Fluorescence and chemiluminescence determination of steroid and bile acid sulphates with lucigenin by flow injection analysis based on ion-pair solvent extraction

Abstract

A flow injection analyser with a membrane phase separator and a chemiluminescence detector was constructed for the determination of steroid sulphates using lucigenin as an extracting and chemiluminescent agent. Steroid sulphates injected into the analyser were extracted into the organic phase through the membrane phase separator with lucigenin. The extracted ion pair of steroid sulphate-lucigenin was mixed with alkaline hydrogen peroxide solution and the resulting chemiluminescence was measured by a home-made chemiluminescence detector equipped with a spiral coil cell. The calibration graphs obtained were linear and reproducible when samples of 3–30 pmol of steroid sulphates were injected. Steroid glucuronides, bile acid 3-O-sulphates and bile acids are also detected by this system.