A method for the determination of oestriol, oestrone and oestradiol-17β in human urine by partition chromatography and colorimetric estimation

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Abstract
A detailed procedure for the extraction, separation and purification of estriol, estrone and estradiol-17[beta] in human urine is described. The method involves: acid hydrolysis; ether extraction; separation of estriol from estrone and estradiol-17[beta] by distribution between benzene and water; purification of estriol by saponification and column partition chromatography; separation and purification of estrone and estradiol-17[beta] by column partition chromatography and saponification. The purified fractions are determined colorimetrically by an improved Kober reaction with a spectrophotometric correction for non-estrogen chromogenic material. The precautions necessary for satisfactory analyses and the efficiency of the various purification procedures are described. The specificity, accuracy, precision and sensitivity of the method are discussed.