Liquid chromatography with multichannel electrochemical detection for the determination of trans-resveratrol in rat blood utilizing an automated blood sampling device

Abstract

A sensitive and selective multichannel liquid chromatography–electrochemistry method was developed for the determination of the natural product trans-resveratrol (resveratrol) in rat blood. After administration of resveratrol, blood samples were periodically collected by an automated blood sampling device. Resveratrol was extracted from 150 μl of diluted blood (blood and saline at a ratio of 1:1) with acetonitrile containing 1% of trichloroacetic acid. Chromatographic separation was achieved within 12 min using a C 18 (100×2.0 mm) 3 μm column with a mobile phase containing 20 m M sodium acetate, 0.5 m M EDTA, pH 4.5 and 21% acetonitrile at a flow-rate of 0.4 ml/min. A multichannel detector with glassy carbon electrodes was used, which can control up to four working electrodes simultaneously with applied potentials of +800, 700, 600, 500 mV vs. Ag/AgCl. The limit of detection was 2 ng/ml at a signal-to-noise ratio of 3:1 and the limit of quantitation was 4 ng/ml. The linearity of the calibration curve was obtained over the analytical range of 5–1000 ng/ml. The intra- and interassay precision was in the range of 2.5–4.4% and 1.2–4.3%, respectively. Using this method it was possible to quantify blood concentration following a single dose of resveratrol to rats with good accuracy and precision. Thus the pharmacokinetic properties of resveratrol in rats can be examined for intraperitoneal, oral and intravenous dosing.