title: Essential Oil from the Resin of Protium heptaphyllum: Chemical Composition, Cytotoxicity, Antimicrobial Activity, and Antimutagenicity.
abstract: BACKGROUND
  Protium heptaphyllum (Aubl.) March is popularly used as an analgesic and anti-inflammatory agent.
  
  
  OBJECTIVE
  This study aimed to evaluate the chemical composition of P. heptaphyllum essential oil, its cytotoxicity in a breast cancer cell line (MCF-7), antimicrobial activity, and its antimutagenicity in vivo.
  
  
  METHODS
  The chemical composition of the essential oil collected in three 3 years was determined by gas chromatography-mass spectrometry. The cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Annexin V conjugated with fluorescein isothiocyanate, caspase-3, and tumor necrosis factor-alpha (TNF-α) assays were performed to evaluate apoptosis and inflammatory events. The antimutagenic activity at doses of 25, 50, and 100 mg/kg was determined using a micronucleus test in murine bone marrow.
  
  
  RESULTS
  The essential oil showed a predominance of monoterpene compounds, being the terpinolene, p-cymene-8-ol, and p-cymene, present in the essential oil extracted in the 3 years. The essential oil showed a protection against cyclophosphamide-induced genotoxicity, and the cytotoxicity index polychromatic erythrocytes/normochromatic erythrocytes ratio in animals treated with oil at all doses (1.34 ± 0.33; 1.15 ± 0.1; 1.11 ± 0.13) did not differ from the negative control animal (1.31 ± 0.33), but from the cyclophosphamide group (0.61 ± 0.12). Cytotoxicity, at a concentration of 40.0 μg/mL, and antimicrobial activity were not observed for the essential oil (minimum inhibitory concentration ≥0.5 mg/mL). The essential oil did not change the levels of caspase-3 in the TNF-α level.
  
  
  CONCLUSIONS
  The essential oil showed antimutagenic activity due to its chemical composition.
  
  
  CONCLUSIONS
  Terpinolene, p-cymene-8-ol, and p-cymene are the main constituents of the essential oil of P. heptaphyllum collected within 3-yearsThe essential oil of P. heptaphyllum did not show antimicrobial activity (MIC >0.5 mg/mL) against E. coli, S. aureus, E. faecalis, and C. albicansThe essential oil of P. heptaphyllum has activity against S. mutans (MIC = 0.5 mg/mL)The essential oil showed a protection against cyclophosphamide-induced genotoxicity in the micronuclei assay. Abbreviations used: GC-MS: Gas Chromatography-Mass Spectrometry, MTT: 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Annexin V-FITC: Annexin V conjugated with fluorescein isothiocyanate, TNF-α: Tumor necrosis factor alpha, MIC: Minimum Inhibitory Concentration.
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Ewelyne Miranda
authors.initials: EM
authors.lastname: de Lima
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Didley Sâmia Paiva
authors.initials: DS
authors.lastname: Cazelli
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Fernanda Endringer
authors.initials: FE
authors.lastname: Pinto
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Renata Alves
authors.initials: RA
authors.lastname: Mazuco
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Ieda Carneiro
authors.initials: IC
authors.lastname: Kalil
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Dominik
authors.initials: D
authors.lastname: Lenz
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Scherer
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Tadeu Uggere
authors.initials: TU
authors.lastname: de Andrade
authors.affiliation: Department of Pharmaceutical Sciences, University Vila Velha, Av. Comissário José Dantas de Melo, n°21, Boa Vista Vila Velha - Espírito Santo. CEP 29102-920. Brazil.
authors.forename: Denise Coutinho
authors.initials: DC
authors.lastname: Endringer
keywords: Antimutagenicity; Protium heptaphyllum; cytotoxicity; essential oil; micronuclei assay