Novel substances of expected doping activity are constantly introduced to the market. β-Methylphenethylamine (BMPEA) is classified as a doping agent by the World Anti-Doping Agency as it is a positional isomer of amphetamine. In this work, the development and application of a simple and rapid analytical procedure that enables discrimination between both isomers is described. The analytes of interest were extracted from urine by a two-step liquid–liquid extraction and then analyzed by UPLC/MS/MS under isocratic conditions. The entire analytical procedure was validated by evaluating its selectivity, discrimination capabilities, carry-over, sensitivity, and influence of matrix effects on its performance. Application of the method resulted in detection of BMPEA in eight anti-doping samples, including the first report of adverse analytical finding regarding its use. Further analysis showed that BMPEA may be eliminated unchanged along with its phase II conjugates, the hydrolysis of which may considerably improve detection capabilities of the method. Omission of the hydrolysis step may therefore, produce false-negative results. Testing laboratories should also carefully examine their LC/MS/MS-based amphetamine and BMPEA findings as both isomers fragment yielding comparable collision-induced dissociation spectra and their insufficient chromatographic separation may result in misidentification. This is of great importance in case of forensic analyses as BMPEA is not controlled by the public law, and its manufacturing, distribution, and use are legal.
Reports of new designer agents banned in sport being detected in supplements widely available for athletes are constantly emerging. The task of anti-doping laboratories is to control athletes for the presence of substances listed by the World Anti-Doping Agency (WADA) and those that are structurally/biologically similar to them. Recently, a new designer stimulant, N,N-dimethyl-2-phenylpropan-1-amine (NN-DMPPA), was detected by the WADA accredited anti-doping laboratory in Warsaw during routine anti-doping control. The urine samples from four athletes were analyzed in the screening method for stimulants and narcotics and the presence of NN-DMPPA was detected. The identity of NN-DMPPA was confirmed by gas chromatography-mass spectrometry using a synthesized reference standard. The measured concentrations of NN-DMPPA were between 0.51 and 6.51 µg/mL. The presence of the NN-DMPPA compound has been detected in the 'nutritional supplement' NOXPUMP that had been purchased in a store in Poland. NN-DMPPA at 121.7 µg/g was indicated in the investigated supplement together with another banned stimulant β-methylphenethylamine. The presence of this new stimulant was not indicated on the labelling of the supplement, a situation which is not unusual within this market. Thus, it is important to make athletes aware of the risk related to the use of supplements. Moreover, specific legistation dealing with the commercialization of drugs banned for sport should be undertaken.
Stimulants, together with anabolic androgenic steroids, are regarded as one of the most popular doping substances in sport. Owing to a great variety of these substances and new designer drugs being introduced to the market, each year the World Anti-Doping Agency (WADA) updates the list of substances and methods prohibited in sport. On 1 January 2014, a new doping agent - trimetazidine (TMZ) - was added to the WADA Prohibited List. TMZ, a substance prohibited in competition, is classified in the S6b Specified Stimulant Group. TMZ is used as a well-known cardiologic drug with confirmed biochemical and clinical activity. According to knowledge of the pharmacology and mechanism of TMZ action, TMZ can be used by athletes to improve physical efficiency, especially in the case of endurance sports. This study presents the phenomena of TMZ use by Polish athletes involved in anti-doping control in the WADA-accredited laboratory in Warsaw (Poland) between 2008 and 2013. Samples were taken from the athletes of such disciplines as cycling, athletics, and triathlon. Moreover, the elimination study of TMZ has been conducted to establish the change of TMZ concentration in urine sample after oral administration of a single or double (during the long-term therapy) dose. TMZ was monitored in urine samples by gas chromatography-mass spectrometry-nitrogen phosphorus detection (GC-MS-NPD).
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