1. The in vivo metabolism of Furazadrol in greyhounds
- Author
-
Steven Karamatic, Andy Pranata, Blake Curtis, Karen Caldwell, Christopher C. Waller, Paul Zahra, and Malcolm D. McLeod
- Subjects
Male ,Anabolism ,Urinary system ,Pharmaceutical Science ,01 natural sciences ,Analytical Chemistry ,Excretion ,03 medical and health sciences ,chemistry.chemical_compound ,Anabolic Agents ,Dogs ,Limit of Detection ,In vivo ,Oral administration ,Enzymatic hydrolysis ,Animals ,Environmental Chemistry ,Isoxazole ,Spectroscopy ,030304 developmental biology ,Doping in Sports ,0303 health sciences ,Chemistry ,010401 analytical chemistry ,Metabolism ,0104 chemical sciences ,3. Good health ,Substance Abuse Detection ,Biochemistry ,Female ,Androstanes - Abstract
Samples of the "dietary supplement" Furazadrol sourced through the internet have been reported to contain the designer anabolic androgenic steroids [1',2']isoxazolo[4',5':2,3]-5α-androstan-17β-ol (furazadrol F) and [1',2']isoxazolo[4',3':2,3]-5α-androstan-17β-ol (isofurazadrol IF). These steroids contain an isoxazole fused to the A-ring and were designed to offer anabolic activity while evading detection, raising concerns over the potential for abuse of this preparation in sports. The metabolism of Furazadrol (F:IF, 10:1) was studied by in vivo methods in greyhounds. Urinary phase II Furazadrol metabolites were detected as glucuronides after a controlled administration. These phase II metabolites were subjected to enzymatic hydrolysis by Escherichia coli β-glucuronidase to afford the corresponding phase I metabolites. Using a library of synthetically-derived reference materials, the identities of seven urinary Furazadrol metabolites were confirmed. Major confirmed metabolites were isofurazadrol IF, 4α-hydroxyfurazadrol 4α-HF, and 16α-hydroxy oxidised furazadrol 16α-HOF, while the minor confirmed metabolites were furazadrol F, 4β-hydroxyfurazadrol 4β-HF, 16β-hydroxyfurazadrol 16β-HF, and 16β-hydroxy oxidised furazadrol 16β-HOF. One major hydroxyfurazadrol and two dihydroxyfurazadrol metabolites remained unidentified. Qualitative excretion profiles, limits of detection, and extraction recoveries were established for furazadrol F and major confirmed metabolites. These investigations identify the key urinary metabolites of Furazadrol following oral administration, which can be incorporated into routine screening by anti-doping laboratories to aid the regulation of greyhound racing.
- Published
- 2021