Sulfate metabolites as alternative markers for the detection of 4-chlorometandienone misuse in doping control.
Sulfate metabolites have been described as long-term metabolites for some anabolic androgenic steroids (AAS). 4-chlorometandienone (4Cl-MTD) is one of the most frequently detected AAS in sports drug testing and it is commonly detected by monitoring metabolites excreted free or conjugated with glucuronic acid. Sulfation reactions of 4Cl-MTD have not been studied. The aim of this work was to evaluate the sulfate fraction of 4Cl-MTD metabolism by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to establish potential long-term metabolites valuable for doping control purposes. 4Cl-MTD was administered to two healthy male volunteers and urine samples were collected up to 8 days after administration. A theoretical selected reaction monitoring (SRM) method working in negative mode was developed. Ion transitions were based on ionization and fragmentation behaviour of sulfate metabolites as well as specific neutral losses (NL of 15 Da and NL of 36 Da) of compounds with related chemical structure. Six sulfate metabolites were detected after the analysis of excretion study samples. Three of the identified metabolites were characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS). Results showed that five out of the six identified sulfate metabolites were detected in urine up to the last collected samples from both excretion studies. Copyright © 2016 John Wiley & Sons, Ltd.
Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Barcelona, Spain.|Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Barcelona, Spain.
Georgina
G
Balcells
Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Barcelona, Spain.|Experimental Asthma and Allergy Research Unit, The National Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.|Unit for Chemistry II, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Cristina
C
Gómez
Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Barcelona, Spain.
Lorena
L
Garrostas
Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Barcelona, Spain.
Óscar J
ÓJ
Pozo
Bioanalysis Research Group, IMIM, Hospital del Mar Medical Research Institute, Barcelona, Spain.|Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Barcelona, Spain.
Rosa
R
Ventura
D000328:Adult; D045930:Anabolic Agents / Q000737:chemistry / Q000378:metabolism* / Q000652:urine*; D002853:Chromatography, Liquid / Q000379:methods; D004300:Doping in Sports; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D020723:Glucuronic Acid / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D054879:Halogenation; D006801:Humans; D008297:Male; D008696:Methandrostenolone / Q000031:analogs & derivatives / Q000378:metabolism* / Q000652:urine*; D015813:Substance Abuse Detection / Q000379:methods*; D013431:Sulfates / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
D045930:Anabolic Agents; D013431:Sulfates; D020723:Glucuronic Acid; D008696:Methandrostenolone
4-chlorometandienone; LC-MS/MS; anabolic steroids; doping analysis; metabolism; sulfate metabolites