Development and Validation of a Novel HPLC-UV Method for the Quantification of Hyaluronic Acid in Anti-Aging Creams

This study introduces a novel, rapid, and cost-effective high-performance liquid chromatography (HPLC-UV) method for the direct quantification of HA in anti-aging creams without requiring derivatization. Chromatographic separation was performed on a reversed-phase C18 (5 μm × 4.6 mm × 150 mm) column, using an acetonitrile-acetate buffer (80:20, v/v) mobile phase at 0.9 mL/min flow rate. UV detection at 230 nm provided high selectivity. The method demonstrated excellent linearity (R² = 0.9997) across 0.1–30 μg/mL, with limits of detection (LOD) and quantification (LOQ) of 0.03 and 0.1 μg/mL, respectively. Validation was conducted per ICH guidelines, confirming high accuracy, precision (RSD < 1.88%), robustness, and repeatability, with minimal matrix interference. Compared to existing HA quantification methods, this optimized HPLC-UV technique offers superior sensitivity, speed, and simplicity, making it an ideal tool for routine analysis in cosmetic formulations.