A sensitive RP-HPLC method for estimation of artemether from polymeric nanoparticles after pre-column acid treatment using UV-visible detector

Abstract

Artemether; a sesquiterpene lactone is widely used for the treatment of malaria as artemisinin-based combination therapy (ACT). The present work involves the development and validation of sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) method for quantification of artemether (ART) in polymeric nanoparticles. ART was transformed to α, β-unsaturated decalones by pre-column acid treatment to enhance the sensitivity of chromophoric group lacking ART for quantification by HPLC-UV. Waters Spherisorb® 5 µm ODS(C18) column (4.6*250 mm) with gradient elution by mobile phase comprising of ACN and PBS (10 mM; pH 6.0) was used to separate acid-treated ART. The analysis was carried at λmax of 253 nm with 20 min and 20 µL run time and injection volume, respectively. The method was found to be linear in the concentration range of 0.5–10 µg mL−1 with 0.09 µg mL−1 and 0.27 µg mL−1 as LOD and LOQ respectively. Further, the method was also found to be specific for ART in presence of blank polymeric nanoparticles, accurate (% average recovery rate 101.7 ± 1.68%), precise (RSD <2%), and robust. The method was successfully used to determine % entrapment efficiency and in vitro release of ART-loaded polymeric nanoparticles with HPLC using a UV-visible detector.