Co-Estimating Estradiol Valerate and Medroxyprogesterone Acetate in a Combined Hormonal Product: A Novel Short Runtime UPLC Method.

Abstract

Background: Hormones regulate growth, development, and reproductive functions. While deficiencies can lead to various health problems, excessive intake may cause metabolic disorders and side effects. Therefore, there is a need to develop new, reliable, and economical methods for quality control and accurate analysis of hormones in tablets used for hormone replacement therapy.

Objective: To develop a novel, reliable, and cost-effective, ultra-performance liquid chromatography (UPLC) method for accurately quantifying estradiol valerate (EV) and medroxyprogesterone acetate (MPG) in hormone replacement therapy tablets.

Method: Chromatographic separation of EV and MPG was achieved using an ACQUITY UPLC BEH C18 (1.7 μm, 2.1 mm × 100 mm) column and a mixture containing water (6%), acetonitrile (44%), methanol (42%), and 0.10 M phosphate buffer (8%, pH = 2.14). Chromatographic detection was accomplished at 280.0 nm for EV and 240.0 nm for MPG, with a flow rate of 0.34 mL/min at a column temperature of 40 °C. The method's validity was assessed using individual and spiked sample analyses.

Results: The validated UPLC method offered an alternative procedure to quantify EV and MPG in commercial hormone tablets. This method demonstrated highly accurate, and precise analytic results with a short retention and low cost. The measured concentrations of EV and MPG in the tablet samples were found within the expected ranges, confirming the method's suitability for reliable hormone quantification in commercial tablets.

Highlights: This study introduces the first UPLC-PDA method for analyzing EV and MPG in a commercial hormone formulation. The method showed excellent assay results in the analysis of real samples.