Gradient HPLC separation of dehydroepiandrosterone (DHEA) from its metabolites and biological congeners: role of tetrahydrofuran in the chromatographic mechanism.
A three-step gradient reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed for the separation of dehydroepiandrosterone (DHEA), its sulfate ester (DHEA-S), its three C7-oxidized metabolites (7alphaOH-DHEA, 7betaOH-DHEA, 7-keto-DHEA), and its biosynthetic congeners (androstenedione, testosterone, estradiol, pregnenolone). This new method allows the quantitative characterization of DHEA metabolism and biosynthetic transformation under given physiological, pathological, or therapeutically influenced circumstances. Tetrahydrofuran probably acts as a proton acceptor coadsorbent, while isopropanol behaves as a proton donor during the separation of testosterone, estradiol, and the stereoisomers of 7-OH-DHEA.
Gergely
András
A
Department of Pharmaceutical Chemistry, Semmelweis University, Hogyes Endre u. 9, Budapest 1092, Hungary.
Horváth
Péter
P
Szász
György
G
Veress
Gábor
G
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003687:Dehydroepiandrosterone / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D005663:Furans; D012680:Sensitivity and Specificity
D005663:Furans; C018674:tetrahydrofuran; D003687:Dehydroepiandrosterone