title: Isolation and characterization of a β-glucuronide of hydroxylated SARM S1 produced using a combination of biotransformation and chemical oxidation.
abstract: In this study, using mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy, it has been confirmed that biotransformation with the fungus Cunninghamella elegans combined with chemical oxidation with the free radical tetramethylpiperidinyl-1-oxy (TEMPO) can produce drug glucuronides of β-configuration. Glucuronic acid conjugates are a common type of metabolites formed by the human body. The detection of such conjugates in doping control and other kinds of forensic analysis would be beneficial owing to a decrease in analysis time as hydrolysis can be omitted. However the commercial availability of reference standards for drug glucuronides is poor. The selective androgen receptor modulator (SARM) SARM S1 was incubated with the fungus C. elegans. The sample was treated with the free radical TEMPO oxidizing agent and was thereafter purified by SPE. A glucuronic acid conjugate was isolated using a fraction collector connected to an ultra high performance liquid chromatographic (UHPLC) system. The isolated compound was characterized by NMR spectroscopy and mass spectrometry and its structure was confirmed as a glucuronic acid β-conjugate of hydroxylated SARM S1 bearing the glucuronide moiety on carbon C-10.
authors.affiliation: Division of Analytical Pharmaceutical Chemistry, Department of Medicinal Chemistry, Uppsala University, Box 574, SE-75123 Uppsala, Sweden. Electronic address: Axel.Rydevik@farmkemi.uu.se.
authors.forename: Axel
authors.initials: A
authors.lastname: Rydevik
authors.affiliation: Bayer CropScience AG, Monheim, Germany. Electronic address: Andreas.Lagojda@bayer.com.
authors.forename: Andreas
authors.initials: A
authors.lastname: Lagojda
authors.affiliation: Institute of Biochemistry and Center for Preventive Doping Research, German Sport University, Cologne, Germany. Electronic address: Thevis@dshs-koeln.de.
authors.forename: Mario
authors.initials: M
authors.lastname: Thevis
authors.affiliation: Division of Analytical Pharmaceutical Chemistry, Department of Medicinal Chemistry, Uppsala University, Box 574, SE-75123 Uppsala, Sweden; National Veterinary Institute (SVA), Department of Chemistry, Environment and Feed Hygiene, SE-75189 Uppsala, Sweden. Electronic address: Ulf.Bondesson@sva.se.
authors.forename: Ulf
authors.initials: U
authors.lastname: Bondesson
authors.affiliation: Division of Analytical Pharmaceutical Chemistry, Department of Medicinal Chemistry, Uppsala University, Box 574, SE-75123 Uppsala, Sweden; National Veterinary Institute (SVA), Department of Chemistry, Environment and Feed Hygiene, SE-75189 Uppsala, Sweden. Electronic address: Mikael.Hedeland@sva.se.
authors.forename: Mikael
authors.initials: M
authors.lastname: Hedeland
mesh_terms: D000818:Animals; D001711:Biotransformation / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D020093:Cunninghamella / Q000737:chemistry*; D003497:Cyclic N-Oxides / Q000737:chemistry; D020723:Glucuronic Acid / Q000737:chemistry / Q000378:metabolism; D020719:Glucuronides / Q000737:chemistry* / Q000378:metabolism*; D006900:Hydroxylation / Q000502:physiology*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D010084:Oxidation-Reduction; D011944:Receptors, Androgen / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D003497:Cyclic N-Oxides; D020719:Glucuronides; D011944:Receptors, Androgen; D020723:Glucuronic Acid; C003959:TEMPO
keywords: Glucuronic acid conjugate; Glucuronide; NMR; SARM S1; TEMPO