Analytical Method Development and Validation for Simultaneous Estimation of Artemether and Lumefantrine in Pure And Pharmaceutical Dosage Form Using Rp-Hplc Method

A simple, rapid, precise and cost effective reversed phase-high performance liquid chromatography (RP-HPLC) method was developed for the simultaneous estimation of Artemether (AT) and Lumefantrine (LU) in pure drug and pharmaceutical dosage forms. The separation was carried out using BDS Hypersil C18 (150 x 4.6 mm i.d. 3 μm particle size) column, with mobile phase comprising of 0.01M tetra butyl ammonium hydrogen sulphate and acetonitrile in the ratio of 20 : 80 (v/ v). The flow rate was 1.0 ml/ min and the detection was carried out using UV-visible detector at 222 nm. The method was validated by evaluation of different parameters such as accuracy, precision, linearity, ruggedness, and robustness, limit of detection (LOD) and limit of quantification (LOQ). The retention time were found to be 4.19 and 5.22 min for AT and LU, respectively. Correlation coefficient (r2) of 0.999 for both over concentration range of 3.2-19.2 μg/ ml and 16-96 μg/ ml for AT and LU, respectively. Parameters like mobile phase ratio, wavelength, flow rate and others were deliberately varied. It was observed that there were no marked changes in chromatograms, which demonstrated that the developed RP-HPLC method is robust. Intra and inter day precision reproducibility study was carried out and it was checked by determining precision on the same instrument, but by a different analyst. The percentage recovery for AT and LU were ranged between 99.18-100.19 and 99.96-100.07, respectively. The LOD for AT and LU were found to be 0.201 and 2.99 μg/ ml and the LOQ were 0.609 and 9.086 μg/ml respectively. Method was found to be reproducible with relative standard deviation (RSD) for intra and inter day precision less than 2 %. (author)