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

Abstract

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.