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Titolo:
Simultaneous determination of testosterone metabolites in liver microsomesusing column-switching semi-microcolumn high-performance liquid chromatography
Autore:
Tachibana, S; Tanaka, M;
Indirizzi:
Daiichi Pharmaceut Co Ltd, Drug Metab & Physiochem Property Res Lab, Edogawa Ku, Tokyo 1348630, Japan Daiichi Pharmaceut Co Ltd Tokyo Japan 1348630a Ku, Tokyo 1348630, Japan
Titolo Testata:
ANALYTICAL BIOCHEMISTRY
fascicolo: 2, volume: 295, anno: 2001,
pagine: 248 - 256
SICI:
0003-2697(20010815)295:2<248:SDOTMI>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
MIXED-FUNCTION PRECOLUMN; 3A4 CATALYTIC ACTIVITIES; CYTOCHROME-P450 3A4; STEROID HYDROXYLATION; RAT-LIVER; ASSAY; OXIDATION; DRUGS; B(5); IDENTIFICATION;
Keywords:
HPLC; column switching; testosterone; cytochrome P450; human liver microsomes; cytochrome b(5 center dot);
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
42
Recensione:
Indirizzi per estratti:
Indirizzo: Tachibana, S Daiichi Pharmaceut Co Ltd, Drug Metab & Physiochem Property Res Lab, Edogawa Ku, 1-16-13 Kitakasai, Tokyo 1348630, Japan Daiichi Pharmaceut Co Ltd 1-16-13 Kitakasai Tokyo Japan 1348630
Citazione:
S. Tachibana e M. Tanaka, "Simultaneous determination of testosterone metabolites in liver microsomesusing column-switching semi-microcolumn high-performance liquid chromatography", ANALYT BIOC, 295(2), 2001, pp. 248-256

Abstract

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 (6 alpha-, 6 beta-, 16 alpha-, 16 beta-, 7 alpha-, 2 alpha-, and 2 beta -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 wasdissolved 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 < 10.7%. This method has been subsequently used to determine the testosterone hydroxylation activities catalyzed by 15 different recombinant CYP isozymes. The results confirmed the formation of stereoselectively hydroxylated metabolites by each CYP isozyme. (C) 2001 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 19/01/20 alle ore 14:30:37