Pasquale Crupi

HPLC-DAD-ESI-MS Analysis of Flavonoid Compounds in 5 Seedless Table Grapes Grown in Apulian Region

UNLABELLED Flavonoids present in skin extracts of red seedless table grape varieties Summer Royal, Autumn Royal, and Crimson, and white seedless varieties Carati and Thompson were analyzed by HPLC-DAD-MS, in 3 y of study (2006 to 2008). The anthocyanins, delphinidin-3-O-glucoside, cyanidin-3-O-glucoside, petunidin-3-O-glucoside (with their corresponding p-coumaroyl derivatives), peonidin-3-O-glucoside, and malvidin-3-O-glucoside (with their corresponding acetyl, caffeoyl, and p-coumaroyl derivatives) were found. In addition the flavonols quercetin-3-O-glucuronide, quercetin-3-O-rutinoside, quercetin-3-O-glucoside, kaempferol-3-O-glucoside, kaempferol-3-O-galactoside, and the flavan-3-ols procyanidin B1, procyanidin B2, and catechin were also detected. Anthocyanins were the main flavonoids in red grapes ranging from 24 (Crimson) to 500 (Summer Royal) mg/kg fresh weight of grapes; consistent levels of flavonols and flavan-3-ols were also quantified in all varieties. To determine the effective climatic influence on flavonoids content in field conditions, viticultural practices have been developed, that could exclude the effects of direct solar radiation from confounding the assessment of those related to thermal conditions alone. A strong positive correlation was determined between flavonoids and temperature data that seem to be responsible for the difference of these metabolites along the years; furthermore, it has been possible to define a linear relationship (R(2) = 0.6871, P = 0.0057) between thermal amplitude and total flavonoids values in the red grapes. PRACTICAL APPLICATION Grapes are economically the most important fruit species in the world and approximately 30% of its production is used as fresh fruit. Because of the very important role of flavonoids in food quality as well as their health-promoting properties, and considering that our experiments were performed along 3 consecutive years, gathered results in this research are quite promising to give a useful information on the flavonoid contents and their evolution in 5 seedless table grapes that are widespread in Mediterranean regions but also in California and South America, and are grown in a viticultural climate (Apulia, South Italy) very close to some regions of Spain, Turkey, Tunisia, and Israel.

Analysis of carotenoids in grapes to predict norisoprenoid varietal aroma of wines from Apulia.

To determine a correlation between carotenoid precursors in grapes and norisoprenoid varietal aroma of wine, carotenoids were identified and quantified by HPLC-DAD-MS (ESI+) from four representative wine grape varieties of the Apulian region (Chardonnay, Merlot, Negroamaro, Primitivo) in two years of study (2006-2007), and C13-norisoprenoid aroma potential, DeltaC (microg/kg), was calculated from the difference of total carotenoid concentration between veraison and maturity. C13-norisoprenoids were analyzed by GC-MS in the obtained wines from 2006 and 2007 vintages. Higher DeltaC values, found in Chardonnay and Merlot grapes, corresponded to higher norisoprenoid contents in the respective wines, particularly characterized by highly flavorant compounds such as beta-damascenone and 3-oxo-alpha-ionol. A linear regression was determined that was significant at the 0.01% level (F=36.12, p=0.00096) with R=0.9261, between grape DeltaC values and total norisoprenoid contents in wine. These findings support the hypothesis that DeltaC could be a useful technological tool to predict norisoprenoid aroma of wine and, consequently, to identify grapes with higher aroma potential.

Antithrombotic activity of 12 table grape varieties. Relationship with polyphenolic profile

The synthesis of tissue factor (TF) by monocytes/macrophages activated by inflammatory agents is of utmost importance in the pathogenesis of thrombotic diseases and substances inhibiting TF synthesis represent novel and promising antithrombotics. We investigated the effect of 12 table grape varieties (white, red and black) on TF synthesis and the possible relation with the phenolic profile. The ability of grape skin extracts (GSEs) to inhibit TF was evaluated in whole blood and isolated mononuclear cells challenged with endotoxin. TF expression was assayed by functional and immunological assays. All GSEs inhibited TF synthesis but with a different efficiency, red grapes being the most active. By correlation analysis, the compounds showing the strongest association with TF-inhibiting activity were quercetin and cyanidin. However, no single polyphenol was able to inhibit TF synthesis as efficiently as the crude grape extracts, unless it was combined with at least another compound, suggesting a synergism.

Free and bound aroma compounds characterization by GC-MS of Negroamaro wine as affected by soil management†

Negroamaro is an autochthonous wine grape variety of Southern Italy, which is becoming very important for the Italian wine market. The wine aroma is primary affected by the chemical composition of grapes, which can be influenced also by agronomic practices such as soil management. In this study, the free and bound aroma characterization was performed by gas chromatography-mass spectrometry analyses, and the influence of two soil managements (cover cropping and soil tillage) was evaluated. A total of 40 volatile compounds were observed in the wine samples. Alcohols (55.7 mg/L), fatty acids (7.0 mg/L) and esters (6.6 mg/L) were found as the main classes in Negroamaro wine. The results showed that the aroma composition of Negroamaro wine was positively affected by soil tillage probably because of the higher water stress (ψ(s)) recorded in the vines from this treatment. Indeed, among the free volatile compounds, higher contents of esters, carboxylic acids, alcohols, phenolics and acetamides together with lower contents of sulfurs compounds were found in soil tillage wine. Conversely, no difference was observed in glycoside volatile compounds.

Comparison Between Different Flavored Olive Oil Production Techniques: Healthy Value and Process Efficiency.

Three different flavoring methods of olive oil were tested employing two different herbs, thyme and oregano. The traditional method consist in the infusion of herbs into the oil. A second scarcely diffused method is based on the addition of herbs to the crushed olives before the malaxation step during the extraction process. The third innovative method is the implementation of the ultrasound before the olive paste malaxation. The objective of the study is to verify the effect of the treatments on the quality of the product, assessed by means of the chemical characteristics, the phenol composition and the radical scavenging activity of the resulting oils. The less favorable method was the addition of herbs directly to the oil. A positive effect was achieved by the addition of herbs to the olive paste and other advantages were attained by the employment of ultrasound. These last two methods allow to produce oils “ready to sell”, instead the infused oils need to be filtered. Moreover, the flavoring methods applied during the extraction process determine a significant increment of phenolic content and radical scavenging activity of olive oils. The increments were higher when oregano is used instead of thyme. Ultrasound inhibited the olive polyphenoloxidase, the endogenous enzyme responsible for olive oil phenol oxidation. This treatment of olive paste mixed with herbs before malaxation was revealed as the most favorable method due to the best efficiency, reduced time consumption and minor labor, enhancing the product quality of flavored olive oil.

Sangiovese and Its Offspring in Southern Italy

This paper demonstrates the importance of different approaches such as ampelography, historical researches, and molecular analysis to reveal direct parent–child relationship. The aim of this paper was to highlight the degree of relationship to five varieties spread in southern Italy, through ampelographic and molecular characterization: Sangiovese, Mantonico di Bianco, Gaglioppo di Cirò, Mantonicone, and Nerello Mascalese. Molecular characterization was carried out through 52 SSR molecular markers, showing that Sangiovese and Mantonico di Bianco are the parents of Gaglioppo di Cirò, Mantonicone, and Nerello Mascalese. Ampelographic description was performed using the method developed by the Organisation Internationale de la Vigne et du Vin. This analysis identifies three distinct groups: the first brings together Sangiovese and the two offspring Nerello Mascalese and Gaglioppo di Cirò, while Mantonico di Bianco and Mantonicone are positioned at a distance from the first and between them. Using molecular characterization, supported by the ampelographic one, we showed that Gaglioppo di Cirò, Mantonicone, and Nerello Mascalese, three varieties recovered in the southern regions of Italy, such as Calabria and Sicily, originated by the cross between a nationally spread grape variety as Sangiovese and a Calabria autochthonous vine as Mantonico di Bianco.

Skin extracts from 2 Italian table grapes (Italia and Palieri) inhibit tissue factor expression by human blood mononuclear cells.

UNLABELLED Grape and its products such as red wine and grape juice have well-known antithrombotic properties, which have been attributed to their high content in polyphenolic compounds. Most studies on the mechanisms underlying these beneficial effects, among which the suppression of tissue factor (TF) synthesis in blood mononuclear cells (MNC) and vascular endothelium is a prominent one, have been performed with purified polyphenols, while little is known about the effect of fresh grapes which contain a multitude of phytochemicals whose interaction may lead to different cell responses. In this study, we investigated the effect of grape skin extracts (GSEs) on TF expression in isolated blood MNC and in whole blood. Alcoholic extracts from skins of 2 grape varieties (Palieri and Italia) inhibited TF expression in lipopolysaccharide (LPS)-stimulated MNC in a concentration-dependent manner with ≥90% inhibition of TF activity and antigen at 6 μg/mL of gallic acid equivalents. Noteworthy, GSEs were also able to inhibit the appearance of TF in whole blood challenged with LPS. The 2 grape varieties displayed a fairly similar TF-inhibiting capacity despite marked differences in phenolic profile. When selected purified polyphenols were tested, their ability to inhibit TF expression was markedly lower as compared to grape extracts, whereas a mixture of some representative polyphenols was much more efficient, supporting the occurrence of a synergistic effect. Given the key role of cell TF in thrombotic diseases, the inhibition of MNC-mediated clotting activation, if confirmed by in vivo studies, might represent an important antithrombotic mechanism. PRACTICAL APPLICATION Our data indicate that the combination of different polyphenols, as in grape extracts, is much more efficient than the single constituents, a finding that might be useful as starting point for the development of new antithrombotic nutraceutics. In addition, our study validated a simple, inexpensive, and physiologically relevant in vitro method on whole blood that allows the evaluation of one of the most important antithrombotic activities of food and food-derived products. The simplicity of the method makes it suitable also for screening purposes in large-scale studies.

Grape seed extracts modify the outcome of oxaliplatin in colon cancer cells by interfering with cellular mechanisms of drug cytotoxicity

Grape seed extracts are commonly utilized as dietary supplements for their antioxidant properties, even from cancer patients. However, whether these natural extracts interfere with chemotherapeutics utilized in colon cancer treatment is still poorly investigated. The cytotoxicity of extracts from Italia and Palieri cultivars either alone or in combination with oxaliplatin was evaluated in colon cancer cells. Grape seed extracts displayed anti-proliferative activity depending on the concentration utilized through apoptosis induction. In combination, they affected the activation of Erk1/2 and counteracted the intrinsic and the extrinsic pathway of apoptosis, the DNA damage and the generation of ROS induced by oxaliplatin. Noteworthy grape seed extracts strongly enhanced the uptake of oxaliplatin into all cells, by affecting the cell transport system of platinum. The addition of these natural extracts to oxaliplatin strongly reduced the cellular response to oxaliplatin and allowed a huge accumulation of platinum into cells. Here, we shed light on the chemical biology underlying the combination of grape seed extracts and oxaliplatin, demonstrating that they might be detrimental to oxaliplatin effectiveness in colon cancer therapy.Grape seed extracts are commonly utilized as dietary supplements for their antioxidant properties, even from cancer patients. However, whether these natural extracts interfere with chemotherapeutics utilized in colon cancer treatment is still poorly investigated. The cytotoxicity of extracts from Italia and Palieri cultivars either alone or in combination with oxaliplatin was evaluated in colon cancer cells. Grape seed extracts displayed anti-proliferative activity depending on the concentration utilized through apoptosis induction. In combination, they affected the activation of Erk1/2 and counteracted the intrinsic and the extrinsic pathway of apoptosis, the DNA damage and the generation of ROS induced by oxaliplatin. Noteworthy grape seed extracts strongly enhanced the uptake of oxaliplatin into all cells, by affecting the cell transport system of platinum. The addition of these natural extracts to oxaliplatin strongly reduced the cellular response to oxaliplatin and allowed a huge accumulation of platinum into cells. Here, we shed light on the chemical biology underlying the combination of grape seed extracts and oxaliplatin, demonstrating that they might be detrimental to oxaliplatin effectiveness in colon cancer therapy.