Development and Validation of High-Performance Thin Layer Chromatographic Method for Simultaneous Estimation of Curcumin and Artemisinin

Abstract

Curcumin and Artemisinin are proposed to be formulated as combination therapy to expand the antimalarial effect in resistant and liable isolates. The present research work was to develop and validate a simple, accurate, precise, high-performance thin layer chromatographic method for simultaneous determination of Curcumin and Artemisinin. The chromatographic separation was carried out on a precoated silica gel aluminum plate, 20 × 20 cm, 100μm thickness, with a mixture of Hexane, Toluene and Ethyl acetate (6:1:3) as mobile phase UV detection was performed at 496nm. The Rf were 0.23 and 0.76 for Curcumin and Artemisinin, respectively. Calibration plots were linear over the concentration range 200-1000ng/band for both Curcumin and Artemisinin. The method was validated in accordance with ICH Q2(R1) guidelines for linearity, sensitivity accuracy, precision, and robustness. The percent recoveries in terms of accuracy for the synthetic mixture were found to be 99.8% to 100.0% and 99.94% to 100.4%, and the pooled percent relative standard deviation for repeatability and intermediate precision studies was found to be 0.015-0.042% and 0.008-0.070% for Curcumin and Artemisinin, respectively. A factorial design was used to optimize the chromatographic conditions for the robustness study. In conclusion, a novel, simple, accurate and reproducible HPTLC method was developed and could be applied for quality control testing of Curcumin and Artemisinin in synthetic mixture.