Rosuvastatinin Kersetin Varlığında Protein Bağlanma Kapasitesinin HPLC ile İncelenmesi

Abstract

Daştan, K. Evaluation of Protein Binding Capacity of Rosuvastatin in the Presence of Quercetin by HPLC, Hacettepe University Graduate School of Health Sciences, Analytical Chemistry Program Master's Thesis, Ankara, 2025. In this study, a high-performance liquid chromatography (HPLC) method was developed to determine the change in plasma protein binding (PPB) capacity of rosuvastatin (ROS), an active ingredient in the statin group, in the presence of quercetin (QUE), a phytochemical compound. The method allows the quantification of free ROS in plasma and the separation of ROS and QUE using an ultrafiltration technique. The developed method was validated according to the validation parameters outlined in the FDA Bioanalytical Method Validation Guidance. All analyses conducted within the scope of this study were performed using an ACE C18 (250 × 4.6 mm i.d., 5 μm) analytical column. For chromatographic separation, a mobile phase consisting of acetonitrile and 20 mM phosphate buffer (pH 4.0) in a 45:55 v/v ratio was used, with a flow rate set at 1 mL/min. A UV detector recorded signals at a wavelength of 250 nm. Under these chromatographic conditions, the retention times for ROS and QUE were determined as 6.84 and 4.29 minutes, respectively. The method was tested for linearity within the range of 0.04–2.00 μg/mL for ROS and 0.10–2.00 μg/mL for QUE. Following validation, the developed method was applied to analyze ROS and QUE in commercial plasma and commercial bovine serum albumin (BSA). The recovery values of ROS from commercial BSA, both in the absence and presence of QUE, were calculated. The recovery value for ROS alone was determined as 85.38±1.44%, while in the presence of QUE, it was found to be 83.79±3.91%. The obtained values statistically demonstrated that the protein binding capacity of ROS was not affected by the presence of QUE.