Fast method for the resolution and determination of sex steroids in urine

The main aim of the study was to develop a simple, fast, sensitive and inexpensive method for the separation and quantification of various steroid hormones in urine. Ultra high performance liquid chromatography was used to analyze estrone, estriol, 17-α-estradiol, 17-β-estradiol, progesterone, pregnenolone, and testosterone. Three columns were chosen for the present study: two octadecyl columns and one octyl column. The best results of separation were obtained for the octadecyl columns. Complete separation of all sex steroids was impossible when methanol was used during the chromatographic studies. The most interesting and valuable result was obtained with regard to the complete separation of isomers. All seven steroids were successfully separated in 10 min, next the time of single analysis was reduced to 5.5 min with gradient elution. Linearity was evaluated over a range of concentrations of 0.08–12.11 ng ml−1. The correlation coefficient ranged from 0.9987 to 0.9998. The LOD values were between 0.02 and 0.33 ng ml−1 and LOQ ranged between 0.10 and 1.10 ng ml−1. The developed method is suitable for routine analysis of these compounds in urine.