Quantitative determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cellular environment of a human colon adenocarcinoma cell line for pharmacokinetic studies

Abstract: A simple, accurate, precise, specific and reproducible high-performance liquid chromatography (HPLC) method was developed for determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cell suspension. The assay procedure involved simple liquid–liquid extraction, the supernatant liquid was added an equal volume of water to avoid solvent effect. The detection of the analyte peak was achieved by monitoring the eluate using a UV detector set at 303nm. The analysis used a Hypersil ODS2 C18 column (5μm, 4.6mm×250mm) and methanol/distilled water as the mobile phase (flow rate=1mL/min). A total analytical run was achieved within 6.0min and calibration curve was linear over a wide concentration range of 0.25–40μg/mL for plasma sample and 1.0–500μM for cell suspension, the coefficients of correlation were 0.9997 and 0.9999 or better, respectively. There was 80.7±7.86%, 96.8±3.20% and 102.7±9.72% recovery from 0.5, 10, and 40μg/mL plasma samples, respectively. Intra- and inter-batch accuracy and precision were acceptable for the both matrices. The RSD of intra- and inter-day assay variations were all less than 10%. Both analyte and IS were stable in the battery of stability studies, freeze–thaw cycles. The described assay method was applied to pharmacokinetic studies in rats and a human colon adenocarcinoma cell line (Caco-2) successfully. The application of the assay to determine the pharmacokinetic is described. [Copyright &y& Elsevier]