A new method for the separation of androgens from estrogens and for the partition of estriol from the estrone-estradiol fraction: with special reference to the identification and quantitative microdetermination of estrogens by ultraviolet absorption spectrophotometry

Abstract

It is recognized generally that a qualitative and quantitative knowledge of the excretion pattern of the urinary estrogens is one index to an understanding of the functional activity of the ovary and adrenal cortex. Obviously, such determinations may be useful also in evaluating the normal and abnormal functions of other physiologically related endocrine glands as well as of organs like the liver and kidneys. The clinical applications of these data are self-evident. Various attempts have been made to circumvent the notoriously inaccurate values which have been obtained for the urinary estrogens by a variety of bioassay methods and calorimetric techniques (1, 2). The acknowledged shortcomings of these methods have led us to investigate the application of ultraviolet absorption spectrophotometry to the quantitative determination of the urinary estrogens in an attempt to develop an objective physical method for their accurate determination. It is known that the infra-red portion of the spectrum yields more differentially characteristic curves, but those of the ultraviolet range are more readily obtainable, and consequently better adapted to clinical use. This communication is concerned with studies of the following aspects of the problem: (1) spectrophotometric identification and quantitative micro determination of crystalline estrogens; (2) detection by spectrophotometric assay of gross errors in current methods for extraction and partition of estrogens; (3) studies on the ultraviolet absorption of substances comprising the background material; (4) separation of the phenolic estrogens from the so called neutral steroid fraction; (5) separation of urinary estrogens from other urinary phenolic substances by steam distillation; (6) micro-Girard separation of estrone from estradiol; (7) an essentially new method for the extraction and partition of crystalline estrone, estradiol, and estriol, and their quantitative assay by ultraviolet spectrophotometry