Development and validation of RP-HPLC-DAD method for determination of progesterone in capsule

Abstract

Progesterone is a hormone produced in the human body that is essential for regulating many vital functions. A reversed phase-High Performance Liquid Chromatography-Diode Array Detection (RP-HPLC-DAD) method for the quantitative determination of progesterone in pure forms and capsule was developed and validated in the present study. The validation parameters such as linearity, precision, accuracy, specificity, sensitivity and stability were studied according to the International Conference on Harmonization Guidelines. Progesterone was eluated by Agilent C18 (5μm, 150 × 4.6 mm) column and detected at 240 nm. The eluation was achieved isocratically with a mobile phase of acetonitrile-Milli-Q grade water (70:30, v:v) at a flow-rate of 1 mL min-1. The injection volume was 10 μL. The range of quantification was 0.25-5 μg mL-1. Intra-day and inter-day precision expressed as the percent relative standard deviation (RSD %) were less than 2.5 %, and accuracy (relative error) was better than 11.0 % (n = 6). LOQ and LOD values were found to be 0.1 and 0.08 μg mL-1, respectively. The developed method was successfully applied to assay of progesterone in capsule.