Peptides are very diverse molecules that can participate
in a wide
variety of biological processes. In this way, peptides are attractive
for doping, since these molecules can activate or trigger biological
processes that can improve the sports performance of athletes. Peptide
molecules are found in the official World Anti-Doping Agency lists,
mainly in sections S2, S4, and S5. In most cases, these molecules
have a very short half-life in the body and/or are identical to natural
molecules in the body, making it difficult to analyze them as performance-enhancing
drugs. This article reviews the role of peptides in doping, with special
emphasis on the peptides used as reference materials, the pretreatment
of samples in biological matrices, the instrumentation, and the validation
of analytical methodologies for the analysis of peptides used in doping.
The growing need to characterize and quantify these molecules, especially
in complex biological matrices, has generated the need to search for
robust strategies that allow for obtaining sensitive and conclusive
results. In this sense, strategies such as solid phase peptide synthesis
(SPPS), seeking to obtain specific peptides, metabolites, or isotopically
labeled analogs, is a key tool for adequate quantification of different
peptide molecules in biological matrices. This, together with the
use of optimal methodologies for sample pretreatment (e.g., SPE or
protein precipitation), and for subsequent analysis by high-resolution
techniques (mainly hyphenated LC-HRMS techniques), have become the
preferred instrumentation to meet the analytical challenge involved
in the analysis of peptides in complex matrices.