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  • Imitation of phase I oxidative metabolism of anabolic steroids by titanium dioxide photocatalysis.

Imitation of phase I oxidative metabolism of anabolic steroids by titanium dioxide photocatalysis.

European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences (2014-09-16)
Miina Ruokolainen, Minna Valkonen, Tiina Sikanen, Tapio Kotiaho, Risto Kostiainen
ABSTRACT

The aim of this study was to investigate the feasibility of titanium dioxide (TiO2) photocatalysis for oxidation of anabolic steroids and for imitation of their phase I metabolism. The photocatalytic reaction products of five anabolic steroids were compared to their phase I in vitro metabolites produced by human liver microsomes (HLM). The same main reaction types - hydroxylation, dehydrogenation and combination of these two - were observed both in TiO2 photocatalysis and in microsomal incubations. Several isomers of each product type were formed in both systems. Based on the same mass, retention time and similarity of the product ion spectra, many of the products observed in HLM reactions were also formed in TiO2 photocatalytic reactions. However, products characteristic to only either one of the systems were also formed. In conclusion, TiO2 photocatalysis is a rapid, simple and inexpensive method for imitation of phase I metabolism of anabolic steroids and production of metabolite standards.

MATERIALS
Product Number
Brand
Product Description

Supelco
Sodium hydroxide concentrate, 0.1 M NaOH in water (0.1N), Eluent concentrate for IC
Supelco
Phosphate Standard for IC, TraceCERT®, 1000 mg/L phosphate in water (nominal concentration)
Testosterone, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Magnesium chloride solution, 0.1 M
Sigma-Aldrich
Methanol, HPLC Plus, ≥99.9%, poly-coated bottles
Sigma-Aldrich
Acetonitrile solution, contains 0.05 % (w/v) ammonium formate, 5 % (v/v) water, 0.1 % (v/v) formic acid, suitable for HPLC
Supelco
Acetic acid, analytical standard
Supelco
17α-Methyltestosterone, VETRANAL®, analytical standard
Supelco
4-Androstene-3,17-dione, VETRANAL®, analytical standard
Adenine, European Pharmacopoeia (EP) Reference Standard
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Acetonitrile solution, contains 10.0% acetone, 0.05% formic acid, 40.0% 2-propanol
Sigma-Aldrich
Sodium hydroxide, BioUltra, for luminescence, ≥98.0% (T), pellets
Sigma-Aldrich
Sodium hydroxide solution, BioUltra, for molecular biology, 10 M in H2O
Sigma-Aldrich
Acetic acid, for luminescence, BioUltra, ≥99.5% (GC)
Supelco
Sodium hydroxide solution, 49-51% in water, eluent for IC
Sigma-Aldrich
Magnesium chloride solution, BioUltra, for molecular biology, ~0.025 M in H2O
Sigma-Aldrich
Testosterone, purum, ≥99.0% (HPLC)
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Acetic acid, ≥99.5%, FCC, FG
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Acetic acid, natural, ≥99.5%, FG
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Magnesium chloride solution, BioUltra, for molecular biology, ~1 M in H2O
Supelco
Methanol, analytical standard
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Ammonium acetate solution, BioUltra, for molecular biology, ~5 M in H2O
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Ammonium acetate, 99.999% trace metals basis
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Magnesium chloride solution, BioUltra, for molecular biology, 2 M in H2O
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Acetic acid-12C2, 99.9 atom % 12C
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Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC
Sigma-Aldrich
Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid
Sigma-Aldrich
Acetonitrile solution, contains 0.1 % (v/v) formic acid, suitable for HPLC
Sigma-Aldrich
Methanol, anhydrous, 99.8%
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Ammonium acetate, BioUltra, for molecular biology, ≥99.0%