title: LC-MS/MS detection of unaltered glucuronoconjugated metabolites of metandienone.
abstract: The aim of this study was to evaluate the direct detection of glucuronoconjugated metabolites of metandienone (MTD) and their detection times. Metabolites resistant to enzymatic hydrolysis were also evaluated. Based on the common mass spectrometric behaviour of steroid glucuronides, three liquid chromatography-tandem mass spectrometry (LC-MS/MS) strategies were applied for the detection of unpredicted and predicted metabolites: precursor ion scan (PI), neutral loss scan (NL), and theoretical selected reaction monitoring (SRM) methods. Samples from four excretion studies of MTD were analyzed for both the detection of metabolites and the establishment of their detection times. Using PI and NL methods, seven metabolites were observed in post-administration samples. SRM methods allowed for the detection of 13 glucuronide metabolites. The detection times, measured by analysis with an SRM method, were between 1 and 22 days. The metabolite detected for the longest time was 18-nor-17β-hydroxymethyl-17α-methyl-5β-androsta-1,4,13-triene-3-one-17-glucuronide. One metabolite was resistant to hydrolysis with β-glucuronidase; however it was only detected in urine up to four days after administration. The three glucuronide metabolites with the highest retrospectivity were identified by chemical synthesis or mass spectrometric data, and although they were previously reported, this is the first time that analytical data of the intact phase II metabolites are presented for some of them. The LC-MS/MS strategies applied have demonstrated to be useful for detecting glucuronoconjugated metabolites of MTD, including glucuronides resistant to enzymatic hydrolysis which cannot be detected by conventional approaches. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Doctor Aiguader 88, 08003, Barcelona, Spain.|Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Doctor Aiguader 88, 08003, Barcelona, Spain.
authors.forename: Argitxu
authors.initials: A
authors.lastname: Esquivel
authors.affiliation: Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Doctor Aiguader 88, 08003, Barcelona, Spain.
authors.forename: Oscar J
authors.initials: OJ
authors.lastname: Pozo
authors.affiliation: Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Doctor Aiguader 88, 08003, Barcelona, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: Garrostas
authors.affiliation: Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Doctor Aiguader 88, 08003, Barcelona, Spain.|Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Doctor Aiguader 88, 08003, Barcelona, Spain.
authors.forename: Georgina
authors.initials: G
authors.lastname: Balcells
authors.affiliation: Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Doctor Aiguader 88, 08003, Barcelona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Gómez
authors.affiliation: Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Doctor Aiguader 88, 08003, Barcelona, Spain.|Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña, Consejo Superior de Investigaciones Científicas ((IQAC-CSIC), Jordi Girona 18-26, 08034, Barcelona, Spain.
authors.forename: Aristotelis
authors.initials: A
authors.lastname: Kotronoulas
authors.affiliation: Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña, Consejo Superior de Investigaciones Científicas ((IQAC-CSIC), Jordi Girona 18-26, 08034, Barcelona, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Joglar
authors.affiliation: Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Doctor Aiguader 88, 08003, Barcelona, Spain.|Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Doctor Aiguader 88, 08003, Barcelona, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Ventura
mesh_terms: D000328:Adult; D045930:Anabolic Agents / Q000378:metabolism / Q000652:urine*; D002853:Chromatography, Liquid / Q000379:methods; D004300:Doping in Sports; D020719:Glucuronides / Q000378:metabolism / Q000652:urine*; D006801:Humans; D008297:Male; D008696:Methandrostenolone / Q000378:metabolism / Q000652:urine*; D008875:Middle Aged; D058573:Performance-Enhancing Substances / Q000378:metabolism / Q000652:urine*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D045930:Anabolic Agents; D020719:Glucuronides; D058573:Performance-Enhancing Substances; D008696:Methandrostenolone
keywords: LC-MS/MS; doping, metabolism; glucuronoconjugates; metandienone