Development and Validation of RP-HPLC Method for Simultaneous Determination of Resveratrol and Curcumin in Pure Form

Cancer chemopreventive agents are designed to reduce the incidence of tumorigenesis by intervening at one or more stages of carcinogenesis. Recently, resveratrol, a natural product found in the diet of humans, has been shown to function as a cancer chemopreventive agent. Resveratrol was first shown to act as an antioxidant and antimutagenic agent, thus acting as an anti-initiation agent. Resveratrol belongs to a class of polyphenolic compounds called stilbenes.(3,5, 4′-trihydroxy-trans-stilbene) is a natural compound found in red grape skin, Japanese knotweed (polygonum cuspidatum), peanuts and blueberries. A naturally occurring polyphenolic phytoalexin Curcumin, [1,7-bis (4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3, 5-dione] is the major yellow pigment extracted from turmeric, a commonly used spice, derived from the rhizome of Curcuma longa Linn. In animal studies, curcumin has been shown to increase the survival of tumor-bearing rodents by inhibiting tumor growth and impeding metastasis. At present study a simple, precise, rapid and accurate reverse phase HPLC method developed for the estimation of Resveratrol and Curcumin in combination as well as individually were developed and validated. The retention time of Resveratrol and Curcumin were found to be 2.90 and 4.11 minutes. Detection was carried out at 424 and 306nm. The regression coefficient value of Resveratrol and Curcumin is 0.9904 and 0.9937 which was found to be linear in the detection range. Limit of detection and limit of quantification of Resveratrol was found to be 0.08μg/ml and 0.32 μg/ml and Curcumin is 0.05 μg/ml and 0.17 μg/ml. Analysis was performed using a C18 column (250 X 4.6 mm) at room temperature in isocratic mode. The mobile phase used was Citric acid (pH adjusted to 3.5): Acetonitrile (40:60) at flow rate of 1.0 ml/min.