow, from advanced sample preparation techniques such as solid-phase extraction and immunoaffinity purification to state-of-the-art separation science using ultra-high-performance liquid chromatography (UHPLC) and nano-LC. We critically evaluate the ascendancy of HRMS platforms, particularly Orbitrap and time-of-flight (TOF) analyzers, which have enabled a paradigm shift from targeted to nontargeted and retrospective screening, fundamentally altering the strategic dynamics of the antidoping “arms race.” Key scientific controversies are explored in depth, including the forensic dilemma posed by the ultra-high sensitivity of HRMS and the resulting challenges in distinguishing deliberate doping from inadvertent contamination, as well as the ongoing debate between traditional collision-induced dissociation (CID) and newer electron-transfer dissociation (ETD) for peptide characterization. Furthermore, this review examines the synergistic integration of HRMS with emerging technologies such as ion mobility spectrometry (IMS) for isomer separation and artificial intelligence (AI) for managing the immense datasets generated. We conclude that while HRMS provides unprecedented analytical power, its implementation is not a panacea but a complex advancement that introduces new scientific, economic, and forensic questions. The future of antidoping lies in the development of integrated, multidimensional analytical platforms, with HRMS at their core, to ensure the integrity of sport.