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An improved reversed-phase HPLC method for simultaneous estimation of curcuminoids in a dietary supplement formulation

Vaibhav Ajit Kekre, Sanjay Gomaji Walode

A reversed phase High Performance Liquid Chromatography method was developed for simultaneous estimation of Curcumin (C), Demethoxy curcumin (DMC) and Bis-demethoxy curcumin (BDMC) in a dietary supplement formulation. The formulation analysed also contained piperine along with curcuminoids. Separation of curcuminoidswas performed on an HQsil HS C18 column using 50 mmol Potassiumdihydrogen phosphate (PH-4.4): acetonitrile in a ratio 45:55 (v/v) as the mobile phase at the flow rate of 1.0 ml/min. UV detection for curcuminoids was done at 421nm. The retention time of BDMC, DMC, C were found to be 11.5 and 12.6 and 13.7 respectively. Linearity for curcuminoids was established at concentrations ranging from12.8 ìg/ml - 38.34 ìg/ml for (C), 3.15 µg/ml - 9.49 µg/ml for (DMC) and 0.61 µg/ml - 1.95 µg/ml for (BDMC). The LOD and LOQ were found to be 0.101 ìg and 0.305 µg for (BDMC), 0.63 µg and 1.89 µg for (DMC) and 2.37 µg and 7.11 ìg for (C). This method was further validated in terms of precision, recovery and robustness as per ICH guidelines giving satisfactory results.