Development of a liquid chromatographic method for the simultaneous quantification of curcumin, β-arteether, tetrahydrocurcumin and dihydroartemisinin. Application to lipid-based formulations.

A liquid chromatographic method was developed for the simultaneous separation of curcumin, β-arteether, tetrahydrocurcumin and dihydroartemisinin based on the design of experiments and the design space methodology. The influence of the percentage of organic modifier, flow rate of the mobile phase and column temperature on the analytes separation was investigated. The optimal chromatographic separation was achieved on a C18 column (125mm×4mm, 5μm) using an isocratic elution with a mobile phase consisting of methanol-ammonium acetate (pH 4; 10mM) (80:20, v/v) at a flow rate of 0.45ml/min and a column temperature of 32.5°C. This method was then validated for simultaneous quantification of curcumin and β-arteether contained in lipid-based formulations taking into account the β-expectation tolerance interval for the total error measurement. Finally, the suitability of the proposed liquid chromatographic method for routine analysis of curcumin and β-arteether loaded in lipid-based formulations has been proven.