This paper describes for the first time a complete characterisation of the phenolic compounds in different anatomical parts of the Albariño grape. The application of high-performance liquid chromatography coupled with two complementary techniques, hybrid quadrupole time-of-flight and triple-quadrupole mass spectrometry, allowed the phenolic composition of the Albariño grape to be unambiguously identified and quantified. A more complete phenolic profile was obtained by product ion and precursor ion scans, while a neutral loss scan at 152 u enabled a fast screening of procyanidin dimers, trimers and their galloylated derivatives. The compounds were confirmed by accurate mass measurements in QqToF-MS and QqToF-MS/MS modes at high resolution, and good fits were obtained for all investigated ions, with errors ranging from 0.2 to 4.5 mDa. To the best of our knowledge, two flavanol monomer hexosides were detected in the grape berry for the first time.

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Flavanones and flavonols in ROOE presented higher bioavailability, suggesting that the processing undergone by the raw tomato improved their absorption.

This study was aimed at investigating the cardiovascular effects of an Olea europea L. leaf extract (OEE), of a Hibiscus sabdariffa L. flower extract (HSE), and of their 13 : 2 w/w mixture in order to assess their cardiac and vascular activity. Both extracts were fully characterized in their bioactive compounds by HPLC-MS/MS analysis. The study was performed using primary vascular endothelial cells (HUVECs) to investigate the antioxidant and cytoprotective effect of the extracts and their mixture and isolated guinea-pig left and right atria and aorta to evaluate the inotropic and chronotropic activities and vasorelaxant properties. In cultured HUVECs, OEE and HSE reduced intracellular reactive oxygen species formation and improved cell viability, following oxidative stress in dose-dependent manner. OEE and HSE exerted negative inotropic and vasorelaxant effects without any chronotropic property. Interestingly, the mixture exerted higher cytoprotective effects and antioxidant activities. Moreover, the mixture exerted an inotropic effect similar to each single extract, while it revealed an intrinsic negative chronotropic activity different from the single extract; its relaxant activity was higher than that of each single extract. In conclusion OEE and HSE mixture has a good potential for pharmaceutical and nutraceutical application, thanks to the synergistic effects of the single phytochemicals.

Virgin olive oil is an important ingredient of the Mediterranean diet highly appreciated for its beneficial health effects due to the presence of mono-unsaturated fatty acids and antioxidant properties. The olive oil quality is defined by many parameters, among which acidity and peroxide index and according to international regulations, these two determinations are carried out in laboratory environment, by means of manual titration. This kind of method, however, cannot be used on oil production sites, where, instead, it would be desirable to know the main characteristics of the oil for proper product classification.In this paper a new technique to measure olive oil acidity by electrical conductance is presented that allows for fast, costeffective and on-site measurements. Such a technique has been validated with a set of 55 olive oil samples. Two different oil emulsions have been used and compared for the measurements: based on a hydro-alcoholic solution (60% ethanol 40% distilled water) and distilled water, respectively. The data show that the electrical conductance of the emulsion with hydro-alcoholic solution is a strong function of the olive oil acidity, that can be estimated with good accuracy (R 2 = 0.9308). The experiments with distilled water, instead, exhibit no significant correlation between oil acidity and emulsion conductance that, however, is found to provide information about peroxide index, polyphenols content and filtration technique.Furthermore, using both types of emulsions to compensate for conductance variations other than free fatty acids the accuracy in acidity determination can be improved reaching R 2 = 0.9786.

Mediterranean diet health benefits have been partially attributed to the dietary consumption of extra virgin olive oil (VOO). Most recent interest has focused on the biologically active phenolic compounds naturally present in VOO. Selected VOO from Italy were investigated for their phenolic profile and inhibitory activity against a-glucosidase and a-amylase, as well as angiotensin-converting enzyme (ACE). These enzymes have been linked to hyperglycemia-associated to hypertension. The total phenolic content (Folin-Ciocalteu) ranged from 109 to 250 mg/kg of oil and was well correlated with the high-performance liquid chromatography (HPLC) data (r 2 = 0.8533). The phenolic profile obtained by HPLC revealed the presence of 3,4-dihydroxyphenylethanol (3,4-DHPEA; 2.9 Ϯ 2 mg/kg) and p-hydroxyphenylethanol (p-HPEA; 2.6 Ϯ 1.3 mg/kg), as the main phenolic alcohols. Secoiridoids and their derivatives were also detected in high concentration in all samples, particularly 3,4-DHPEA-EDA (55 Ϯ 38 mg/kg) and p-HPEA-EDA (42 Ϯ 14 mg/kg). Vanillic acid (1.8 Ϯ 1.1 mg/kg), vanillin (1.2 Ϯ 0.5 mg/kg), p-coumaric acid (0.94 Ϯ 0.7 mg/kg), apigenin (1.0 Ϯ 0.2 mg/kg) and luteolin (2.4 Ϯ 0.5) were also identified. All samples demonstrated a stronger activity against a-glucosidase with respect to a-amylase. The sample that was richer in phenolics showed a half maximal 3 Corresponding inhibitory concentration of 184 and 258 mg/mL against a-glucosidase and a-amylase, respectively. The same sample exhibited the highest activity against the ACE. In conclusion this in vitro study evidenced the hypoglycemic and ACE inhibitory activities of VOOs phenolic extracts. The multivariate statistical model pointed out that the effect of secoiridoids was more pronounced than flavonoids. PRACTICAL APPLICATIONSIn this study potential health benefits of extra virgin olive oil micronutrients (phenolic compounds) were confirmed through assays for the two major carbohydrate digestive enzymes, a-glucosidase and a-amylase as well as for the angiotensin I-converting enzyme. These enzymes have been linked to hyperglycemia-associated to hypertension that is a common condition especially in a late diabetic stage. The in vitro inhibitory activities that were encountered for these enzymes provide a strong biochemical rationale for further in vivo studies and dietary management strategy for patients affected by diabetes type 2 and/or hypertension. 382 M.R. LOIZZO ET AL.

The purpose of this work was to find a simple, cheap, and suitable method, among the most widely employed, able to guarantee a proper determination and quantification of the phenolic content of extra virgin olive oils (EVOOs), in order to satisfy the requirements of the specific health claim (EU Reg. 432/2012). Total phenolic content by Folin-Ciocalteu (FC) was used and compared versus phenolic profile by HPLC-UV, considering this latter as the most sensitive and specific method for evaluating the phenolic content. Both protocols were performed before and after an acid hydrolysis of the polar phenolic fraction that involves a break of the bound forms of hydroxytyrosol (HTyr) and tyrosol (Tyr), with a simplification of the phenolic profile, and quantification of their total free forms. Results of the phenolic compounds of twelve EVOOs, determined by the different analytical approaches, were statistically compared by means of two-tailed paired t-tests: data obtained by the FC assay (expressed as HTyr) before and/or after acid hydrolysis were statistically comparable with results obtained by acid hydrolysis-HPLC (as sum of HTyr and Tyr).Practical applications: The promising results obtained in this study show that the simple and cheap colorimetric assay based on the use of the FC reagent, commonly used for the evaluation of phenolic compounds in hydro-alcoholic extracts of EVOO, can be also efficiently applied, without acid hydrolysis of extracts and HPLC analysis, to verify the compliance to the polyphenols health claim introduced by EU Reg. 432/2012. In fact, in order to preserve the positive image of EVOO due to its healthy properties, it is necessary i) to share an analytical protocol to determine the amount of hydroxytyrosol and its derivatives having a demonstrated effect of protection of blood lipids from oxidative stress ii) to check by this protocol if EVOOs satisfy the EU requirement for including the specific health claim on the oil label.Abbreviations: Api, apigenin; DLA, dialdehydic form of decarboxymethylelenolic acid linked to tyrosol; DOA, dialdehydic form of decarboxymethylelenolic acid linked to hydroxytyrosol; EVOOs, extra virgin olive oils; FC, Folin-Ciocalteu; GA, gallic acid; HTyr, hydroxytyrosol; LA, ligstroside aglycon; Lut, luteolin; OA, oleuropein aglycon; Tyr, tyrosol; VOOs, virgin olive oils Eur.