Identification and characterization of urinary prenylamine metabolites by means of liquid chromatography-tandem mass spectrometry

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Deutscher übersetzter Titel:Identifizierung und Charakterisierung von Prenylamin-Metaboliten im Urin durch Flüssigkeitschromatographie-Tandem-Massenspektrometrie
Autor:Beuck, S.; Sigmund, G.; Koch, A.; Schänzer, W.; Pokrywka, A.; Kwiatkowska, D.; Thevis, Mario
Erschienen in:Drug testing and analysis
Veröffentlicht:4 (2012), 9-10 (30th Cologne Workshop: Advances in sports drug testing), S. 701-716, Lit.
Format: Literatur (SPOLIT)
Publikationstyp: Zeitschriftenartikel
Medienart: Gedruckte Ressource Elektronische Ressource (online)
Sprache:Englisch
ISSN:1942-7603, 1942-7611
DOI:10.1002/dta.1388
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Erfassungsnummer:PU201301000880
Quelle:BISp

Abstract

Prenylamine is a vasodilator of phenylalkylamine structure and was used for the treatment of angina pectoris, until reports of undesirable effects including ventricular tachycardia led to a decreasing use of the drug in the 1980s. Metabolic N-dealkylation of orally ingested prenylamine can liberate amphetamine in humans and cause positive findings for amphetamine in doping and forensic analysis. In 2010, the World Anti-Doping Agency (WADA) classified prenylamine as a non-specified stimulant according to the 2010 Prohibited List, thus banning its use in sports in-competition. Supporting the development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) based detection method, a post-administration urine sample following a single oral prenylamine ingestion (Segontin® 60 mg) was analyzed for urinary metabolites. The LC-separated analytes were ionized in positive electrospray ionization (ESI) mode and detected as protonated ions using an AB Sciex TripleTOF 5600 quadrupole-time-of-flight hybrid mass spectrometer. Over 40 phase I metabolites were detected, including previously unknown mono- bis-, tris- and tetra-hydroxylated prenylamine, several hydroxylated and methoxylated prenylamine metabolites and (hydroxylated) diphenylpropylamine. Investigation of the collision-induced dissociation behaviours of the metabolites by high resolution/high accuracy mass spectrometry allowed for the assignment of the nature and the site of observed metabolic transformations. The most abundant phase I metabolite was confirmed as p-hydroxy-prenlyamine by chemical synthesis and stable isotope labelling of reference material. An existing routine screening assay based on direct injection and LC-MS/MS analysis of urine was modified and validated according to common guidelines, in order to allow for the detection of p-hydroxy-prenylamine in sports drug testing. The assay demonstrated the ability to detect the target metabolite at 0.1 ng/ml at intra- and inter-day imprecisions below 10%. Verf.-Referat