title: Enzyme-assisted synthesis and structure characterization of glucuronide conjugates of eleven anabolic steroid metabolites.
abstract: Enzyme-assisted in vitro synthesis of eleven glucuronide-conjugated anabolic androgenic steroid (AAS) metabolites was performed using biphenyl-induced rat liver microsomal enzymes. The substrates within the study were the main compounds and metabolites detected in human urine after dosing of, e.g. metandienone, metenolone, methyltestosterone, nandrolone, and testosterone. Yields of glucuronidation reactions were 13-28% for most compounds, but significantly higher (77-78%) for the substrates with 4-ene-3-one double bond system of the steroid A-ring. Characterization of glucuronide-conjugated AAS structures was based on nuclear magnetic resonance spectroscopy ((1)H NMR) and on liquid chromatographic-mass spectrometric (LC-MS) and tandem mass spectrometric (LC-MS/MS) analyses in positive and negative ion mode electrospray ionization (ESI). Only minor differences were observed in optimal synthesis conditions between various substrates, which offer a potential to apply this in vitro assay as a default method for glucuronidation of new AAS substrates. The method allowed for a rapid production pathway of stereochemically pure AAS glucuronides in milligram amount, such as needed, e.g. in the development of analytical methods in forensic or pharmaceutical sciences, as well as in doping control.
authors: Division of Pharmaceutical Chemistry, Faculty of Pharmacy, Viikinkaari 5E, University of Helsinki, Finland.
authors: Laura
authors: L
authors: Hintikka
authors: Tiia
authors: T
authors: Kuuranne
authors: Olli
authors: O
authors: Aitio
authors: Mario
authors: M
authors: Thevis
authors: Wilhelm
authors: W
authors: Schänzer
authors: Risto
authors: R
authors: Kostiainen
mesh_terms: D000818:Animals; D001711:Biotransformation; D002853:Chromatography, Liquid; D020719:Glucuronides / Q000096:biosynthesis* / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization; D013256:Steroids / Q000096:biosynthesis* / Q000737:chemistry
chemical_list: D020719:Glucuronides; D013256:Steroids