Development and Validation of a Stability-indicating High-performance Liquid Chromatographic Method for...

The objective of this study was to develop a validated stability-indicating high-performance liquid chromatographic method that quantifies progesterone in compounded glycerinated gelatin troches. The mobile phase was composed of methanol and water (75:25 v/v), while the stationary phase was a Waters Nova-Pak C18 column (3.9 mm Å~ 15 cm Å~ 4.0 µm) with the column's temperature set to 40°C. The injection volume was 20 µL, while the gradient flow rate was maintained at 0.75 mL/min for a run time of 15 minutes. The detection wavelength for progesterone was set to 245 nm. In the forced degradation study, there was significant hydrolytic, oxidative, ultraviolet, and thermal degradation but insignificant photodegradation. However, no degradants co-eluted with progesterone. All method validation parameters met the respective acceptance criteria established by the International Conference on Harmonisation guidelines. This developed and validated method is suitable for both routine potency/strength testing as well as stability testing of progesterone in compounded glycerinated gelatin troche dosage forms. The method was utilized to successfully quantify progesterone in multiple compounded preparations from two different compounding pharmacies.