Pentafluorophenyl bonded phase separation: the assay of Australian antioxidant formulation of ascorbic acid, rutin and hesperidin

Abstract

This work describes two novel validated HPLC methods for the separation and determination of potent antioxidant formulation. The compounds of interest are ascorbic acid (ASC), rutin (Rut) and hesperidin (HES). Two pentafluorophenyl bonded phases (PFP) of different geometries and particle-type (fully porous and core–shell) were used and both methods yielded linear response over the ranges 80–800, 20–220 and 20–200 μg mL⁻¹ for ascorbic acid, rutin and hesperidin, respectively. UV detection was accomplished at 260 nm using DAD. The developed methods were validated using ICH guidelines and were successfully applied to the determination of the ternary mixture in their pharmaceutical formulation in the Australian market with acceptable percent recoveries. Results obtained by the proposed methods were statistically compared to those of the reported methods regarding the mean percent recoveries and variances showing no significant difference. However, the analyses undertaken on core shell particles provided separations that were complete within 36 seconds.