Determination Of Steroids In Urine Research Articles

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 10min, next the time of single analysis was reduced to 5.5min with gradient elution. Linearity was evaluated over a range of concentrations of 0.08-12.11ngml(-1). The correlation coefficient ranged from 0.9987 to 0.9998. The LOD values were between 0.02 and 0.33ngml(-1) and LOQ ranged between 0.10 and 1.10ngml(-1). The developed method is suitable for routine analysis of these compounds in urine.

Determination of Steroids in Human Urine by Micellar Liquid Chromatography

A simple and sensitive method for the determination of steroids using micellar liquid chromatography is described. The steroids, including hydroxycorticosterone. corticosterone, northisterone, testosterone, mexdroprogesterone acetate and progesterone, were separated by reversed-phase using a micelles mobile phase following UV detection at 245 nm. The parameters affecting retention of the test solutes such as the concentration of sodium dodecyl sulfate (SDS) and n-butanol-1 in the mobile phase were investigated. It was found that the retention of the solutes was dependent on the composition of mobile phase. The linear calibration plots range from 0.1 to 10 μg ml−1 in mobile phase containing 5.0 × 10−2 mol l−1 SDS/9 % n-butanol-1 at pH 6.0, and the detection limit in order of 0.1 μg ml−1 was obtained. The proposed method was used for the determination of steroids in urine using direct injection of samples without previous treatment.

Determination of steroids in urine by micellar HPLC with detection by sensitized terbium fluorescence

Efficient Förster energy transfer from steroid donors to a terbium ion acceptor occurs in aqueous solution when the two species are sequestered in sodium dodecyl sulfate micelles. The microorganized medium provides both proximity between donor and acceptor and protection from quenching of terbium fluorescence by water. Detection limits down to 100 pg have been achieved for steroids with an alpha,beta-unsaturated carbonyl group in the A-ring. This detection strategy has been used in HPLC separations, using either normal- or reversed-phase chromatography with postcolumn addition of detergent and/or terbium, or micellar chromatography with the detergent solution as the mobile phase. This last method proved to be superior, requiring no sample preparation and allowing direct injection of urine onto the HPLC column.

Determination of steroids using rapid high-performance liquid chromatography with diode array detection.

The recent development of small, high-efficiency columns for rapid HPLC was investigated for application to the determination of steroids. Two applications are described, the determination of estrogenic steroids in urine and the determination of progesterone and estradiol benzoate in an injection formulation. Methods were developed for both these applications, each with over-all run times of less than 6 min, representing a considerable reduction over previously used methods. These methods were extensively validated and were found to be accurate, robust and reliable. The problem of reduced sample loadability is considered and the practical solution of peak compression illustrated.

The determination of steroids in urine

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