title: Macroalga Padina pavonica water extracts obtained by pressurized liquid extraction and microwave-assisted extraction inhibit hyaluronidase activity as shown by capillary electrophoresis.
abstract: Hyaluronidase degrades hyaluronic acid, the principal component of the extracellular matrix. Inhibition of this enzyme is thus expected to hinder skin aging. Brown alga Padina pavonica activity toward hyaluronidase was evaluated using capillary electrophoresis (CE)-based enzymatic assays. This green technique allows evaluation of the biological activity of the natural material in an economic manner. Pressurized liquid extraction (PLE), microwave assisted extraction (MAE), supercritical fluid extraction and electroporation extraction techniques were used. Extraction conditions were optimized to obtain cosmetically acceptable Padina pavonica extracts with the best inhibition activity. CE-based assays were conducted using only a few nanoliters of reactants, a capillary of 60cm total length and of 50μm internal diameter, +20kV voltage for separation in 50mM ammonium acetate buffer (pH 9.0) and 200nm wavelength for detection. The reaction mixture was incubated for 1h and CE analysis time was about 11min. A novel online CE-assay using transverse diffusion of laminar flow profiles for in-capillary reactant mixing allowed efficient monitoring of hyaluronidase kinetics with Kmand Vmaxequal to 0.46±0.04mgmL-1and 137.1±0.3nMs-1(r2=0.99; n=3), respectively. These values compared well with literature, which validates the assay. Water extracts obtained by PLE (60°C; 2 cycles) and MAE (60°C; 1000W; 2min) presented the highest anti-hyaluronidase activity. The half maximal effective concentration (IC50) of water PLE extract was 0.04±0.01mgmL-1(r2=0.99; n=3). This value is comparable to the one obtained for Einsenia bicyclis phlorotannin fractions (IC50=0.03mgmL-1), which makes Padina pavonica bioactivity very promising.
authors.affiliation: Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans - CNRS FR 2708 - UMR 7311, Orléans, France.
authors.forename: Syntia
authors.initials: S
authors.lastname: Fayad
authors.affiliation: Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans - CNRS FR 2708 - UMR 7311, Orléans, France. Electronic address: reine.nehme@univ-orleans.fr.
authors.forename: Reine
authors.initials: R
authors.lastname: Nehmé
authors.affiliation: Faculty of Science II, Department of Biology, Lebanese University, 90656 Jdeidet El Metn, Fanar, Lebanon.
authors.forename: Mona
authors.initials: M
authors.lastname: Tannoury
authors.affiliation: Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans - CNRS FR 2708 - UMR 7311, Orléans, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Lesellier
authors.affiliation: Centre de Biophysique moléculaire, CNRS UPR4301 and Université d'Orléans, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Pichon
authors.affiliation: Institut de Chimie Organique et Analytique (ICOA), Université d'Orléans - CNRS FR 2708 - UMR 7311, Orléans, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Morin
mesh_terms: D025924:Chromatography, Supercritical Fluid; D004058:Diffusion; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D018274:Electroporation; D006821:Hyaluronoglucosaminidase / Q000037:antagonists & inhibitors*; D020128:Inhibitory Concentration 50; D007700:Kinetics; D059625:Liquid-Liquid Extraction / Q000379:methods*; D008872:Microwaves; D000459:Phaeophyceae / Q000737:chemistry*; D011312:Pressure; D015203:Reproducibility of Results; D012624:Seaweed / Q000737:chemistry*; D014867:Water / Q000737:chemistry*
chemical_list: D014867:Water; D006821:Hyaluronoglucosaminidase
keywords: Algae; Capillary electrophoresis; Electroporation; Enzyme kinetics; Hyaluronidase