A Validated RP-HPLC Method for the Determination of Piperidone Analogue of Curcumin

Abstract

Curcumin (Diferuloylmethane) is a natural product extracted from the root of Curcuma longa. 5-Bis (4-hydroxy-3-methoxybenzylidene)-Nmethyl-4-piperidone, the piperidone analogue of curcumin (PAC), was one of the analogues that, demonstrated potential anticancer effects against breast and colon cancers compared with native curcumin. A simple, accurate, and rapid isocratic reverse phase HPLC analytical method utilizing UV detection was developed and validated for the determination of PAC. A C18 column (4.6 mm × 150 mm, 3 μm spherical particles) was used with acetonitrile: 5% acetic acid (50:50, v/v) as a mobile phase at a flow rate of 1.0 mL/min and monitored at 392 nm. The run time was 7 min. Chromatogram showed a peak of PAC at retention time of 5.8 ± 0.92 min. The method was validated for linearity, accuracy, precision, limit of detection, limit of quantitation and robustness. Linear relationship (r>0.99) was observed between AUP of PAC and the corresponding concentrations over 100-10000 µg/mL. The mean linear regression equation was Y=52.42 X - 2562.7 for PAC. The LOQ of this assay was 3.9 ng/mL with corresponding relative standard deviation of 4.8 and 4.0%. The LOD was 13.1 ng/mL at a signal-to-noise ratio of >3.