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LC-MS identification and preparative HPLC isolation of Frankenia pulverulenta phenolics with antioxidant and neuroprotective capacities in PC12 cell line
Authors:Rim Ben Mansour  Megdiche Ksouri Wided  Stéphanie Cluzet  Stéphanie Krisa  Tristan Richard  Riadh Ksouri
Institution:1. Laboratory of Aromatic and Medicinal Plants, Center of Biotechnology, Technopark of Borj-Cedria (CBBC), Hammam-Lif, Tunisia;2. ISVV, GESVAB EA3675, University of Bordeaux, Villenave d‘Ornon, France
Abstract:Context: Frankenia pulverulenta L. (Frankeniaceae) is a medicinal species with carminative, analgesic and antiviral properties. However, phytochemical investigations, antioxidant and neuroprotective capacities of this plant remain unclear.

Objective: This work assesses the phenolic composition of F. pulverulenta shoot and root and evaluates their antioxidant and neuroprotective capacities.

Materials and methods: Successive fractionation of F. pulverulenta shoot and root using 6 solvents were used. Antioxidant capacity of these fractions was assessed through four in vitro tests (DPPH, ABTS, Fe-chelating activity and ORAC). Phenolic identification, purification as well as neuroprotective activity of ethyl acetate (EtOAc) fraction and purified molecules were assessed.

Results: Among the tested fractions, EtOAc shoot and root fractions possessed considerable phenolic contents (383 and 374?mg GAE/g E, respectively) because of their important ORAC (821 and 1054?mg of TE/g E), DPPH (586 and 750?mg of TE/g) and ABTS (1453 and 1319?mg of TE/g) results. Moreover, gallic acid, quercetin, quercetin galloyl glucoside, trigalloyl hexoside, procyanidin dimers and sulfated flavonoids were identified by LC-DAD-ESI-MS for the first time in this species. The relevant cytoprotective capacity (at 300?μg/mL) against β-amyloid peptide induced toxicity in PC12 cells of EtOAc fractions were corroborated with the chemical composition. In addition, purified molecules were tested for their ORAC and neuroprotective activity. Quercetin showed the best ORAC value (33.55?mmol TE/g polyphenols); nevertheless, procyanidin dimer exhibited an exceptionally efficient neuroprotective activity (100% of viability at 50?μg/mL).

Discussion and conclusions: These findings suggest that this halophyte is a promising source of antioxidant and neuroprotective molecules for pharmaceutical purposes.
Keywords:β-Amyloid peptide  LC-DAD-ESI-MS  fractionation  MTT test  ORAC
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