Separation of oestrogens in biological fluids and synthetic mixtures on Sephadex G-type gels.
The separations achieved when mixtures of both free and conjugated oestrogens from a variety of sources are chromatographed on columns of Sephadex gel are reviewed. The molecular identities of oestrogen conjugates which have been separated from human urine by these methods are listed in Table 1. Tables 2 and 3 contain the key experimental details for a total of 26 separations of oestrogen mixtures, abstracted from the total of 20 papers which were published during the period 1961-82. Table 4 details corresponding experimental data for the separation of free oestriol (in human blood) achieved by methods involving a combined Sephadex gel and immunochemical procedure abstracted from a further two papers. A careful analysis of the separation data given in the tables leads to the initial conclusion that the elution profile depends on the expected chromatographic variables for gel filtration chromatography, namely, type of Sephadex gel, length of column, nature and amount of sample applied and the sensitivity of detection methods. However, the separation achieved by the Sephadex columns is also shown to be critically dependent on the column temperature and the pH and chemical composition of the eluent and wash solvents. These latter effects, together with the realization that the molecular weights of the oestrogens being separated are very similar, leads to the conclusion that the separations summarized in Tables 2 and 3 are all being achieved by an absorption process. This being the case, it is suggested that the time-consuming methods of gel filtration chromatography need not be used. Confirmation of this proposal is afforded by a discussion of a recent paper in which the rapid separation of the oestrogens from other components in the biological matrix (urine) was achieved by an adsorption procedure. It is suggested that in the future, separations of oestrogens in biological materials may be most rapidly achieved in combining this type of adsorption procedure with HPLC.
D001826:Body Fluids / Q000032:analysis*; D002850:Chromatography, Gel / Q000379:methods; D004964:Estriol / Q000302:isolation & purification; D004967:Estrogens / Q000302:isolation & purification*; D006801:Humans; D007120:Immunochemistry
D004967:Estrogens; D004964:Estriol