Pharmaceutical Analytical Chemistry: Open Access
Open Access

Abstract

Method Development and Validation of Stability Indicating RP-HPLC Method for the Determination of Female Hormones in Hormone Concentrates Creams

Pradeep Gautam

A rapid, novel, and efficient stability indicating reverse-phase high performance liquid chromatographic (RPHPLC) analytical method was developed and validated for the determination of four female hormones: Progesterone, Estrone, Estradiol, and Estriol in Hormones Concentrate compounding bases. Separation was achieved with a RP- C18 Phenomenex® Gemini 150 × 4.6 mm C18 5µm column using a phosphate buffer pH 6.4 and acetonitrile. The developed HPLC method was validated with method validation components: linearity, accuracy, precision, intermediate precision, and robustness. Limit of detection (LOD) and limit of quantification (LOQ) were also obtained for all four hormones for this method. Photodiode array (PDA) detector was set up at 225 nm wavelength for the simultaneous analysis of all four peaks. Hormone concentrate samples, placebo, and hormone actives were subjected to stress conditions of hydrolysis (acid and base), oxidative, and thermal stress degradation. Standard solution stability was also performed. This female hormones HPLC method was validated as per the International Conference on Harmonization (ICH) guidelines. The proposed validated method was successfully used for the quantitative analysis of bulk, stability, and finished hormone concentrate and hormone compounding formulations.