title: In vitro and in vivo toxicological evaluations of methyl ferulate, methyl p-coumarate, and pulegone 1,2-epoxide.
abstract: BACKGROUND
  Toxicological screening of natural compounds for medicinal purposes.
  
  
  OBJECTIVE
  The objective of this study is to evaluate the toxicity of methyl ferulate (MF), methyl p-coumarate (MpC), and pulegone 1,2-epoxide (PE) with in vitro and in vivo assays.
  
  
  METHODS
  The in vitro toxicity of MF, MpC, and PE was assessed at a concentration of 10 mg/ml with the Ames assay using two strains of Salmonella typhimurium TA98 and TA100. Human red blood cells (RBC) were used to determine the hemolytic activity of these compounds. The cytotoxicity of above compounds was determined with brine shrimp lethality bioassay (BSLB) at the concentrations of 0.1-20 mg/ml. While dermal and ocular irritation studies were conducted on healthy rabbits (n = 8) for 96 and 12 h post-topical application of test compounds, respectively.
  
  
  RESULTS
  PE produced 6-8% hemolysis of RBCs at all the tested concentrations while MF and MpC produced 10-5% hemolysis up to 20 mg/ml, and 50-85% hemolysis at concentrations of 40 and 80 mg/ml, respectively. The Ames assay indicated that MF, MpC, and PE were non-mutagenic as the test values were not significantly higher as compared with background values of the assay. BSLB suggested the lethal concentration (LC50) values of MF, MpC, and PE as 4.38, 6.74, and 25.91 mg/ml, respectively. In vivo ocular and dermal irritation scores of MF, MpC, and PE were comparable with ethanol (control) in rabbits indicating the non-irritant nature of these natural compounds.
  
  
  CONCLUSIONS
  The present studies suggest that these compounds are non-toxic/non-irritant and might be used for medicinal purposes.
authors.affiliation: a Institute of Pharmacy, Physiology and Pharmacology, University of Agriculture , Faisalabad , Pakistan .
authors.forename: Ahmad
authors.initials: A
authors.lastname: Raza
authors.affiliation: a Institute of Pharmacy, Physiology and Pharmacology, University of Agriculture , Faisalabad , Pakistan .
authors.forename: Faqir
authors.initials: F
authors.lastname: Muhammad
authors.affiliation: b Department of Pharmaceutical Sciences , Federal University of Paraíba , João Pessoa, PB , Brazil .
authors.forename: Damião Pergentino
authors.initials: DP
authors.lastname: de Sousa
authors.affiliation: a Institute of Pharmacy, Physiology and Pharmacology, University of Agriculture , Faisalabad , Pakistan .
authors.forename: Tanweer
authors.initials: T
authors.lastname: Khaliq
authors.affiliation: a Institute of Pharmacy, Physiology and Pharmacology, University of Agriculture , Faisalabad , Pakistan .
authors.forename: Bilal
authors.initials: B
authors.lastname: Aslam
authors.affiliation: c Department of Pharmacy , Federal University of Sergipe , São Cristóvão , Brazil .
authors.forename: Luciana
authors.initials: L
authors.lastname: Andrade
authors.affiliation: d Industrial Biotechnology, National Institute for Biotechnology and Genetic Engineering (NIBGE) , Faisalabad , Pakistan .
authors.forename: Saira
authors.initials: S
authors.lastname: Bashir
authors.affiliation: e Poultry Research Institute, Office of Deputy District Livestock Officer (Poultry) , Faisalabad , Pakistan , and.
authors.forename: Muhammad Irfan
authors.initials: MI
authors.lastname: Anwar
authors.affiliation: f Department of Biochemistry , University of Agriculture , Faisalabad , Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Shahid
authors.affiliation: a Institute of Pharmacy, Physiology and Pharmacology, University of Agriculture , Faisalabad , Pakistan .
authors.forename: Mahwish
authors.initials: M
authors.lastname: Qamar
mesh_terms: D000818:Animals; D001156:Artemia / Q000187:drug effects / Q000502:physiology; D002109:Caffeic Acids / Q000633:toxicity*; D002934:Cinnamates / Q000633:toxicity*; D000081005:Cyclohexane Monoterpenes; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004912:Erythrocytes / Q000187:drug effects* / Q000502:physiology; D006461:Hemolysis / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D039821:Monoterpenes / Q000633:toxicity*; D011817:Rabbits; D018675:Toxicity Tests / Q000379:methods
chemical_list: D002109:Caffeic Acids; D002934:Cinnamates; D000081005:Cyclohexane Monoterpenes; D039821:Monoterpenes; C478851:methyl-p-coumarate; C039648:pulegone; C530806:methyl ferulate
keywords: Ames assay; brine shrimp lethality bioassay; hemolytic assay; natural compounds; rabbits