Application of solid-phase extraction for the quantification of several abused drugs in sports in human urine using UHPLC-MS/MS method

Abstract

A procedure for the determination of several selective androgen receptor modulators in human urine, namely, Andarine (S-4, GTx-007), Ostarine (S-22, GTx-024, MK-2866) and Ligandrol (LGD-4033, VK-5211), as well as a phytosteroid Laxogenin and Ibutamoren (MK-677), related to non-peptide growth hormone secretagogues, has been developed. Most of these are prohibited by the World Anti-Doping Agency due to their anabolic effects. With the aim of the analytes purification and concentration, a procedure for solid-phase extraction with the use of Varian Bond Elut C8 (100 mg, 1 mL), Biotage Isolute C18 (EC) (100 mg, 1 mL) and Waters Oasis HLB (30 mg, 1 mL) cartridges was applied. To achieve good resolution of the analytes, reversed-phase ultra-high-performance liquid chromatography coupled with the triple quadrupole mass spectrometer as a detector in positive and negative ion detection modes was used. The limits of detection were in the range of 0.1–0.5 ng/mL, limits of quantification were between 0.25–0.5 ng/mL. Calibration curves were examined to be linear in the wide range of concentrations. Matrix effects were also evaluated and were found not to influence the results of the analytes quantification. The proposed method was applied for the analysis of real samples after single 15 mg oral administration of the analytes. All the analytes were positively detected in the human urine samples in 12h, thus, proving the applicability of the procedure for the analytes quantification in the urine. Key words : UHPLC-MS/MS, SARM, Laxogenin, Ibutamoren, solid-phase extraction, doping control DOI: http://dx.doi.org/10.15826/analitika.2018.22.3.008 (Russian) E.V. Dmitrieva, A.Z. Temerdashev, A.A. Azaryan, E.M. Gashimova Kuban State University, Stavropolskaya st., 149, Krasnodar, 350040, Russian Federation