The Development and Validation of the HPLC Method for Determination of Artesunate and Amodiaquine in Novel Antimalarial Formulations

Abstract

ASAQ combination drug used in treating malaria is bitter in taste, resulting in poor adherence and difficulty in swallowing by children. To mitigate this issue, taste masking is achieved by coating the drug with polymers to improve the taste of the dosage form. However, this produces a novel drug product which must undergo analytical evaluation to ensure that it is safe and drug content matches the label claim. The aim of this study is to develop and validate a novel analytical methodology which is suitable for this purpose. Thus, a reversed phase high performance liquid chromatography for the quantitative determination of AS and its impurities was developed. The method was validated according to ICH guidelines for linearity, precision, accuracy, specificity, limit of detection (LOD) and limit of quantification (LOQ). The method was found accurate and precise with an average retention time of 8.2 min. Good linearity was observed in the concentration range of 0.25 – 0.75 mg/ml with regression coefficient of r2 =0.999. The assay of the proposed method was found to be 101.5%. The recoveries of artesunate were found to be within 99% - 102%. The % RSD for precision was found to be <2%. Specificity was done to check potential interferences with excipients and degradation products and none of the retention times were interfering with artesunate peak. A gradient method was developed for quantification of amodiaquine and its impurities. The method was validated according to ICH guidelines for linearity, precision, accuracy, specificity, limit of detection (LOD) and limit of quantification (LOQ). The method was found accurate and precise with an average retention time of 13.75min. Good linearity was observed in the range of 0.075 – 0.225 mg/ml with regression coefficient of r2 =0.999. The assay of the proposed method was found to be 106.5%. The recoveries of artesunate were found to be within 98% - 102%. The % RSD from repeatability, intra and inter day precisions was found to be <2.0%. Specificity was done to check potential interferences with excipients and degradation products.