Estimation of testosterone in and isolation of progesterone from human urine

A method for the estimation of urinary testosterone is described. This depends on acid hydrolysis, a modified form of the Girard separation and chromatography on an alumina column and on paper. The final method of detection is by gas -liquid chromatography.The reliability critlria of the method have been investigated. The recovery of added testosterone was approximately 80%. The method is reasonably precise, the coefficients of variation being 6.4% and 8.0% for male and female urine respectively. The chromatographic, chemical and physical behaviour of the compound estimated in the final fraction is similar to that of authentic testosterone. Accordingly, the method which is also practicable, appears to be specific.When testosterone assays were performed in normal men and in normally menstruating women, considerable variations were found from one subject to another. However, levels in males were consistently higher than those in females, there being no overlap between the two groups.Serial assays of urinary testosterone in normal male subjects showed the presence of peaks of excretion at fairly regular intervals. In two cases studied sekual intercourse had no effect on the occurrence of such peaks but caused an overall rise in testosterone output.Testosterone excretion values in normal young men aged from 16 to 20 were generally higher than those of the elder group of subjects aged from 21 to 63 years.Serial assays of urinary testosterone in normally menstruating women have shown an elevation of levels during the luteal phase probably associated with the presence of a functioning corpus luteum. A second rise was also noted at the time of ovulation, and a slight increase was observed during the follicular phase in two of the cycles studied.The assay method for urinary testosterone is now being applied to conditions such as acne vulgaris, sex chromosome abnormalities, athersclerosis in males and hirsutism in females.Progesterone has been isolated from the urine of a woman in late pregnancy. A modification of the Girard reaction was used by which it was :possible to separate the 'conjugated ketone' fraction both from non-ketonic fraction and from the bulk of the saturated ketones. The final separation was made using gas-liquid chromatography with a stream splitter.Work is proceeding on the use of the modified Girard separation technique to estimate plasma steroids with a Δ⁴-3-ketone structure. By this method the majority of the biologically active neutral steroid can be estimated simultaneously in one sample