Olive Biophenols Research Articles

Strategies to Broaden the Applications of Olive Biophenols Oleuropein and Hydroxytyrosol in Food Products.

Oleuropein (OLE) and hydroxytyrosol (HT) are olive-derived phenols recognised as health-promoting agents with antioxidant, anti-inflammatory, cardioprotective, antifungal, antimicrobial, and antitumor activities, providing a wide range of applications as functional food ingredients. HT is Generally Recognised as Safe (GRAS) by the European Food Safety Authority (EFSA) and the Food and Drug Administration (FDA), whereas OLE is included in EFSA daily consumptions recommendations, albeit there is no official GRAS status for its pure form. Their application in food, however, may be hindered by challenges such as degradation caused by processing conditions and undesired sensorial properties (e.g., the astringency of OLE). Among the strategies to overcome such setbacks, the encapsulation in delivery systems and the covalent and non-covalent complexation are highlighted in this review. Additionally, the synthesis of OLE and HT derivatives are studied to improve their applicability. All in all, more research needs however to be carried out to investigate the impact of these approaches on the sensory properties of the final food product and its percussions at the gastrointestinal level, as well as on bioactivity. At last limitations of these approaches at a scale of the food industry must also be considered.

Structure-Biological Activity Relationships of Extra-Virgin Olive Oil Phenolic Compounds: Health Properties and Bioavailability.

Extra-virgin olive oil is regarded as functional food since epidemiological studies and multidisciplinary research have reported convincing evidence that its intake affects beneficially one or more target functions in the body, improves health, and reduces the risk of disease. Its health properties have been related to the major and minor fractions of extra-virgin olive oil. Among olive oil chemical composition, the phenolic fraction has received considerable attention due to its bioactivity in different chronic diseases. The bioactivity of the phenolic compounds could be related to different properties such as antioxidant and anti-inflammatory, although the molecular mechanism of these compounds in relation to many diseases could have different cellular targets. The aim of this review is focused on the extra-virgin olive oil phenolic fraction with particular emphasis on (a) biosynthesis, chemical structure, and influence factors on the final extra-virgin olive oil phenolic composition; (b) structure–antioxidant activity relationships and other molecular mechanisms in relation to many diseases; (c) bioavailability and controlled delivery strategies; (d) alternative sources of olive biophenols. To achieve this goal, a comprehensive review was developed, with particular emphasis on in vitro and in vivo assays as well as clinical trials. This report provides an overview of extra-virgin olive oil phenolic compounds as a tool for functional food, nutraceutical, and pharmaceutical applications.

Mediterranean and MIND Diets Containing Olive Biophenols Reduces the Prevalence of Alzheimer's Disease.

The risk of Alzheimer’s disease (AD) increases with nonmodifiable conditions including age and lack of effective efficacious pharmacotherapy. During the past decades, the non-pharmacotherapy mode of treatment of dietary modification received extensive attention in AD research. In order to reduce the AD pathology and cognitive decline, various dietary patterns have been attempted including caloric restriction (CR), dietary approaches to stop hypertension (DASH), ketogenic diets (KD), Mediterranean diet (MedDi) and Mediterranean-DASH diet Intervention for Neurological Delay (MIND) diet. Higher adherence to the MedDi diet was associated with decreases in cardiovascular and neurological disorders including AD and related cognitive decline. However, another emerging healthy dietary pattern MIND diet has also been associated with slower rates of cognitive decline and significant reduction of AD rate. Olive serves as one of the building block components of MedDi and MIND diets and the exerted potential health beneficial might be suggested due to the presence of its bioactive constituents such as oleic acids and phenolic compounds (biophenols). A few trials using medical food showed an optimal result in presymptomatic or early stages of AD. The review supports the notion that MedDi and MIND diets display potential for maintaining the cognitive function as nonpharmacological agents against AD pathology and proposed preventative mechanism through the presence of olive biophenols and presents the gaps along with the future directions.

Olive Biophenols Reduces Alzheimer's Pathology in SH-SY5Y Cells and APPswe Mice.

Alzheimer’s disease (AD) is a major neurodegenerative disease, associated with the hallmark proteinacious constituent called amyloid beta (Aβ) of senile plaques. Moreover, it is already established that metals (particularly copper, zinc and iron) have a key role in the pathogenesis of AD. In order to reduce the Aβ plaque burden and overcome the side effects from the synthetic inhibitors, the current study was designed to focus on direct inhibition of with or without metal-induced Aβ fibril formation and aggregation by using olive biophenols. Exposure of neuroblastoma (SH-SY5Y) cells with Aβ42 resulted in decrease of cell viability and morphological changes might be due to severe increase in the reactive oxygen species (ROS). The pre-treated SH-SY5Y cells with olive biophenols were able to attenuate cell death caused by Aβ42, copper- Aβ42, and [laevodihydroxyphenylalanine (l-DOPA)] l-DOPA-Aβ42-induced toxicity after 24 h of treatment. Oleuropein, verbascoside and rutin were the major anti-amyloidogenic compounds. Transgenic mice (APPswe/PS1dE9) received 50 mg/kg of oleuropein containing olive leaf extracts (OLE) or control diet from 7 to 23 weeks of age. Treatment mice (OLE) were showed significantly reduced amyloid plaque deposition (p < 0.001) in cortex and hippocampus as compared to control mice. Our findings provide a basis for considering natural and low cost biophenols from olive as a promising candidate drug against AD. Further studies warrant to validate and determine the anti-amyloid mechanism, bioavailability as well as permeability of olive biophenols against blood brain barrier in AD.

Fructosylation of Hydroxytyrosol by the β‐Fructofuranosidase from Xanthophyllomyces dendrorhous: Insights into the Molecular Basis of the Enzyme Specificity

AbstractThis work investigates the ability of the β‐fructofuranosidase pXd‐INV from the yeast Xanthophyllomyces dendrorhous to glycosylate the olive biophenol hydroxytyrosol (HT). Two fructosylated derivatives (Fru‐HT1 and Fru‐HT2) were synthesized. Under the best conditions (300 mg/mL sucrose, 25 mg/mL HT), the maximum yield was 45.6 %. MS and 2D‐NMR analyses showed that the major product (Fru‐HT1) was fructosylated at the primary OH of HT. The structure of the complexes with the substrates and the product analyzed by crystallography led to the understanding of the molecular determinants regulating the enzymatic mechanism. Product‐soaked crystals revealed that the minor derivative (Fru‐HT2) was fructosylated at the phenolic p‐OH group. The two binding modes for HT at pXd‐INV active site are governed almost exclusively by packing to Trp105 (Fru‐HT1) and polar interactions with the loop Glu334‐Asn342 (Fru‐HT2), respectively. Specific mutagenesis at these residues was accomplished to tune the enzyme regiospecificity. One of the studied mutants (N342Q) was notably more specific for the fructosylation at the phenolic OH than the wild‐type.

The Olive Biophenols Oleuropein and Hydroxytyrosol Selectively Reduce Proliferation, Influence the Cell Cycle, and Induce Apoptosis in Pancreatic Cancer Cells.

Current chemotherapy drugs for pancreatic cancer only offer an increase in survival of up to six months. Additionally, they are highly toxic to normal tissues, drastically affecting the quality of life of patients. Therefore, the search for novel agents, which induce apoptosis in cancer cells while displaying limited toxicity towards normal cells, is paramount. The olive biophenols, oleuropein, hydroxytyrosol and tyrosol, have displayed cytotoxicity towards cancer cells without affecting non-tumorigenic cells in cancers of the breast and prostate. However, their activity in pancreatic cancer has not been investigated. Therefore, the aim of this study was to determine the anti-pancreatic cancer potential of oleuropein, hydroxytyrosol and tyrosol. Pancreatic cancer cells (MIA PaCa-2, BxPC-3, and CFPAC-1) and non-tumorigenic pancreas cells (HPDE) were treated with oleuropein, hydroxytyrosol and tyrosol to determine their effect on cell viability. Oleuropein displayed selective toxicity towards MIA PaCa-2 cells and hydroxytyrosol towards MIA PaCa-2 and HPDE cells. Subsequent analysis of Bcl-2 family proteins and caspase 3/7 activation determined that oleuropein and hydroxytyrosol induced apoptosis in MIA PaCa-2 cells, while oleuropein displayed a protective effect on HPDE cells. Gene expression analysis revealed putative mechanisms of action, which suggested that c-Jun and c-Fos are involved in oleuropein and hydroxytyrosol induced apoptosis of MIA PaCa-2 cells.

Biophenols: Enzymes (β-secretase, Cholinesterases, histone deacetylase and tyrosinase) inhibitors from olive (Olea europaea L.)

The focus of this study was on inhibition of enzymes involved in the pathogenesis Alzheimer's disease (AD) including prime amyloid beta (Aβ) producing enzyme (β-secretase: BACE-1) and disease progression enzymes including acetylcholinesterase (AChE), butyrylcholinesterase (BChE), histone deacetylase (HDAC), and tyrosinase along with the catecholamine L-DOPA, by using olive biophenols. Here we report the strongest inhibition of BACE-1 from rutin (IC50: 3.8 nM) followed by verbascoside (IC50: 6.3 nM) and olive fruit extract (IC50: 18 ng), respectively. Olive biophenol, quercetin exhibited strongest enzyme inhibitory activity against tyrosinase (IC50: 10.73 μM), BChE (IC50: 19.08 μM), AChE (IC50: 55.44 μM), and HDAC (IC50: 105.1 μM) enzymes. Furthermore, olive biophenol verbascoside (IC50: 188.6 μM), and hydroxytyrosol extreme extract (IC50: 66.22 μg) were showed the highest levels of inhibition against the HDAC enzyme. Neuroprotective capacity against levodopa-induced toxicity in neuroblastoma (SH-SY5Y) cells of olive biophenols were assessed, where rutin indicated the highest neuroprotection (74%), followed by caffeic acid (73%), and extract hydroxytyrosol extreme (97%), respectively. To the best of our knowledge, this is the first in vitro report on the enzymes inhibitory activity of olive biophenols. Taken together, our in vitro results data suggest that olive biophenols could be a promising natural inhibitor, which may reduce the enzyme-induced toxicity associated with the oxidative stress involved in the progression of AD. Chemical compounds used in the studyAcetylthiocholine iodide (PubChem CID: 74629); S-Butyrylthiocholine chloride (PubChem CID: 3015121); Caffeic acid (PubChem CID: 689043); Dimethyl sulfoxide (DMSO) (PubChem: 679); L-3,4-Dihydroxyphenylalanine (L-DOPA) (PubChem CID: 6047); 5,5′-Dithiobis (2-nitrobenzoic acid) (DTNB) (PubChem CID: 6254); Epigallocatechin gallate (EGCG) (PubChem CID: 65064); Ethylenediamine tetraacetic acid (EDTA) (PubChem CID: 6049); Galantamine hydrobromide (PubChem CID: 121587); l-Glutamine (PubChem CID: 5961); Hydroxytyrosol (PubChem CID: 82755); Kojic acid (PubChem CID: 3840); Luteolin (PubChem CID: 5280445); Oleuropein (PubChem CID: 5281544); Penicillin-streptomycin (PubChem CID: 131715954); Quercetin (PubChem CID: 5280343); Rutin (PubChem CID: 5280805); Tris-HCl buffer (PubChem: 93573); Trypan blue (PubChem: 9562061).

Development and Validation of an Analytical Methodology Based on Liquid Chromatography-Electrospray Tandem Mass Spectrometry for the Simultaneous Determination of Phenolic Compounds in Olive Leaf Extract.

A simple method was validated for the analysis of 31 phenolic compounds using liquid chromatography-electrospray tandem mass spectrometry. Proposed method was successfully applied to the determination of phenolic compounds in an olive leaf extract and 24 compounds were analyzed quantitatively. Olive biophenols were extracted from olive leaves by using microwave-assisted extraction with acceptable recovery values between 78.1 and 108.7%. Good linearities were obtained with correlation coefficients over 0.9916 from calibration curves of the phenolic compounds. The limits of quantifications were from 0.14 to 3.2 μg g-1. Intra-day and inter-day precision studies indicated that the proposed method was repeatable. As a result, it was confirmed that the proposed method was highly reliable for determination of the phenolic species in olive leaf extracts.

Oleuropein: Molecular Dynamics and Computation.

Olive oil and table olive biophenols have been shown to significantly enrich the hedonic-sensory and nutritional quality of the Mediterranean diet. Oleuropein is one of the predominant biophenols in green olives and leaves, which not only has noteworthy freeradical quenching activity but also putatively reduces the incidence of various cancers. Clinical trials suggest that the consumption of extra virgin olive oil reduces the risk of several degenerative diseases. The oleuropein-based bioactives in olive oil could reduce tumor necrosis factor α, interleukin-1β and nitric oxide. Therefore, the quality of olive biophenols should be preserved and even improved due to their disease-fighting properties. Understanding the molecular dynamics of oleuropein is crucial to increase olive oil and table olive quality. The objective of this review is to provide the molecular dynamics and computational mapping of oleuropein. The oleuropein molecular bond sequential breaking mechanisms were analyzed through unimolecular reactions under electron spray ionization, collision activated dissociations, and fast atom bombardment mass spectrometry. Oleuropein is a biophenol-secoiridoid expressing different functionalities such as two π-bonds, two esters, two acetals, one catechol, and four hexose hydroxyls within 540 mw. The oleuropein solvent-free reactivity is leading to glucose loss and bioactive aglycone-dialdehydes via secoiridoid ring opening. Oleuropein electron distribution revealed that the free-radical non-polar processes occur from its highest occupied molecular orbital, while the lowest unoccupied molecular orbital is clearly devoted to nucleophilic and base site reactivity. This molecular dynamics and computational mapping of oleuropein could contribute to the engineering of olive-based biomedicine and/or functional food.

Olive (Olea europaea L.) Biophenols: A Nutriceutical against Oxidative Stress in SH-SY5Y Cells

Plant biophenols have been shown to be effective in the modulation of Alzheimer’s disease (AD) pathology resulting from free radical-induced oxidative stress and imbalance of the redox chemistry of transition metal ions (e.g., iron and copper). On the basis of earlier reported pharmacological activities, olive biophenols would also be expected to have anti-Alzheimer’s activity. In the present study, the antioxidant activity of individual olive biophenols (viz. caffeic acid, hydroxytyrosol, oleuropein, verbascoside, quercetin, rutin and luteolin) were evaluated using superoxide radical scavenging activity (SOR), hydrogen peroxide (H2O2) scavenging activity, and ferric reducing ability of plasma (FRAP) assays. The identification and antioxidant activities in four commercial olive extracts—Olive leaf extractTM (OLE), Olive fruit extractTM (OFE), Hydroxytyrosol ExtremeTM (HTE), and Olivenol plusTM (OLP)—were evaluated using an on-line HPLC-ABTS•+ assay, and HPLC-DAD-MS analysis. Oleuropein and hydroxytyrosol were the predominant biophenols in all the extracts. Among the single compounds examined, quercetin (EC50: 93.97 μM) and verbascoside (EC50: 0.66 mM) were the most potent SOR and H2O2 scavengers respectively. However, OLE and HTE were the highest SOR (EC50: 1.89 μg/mL) and H2O2 (EC50: 115.8 μg/mL) scavengers among the biophenol extracts. The neuroprotection of the biophenols was evaluated against H2O2-induced oxidative stress and copper (Cu)-induced toxicity in neuroblastoma (SH-SY5Y) cells. The highest neuroprotection values (98% and 92%) against H2O2-induced and Cu-induced toxicities were shown by the commercial extract HTETM. These were followed by the individual biophenols, caffeic acid (77% and 64%) and verbascoside (71% and 72%). Our results suggest that olive biophenols potentially serve as agents for the prevention of neurodegenerative diseases such as AD, and other neurodegenerative ailments that are caused by oxidative stress.

Olive biophenol integral extraction at a two-phase olive mill

The biophenol integral extraction protocol from vegetation water developed in the early 2000s for the three-phase olive mill was adapted to a large two-phase mill operating in Sicily during the 2016/2017 season. The new set-up allows extensive recovery of olive phenolics, affording after nine months natural extracts with hydroxytyrosol concentration exceeding 1500 ppm. Previous toxic waste is transformed into a source of additional revenues for the milling company, and of valued bioproducts for the bioeconomy partner, eliminating altogether a source of potential pollution.

Biophenols-loaded solid lipid particles (SLPs) development by membrane emulsification

The present study investigates the preparation of biophenols-loaded solid lipid particles (SLPs) using the membrane emulsification process. This study involves two main novelties: the first is the encapsulation of amphiphilic/hydrophilic bioactive compounds in the hydrophobic matrix and the second is the use of cold membrane emulsification for the production of SLPs. The main objectives of this work were: i) to investigate the process parameters influencing the encapsulation efficiency in SLPs, (ii) to compare the membrane emulsification technique with conventional emulsification methods and (iii) to identify the operative conditions that allow to produce highly concentrated SLPs with improved encapsulation efficiency and low energy consumption. The influence of lipid phase compositions (cocoa butter, cocoa butter/ethyl acetate (4:1)) and the temperature of operation on the SLPs size, encapsulation efficiency and energy consumption was investigated. The membrane emulsification process allowed producing uniform particles with mean size corresponding to 3.6 times the pore size of the membrane. The encapsulation efficiency of hydroxytyrosol, an amphiphilic compound with antioxidant properties of the olive biophenol family, was about 2 times higher than with the homogenizer when the process was carried out at 40°C. The high solidification rate obtained when the process was carried out at room temperature determined an increase of the encapsulation efficiency up to 90%. The energy consumption associated with membrane emulsification process was two order of magnitude lower than the homogenizer.

Abstract 2177: The olive biophenol oleuropein selectively induces apoptosis in pancreatic cancer cells in vitro

Abstract Pancreatic cancer is a devastating disease with a 5-year survival rate of less than 5%. Resistance to conventional treatment options and toxicity of current chemotherapy agents (gemcitabine) makes pancreatic cancer a target for the development of novel therapeutic agents. Oleuropein is the most abundant biophenol found in olive products; it has anti-atherogenic and anti-inflammatory properties as well as activity against cancers of the breast, colon and prostate. However, there has yet to be any investigation into the effects of oleuropein on pancreatic cancer cells. Consequently, this study aimed to assess the anti-pancreatic cancer activity of oleuropein in vitro. Two cell lines were investigated; a pancreatic cancer cell line (MiaPaCa-2) and a normal pancreas cell line (HPDE). The viability of cells after treatment with 0-200μM oleuropein was assessed using the Dojindo CCK-8 viability assay and compared to gemcitabine. The induction of apoptosis was measured by way of caspase 3/7 activation, using a MUSE flow cell analyser, and expression of Bcl-2, Bax and Bac using Western blot. Cell cycle analysis was conducted using the MUSE flow cell analyser. RNA expression was assessed using Affymetrix GeneChip® Whole Transcript (WT) expression arrays. The IC50 values for oleuropein against MiaPaCa-2 cells was148μM. However, importantly, oleuropein did not decrease the viability of HPDE cells within the treatment range. In comparision, 20nM of gemcitabine did not show selectivity; it reduced the viability of MiaPaCa-2 cells to 21% and of HPDE cells to 2%. An increase in the expression of caspase 3/7 was seen in MiaPaCa-2 cells when treated with oleuropein but it had no effect on the HPDE cells. Furthermore, when treated with oleuropein, an increase in the expression of genes involved in the NRF-2 (oxidative stress) pathway was observed in MiaPaCa-2 cells, an effect not observed in HPDE cells. Conclusion Oleuropein selectively induced apoptosis in the pancreatic cancer cells (MiaPaCa-2), appearing non-toxic to normal pancreas cells (HPDE) within the treatment ranges; this is significant, since gemcitabine was comparatively more toxic to HPDE cells. Furthermore, the link between oleuropein and the NRF-2 pathway in MiaPaCa-2 cells justifies further study into the mechanisms of action of oleuropein and its potential as a novel therapeutic approach for pancreatic cancer. Note: This abstract was not presented at the meeting. Citation Format: Chloe D. Goldsmith, Helen Jankowski, Danielle Bond, Judith Weidenhofer, Costas Stathopoulos, Paul Roach, Christopher Scarlett. The olive biophenol oleuropein selectively induces apoptosis in pancreatic cancer cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2177. doi:10.1158/1538-7445.AM2017-2177

Soft-MS and Computational Mapping of Oleuropein.

Olive oil and table olives are rich sources of biophenols, which provides a unique taste, aroma and potential health benefits. Specifically, green olive drupes are enriched with oleuropein, a bioactive biophenol secoiridoid. Olive oil contains hydrolytic derivatives such as hydroxytyrosol, oleacein and elenolate from oleuropein as well as tyrosol and oleocanthal from ligstroside. Biophenol secoiridoids are categorized by the presence of elenoic acid or its derivatives in their molecular structure. Medical studies suggest that olive biophenol secoiridoids could prevent cancer, obesity, osteoporosis, and neurodegeneration. Therefore, understanding the biomolecular dynamics of oleuropein can potentially improve olive-based functional foods and nutraceuticals. This review provides a critical assessment of oleuropein biomolecular mechanism and computational mapping that could contribute to nutrigenomics.

C18 alkyl-modified silica: A suitable tool for olive biophenol green extraction

A clean and straightforward methodology to assess the value of olive mill wastewater (OMWW) samples as well as to obtain biophenol-rich oils that can find employment in a variety of cosmetic, nutraceutical and pharmaceutical products is described. The method employs chromatographic spherical organosilica microparticles, coupled to advanced analytical techniques. OMWW samples obtained in Sicily from two different organically grown cultivars during the exceptional 2015/2016 olive oil season were analyzed, leading us to establish the conditions for optimal extraction of hydroxytyrosol and tyrosol.

Olive Biophenols as New Antioxidant Additives in Food and Beverage

AbstractReplacing synthetic phenols, sulfites and other synthetic antioxidants and antimicrobials, olive biophenols can be successfully used as multipurpose additives in foodstuffs and beverages. Are these natural phenolics suitable for large‐scale replacement of synthetic additives? This study aims to provide an answer to this and related questions of significant health and environmental significance.

Quick assessment of the economic value of olive mill waste water.

BackgroundOlive biophenols are emerging as a valued class of natural products finding practical application in the food, pharmaceutical, beverage, cosmetic and nutraceutical industries due to their powerful biological activity which includes antioxidant and antimicrobial properties. Olive mill waste water (OMWW), a by-product in olive oil manufacturing, is rich in biophenols such as hydroxytyrosol and tyrosol. The amount of biophenols depends on the cultivar, the geographical area of cultivation, and the seasonal conditions. The goal of this study was to develop a straightforward method to assess the economic value of OMWW via quantification of hydroxytyrosol and tyrosol.ResultsThe amount of hydroxytyrosol and tyrosol phenolic compounds in the OMWW from four different cultivars grown in four different regions of Sicily was analyzed using liquid–liquid and solid–liquid analytical protocols developed ad hoc. Results showed significant differences amongst the different cultivars and their geographical origin. In all samples, the concentration of hydroxytyrosol was generally from 2 to 10 times higher than that of tyrosol. In general, the liquid–liquid extraction protocol gave higher amounts of extracted biophenols. The cultivar Cerasuola had the highest amount of both hydroxytyrosol and tyrosol. The cultivar Nocellara Etnea had the lowest content of both biophenols.ConclusionsA quick method to assess the economic value of olive mill waste water via quantification of hydroxytyrosol and tyrosol in olive phenolic enriched extracts is now available.

DNA diversity in olive (Olea europaea L.) and its relationships with fatty acid, biophenol and sensory profiles of extra virgin olive oils

Olive (Olea europaea L.) is gaining importance worldwide primarily for the production of virgin olive oil (VOO), the main lipid source of the Mediterranean diet.We evaluated the genetic diversity of 20 traditional varieties of Southern Italy using 10 SSR markers. We also produced 382 monovarietal extra virgin olive oils (EVOOs) from the same varieties and studied their chemical profiles by analysing 13 fatty acids (FAs) and 6 olive biophenols (OBPs). In addition, the sensory profile of EVOOs was also assessed.Significant differences among the 20 varieties were found in terms of DNA polymorphism, chemical composition and sensory profile of the EVOOs, highlighting that the genetic variability in olive is detected also in different EVOO features. The chemical variation of EVOOs throughout different years suggested a genotype-specific level of environmental stability and adaptation. The comparison of pairwise distances between varieties obtained by genetic, chemical and sensory data indicated a significant correlation of sensory attributes with the other parameters.The genetic and EVOO composition data underlined the significant diversity present in the varieties under investigation, suggesting that traditional varieties specialized to particular environmental conditions have distinctive, unexplored features. Our work also shows the efficacy of establishing integrated compositional databases as a means to make evident differences among VOOs from traditional varieties. This multidisciplinary information is highly useful to differentiate origin-linked products from others of the same category.

Effect of olive mill wastewater phenolic extract, whey protein isolate and xanthan gum on the behaviour of olive O/W emulsions using response surface methodology

Spray-dried polyphenols extracted from olive mill wastewater (OMW) were used to formulate 20% olive oil-in-water (O/W) emulsions stabilised with whey protein isolate (WPI) and xanthan gum at pH 7. The effects of olive biophenol extract (0.9–4.4 mM), whey protein isolate (0.13–0.5%) and xanthan gum (0.06–0.2%) on the physical stability (creaming index), viscosity, emulsion droplet size and distribution, primary and secondary oxidation products were assessed over accelerated storage of emulsions. Response surface methodology (RSM) was applied for experimental design using three-factor central composite design (CCD) with 3 central points.The effects of OMW phenolics, WPI and xanthan gum were statistically significant on emulsion creaming rate, viscosity, PV, TBARS and cloudiness. Second-order polynomial models were obtained for predicting these responses, resulting in good performances especially for the first two responses. High coefficient of determination (R2) values ranging from 0.847 to 0.973 were obtained for creaming and viscosity, acceptable R2 values were obtained for lipid oxidation parameters (peroxide value and TBARS) and cloudiness value, while non-significant results were obtained for mean droplet size. The performance of the model was particularly good for creaming rate and viscosity. It has been highlighted that the interactions between OMW phenolic extract and the ingredients studied should be considered for the formulation of olive O/W emulsions. This research could be useful for the formulation of functional food products based on O/W emulsions, as well as understanding the physico-chemical interactions among olive biophenols, whey proteins and xanthan gum.

Selected bioactives from callus cultures of olives (Olea europaea L. Var. Coratina) by LC-MS

Olive biophenols, OBPs, and biophenol-secoiridoids, OBPsecos, soluble bioactives from olives, and then in olive oil and table olives, were biosynthesized from Coratina olive callus cultures, under bio- and techno-mimetic experimental conditions. We detected major production of OBPs, hydroxytyrosol, tyrosol, caffeic acid and verbascoside, and of OBP-secos, oleuropein and ligstroside, together with minor amounts of oleuroside, and of demethyl and deglucosil bioactives, by high-performance liquid chromatography, HPLC, coupled with electrospray ionization mass spectrometry, ESI-MS. The olive callus system, in an experimental non-pathogenic environment, produced a nearly model balance of OBPs and OBPsecos in suitable amount comparable to drupes, leaves and seeds. The bioreactor scaling-up of more homogeneous and rapidly growing systems, processed by callus shaking to suspension cell cultures, should produce nutraceutical and cosmeceutical bioactives.