BIOANALYTICAL OF UPLC METHOD DEVELOPMENT AND VALIDATION OF XANTHORRIZOL AND ITS APPLICATION TO PHARMACOKINETIC STUDY

Abstract

Objective: A simple, straightforward, ultra-performance liquid chromatography (UPLC) with a fluorescence detector method was developed and validated to determine xanthorrhizol in rat plasma. This method was successfully applied to an oral pharmacokinetic study. Methods: Xanthorrhizol was separated using a C18 column in an isocratic mode using a mobile phase of acetonitrile: water (85:15 v/v) at a 0.4 ml/min flow rate. The fluorescence detector was set at 230 nm excitation and 320 nm emission wavelengths. The method was then applied in the pharmacokinetic study involving 12 Sprague-Dawley rats. Results: The developed bioanalytical methods were found to be linear in the range of 0.078–5 mg/ml with a correlation coefficient of r2=0.999. The percentage recovery of xanthorrhizol was more than 95%, and the relative standard deviation was less than 2. These results indicate that the method is accurate and precise. The limit of detection (LOD) and limit of quantification (LOQ) of the technique were 0.123 µg/ml and 0.373 µg/ml, respectively. Furthermore, the stability studies demonstrated that xanthorrhizol is stable under various analytical conditions. The pharmacokinetic study revealed that the area under the curve (AUC) was 27.23±19.65 (µg. h/ml), the half-life (t 1/2) was 7.71±2.89 h, the mean residence time (MRT) was 13.86±4.06 h while the maximum concentration (Cmax) was 1.58±0.62 µg/ml, and the time to reach the maximum concentration (Tmax) was 1.33±0.20 h. Conclusion: The developed bioanalytical method was reliable and successfully met all validation criteria, making it a robust choice for quantifying xanthorrhizol. Therefore, it may be effectively utilized to determine xanthorrhizol in rat plasma following a pharmacokinetic study.