UPLC-QTOF-MS and NMR analyses of graviola (Annona muricata) leaves

Abstract

Graviola leaves (Annona muricata L., (ceae) are used by some people to try to treat or even cure cancer, even though over-consumption of the fruit, which contains the neurotoxins annonacin and squamocin has caused an atypical form of Parkinson’s disease. In previous analyses, the fruits were extracted with methanol under ambient conditions before analyses. In the present study, UPLC-QTOF-MS and NMR were used to analyze freeze-dried graviola leaves that were extracted using dry methanol and ethanol at 100° C and 10MPa (100atm) pressure in a sealed container. Methanol solubilized 33% of the metabolites in the lyophilized leaves. Ethanol solubilized 41% of metabolites in the lyophilized leaves. The concentrations of total phenolic compounds were 100.3 ±2.8 and 93.2 ± 2.0 mg gallic acid equivalents per g of sample, for the methanolic and ethanolic extracts, respectively. Moreover, the toxicophore (unsaturated 7-lactone) that is present in neurotoxic acetogenins was found in the lipophilic portion of this extract. The concentrations of the neurotoxins annonacin and squamocin were found by UPLC-QTOF-MS to be 305.6 ± 28.3 and 17.4 ± 0.89 (xg/g-dw, respectively, in the dried leaves. Pressurized methanol solubilized more annonacin and squamocin than ethanol. On the other hand, a hot, aqueous infusion solubilized only 0.213% of the annonacin and too little of the squamocin to be detected. So, graviola leaves contain significant amounts of the neurotoxins annonacin and squamocin, as well as some potentially healthy phenolic compounds. Finally, the potential neurotoxicity of whole leaves in dietary supplements could be much higher than that of a tea (hot aqueous infusion) that is made from them.

References

  1. Adewole, S.O., Ajewole, J.A.O., 2009. Protective effects of (Annona muricata Linn. (ceae) leaf aqueous extract on serum lipid profiles and oxidative stress in hepatocytes of streptozotocin-treated diabetic rats. Afr. J. Compi. Alt. Med. 6, 30–41.

    Google Scholar 

  2. Akihisa, T., Matsumoto, K., Tokuda, H., Yasukawa, K., Seine, K., Nakamoto, K., Kun-inaga, H., Suzuki, T., Kimura, Y., 2007. Anti-inflammatory and potential cancer chemopreventive constituents of the fruits of Morinda citrifolia (Noni). J. Nat. Prod. 70, 754–757.

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Badrie, N., Schauss, A.C., 2009. Soursop (Annona muricata L.): composition, nutritional value, medicinal uses, and toxicology. In: Watson, R.R., Preddy, V.R. (Eds.), Bioactive Foods in Promoting Health. Fruits and Vegetables. Academic Press, Oxford, pp. 621–643.

    Google Scholar 

  4. Caparros-Lefebvre, D., Elbaz, A., 1999. Possible relation of atypical parkinsonism in the French West Indies with consumption of tropical plants: a case-control study. Lancet 354, 281–285.

    CAS  Article  PubMed  Google Scholar 

  5. Champy, P., Melot, A., Guerineau, V., Gleye, C., Fall, D., Höglinger, G.U., Ruberg, M., Lannuzel, A., Laprevote, O., Laurens, A., Hocquemiller, R., 2005. Quantification of acetogenins in (Annona muricata linked to atypical parkinsonism in Guadeloupe. Movement Disorders 20, 1629–1633.

    Article  PubMed  Google Scholar 

  6. Florence, N.T., Benoit, M.Z., Jonas, It, Alexandra, T., Desire, D.D.P., Pierre, K., Theophile, D., 2014. Antidiabetic and antioxidant effects of (Annona muricata (Annonaceae), aqueous extract on streptozotocin-induced diabetic rats. J. Ethnopharmacol. 151, 784–790.

    Article  PubMed  Google Scholar 

  7. Foong, C.P., Hamid, R.A., 2012. Evaluation of anti-inflammatory activities of ethanolic extract of Annona muricata leaves. Rev. Bras. Farmacogn. 22, 1301–1307.

    Article  Google Scholar 

  8. Gajalakshmi, S., Vijayalakshmi, S., Devi, R.V., 2012. Phytochemical and pharmacological properties of (Annona muricata: a review. Intl. J. Pharm. Pharmaceut. Sci. 4, 3–6.

    Google Scholar 

  9. Hamizah, S., Roslida, A.H., Fezah, O., Tan, K.L., Tor, Y.S., Tan, C.I., 2012. Chemopreventive potential of (Annona Muricata L leaves on chemically-induced skin papillomagenesis in mice. Asian Pacific J. Cancer Prevention 13, 2533–2539.

    Article  Google Scholar 

  10. Hansra, D.M., Silva, O., Mehta, A., Ahn, E., 2014. Patient with metastatic breast cancer achieves stable disease for 5 years on graviola and Xeloda after progressing on multiple lines of therapy. Adv. Breast Cancer Res. 3, 84–87.

    Article  Google Scholar 

  11. Höllerhage, M., Matusch, A., Champy, P., Lombes, A., Ruberg, M., Oertel, W.H., Höglinger, G.U., 2009. Natural lipophilic inhibitors of mitochondrial complex I are candidate toxins for sporadic neurodegenerative tau pathologies. Exp. Neurol. 220, 133–142.

    Article  PubMed  Google Scholar 

  12. Kim, H.-It, 2010. Identification of novel fatty acid glucosides from the tropical fruit Morinda citrifolia L. Phytochem. Lett. 3, 238–241.

    CAS  Article  Google Scholar 

  13. Kim, G.S., 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 49, 565–571.

    CAS  Article  PubMed  Google Scholar 

  14. Luo, R., Maia, J.G., de Moraes, M.R., Godoy, H.T., Sabaa-Srur, A.U.O., Tran, K., Richards, K.M., Monroe, D.M., Vocque, R.H., Smith, R.E., 2013. NMR analysis of potentially neurotoxic annonaceous fruits. Nat. Prod. J. 3, 230–241.

    CAS  Google Scholar 

  15. Lannuzel, A., Michel, P.P., 2009. Atypical parkinsonism in the French West Indies: the plant toxin annonacin as a potential etiological factor. In: Tseng, K-Y. (Ed.), Cortico-Subcortical Dynamics in Parkinson’s Disease. Humana Press, New York, pp. 283–290.

    Google Scholar 

  16. Levine, R.A., Richards, K.M., Tran, K., Luo, R., Thomas, A.L., Smith, R.E., 2015. Determination of neurotoxic acetogenins in pawpaw (Asimina triloba) fruit by LC-HRMS. J. Agric. Food Chem. 63, 1053–1056.

    CAS  Article  PubMed  Google Scholar 

  17. Machado, A.R.T., Lage, G.A., Medeiros, F.S., Filho, J.D.S., Pimenta, L.P.S., 2015. Total α, β-unsaturated-γ-lactone acetogenins in Annona muricata by proton NMR spectroscopy. Appi. Mag. Res. 46, 153–160.

    CAS  Article  Google Scholar 

  18. Port’s, P.S., Chisté, R.C., Godoy, H.T., Prado, M.A., 2013. The phenolic compounds and the antioxidant potential of infusion of herbs from the Brazilian Amazonian region. Food Res. Intl. 53, 875–883.

    Article  Google Scholar 

  19. Potts, L.F., Luzzio, F.A., Smith, S.C., Hetman, M., Champy, P., Litvan, I., 2012. cin in Asimina triloba fruit: Implication for neurotoxicity. NeuroToxicol. 33, 53–58.

    CAS  Article  Google Scholar 

  20. Singleton, V.L., Orthofer, R., Lamuela-Raventos, R.M., 1999. Analysis of total phenols and others oxidation substrates and antioxidants by Folin—Ciocauteau reagent. Methods Enzymol. 299, 152.

    CAS  Article  Google Scholar 

  21. Smith, R.E., 2014. Medicinal Chemistry - Fusion of Traditional and Western Medicine, Second Edition. Bentham Science, Sharjah, U.A.E, pp. 316–326.

    Google Scholar 

  22. Smith, R.E., Tran, K., Richards, K.M., 2014. Bioactive (Annonaceous Acetogenins, Chapter 4, in Studies in Natural Products Chemistry, vol. 41. Elsevier, New York, pp. 93–117.

    Google Scholar 

  23. Sun, S., Liu, J., Kadouh, H., Sun, X., Zhou, It, 2014. Three new antiproliferative (Annonaceous acetogenins with mono-tetrahydrofuran ring from graviola fruit (Annona muricata). Bioorg. Med. Chem Lett. 24, 2773–2776.

    CAS  Article  Google Scholar 

  24. Torres, M.P., Rachagani, S., Purohit, V., Pandey, P., Joshi, S., Moore, E.D., Johansson, S.L., Singh, P.K., Ganti, A.K., Batra, S.K., 2012. Graviola: a novel promising natural-derived drug that inhibits tumorigenicity and metastasis of pancreatic cancer cells in vitro and in vivothrough altering cell metabolism. Cancer Lett. 323, 29–40.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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Authors

Contributions

IVMM, PRVR, FLS, KMC, GJZ, ESB and RES conceived and designed the experiments. IVMM, PRVR, FLS, KMC, GJZ and ESB did the LC-MS and total phenolics analyses as well as data interpretation. RL did the NMR analyses. IVMM, KMR and KT did the pressurized liquid extractions. RES wrote the article. Everyone read the article and suggested some changes.

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Correspondence to Robert E. Smith.

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de Moraes, I.V.M., Ribeiro, P.R.V., Schmidt, F.L. et al. UPLC-QTOF-MS and NMR analyses of graviola (Annona muricata) leaves. Revista Brasileira de Farmacognosia 26, 174–179 (2016). https://doi.org/10.1016/j.bjp.2015.12.001

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Keywords

  • Graviola
  • Annona muricata
  • UPLC-QTOF-MS
  • Parkinson’s disease
  • NMR