Optimization and validation of conventional and micellar LC methods for the analysis of methyltestosterone in sugar-coated pills

Abstract

Two isocratic liquid chromatographic methods (conventional and micellar) for the determination of methyltestosterone in sugar-coated pills using fluoxymesterone as internal standard have been developed and validated. In conventional liquid chromatography a mobile phase 45% water:acetonitrile 55% (v:v), a flow-rate 1 ml min −1 and a C 18 Hypersil ODS (250×4.6 mm, 5 μm) column (25 °C) were used. In micellar liquid chromatography the conditions were: mobile phase 40 mM sodium dodecyl sulfate: 10% propanol, flow-rate 0.5 ml min −1 and C 18 Hypersil ODS (150×3.0 mm, 5 μm) column (60 °C). For both methods, UV absorbance detection at 245 nm was used and a separation up to base line was achieved. Prior to HPLC analysis a simple sample preparation was required.