title: Biological activity of HPLC-characterized ethanol extract from the aerial parts of Haplophyllum tuberculatum.
abstract: BACKGROUND
  The search for new sources of natural antioxidants from plant material may have beneficial therapeutic potential for those diseases associated with oxidative stress. The medicinal plant Haplophyllum tuberculatum (Forsskal) A. Juss. (Rutaceae) contains phenolic compounds as main phytochemicals; however, there are no reports on its antioxidant properties.
  
  
  OBJECTIVE
  To evaluate antioxidant and cytoprotective potential of ethanol extract of Haplophyllum tuberculatum aerial parts.
  
  
  METHODS
  Total phenol content was determined using Folin-Ciocalteu reagent; antiradical activity was measured using ORAC assay and the analysis of the major polyphenols was carried out using a HPLC-MS method. The antioxidant and cytoprotective effect were also investigated by the MTT assay and DCFH-DA method. The human astrocytoma U373-MG cell line was pretreated with ethanol extract (from 0.025 to 250 µg/mL) for 24 h, prior to 1 mM H2O2 exposure (30 min).
  
  
  CONCLUSIONS
  Total phenol content was 46.2 mg gallic acid/g sample and ORAC value was 1.283 µmol TE/mg sample. Chemical constituents were methoxyflavones, flavonols (mainly quercetin derivatives), cinnamic acids and benzoic acids. In cell system model of oxidative stress, pretreatments with ethanol extract at the concentrations of 2.5, 0.25 and 0.025 µg/mL significantly attenuated H2O2-induced loss in viability by 13.5, 17 and 20.5%, respectively. Furthermore, these ethanol extract concentrations markedly inhibited intracellular ROS production with IC50 0.026 µg/mL. These findings demonstrate the beneficial properties of ethanol extract of Haplophyllum tuberculatum aerial parts, rich in phenolic compounds, as antioxidant and radical scavenger ameliorating ROS-related processes and diseases such as several neurodegenerative disorders.
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, University Complutense of Madrid , Spain.
authors.forename: T F
authors.initials: TF
authors.lastname: Eissa
authors.forename: E
authors.initials: E
authors.lastname: González-Burgos
authors.forename: M E
authors.initials: ME
authors.lastname: Carretero
authors.forename: M P
authors.initials: MP
authors.lastname: Gómez-Serranillos
mesh_terms: D000975:Antioxidants / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D001254:Astrocytoma / Q000378:metabolism; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000737:chemistry; D016166:Free Radical Scavengers / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D020128:Inhibitory Concentration 50; D013058:Mass Spectrometry / Q000379:methods; D018384:Oxidative Stress / Q000187:drug effects; D010636:Phenols / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D035261:Plant Components, Aerial; D010936:Plant Extracts / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D059808:Polyphenols / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D027821:Rutaceae / Q000737:chemistry*
chemical_list: D000975:Antioxidants; D016166:Free Radical Scavengers; D010636:Phenols; D010936:Plant Extracts; D059808:Polyphenols; D017382:Reactive Oxygen Species; D000431:Ethanol; D006861:Hydrogen Peroxide