XXII International Mass Spectrometry Conference, IMSC 2018, Florence, Italy

Emiliano attended the XXII International Mass Spectrometry Conference, IMSC 2018 held in Florence on August 2018 with an oral presentation namely “A fit-for-purpose UHPLC-MS/MS method for routine screening of SARM residues in equine plasma and bovine serum”.

Oral presentation abstract: selective androgen receptor modulators (SARMs) are an attractive alternative to anabolic-androgenic steroids despite the fact they are not yet approved as pharmaceutical drugs. Consequently, there is much evidence of their availability through black and grey market sources. In this study, a semi-quantitative UHPLC-MS/MS method was developed to monitor the abuse of 15 compounds belonging to eight different SARM families, in bovine and equine animals, respectively. Serum samples underwent protein crash (0.5 mM NH4OH in MeCN), induced phase separation (NaCl) and defatting (n-hexane) before concentration under nitrogen. Extracted SARM residues were subsequently analysed by UHPLC-MS/MS operating in ESI+/ESI-. Chromatographic separation was performed using a Luna Omega Polar C18 column at 45 °C, and a binary gradient elution of 14 minutes employing H2O and MeOH, both containing 0.1% (v/v) CH3COOH as mobile phases at a flow rate of 0.4 mL/min. Validation of the semi-quantitative assay was carried out according to the Community Reference Laboratories Residues (CRLs) 20/1/2010 guidelines. The following performance studies were carried out during the validation process: specificity, selectivity, detection capability (CCβ), absolute recovery as well as applicability, ruggedness and matrix effects. The detection capability was calculated by assessing threshold value (T) and cut-off factor (Fm) and CCβ values were determined at the following concentrations: 0.5 ng mL-1(CDS-025139), 1 ng mL-1 (AC-252636, PF-06260414), 2 ng mL-1 (bicalutamide, GLPG-0492, LGD-2226, S-1, S-6, S-22, S-23) and  5 ng mL-1 (BMS-564929, LGD-4033, , RAD140, S-4, S-9), respectively. Moreover, ongoing work aims to identify in vivo metabolites of selected SARMs compounds following animal administration studies, which could serve as additional marker residues to be included within the current test method. SARMs are widely recognised as emerging drugs of abuse in animal sport, and also as potent candidates for misuse in food-producing animals. The developed method is currently being employed as a rapid, simple, environmental friendly and cost-effective tool for the screening of serum samples to evaluate the abuse of SARM drugs in stock farming and to monitor for doping practices in racing horses, ensuring customer safety and fair play in animal sport competitions. The developed high-throughput assay allows, through a relatively short run and a small test sample volume (400 µL) of serum, the analysis of a total of 50 samples in a single day.