title: An integrated process for the recovery of high added-value compounds from olive oil using solid support free liquid-liquid extraction and chromatography techniques.
abstract: An integrated extraction and purification process for the direct recovery of high added value compounds from extra virgin olive oil (EVOO) is proposed by using solid support free liquid-liquid extraction and chromatography techniques. Two different extraction methods were developed on a laboratory-scale Centrifugal Partition Extractor (CPE): a sequential strategy consisting of several "extraction-recovery" cycles and a continuous strategy based on stationary phase co-current elution. In both cases, EVOO was used as mobile phase diluted in food grade n-hexane (feed mobile phase) and the required biphasic system was obtained by adding ethanol and water as polar solvents. For the sequential process, 17.5L of feed EVOO containing organic phase (i.e. 7L of EVOO treated) were extracted yielding 9.5g of total phenolic fraction corresponding to a productivity of 5.8g/h/L of CPE column. Regarding the second approach, the co-current process, 2L of the feed oil phase (containing to 0.8L of EVOO) were treated at 100mL/min yielding 1.03g of total phenolic fraction corresponding to a productivity of 8.9g/h/L of CPE column. The total phenolic fraction was then fractionated by using stepwise gradient elution Centrifugal Partition Chromatography (CPC). The biphasic solvent systems were composed of n-hexane, ethyl acetate, ethanol and water in different proportions (X/Y/2/3, v/v). In a single run of 4h on a column with a capacity of 1L, 910mg of oleocanthal, 882mg of oleacein, 104mg of hydroxytyrosol were successfully recovered from 5g of phenolic extract with purities of 85%, 92% and 90%, respectively. CPC fractions were then submitted to orthogonal chromatographic steps (adsorption on silica gel or size exclusion chromatography) leading to the isolation of additional eleven compounds belonging to triterpens, phenolic compounds and secoiridoids. Among them, elenolic acid ethylester was found to be new compound. Thin Layer Chromatography (TLC), Nuclear magnetic Resonance (NMR) and High Performance Liquid Chromatography - Diode Array Detector (HPLC-DAD) were used for monitoring and evaluation purposes throughout the entire procedure.
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece; UMR CNRS 7312, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, BP 1039, 51687 Reims, Cedex 2, France.
authors.forename: Apostolis
authors.initials: A
authors.lastname: Angelis
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Hamzaoui
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
authors.forename: Nektarios
authors.initials: N
authors.lastname: Aligiannis
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
authors.forename: Theodora
authors.initials: T
authors.lastname: Nikou
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
authors.forename: Dimitris
authors.initials: D
authors.lastname: Michailidis
authors.affiliation: PharmaGnose Natural Products Approaches, Papathanasiou 24, Eyboia 34100, Greece.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Gerolimatos
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
authors.forename: Aikaterini
authors.initials: A
authors.lastname: Termentzi
authors.affiliation: UMR CNRS 7312, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, BP 1039, 51687 Reims, Cedex 2, France.
authors.forename: Jane
authors.initials: J
authors.lastname: Hubert
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
authors.forename: Maria
authors.initials: M
authors.lastname: Halabalaki
authors.affiliation: UMR CNRS 7312, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, BP 1039, 51687 Reims, Cedex 2, France. Electronic address: jh.renault@univ-reims.fr.
authors.forename: Jean-Hugues
authors.initials: JH
authors.lastname: Renault
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece. Electronic address: skaltsounis@pharm.uoa.gr.
authors.forename: Alexios-Léandros
authors.initials: AL
authors.lastname: Skaltsounis
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D059625:Liquid-Liquid Extraction / Q000379:methods*; D000069463:Olive Oil / Q000737:chemistry*; D010636:Phenols / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000069463:Olive Oil; D010636:Phenols
keywords: Centrifugal partition extraction (CPE)/chromatography (CPC); Co-current; Extra virgin olive oil (EVOO); Hydroxytyrosol; Oleacein; Oleocanthal