RP-HPLC Method Development and Validation for the Estimation of Lumefantrine in Bulk Drug

Abstract

Lumefantrine is an antimalarial agent used to treat acute uncomplicated malaria. It is administered in combination with artemether for improved efficacy. This combination therapy exerts its effects against the erythrocytic stages of Plasmodium spp. and may be used to treat infections caused by P. falciparum and unidentified Plasmodium species, including infections acquired in chloroquine-resistant areas. A reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the estimation of lumefantrine in bulk drug. The separation was achieved on Thermo C18 analytical column (250 mm × 4.6 mm i.d., 5.0 μm) using 10mM KH2PO4: acetonitrile  (pH adjust 3.0 with OPA) in the ratio 20:80 v/v as mobile phase and at a flow rate of 1.0 ml/min. Detection was carried out using a UV detector at 240nm. The total chromatographic analysis time per sample was about 6.0 min with lumefantrine eluting at retention time of about 3.225 ± 0.001min. The method was validated for accuracy, precision, specificity, linearity and sensitivity. Validation studies demonstrated that this HPLC method is simple, specific, rapid, reliable and reproducible. The standard curve was linear over the concentration range of 5-25 μg/ml with r2 close to one (0.999). The limit of detection (LOD) and limit of quantitation (LOQ) obtained for lumefantrine were 0.25 μg/ml and 0.75μg/ml respectively. The high recovery and low relative standard deviation confirm the suitability of the proposed method for the determination of lumefantrine in bulk drugs.