The multicomponent analysis of estrogens in urine by ion exchange chromatography and GC-MS--I. Quantitation of estrogens after initial hydrolysis of conjugates.

A method for the multicomponent analysis of estrogens in urine after initial hydrolysis of the conjugates is described. Following protection of the carbonyl functions by ethoximation, estrogen conjugates were extracted on Sep-Pak C18 cartridges and purified on the acetate form of DEAE-Sephadex. The samples were subsequently hydrolysed by Helix pomatia juice and the hydrolysate was purified on the acetate form of QAE-Sephadex. Estrogens with vicinal cis-hydroxyls and diphenolic compounds were fractionated on the borate and bicarbonate form of QAE-Sephadex, respectively. Neutral steroids were removed by the free base form of DEAE-Sephadex after which estrogens were separated into two groups using Lipidex 5000 in a straight phase system. Following trimethylsilyl ether derivatization estrogens were analysed by selected ion monitoring (SIM). The method allows the quantitation of all the important estrogen metabolites including catechol estrogens. It is precise, accurate and sensitive permitting the quantitation of estrogens in urine of males and non-pregnant females.