Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Mar 15:4:100103.
doi: 10.1016/j.crtox.2023.100103. eCollection 2023.

Influence of methanolic extracts from seeds and pulp of Annona squamosa L. on osmotic and morphological fragility in human erythrocytes

Débora Odília Duarte Leite  1 , Cicera Janaíne Camilo  2 , José Walber Gonçalves Castro  2 , Alexandro Rodrigues Dantas  2 , Fabíola Fernandes Galvão Rodrigues  2 , Pablo Antonio Maia de Farias  3 , Edlane Martins de Andrade  3 , Henrique Douglas Melo Coutinho  2 , José Galberto Martins da Costa  1   2
Affiliations

Influence of methanolic extracts from seeds and pulp of Annona squamosa L. on osmotic and morphological fragility in human erythrocytes

Débora Odília Duarte Leite et al. Curr Res Toxicol. .

Abstract

Medicinal plants have been used for many years by communities to treat illnesses. The need for scientific proof of these vegetable's curative effects is as necessary as the proof of the inexistence of toxicity related to the use of extracts with therapeutic potential. Annona squamosa L. (Annonaceae), popularly known as "pinha", "ata" or "fruta do conde", has been used in traditional medicine for its analgesic and antitumor activities. The toxic effects attributed to this plant have also been explored as a pesticide and an insecticide. The aim of the present study was to investigate the toxicity of the methanolic extract of A. squamosa seeds and pulp against human erythrocytes. Blood samples were treated with methanolic extract at different concentrations, osmotic fragility was determined using saline tension assays and morphological analyzes were performed using optical microscopy. The extracts were analyzed using high performance liquid chromatography with diode array detection (HPLC-DAD) for phenolic quantification. The seed's methanolic extract showed toxicity above 50% from a concentration of 100 µg/mL, while also presenting echinocytes in the morphological analysis. The pulp's methanolic extract did not show toxicity to red blood cells or morphological changes at the concentrations tested. HPLC-DAD analysis revealed the presence of caffeic acid in the seed extract and gallic acid in the pulp extract. The seed's methanolic extract is toxic and the pulp's methanolic extract showed no toxicity against human erythrocytes.

Keywords: Annona squamosa; Cytotoxicity; Erythrocytes.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

None
Graphical abstract
Fig. 1
Fig. 1
Erythrotoxicity of methanolic extracts of Annona squamosa seed and pulp. As a negative control, a sodium chloride solution (0.9% NaCl) was used. * = p < 0.05, ** = p < 0.01 and **** = p < 0.0001.
Fig. 2
Fig. 2
Osmotic fragility of blood samples treated with 100 µg/mL of the seed extract and 250 µg/mL of the pulp extract of Annona squamosa at different concentrations of NaCl. Where * = p < 0.05, ** = p < 0.01 and **** = p < 0.0001.
Fig. 3
Fig. 3
(a) Morphology of red cells in the control group − 0.9% NaCl. (b) Morphology of red cells treated with methanolic extract of A. squamosa pulp at a concentration of 250 µg/mL. (c) Morphology of red cells treated with methanolic extract of A. squamosa seed at a concentration of 100 µg/mL. The arrow indicates the presence of echinocytes. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 4
Fig. 4
Elution profiles of HPLC/DAD of Annona squamosa extract (a) seed and (b) pulp. Caffeic acid (peak 4, tR = 16,773); Gallic acid (peak 10, tR = 9,439); Unidentified (peaks 1, 2, 3, 5, 6, 7, 8, 9 and 11). Calibration curve for caffeic acid y = 110754x + 267.5 (r = 0.9902); gallic acid y = 13478x + 94.75 (r = 0.9911).
Fig. 5
Fig. 5
Chemical structures of phenolic acids identified in A. squamosa extracts. (a) caffeic acid;(b) gallic acid.
See this image and copyright information in PMC

References

    1. Ali S.H., Jafar Z.J. Comparison of Antibacterial Efficacy of Annona Squamosa Mouthwash with Chlorhexidine for Children. J Res Med Dent Sci. 2022;10:87–94.
    1. Anand U., Tudu C.K., Nandy S., Sunita K., Tripathi V., Loake G.J., Dey A., Prockow J. Ethnodermatological use of medicinal plants in India: From ayurvedic formulations to clinical perspectives – A review. J Ethnopharmacol. 2022;284:1–28. doi: 10.1016/j.jep.2021.114744. - DOI - PubMed
    1. Barros F.J., Costa R.J.O., Cesário F.R.A.S., Rodrigues L.B., Costa J.G.M., Coutinho H.D.C., Galvao H.B.F., Menezes I.R.A. Activity of essential oils of Piper aduncum anf and Cinnamomum zeylanicum by evaluating osmotic and morphologic fragility of erythrocytes. Eur. J. Integr. Med. 2016;8:505–512. doi: 10.1016/j.eujim.2016.02.011. - DOI
    1. Baskaran R., Pullencheri D., Somasundaram R. Characterization of free, esterified and bound phenolics in custard apple (Annona squamosa L) fruit pulp by UPLC-ESI-MS/MS. Food Res Int. 2016;82:121–127. doi: 10.1016/j.foodres.2016.02.001. - DOI
    1. Chen Y., Cao Y., Li F., Zhu X., Peng C., Lu J., Chen J., Li X., Chen Y. Studies on anti hepatoma activity of Annona squamosa L. pericarp extract. Bioorg Med Chem Lett. 2017;27:1907–1910. doi: 10.1016/j.bmcl.2017.03.036. - DOI - PubMed