Analysis of isomeric ethynylestradiol glucuronides in urine

A method for separation and analysis of conjugates of ethynylestradiol in urine is described. Steroid conjugates are separated on a lipophilic strong anion exchanger (triethylaminohydroxypropyl Sephadex LH-230), and phenolic steroids released by enzyme hydrolysis or solvolysis ar isolated by chromatography on the same ion exchanger. Steroids carrying an ethynyl group are isolated by chromatography on SP-Sephadex (Ag+). Ethynylestradiol is analyzed by gas chromatography-mass spectrometry of the trimethylsilyl ether, using [9,11,11,12,12-(2)H5] ethynylestradiol and internal standard.