Simultaneous determination of testosterone metabolites in liver microsomes using column-switching semi-microcolumn high-performance liquid chromatography
| Autor: | Shuko Tachibana, Makoto Tanaka |
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| Rok vydání: | 2001 |
| Předmět: |
Analyte
Residue (complex analysis) Chromatography Elution Biophysics Ethyl acetate Reproducibility of Results Cell Biology Hydroxylation Biochemistry High-performance liquid chromatography Isozyme Rats chemistry.chemical_compound chemistry Microsome Microsomes Liver Animals Humans Hydroxytestosterones Testosterone Molecular Biology Chromatography High Pressure Liquid |
| Zdroj: | Analytical biochemistry. 295(2) |
| ISSN: | 0003-2697 |
| Popis: | A sensitive and selective column-switching semi-microcolumn high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of testosterone and eight of its metabolites (6alpha-, 6beta-, 16alpha-, 16beta-, 7alpha-, 2alpha-, and 2beta-hydroxytestosterone, and androstenedione) in liver microsomes. After incubation for 10 min, testosterone and its metabolites were extracted from the microsomes with ethyl acetate, and the extract was evaporated to dryness. The residue was dissolved in the mobile phase and loaded onto the HPLC system. The analytes were first concentrated in a precolumn and subsequently transferred to the analytical column, where they were separated using linear gradient elution. A UV detector set at 254 nm was used to detect the analytes. This newly developed method clearly separated TES and the metabolites with high resolution and was found to be reproducible with intra- and interday variability of |
| Databáze: | OpenAIRE |
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