High-Performance Liquid Chromatography of Estrogen 3-Sulfates with Electrochemical Detection Using an Immobilized Enzyme Reactor

Abstract

A simple and sensitive method for the determination of estrogen 3-sulfates by highperformance liquid chromatography with electrochemical detection is described. The sulfates were separated on a reversed-phase column and subsequently hydrolyzed with an immobilized sulfatase reactor in a continuous flow mode. Estrogens liberated were then detected with a coulometric detector. A linear response was observed for four sulfates, estrone sulfate, 16 a -hydroxyestrone 3-sulfate, estradiol 3-sulfate and estriol 3-sulfate in the range 25-1000 ng as an injected amount. The detection limits (S/N=10) were 20 ng for estriol 3-sulfate and 16 a -hydroxyestrone 3-sulfate, and 50 ng for estrone sulfate and estradiol 3-sulfate, respectively.