QUANTIFICATION OF TRANS-RESVERATROL IN RAT PLASMA BY A SIMPLE AND SENSITIVE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHOD AND ITS APPLICATION IN PRE-CLINICAL STUDY

Abstract

The intention of this study was to develop and validate a simple, sensitive, and accurate high performance liquid chromatography (HPLC) method for quantification of trans-resveratrol in rat plasma. Trans-resveratrol was recovered from plasma samples by liquid-liquid extraction. A reverse phase HPLC column was used and the assay was performed at 35°C through isocratic delivery (1 mL · min−1) of acetonitrile and phosphate buffer solution, pH 7.0 (30:70 v/v). UV absorbance was monitored at 320 nm. Plasma samples were stable (>99%) at −80°C for 3 months. The recovery of trans-resveratrol from plasma was >95%. The HPLC method was validated. The retention times of trans-resveratrol and carbamazepine were 5.5 and 10.4 min, respectively. The calibration curve was linear (R2 = 0.9999) within the range of 5–1,000 ng · mL−1. The lower limits of detection and quantification of trans-resveratrol were 2.5 and 5 ng · mL−1, respectively. The intra-day and inter-day variations were <5% and accuracy was >95%. This simple HPLC method was subsequently applied to measure trans-resveratrol concentration in plasma following oral administration of trans-resveratrol suspension in rats. The plasma concentration of trans-resveratrol at 30 min after oral administration was quantified as 151.66 ± 120.53 ng · mL−1.