β-Hematin Formation Assay for Screening of Potential Antimalarial Activity of The Extracts and Fractions of three Artemisia Species

Abstract

In this study, In-vitro β-hematin formation assay, a spectrophotometric assessment is used to screen potential antimalarial effects of Artemisia extracts as well as their fractions. The n-hexane (n-hex), dichloromethane (DCM) and methanolic (MeOH) extracts obtained from aerial parts of A. splendens and A. austriaca by soxhlet apperatus, also DCM fractions, which were acquired by VLC method using n-hex, ethyl acetate (EtOAC) and MeOH, of A. spicigera, A. splendens and A. austriaca were examined by the mentioned method. N-hex and MeOH extracts of the herbs indicated weak anti-malarial activities, whereas, the DCM extracts of A. spicigera (IC50= 0.99±0.01mg/ml, which had been reported in previous manuscript), A. splendens (IC50= 1.93±0.09 mg/ml) and A. austriaca (IC50= 1.15±0.04mg/ml) showed potent antimalarial effects. Among the all examined DCM fractions with different polarities, DCM F4΄΄ (60% EtOAC/n-hex, the DCM fraction of A. austriaca) with IC50 value of 1.02 ± 0.03 mg/ml, DCM F5 (80% EtOAC/n-Hex, the DCM fraction of A. spicigera) with IC50 value of 1.21± 0.06 mg/ml, DCM F6 (100% EtOAC, the DCM fraction of A. austriaca) and DCM F7΄΄ (100% MeOH, the DCM fraction of A. austriaca) with IC50 values of 1.44 ± 0.04 mg/ml for both, as well as DCM F7΄ (100% MeOH, the DCM fraction of A. splendense) with IC50 value of 1.58 ± 0.03 mg/ml indicated higher activities than the other DCM fractions. The mentioned fractions could be useful to bioassay-guided isolation of bioactive plant constituents.