Antioxidant and in vitro cytogenotoxic properties of Amburana cearensis (Allemão) A.C.Sm. leaf extract

Abstract

Amburana cearensis leaves have been used in folk medicine to treat respiratory diseases and inflammations. This study aimed to evaluate the biological potential of A. cearensis leaves by antioxidant and in vitro cytogenotoxic analyses of ethanolic crude extract (EE) and its fractions in healthy human cells. The EE was obtained by percolation, followed by fractionation using dichloromethane, cyclohexane, ethyl acetate (EtOAc), and methanol (MeOH) as organic solvents. Extract and all fractions were evaluated for their antioxidant potential by DPPH and reducing power tests. In vitro cytotoxic activity was determined in human peripheral blood mononuclear cells by MTT assay for the extract, EtOAc and MeOH fractions. In turn, the genotoxic activity was determined in human lymphocytes by the Cytokinesis Block Micronucleus assay only for the EtOAc fraction. Only EtOAc fraction was analyzed via gas chromatography coupled to mass spectrometry due to its higher biological activity. Considering the antioxidant potential, the EtOAc fraction was most effective in DPPH (EC₅₀ 43.37 µg/mL) and reducing power (EC₅₀ 89.80 µg/mL) assays. GC-MS analysis of the EtOAc fraction led to the identification of guaiacol, 2,3-dihydro-benzofuran, 2-methoxy-4-vinylphenol, isovanillic acid methyl ester, 4-hydroxybenzaldehyde, and 4-(ethoxymethyl)-phenol. The EE (400–1000 µg/mL), EtOAc (≤150 µg/mL) and MeOH (50 and 150–600 µg/mL) fractions were not cytotoxic by MTT test. Additionally, the EtOAc fraction (100–400 µg/mL) did not induce significant genotoxic damage. Concentrations of the EtOAc fraction with antioxidant activity showed no cytotoxicity, nor genotoxicity potential, indicating them as a nontoxic natural antioxidant source.

Keywords:

• Amburana cearensis
• cumaru
• cytotoxicity
• genotoxicity
• lymphocytes
• free radical
• ROS

Author contributions

José Rafael da Silva Araujo carried out the cytogenotoxic and antioxidant experiments, analyzed the data, and wrote the paper. Juliana Vieira de Barros and Marília Grasielly de Farias Silva helped to perform the cytogenotoxic and antioxidant experiments, respectively. Edson Renan Barros de Santana, Rafael Jorge Santos Aracati Padilha, and Persio Alexandre da Silva performed the phytochemical analyses. Marccus Alves performed the botanical identification. Neide Santos and Ana Maria Benko-Iseppon planned the experiments and discussed the results. Pedro Marcos de Almeida and Cláudia Sampaio de Andrade Lima were doctorate cosupervisors of José Rafael da Silva Araujo, planned the experiments, discussed the results, and helped writing the paper. Ana Christina Brasileiro-Vidal designed and directed the research and helped to write the paper. All authors read, discussed, and approved the final version of the paper.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The authors thank CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) for financial support and fellowships. We are also grateful to Federal University of Pernambuco for financial and structural support.