Journal of Bioequivalence & Bioavailability

Reversed-phase liquid chromatographic method with UV detection for the determination of trans-resveratrol in human plasma using Catechin as an internal standard

3^rd World Congress Bioavailability & Bioequivalence

March 26-28, 2012 Marriott Hotel & Convention Centre, Hyderabad, India

Gurinder Singh, Roopa S. Pai and Vinay Pandit

Posters: J Bioequiv Availab

Abstract:

A simple and sensitive high-performance liquid chromatographic (HPLC) method was developed for quantification of trans- resveratrol in spiked human plasma employing liquid-liquid extraction. Catechin was used as an internal standard (IS). The present method used protein precipitation for extraction of trans-resveratrol from human plasma. Chromatographic Separation was achieved by employing a Phenomenex C18 column (250mm x 4.6 mm, 5 μm) and the column effluent was monitored by UV detector at 306 nm. The mobile phase consisted of a mixture of methanol and 0.01M pH being adjusted to 6.8 phosphate buffer (63:37%, v/v in Milli-Q water) with 0.5% (v/v) ortho phosphoric acid solution at a flow rate of 1.0 mL min -1 . This method was linear over the range of 50.0-6400.0 ng mL −1 with regression coefficient (>0.998). Nominal retention times of trans-resveratrol and IS were 3.94 and 5.9 min, respectively. Limits of detection (LOD) and Limits of quantification (LOQ) of trans-resveratrol were 9 ng mL -1 and 10 ng mL- 1 , respectively. The method has been validated through a spiking/recovery procedure at three concentration levels. Results obtained are highly satisfactory, with recovery values around 98.372% for trans-resveratrol. Resveratrol was found to be stable for a period of 15 days on storage at -20?C. The method was found to be precise, accurate, and specific during the study. Keywords: Catechin, human plasma, high-performance liquid chromatography, trans-resveratrol.