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.

Acetogenin, Annona muricata, Annonacin, Extraction

Aromdee, C., Wichitchote, P., Jantakun, N., 2005. Spectrophotometric Determination of Total Lactones in Andrographis Paniculata Nees. Songklanakarin Journal of Science and Technology, Volume 27(6), pp. 1227–1231

Asif, M., Acharya, M., Lakshmayya, Singh, A., 2013. Bioactivity of 6-Aryl-4,5-dihydropyridazin-3(2H)-ones using Brine Shrimp (Artemia salina) Lethality Assay. International Journal of Pharmaceutical Chemistry, Volume 3(2), pp. 40–44

Champy, P., Hoglinger, G.U., Feger, J., Gleye, C., Hocquemiller, R., Laurens, A., Guerineau, V., Laprevote, O., Medja, F., Lombes, A., Michel, P.P., Lannuzel, A., Hirsch, E.C., Ruberg, M., 2004. Annonacin, a Lipophilic Inhibitor of Mitochondrial Complex I, Induces Nigral and Striatal Neurodegeneration in Rats: Possible Relevance for Atypical Parkinsonism in Guadeloupe. Journal of Neurochemistry, Volume 88, pp. 63–69

Kim, G.S., Zeng, L., Alali, F., Rogers, L.L., Wu, F.E., Sastrodihardjo, S., McLaughlin, J.L., 1998. Muricoreacin and Murihexocin C, Mono-tetrahydrofuran acetogenins, from the Leaves of Annona muricata. Phytochemistry, Volume 49(2), pp. 565–571

Kreuger, M.R.O., Grootjans, S., Biavatti, M.W., Vandenabeele, P., Herde, K.D., 2012. Sesquiterpene Lactones as Drugs with Multiple Targets in Cancer Treatment: Focus on Parthenolide. Anticancer Drugs, Volume 23(9), pp. 883–896

Luna, J.D.S., Carvalho, J.M.D., Lima, M.R.F.D, Bieber, L.W., Bento, E.D.S., Franck, X., Santana, A.E.G., 2006. Acetogenins in Annona muricata L. (annonaceae) Leaves are Potent Molluscicides. Natural Product Research, Volume 20(3), pp. 253–257

McLaughlin, J.L., 2008. Paw Paw and Cancer: Annonaceous Acetogenins from Discovery to Commercial Products. Journal of Natural Product, Volume 71(7), pp. 1311–1321

Meyer, B.N., Ferrigni, N.R., Putnam, J.E., Jacobsen, L.B., Nichols, D.E., McLaughlin, J.L., 1982. Brine Shrimp: a Convenient General Bioassay for Active Plant Constituents. Planta Medica, Volume 45, pp. 31–34

Moshi, M.J., Innocent, E., Magadula, J.J., Otieno, D.F., Weisheit, A., Mbabazi, P.K., Nondo, R.S.O., 2010. Brine Shrimp Toxicity of Some Plants used as Traditional Medicines in Kagera Region, North Western Tanzania. Tanzania Journal of Health Research, Volume 12(1), pp. 63–67

Oberlies, N.H., Jones, J.L., Corbett, T.H., Fotopoulos, S.S., McLaughlin, J.L., 1995. Tumor Cell Growth Inhibition by Several Annonaceous Acetogenins in an in Vitro Disk Diffusion Assay. Cancer Letters, Volume 96(1), pp. 55–62

Oberlies, N.H., Croy, V.L., Harrison, M.L., McLaughlin, J.L., 1997. The Annonaceous Acetogenin Bullatacin is Cytotoxic Against Multidrug-resistant Human Mammary Adenocarcinoma Cells. Cancer Letters, Volume 115(1), pp. 73–79

Villo, P., 2008. Synthesis of Acetogenin Analogues. M.Sc. Thesis, Tartu University

Yuan, S.S.F., Chang, H.L., Chen, H.W. Yeh, Y.T., Kao, Y.H., 2003. Annonacin, a Mono-tetrahydrofuran Acetogenin, Arrests Cancer Cells at the G1 Phase and Causes Cytotoxicity in a Bax- and Caspase-3-related Pathway. Life Sciences, Volume 72(25), pp. 2853–2861

Zafra-Polo, M.C., Gomzales, M.C., Estonell, E., Sahpaz, S., Cortes, D., 1996. Acetogenins from Annonaceae, Inhibitors of Mitochondrial Complex I. Phytochemistry, Volume 42(2), pp. 253–271

Zeng, L., Wu, F.E., McLaughlin, J.L., 1995. Annohexocin, a Novel Mono-THF Acetogenin with Six Hydroxyls, from Annona muricata (Annonaceae). Bioorganic & Medicinal Chemistry Letters, Volume 5(16), pp. 1865–1868

Zeng, L., Ye, Q., Oberlies, N.H., Shi, G., Gu, Z.-M., He, K., McLaughlin, J.L., 1996. Recent Advances in Annonaceous Acetogenins. Natural Product Reports, Volume 13(4), pp. 275–306