A modification of the Detter and Klingmüller's method for urinary alpha- and beta-pregnanediol and pregnanetriol determination: excretion patterns in men and in women during the normal menstrual cycle.

This report describes the modification of Detter and Klingmüller's method for the determination of urinary alpha- and beta-pregnanediol and pregnanetriol by means of thin -layer chromatography. The modifications were as follows: 1. The addition of formol prior to hydrolysis to prevent pigments penetration into urinary extracts. 2. The use of Kieselgel HF 254+366 nach Stahl (E. Merck, Darmstadt) which allows to localize the spots under UV-light at 366 nm without the use of colour reagents. The results concerning accuracy, sensitivity, precision and specificity in the modification described are profitable. Using this method in 7 healthy women (aged 28-37) with normal menstrual cycles the urinary excretion of alpha- and beta-pregnanediol and pregnanetriol were evaluated every second day (starting the 6th day of the cycle). The maximum of excretion of alpha- and beta-pregnandiol appeared on day 20 of the cycle, with the mean (+/- S.D.) 1.27 +/- 0.37 mg/24hr and 2.97 +/- 0.80 mg/24hr for alpha- and beta-pregnanediol, respectively. Mean values of pregnantriol were at the same level and ranged from 0.25 to 1.42 mg/24 hr. Single determination of these compounds in 8 healthy men (aged 19-42) revealed the mean excretion values (+/- S.D.) 0.96 +/- 0.17 mg/24 hr, 1.24 +/- 0.40 mg/24 hr, 1.12 +/- 0.65 mg/24 hr for alpha- and beta-pregnanediol and pregnanetriol, respectively.