A rapid method for determination of progesterone by reversed-phase liquid chromatography from aqueous media

A simple reversed-phase liquid chromatographic method to assay progesterone in aqueous receiving medium, following in vitro skin permeation studies is presented. Progesterone was analysed using 5 mu m LichroCART(R) RP-18 column (125 x 4 mm i.d.), after extraction with chloroform. The mobile phase used was methanol water (70:30) at a flow rate of 1 ml/min. Detection was carried out at 254 nm at room temperature. The method showed a mean recovery of 99% for progesterone, and intra-assay and inter-assay coefficients of variation were below 2%. The proposed method was validated for linearity, specificity, precision and accuracy and showed to be useful for analysis of progesterone in in vitro skin permeation studies.