Development and validation of a LC-MS/MS analytical procedure for the combined detection of prohibited peptides in biological fluids

Recently a lot of peptide hormones were included in the section 2 “Peptide hormones, growth factors and related substances” of the World Anti-Doping Agency list of prohibited substances and methods [1]. Traditionally, the analytical procedures used to detect macromolecules in the anti-doping field were based on the use of immunological techniques. The increasing number of peptide hormones and other macromolecules with minimal differences in the molecular structure with respect to those produced endogenously has imposed the development of more selective techniques, also based on mass spectrometric. Several analytical procedures were already published to detect antidiuretic peptides [2-3], growth hormone releasing factors [4-5], ACTH and its analogues, IGF1 and its analogues and insulin analogues [6] in biological fluids. Here we present an analytical method for the combined screening of 14 prohibited peptides and metabolites by means of liquid chromatography/tandem mass spectrometry in selected reaction monitoring acquisition mode after solid phase extraction.