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An automated direct assay for the simultaneous determination of unconjugated estetrol, estriol, cortisone and cortisol in serum and amniotic fluid, using high-performance liquid chromatography with electrochemical detection and ultraviolet detection, has been developed. The analysis time is ca. 1 h. This system offers good reproducibility with low coefficients of variation (estetrol, 2.3%; estriol, 2.3%; cortisone, 2.6%; cortisol, 1.9%). Detection limits are low enough for routine determinations (estetrol and estriol, 150 pg; cortisone and cortisol, 5 ng). Comparison of the values measured by the present method and by radioimmunoassay revealed significant correlations for estetrol (r = 0.787, p less than 0.01), estriol (r = 0.957, p less than 0.01), cortisone (r = 0.956, p less than 0.01) and cortisol (r = 0.865, p less than 0.01). This system proved to be valuable in monitoring feto-placental function.

A high-performance liquid chromatographic method for the determination of estriol 16-glucuronide and 17-glucuronide in bile and urine has been developed. The electrochemical detector was found to be more superior than the UV detector with respect to selectivity and sensitivity and therefore more suitable for the determination of estrogen conjugates in biological fluids. The use of the present method revealed that both estriol 16-glucuronide and 17-glucuronide were excreted in rat bile, while only the former was present in human pregnancy urine.

There are different national food regulations concerning the use of hormoneactive substances in the breeding of domestic animals that serve as human food. This situation requires specific and sensitive methods for the detection of any residues in food. The hormones used in practice can be determined with improved sensitivity by coupling to the diazonium compound Fast Dark Blue R Salt. After thin-layer chromatography on high-performance thin-layer chromatographic plates with two different solvent mixtures, zeranol, estradiol, estriol and estrone are analysed as azo dyestuffs. The detection limit of these hormones is below 10 ng.