Abstract
ABSTRACT A method is described for the extraction and separation of free and conjugated oestrogens in urine. Celite 545, a diatomaceous earth, is coated with a high molecular weight secondary amine (Amberlite LA-1, Rohm and Haas, Phila., Pa.) and is used as a chromatographic support for the separation of various free and conjugated oestrogens in urine and in extracts of urine. Elution is carried out with extremely dilute acids. Optimum separation was obtained with a column of Celite coated with LA-1 (formate) in which the conjugated oestrogens in a concentrated extract of urine were eluted with a linear gradient (in the mixing chamber) of formic acid (pH 3.10 to 5.04).
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