A rapid HPLC method for the quantification of 3,5,4′-trimethoxy- trans-stilbene (TMS) in rat plasma and its application in pharmacokinetic study

Abstract

A rapid HPLC-UV method had been developed and validated to quantify 3,5,4′-trimethoxy- trans-stilbene (TMS), a naturally occurring and pharmacologically active analog of resveratrol in rat plasma. The samples were mixed with three volumes of acetonitrile to precipitate protein. Chromatographic separation was achieved on a RP-HPLC column (Agilent ZORBAX Eclipse Plus C18: 250 mm × 4.6 mm i.d., 5 μm), which was protected by a guard column (Agilent ZORBAX Eclipse Plus C18: 12.5 mm × 4.6 mm i.d., 5 μm) through isocratic delivery of a mobile phase of acetonitrile: water (75:25, v/v) at a flow rate of 1.2 ml/min. The assay was executed at 30 °C and the UV absorbance at 320 nm was monitored. The retention time of TMS and trans-stilbene (internal standard) was 6.5 and 8.3 min, respectively. The calibration curve was linear within the range of 15–1000 ng/ml ( R 2 > 0.998) and 15 ng/ml was the lower LOQ. The intra- and inter-day precisions were good and the RSD was all lower than 7.3%. The mean absolute recovery of TMS in plasma ranged from 99.2 to 104.1%. This HPLC method had been successfully applied to study the pharmacokinetics of TMS, which was fully dissolved with hydroxypropyl-β-cyclodextrin (HP-β-CyD). In comparison with resveratrol, TMS had greater plasma exposure, longer elimination half-life and lower clearance. As TMS had superior pharmacokinetic characteristics, its potential as a preventive or therapeutic agent in resveratrol-effective conditions or diseases should be considered.