Simultaneous Determination of Curcumin and Resveratrol in Lipidic Nanoemulsion Formulation and Rat Plasma Using HPLC: Optimization and Application to Real Samples

Abstract

Background: Curcumin and resveratrol are naturally occurring polyphenols that are highly effective in inhibiting the growth of cancer cells. A robust reversed-phase HPLC method has been developed for the simultaneous determination of these two natural drugs. Objective: The method was adapted to analyze both drugs in pure forms, in lipidic nanoemulsion formulation as well as in rat plasma. The method was applied to real samples after intravenous (IV) injection of rats. Method: Analysis utilized C18 column using acetonitrile (ACN)-water (pH adjusted to 4.6 by 1% orthophosphoric acid) in the ratio of 55+45 (v/v) at a flow rate of 0.8 mL/min with detection at 425 and 304 nm for curcumin and resveratrol, respectively. Results: Extraction efficiency of curcumin and resveratrol using ACN-methanol was 96.10-101.00% (RSD 2.49) and 95.00-99.87% (RSD 2.59), respectively. The assay was linear from 0.05 to 4.00 μg/mL (correlation coefficient of 0.9989 and 0.9981, respectively) and precise [average interday and intraday precision for curcumin RSD% (0.45, 2.04) and resveratrol RSD% (2.25, 1.71)] in spiked rat plasma. The LOD and LOQ were found to be (0.0085 μg/mL, 0.025 μg/mL) and (0.02, 0.06), respectively. Conclusions: The data presented demonstrate that the method provides rapid, sensitive, and precise determination of curcumin and resveratrol in spiked rat plasma and in nanoemulsion dosage form without tedious cleanup procedure, which was successfully applied for quantitation of both drugs following their IV administration to albino rats. Highlights: Validated chromatographic method has been developed for simultaneous determination of curcumin and resveratrol. Optimization of chromatographic conditions was achieved. Application of the method on nanoemulsion formula, on spiked rat plasma, and pharmacokinetics study.