4-hydroxyestrone, isolation and identification in human urine.
Portions of pregnancy and midcycle urines were submitted to hot acid hydrolysis, extracted with benzene/ethyl acetate and the extracts washed with ascorbic acid buffer. From the remaining organic phase the catecholestrogens were removed with borate buffer and further purified on Sephadex LH-20 columns. After derivatisation 4-hydroxyestrone was separated from the isomeric 2-hydroxyestrone peak were identical with that of authentic 4-hydroxyestrone. After treatment of the extracts with sodium borohydride 4-hydroxyestradiol-17 beta was identified by GC-MS. By the addition of trace amounts of tritiated 4-hydroxyestrone a recovery of 40% was calculated. On the basis of this recovery and the peak heights of the gas chromatograms an excretion of 4 microgram (midcycle) and 40 microgram (pregnancy) of 4-hydroxyestrone/24 h was estimated.
D002852:Chromatography, Ion Exchange; D004958:Estradiol / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000652:urine; D002393:Estrogens, Catechol; D004970:Estrone / Q000031:analogs & derivatives*; D005260:Female; D006801:Humans; D006894:Hydroxyestrones / Q000302:isolation & purification / Q000652:urine*; D011247:Pregnancy; D013057:Spectrum Analysis
D002393:Estrogens, Catechol; D006894:Hydroxyestrones; D004970:Estrone; C026418:4-hydroxyestrone; D004958:Estradiol; C014036:4-hydroxyestradiol