Selective polarity-guided extraction and purification of acetogenins in Annona muricata L. leaves

Herein is described the selective polar-guided extraction and column chromatography separation of annonaceous acetogenins from the leaves of Annona muricata L., commonly known as soursop. The objective of this study is to obtain an acetogenin-rich fraction, containing mainly annonacin, known to be present in the highest amount in soursop leaves. Solvents with various polarities (ethyl acetate, hexane, chloroform, ethanol, water) and their mixtures were used in the extraction, fractionation, and isolation steps. The liquid chromatography-mass spectrometry analysis of the isolate sample showed that annonacin, having a molecular mass of 597.23, is the dominant acetogenin present. The amount of acetogenin isolated from the soursop leaf samples was 242 mg/g, based on the total lactone determination using the Kedde reagent. The cytotoxicity activity of the isolate sample was determined using the brine shrimp test. This study shows that there is a positive correlation between lactone content due to acetogenins and the level of cytotoxicity in the fractions obtained. The low LC₅₀ value of 1 ppm showed a significant cytotoxicity of the ethyl acetate fraction obtained, higher than cytotoxicity of cancer drug cyclophosphamide. High lactone content and a high cytotoxicity of the ethyl acetate fraction indicate the potential of A. muricata leaves as the source of bioactive compounds for anti-cancer treatment.