Quantification of Agnuside in Human Plasma with a Novel High-Performance Liquid Chromatographic Method and Pharmacokinetic Study

This study presents a combination of High Performance Liquid Chromatography (HPLC) and ultraviolet (UV) detection that provides the quantification of agnuside in human plasma specimens. Reverse-phase chromatographic separation was carried out with C18 column (150 mm × 4.6 mm × 5 μm), at 25°C with isocratic elution of the mobile phase containing methanol: 0.1% formic acid (35:65 v/v) at 0.6 mL/min flow rate. Experiments were carried out at a wavelength of 258 nm. The retention time of the analyte is 9.70 ± 0.01 min. The developed technique was validated based on the International Conference on Harmonization guideline. The correlation coefficient of the technique was 0.9915, and the calibration range was 5-125 μg/mL. The recovery value of the proposed method was found to be 101.4%, and the precision of the method was calculated as 6.35 with the highest RSD% value. A pharmacokinetic study was performed by administering agnuside to a healthy volunteer.