Journal of Applied Botany and Food Quality 90, 197 - 204 (2017), DOI:10.5073/JABFQ.2017.090.025 University of Perugia, Department of Pharmaceutical Sciences, Section of Food Science and Nutrition, Perugia, Italy Phenol composition and antioxidant capacity of red wines produced in Central Italy changes after one-year storage Germana Lombardi, Lina Cossignani*, Laura Giua, Maria S. Simonetti, Angela Maurizi, Giovanni Burini, Roberto Coli, Francesca Blasi (Received January 31, 2017; Accepted March 27, 2017) * Corresponding author Summary Much interest is currently concentrated on phenol compounds and antioxidants of wine. The aim of this study was to characterize and evaluate controlled designation of origin (CDO) and typical geo- graphical indications (TGI) red wines from Central Italy and to evalu- ate possible modifications after one year of storage. The total phenol content and antioxidant activity by ORAC method were determined, while phenolic qualitative and quantitative profiles were evaluated by HRGC-FID or HPLC-DAD. All wines showed a good content of total phenols and an obvious antioxidant effect. After a one-year storage in the bottle, a significant decrease (P<0.05) of the ORAC values was observed for TGI wines. Interesting correlations between phenol and ORAC values for CDO wines were found. It can be con- firmed that one-year storage in the bottle has not significantly affected the quality of the wines analyzed, in particular the CDO category. Keywords: chromatography, ORAC, phenol, red wine, storage. Introduction Red wine is an important source of phenol compounds, natural anti- oxidants known for their biological properties and associated with the prevention of oxidative damage (BECKER et al., 2004). Several clinical studies and epidemiological studies have shown a correla- tion between the intake of phenol compounds from red wine and a decrease in the incidence of many diseases (ARRANZ et al., 2012). Many studies have been carried out over the years and numerous analytical techniques have been developed to analyze phenolic com- pounds of grapes and wine (LORRAIN et al., 2013; KHODDAMI et al., 2013). Complete analysis of phenolic compounds requires analytical techniques such as HRGC and HPLC associated with UV-Vis and diode array detectors (DAD) often coupled to other detection sys- tems. A broad bibliography is now available for the study of red or white wines produced in various parts of the world (GRANATO et al., 2011; TOURTOGLOU et al., 2014; IVANOVA-PETROPULOS et al., 2015) al- though few results relate to the wines of Central Italy. SATO et al. (1996) investigated varietal differences in the phenol content of thirty-one wines from various sources, including a Sangiovese Um- brian sample from 1982. VERSARI et al. (2007), characterized by UV- Vis spectrophotometry, the color components and polymer pigments of commercially available red wines, including a Merlot-Umbrian sample. Recently, ESTI et al. (2010) made an explorative sensory study of Grechetto’s wine tasting, some of which were produced in Umbria. The region of Umbria (Central Italy) plays an important role in the production of red wines, including Montefalco Sagrantino known worldwide. To the best of our knowledge, there are no works that focus solely on phenolic composition and antioxidant capacity of Umbrian wines. In this research, twelve Umbrian red wines, six CDO (controlled designation of origin) and six TGI (typical geographical indication) samples were characterized. Total phenol content, anti- oxidant capacity, HRGC and HPLC phenol profiles were determined. The same analyzes were repeated after one year of storage in the bottle to evaluate possible variations. Materials and methods Wine samples Commercial wines, six CDO and six TGI, all collected in 750 ml bottle of cork (eight bottles for each wine) were collected from wineries in the province of Perugia, the capital of Umbria region (Central Italy). All wines were produced with varieties of Vitis la- brusca L. grape varieties. Wine samples were selected to obtain a broad sampling of vine varieties and production area. All the wine samples were purchased at the market of Umbrian wines in 2010, where the wines are available on the market. It should be noted that the year of release for consumption does not correspond to the same vintage for different wines, on the individual production rules. The samples tested in this work are shown in Tab. 1, while some chemical parameters determined according to the official methods described in Commission Regulation (CEE) No 2676/90 of 17 September 1990 are shown in Tab. 2. Wines were produced according to Italian legis- lation. The wines were stored in the dark at 10 ± 1 °C until analysis. The samples, representative aliquots from four bottles, were tested shortly after opening. The same wines stored in unopened bottles at 10 ± 1 °C were analyzed in 2011, one year after marketing. Chemicals and reagents Rutin, kaempferol-3-O-glucoside, isorhamnetin-3-O-glucoside, ka- empferol, rhamnetin and isorhamnetin were obtained from Extra- sythese (Genay, France). cis-Resveratrol was from Cayman Chemical Company (Ann Arbor, MI, USA). The other phenol compounds were purchased from Sigma-Aldrich (Milan, Italy). They were stored in the dark at temperature recommended by the producer. Folin & Cio- calteu’s phenol reagent, BSTFA [N,O-bis(trimethylsilyl)trifluoracet- amide] and Trolox (6-hydroxy-2,5,7,8-tetramethyl-chroman-2-car- boxyl acid) were purchased from Sigma-Aldrich. All analytical grade solvents and reagents were from J.T. Baker (Deventer, Netherlands), Carlo Erba (Milan, Italy), Panreac (Barcellon, Spain) and Supelco (Bellafonte PA, USA). HPLC-grade acetonitrile and water were purchased from J.T. Baker. Determination of total phenol content The TP content was determined spectrophotometrically according to the method of SINGLETON and ROSSI (1965), with slight modifications (BLASI et al., 2016). The wine samples were diluted (1:20 or 1:30, v/v) with distilled water. Diluted red wine (1.0 mL), 20% sodium carbonate (4.0 mL) and Folin & Ciocalteu’s reagent (1.0 mL) were mixed and brought up to a volume of 20 mL with distilled water. 198 G. Lombardi, L. Cossignani, L. Giua, M.S. Simonetti, A. Maurizi, G. Burini, R. Coli, F. Blasi The solution was mixed, kept at 25 °C and incubated at the dark for 90 min and then the absorbance was measured at 750 nm using a Jasco 7850 UV-Vis spectrophotometer (Jasco Inc., Easton, MD, USA). The TP content, expressed as milligrams of gallic acid equi- valent per liter of wine (mg GAE/L), was determined using a calibra- tion curve built using 15% ethanol gallic acid as standard solution (2.5-12.5 mg/L) that was analyzed in the same mode of wine samples. Extraction and derivatization of phenol compounds Phenols were extracted from red wine according to the method of MINUTI et al. (2006). Briefly, sodium metabisulphite (20 mg) and sodium chloride (20 mg) were added to the red wine (1 mL) and then ethyl acetate (3 mL × 3) was used for the extraction. After anhydri- fication with Na2SO4, the extracts were evaporated to dryness under nitrogen stream at 25 °C. The solid residue was dissolved in acetone (50 μL) and derivatized with BSTFA (100 μL). The obtained solution was held in the dark, at room temperature, for 1 h. Then the volume was made up to 1 mL. High-resolution gas-chromatography analysis High-resolution GC analyses were carried out using a Perkin-Elmer Autosystem apparatus (Norwalk, CT, USA), equipped with a split/ splitless injection port, interfaced to a flame ionization detector (FID). The separation was obtained using the HP1-MS fused silica capillary column (30 m × 0.25 mm i.d., 0.25 μm f.t.). The injector and detector temperature was 300 and 320 °C, respectively. The initial oven temperature was 120 °C, held for 3 min and raised to 320 °C at 5 °C/min; the final temperature was held for 15 min. Carrier gas (He) flow rate was 1 mL/min; the injection volume was 1 μL with a split ratio of 1:50. A standard solution in acetone, suitably derivatized as reported in the previous paragraph, containing twenty compounds (vanillin, trans-cinnamic acid, tyrosol, veratric acid, va- nillic acid, homovanillic acid, hydroxytyrosol, 3,4-dihydroxyphenil- acetic acid, homogentisic acid, syringic acid, p-coumaric acid, gal- lic acid, ferulic acid, caffeic acid, sinapinic acid, cis-resveratrol, trans-resveratrol, epicatechin, catechin, fisetin), was used to iden- tify and to quantify the analytes. Calibration curves were obtained by three injections of four different concentrations ranging from 2.5 to Tab. 1: Characteristics of Umbrian (Central Italy) red wines Code Wine name Grape variety Production area Vintage year Wines of Controlled Designation of Origin (CDO) CDO1 Sangiovese Colli Martani Sangiovese (85% min.) Bettona 2007 Ciliegiolo (43°0‘48.96“N-12°29‘11.04“W) Cannaiolo nero Montepulciano Merlot CDO2 Óscano Colli del Trasimeno Gamay Colle Umberto 2009 Sangiovese (43°07‘59.99“N-12°21‘59.98“W) CDO3 Rosso Colli Perugini Sangiovese Sant’Enea 2008 Merlot (43°06‘43.56“N-12°23‘19.68“W) Cabernet Sauvignon CDO4 Montefalco Rosso Sangiovese 65% Montefalco 2007 Merlot 20% (42°55‘59.99“N-12°35‘60“W) Sagrantino 15% CDO5 Baccio del Rosso Colli del Sangiovese Castiglione del Lago 2008 Trasimeno Trasimeno Gamay (43°06‘59.98“N-12°03‘00“W) CDO6 Montefalco Sagrantino Sagrantino 100% Montefalco 2007 (42°55‘17.98“N-12°38‘31.99“W) Wines of Typical Geographical Indication (TGI) TGI1 Sangiovese Umbria Sangiovese (85% min.) Bettona 2009 and other red grapes (43°0‘48.96“N-12°29‘11.04“W) typical of Umbria TGI2 Campiglione Rosso Merlot Colle Umberto 2009 Cabernet Sauvignon (43°07‘59.99“N-12°21‘59.98“W) Sangiovese Gamay TGI3 Garbino dell’Umbria Sangiovese Sant’Enea 2009 Cabernet Sauvignon (43°06‘43.56“N 12°23‘19.68“W) Merlot TGI4 Umbria Rosso Sangiovese 50% Montefalco 2009 Merlot 50% (42°55‘59.99“ N-12°35‘60“W) TGI5 Corio Rosso Sangiovese (80% min.) Castiglione del Lago 2009 Gamet (43°06‘59.98“N-12°03‘00“W) Cabernet TGI6 Rosso Sangiovese Marsciano 2009 Cabernet (42°91‘66.67“N-42°55‘12.20“W) Antioxidant properties of Italian red wines 199 40 mg/L. The least square method was used to calculate the regres- sion equations. High-performance liquid chromatography analysis of phenol compounds Before HPLC-DAD analysis, wine samples (100 mL) were de-alco- holized and dried under vacuum at 40 °C. The residue was diluted in distilled water (50 mL) and then filtered through a 0.20 μm nylon syringe filter (Corning Incorporated, Corning, Germany). The HPLC analyses were performed using a Shimadzu GT-154 system equipped with a Thermo Spectra Series pump, an Hypersil GOLD column (5 μm particle size, 250 × 4.6 mm i.d., Thermo Sci- entific, Rockford, IL, USA) and a Spectra System UV6000LP DAD. Detection was performed on line in a range of wavelength between 240 and 390 nm. The chromatograms were acquired and the data handled using Xcalibur software version 1.2 (Finnigan Corporation 1998-2000, San Jose, CA, USA). The solvents were (A) acetonitrile and (B) water/acetic acid (40:1, v/v). The samples were analyzed by gradient elution at a flow rate of 0.8 mL/min. The elution conditions were: 0-15 min at 15% A; 15-30 min from 15 to 35% A; 30-40 min from 35 to 55% B; 40-44 min from 55 to 100% A with re-equilibration of the column at 44-47 min 100% A to 15% A and 47-60 min at 15% A. A standard solution solubilized in water/acetic acid/methanol (8:2:90, v/v/v), containing ten compounds (rutin, ethyl gallate, quercetin-3- β-D-glucoside, kaempferol-3-O-glucoside, isorhamnetin-3-O-gluco- side, myricetin, quercetin, kaempferol, isorhamnetin, rhamnetin), was used to identify and to quantify the analytes. Calibration curves were obtained by three injections of five different concentrations ranging from 0.5 mg/L to 60 mg/L. Antioxidant capacity The total antioxidant capacity was assessed using oxygen radical absorbance capacity (ORAC) method as reported by MAURIZI et al. (2015). The assay was conducted with FLUOstar Optima fluorescent microplate reader (BMG LABTH GmbH, Germany), provided with a pump, set at wavelengths of excitation and emission at 485 and 520 nm respectively, and interfaced with a computer provided with a MARS Data Analysis software ver. 2.00 for data acquisition and pro- cessing. Costar 96 well black opaque plates (Corning Costar Corpo- ration, Cambridge, MA, USA) were used. The values were expressed as micromoles of Trolox Equivalents per liter of wine (μM TE/L). The ORAC values were calculated using the following formula: ORAC = [Ct × (AUCs – AUC0) × k]/(AUCt – AUC0) where: AUCs, Area under curve in the presence of wine sample AUC0, Area under curve in the presence of blank AUCt, Area under curve in the presence of Trolox Ct, Trolox concentration k, dilution factor Statistical analysis All analytical determinations were carried out in triplicate and the results were expressed as mean value and standard deviation (SD). Student’s t test (type: 3, heteroscedastic; tails: 2), calculated using Excel 2003 (Microsoft Corporation, Redmond, WA, USA), was used to evaluate the differences between the results obtained for the diffe- rent wine categories in different years. The probability of P<0.05 was considered statistically significant. Correlation analyses have been performed using Microcal OriginTM, version 5.0 (Microcal Software Inc., Northampton, MA, USA). Results and discussion Total phenol content and antioxidant capacity Fig. 1 shows the TP content of red wines analyzed in 2010 and after one-year storage (2011). The present study established that in 2010 the highest concentration of phenol compounds was detected in CDO6 wine (3684 mg GAE/L), even if TGI wines showed TP values (from 1341 to 2436 mg GAE/L) comparable with those of CDO ca- tegory (from 1968 to 3684 mg GAE/L). In fact there were no signifi- cant differences (P>0.05) between the TP content of CDO and TGI wine categories. After one-year storage in bottle, a slight decrease of TP content was observed in all samples, with the exception of CDO3 Tab. 2: Some chemical parameters of Umbrian red wines analyzed in 2010 and after one-year storage Code Total acidity Volatile acidity Total SO2 Free SO2 pH Total dry Alcohol content (g/L) (g/L) (mg/L) (mg/L) extract (g/L) (% vol) 2010 2011 2010 2011 2010 2011 2010 2011 2010 2011 2010 2011 2010 2011 Wines of Controlled Designation of Origin (CDO) CDO1 5.17 5.32 0.35 0.34 86.60 96.00 25.60 32.00 3.41 3.49 25.15 26.30 12.30 12.34 CDO2 5.02 5.10 0.41 0.56 76.80 70.40 25.60 16.00 3.55 3.63 27.50 27375 12.65 12.52 CDO3 5.55 5.17 0.58 0.54 64.00 38.40 38.40 19.20 3.73 3.79 30.60 30.50 14.15 14.11 CDO4 5.62 5.55 0.57 0.60 51.20 57.60 19.20 19.20 3.67 3.71 32.95 32.80 13.83 13.65 CDO5 4.42 4.27 0.42 0.45 89.60 89.60 25.60 44.80 3.91 3.99 31.00 31.00 13.54 13.83 CDO6 4.95 5.02 0.32 0.41 76.80 70.40 25.60 19.20 3.67 3.71 29.70 29.55 13.83 13.79 Wines of Typical Geographical Indication (TGI) TGI1 5.10 5.17 0.60 0.63 64.00 76.80 19.20 9.60 3.60 3.65 26.70 28.25 13.65 13.61 TGI2 5.40 5.32 0.53 0.55 70.40 83.20 25.60 32.00 3.63 3.66 31.93 33.10 13.93 14.02 TGI3 5.25 5.55 0.66 0.76 64.00 57.60 25.60 25.60 3.67 3.67 31.50 32.15 14.91 15.59 TGI4 4.95 5.17 0.32 0.40 70.40 108.8 44.80 51.20 3.66 3.67 29.40 29.70 13.56 13.56 TGI5 5.17 5.40 0.28 0.26 102.40 102.40 38.40 51.20 3.55 3.54 29.30 28.70 14.11 14.11 TGI6 5.25 5.40 0.41 0.38 70.40 70.40 38.40 25.60 3.59 3.58 27.75 29.85 12.48 12.73 200 G. Lombardi, L. Cossignani, L. Giua, M.S. Simonetti, A. Maurizi, G. Burini, R. Coli, F. Blasi and CDO6 samples. The decrease could be due to the transformation of phenol compounds into condensed forms, but also to the enzyma- tic activity from residual microorganisms of wine (MONAGAS et al., 2006). No significant differences (P>0.05) in TP content of the two wine categories were also found in 2011. Neither TP content obtained in the two years was different significantly (P>0.05). The results ob- tained in this paper for TP content were in accordance with those re- ported by other authors. For example, SATO et al. (1996) found a value of 2282.5 ppm for a Sangiovese Umbrian sample, while MILANO et al. (2009) reported a value of 2550 mg GAE/L for an Umbrian red wine (Assisi Rosso 2006, a CDO wine produced in Umbria). All wine samples tested in 2010 showed an evident antioxidant effect (Fig. 2), evaluated by ORAC assay, with values ranging from 22382 μM TE/L to 43801 μM TE/L for CDO and from 21555 μM TE/L to 37111 μM TE/L for TGI wine category. No significant differences (P>0.05) be- tween ORAC values of CDO and TGI wines were found. After one year, a significant (P<0.05) decrease of ORAC values was observed for all samples (with the exception of CDO6 sample). As regards the correlation between the TP content of red wines and their antioxidant capacity (Fig. 3), positive results have been obtained for all wines and in particular for CDO category, in both years. In literature, conflicting and confused data exist about the correlation between the TP content and antioxidant capacity in wines (ARNOUS et al., 2002; KATALINIĆ et al., 2004). Phenol profile by chromatographic analyses In order to determine the polyphenol composition of wine samples, the analysis was initially performed by HRGC-FID (Tab. 3 and 4). The most abundant compounds in both wine categories were tyrosol, gallic acid, epicatechin, and catechin. It was interesting to note that there were no significant differences (P>0.05) between the content of the single compounds in CDO and TGI wines analyzed in 2010. Af- ter one-year storage only syringic acid content changed significantly (P<0.05) between CDO and TGI wines. As regards CDO category, it was verified a significant difference (P<0.05) in the content of some phenol acids, but also of cis-resveratrol and fisetin. As regards TGI category, it was verified a significant difference (P<0.05) also Fig. 1: Total phenol (TP) content (mg GAE/L) of Umbrian red wines analyzed in 2010 and after one-year storage. Error bars represent the SD of the mean values (n = 3). Fig. 2: ORAC values (μM TE/L) of Umbrian red wines analyzed in 2010 and after one-year storage. Error bars represent the SD of the mean values (n = 3). Antioxidant properties of Italian red wines 201 Fig. 3: Correlations between TP content (mg GAE/L) and ORAC value (μM TE/L) obtained for all wines and CDO or TGI categories, in 2010 and after one- year storage. (a1), 2010 – all wines; (a2), 2011 – all wines; (b1), 2010 – CDO wines; (b2), 2011 – CDO wines; (c1), 2010 – TGI wines; (c2), 2011 – TGI wines; TP, total phenol; ORAC, oxygen radical absorbance capacity; CDO, Controlled Designation of Origin; TGI, Typical Geographic Indication.         Fig. 3   (a1) R2 = 0.84059 (b1) R2 = 0.93506 (c1) R2 = 0.54647 (a2) R2 = 0.89433 (b2) R2 = 0.89791 (c2) R2 = 0.52124 in the content of homogentisic acid. As regards the comparison with previous literature data, MILANO et al. (2009) found values much lower for ferulic acid (0.16 mg/L), gallic acid (22.71 mg/L) and trans-resveratrol (1.24 mg/L), while reported much higher concen- trations for caffeic and cinnamic acids, catechin and epicatechin. In order to quantify some flavonols (myricetin, quercetin, kaempferol, rhamnetin/isorhamnetin, rutin), both as aglycones and glycosides, an HPLC-DAD analysis was performed (Tab. 5 and 6). The most abun- dant compounds were ethyl gallate for CDO wines and quercetin-3- β-D-glucoside for TGI wines, while the most represented flavonol was quercetin, both in CDO (on average 6.78 mg/L) and TGI (on average 4.35 mg/L) wines. In all wines, the glycosidic form of quer- cetin, kaempferol and isorhamnetin had always a higher concentra- tion (P<0.05) than the respective aglycones, both in wines analyzed in 2010 and in 2011. The results obtained in this study showed a high variability in the phenol composition in wine samples, although 202 G. Lombardi, L. Cossignani, L. Giua, M.S. Simonetti, A. Maurizi, G. Burini, R. Coli, F. Blasi Tab. 3: Content§ of phenol compounds in red wines analyzed by high-resolution gas chromatography in 2010 Phenols CDO1 CDO2 CDO3 CDO4 CDO5 CDO6 TGI1 TGI2 TGI3 TGI4 TGI5 TGI6 (mg/L) Vanillin 2.4±1.8 1.6±0.5 1.6±0.1 3.3±0.1 1.8±0.2 1.8±0.4 2.4±0.3 2.0±0.5 1.4±0.0 0.9±0.1 2.2±0.7 3.5±2.0 trans-Cinnamic acid 1.5±0.1 1.3±0.0 9.6±1.4 7.8±5.2 12.4±0.2 10.0±0.7 5.7±0.4 1.5±0.0 1.0±0.1 5.6±0.5 9.7±1.4 6.5±1.0 Tyrosol 21.9±1.8 30.6±2.0 27.1±4.2 21.6±1.2 21.2±6.3 26.6±2.2 10.2±1.0 25.5±3.6 13.0±0.4 20.7±1.9 26.9±3.5 10.7±0.7 Veratric acid 1.7±0.2 1.7±0.0 2.1±0.2 2.1±0.3 1.9±0.2 2.3±0.1 1.7±0.1 1.5±0.1 1.7±0.0 1.1±0.1 2.3±0.3 1.4±0.1 Vanillic acid 2.3±0.1 3.1±0.1 4.7±0.4 4.1±0.1 5.8±0.1 4.0±1.0 3.6±0.2 2.7±0.2 4.4±0.1 3.6±0.3 3.3±0.8 5.1±0.3 Homovanillic acid 2.6±4.6 3.8±0.3 1.2±0.0 4.9±0.3 1.2±0.2 1.4±0.1 1.3±0.0 3.9±0.5 3.0±0.8 1.1±0.1 4.6±0.5 1.2±0.1 Hydroxytyrosol 3.2±0.2 3.8±0.1 3.3±0.4 4.3±0.1 3.8±0.3 4.3±0.4 2.7±0.1 3.3±0.3 2.7±0.1 1.4±0.0 4.0±0.4 3.3±0.2 3,4-DIHY acid 1.6±2.2 1.5±0.0 1.4±0.0 1.8±0.2 1.5±0.1 1.3±0.0 1.3±0.1 1.7±0.1 1.4±0.0 1.5±0.0 2.0±0.1 1.5±0.1 Homogentisic acid 1.6±2.3 1.8±0.1 1.4±0.1 1.4±0.1 2.3±0.3 1.4±0.0 1.2±0.1 1.7±0.1 1.5±0.0 2.0±0.2 1.5±0.0 1.4±0.1 Syiringic acid 3.4±0.2 4.2±0.1 4.4±0.3 4.1±0.2 4.3±0.2 5.3±0.2 3.0±0.0 4.0±0.2 3.9±0.0 3.1±0.4 4.3±0.0 3.8±0.2 p-Coumaric acid 6.5±0.4 6.6±0.1 5.5±0.6 7.7±0.4 8.9±0.6 8.0±0.4 5.6±0.2 5.4±0.8 4.9±0.3 5.8±0.5 7.8±0.4 7.4±0.5 Gallic acid 51.3±4.6 13.9±1.7 40.6±5.5 52.5±3.5 34.5±1.3 53.1±3.6 28.3±2.2 36.5±5.1 18.9±1.8 32.1±3.8 38.9±1.5 29.9±4.2 Ferulic acid 3.3±0.7 1.9±0.2 2.0±0.3 2.2±0.6 1.7±0.1 2.5±0.1 1.8±0.2 1.8±0.0 2.2±0.2 2.0±0.1 2.1±0.7 1.7±0.1 Caffeic acid 6.4±0.4 7.3±0.2 6.0±0.5 12.8±0.8 7.3±0.6 15.7±0.9 5.4±0.3 6.5±0.6 17.0±1.7 9.9±0.9 8.9±4.2 9.1±1.0 Sinapinic acid 2.8±0.0 3.6±0.7 2.2±0.0 2.4±0.2 2.3±0.1 4.4±0.2 2.2±0.1 3.6±0.7 2.5±0.0 2.2±0.1 2.4±0.2 2.6±0.3 cis-Resveratrol 1.7±0.2 2.6±0.1 2.5±0.2 1.5±0.1 3.2±0.1 1.3±0.0 1.5±0.0 2.1±0.2 2.1±0.1 1.2±0.1 2.2±0.6 2.0±0.1 trans-Resveratrol 2.1±0.5 2.9±0.3 2.9±0.3 1.8±0.0 3.7±0.2 1.2±0.0 1.8±0.1 2.1±0.2 2.2±0.1 1.2±0.0 2.0±0.2 3.0±0.3 Epicatechin 25.0±2.2 15.2±1.0 40.5±5.1 37.1±2.3 30.3±1.8 16.3±1.1 26.4±0.9 32.4±4.9 41.9±2.8 40.8±2.6 34.9±4.2 21.5±3.2 Catechin 25.7±2.3 28.7±4.3 33.1±4.9 42.3±2.6 35.5±2.0 33.4±1.0 22.4±1.0 33.7±5.1 35.1±2.4 16.9±1.2 38.5±5.8 29.7±3.2 Fisetin 5.3±2.1 2.9±0.2 6.6±6.0 2.7±0.0 2.9±0.2 3.5±0.8 4.6±1.6 3.2±0.3 3.1±0.6 2.7±0.1 3.0±0.0 3.2±0.1 Tab. 4: Content§ of phenol compounds in red wines analyzed by high-resolution gas chromatography in 2011 Phenols CDO1 CDO2 CDO3 CDO4 CDO5 CDO6 TGI1 TGI2 TGI3 TGI4 TGI5 TGI6 (mg/L) Vanillin 3.2±0.1 2.2±0.0 1.9±0.0 2.7±0.1 0.9±0.0 3.3±0.2 3.1±0.1 1.7±0.0 0.9±0.0 2.6±0.1 2.1±0.0 0.6±0.0 trans-Cinnamic acid 1.3±0.0 1.5±0.0 0.9±0.0 1.8±0.1 1.0±0.0 1.7±0.0 1.5±0.0 1.0±0.0 1.1±0.0 1.2±0.1 1.1±0.0 0.9±0.0 Tyrosol 24.9±0.2 30.8±0.9 22.8±0.3 16.4±0.4 28.0±0.3 25.8±0.9 12.5±0.4 19.2±0.0 8.3±0.0 19.5±0.3 28.0±0.2 20.7±0.3 Veratric acid 1.6±0.0 1.1±0.0 1.5±0.0 0.7±0.0 1.4±0.0 0.9±0.0 0.7±0.0 0.8±0.0 1.0±0.0 2.2±0.0 1.7±0.1 1.0±0.0 Vanillic acid 1.1±0.0 4.9±0.6 2.6±0.1 7.1±0.3 0.1±0.0 4.6±0.4 0.4±0.0 12.8±0.2 1.5±0.1 16.0±0.4 1.4±0.2 2.4±0.0 Homovanillic acid 1.7±0.0 1.9±0.0 1.8±0.0 1.9±0.0 2.1±0.0 1.8±0.0 1.6±0.0 1.9±0.0 1.5±0.0 1.6±0.0 1.9±0.0 2.0±0.0 Hydroxytyrosol 3.3±0.0 3.9±0.0 2.7±0.1 3.3±0.1 3.4±0.1 5.9±0.4 2.9±0.1 2.4±0.2 1.7±0.0 2.6±0.1 3.7±0.2 2.7±0.0 3,4-DIHY acid 1.4±0.0 1.3±0.0 1.2±0.0 1.3±0.0 1.3±0.0 1.5±0.0 1.2±0.0 1.2±0.0 1.3±0.0 1.2±0.0 1.3±0.0 1.3±0.0 Homogentisic acid 1.3±0.0 0.8±0.0 2.4±0.0 1.0±0.0 1.4±0.0 1.0±0.1 1.0±0.0 1.9±0.0 1.0±0.0 0.7±0.0 1.3±0.0 1.0±0.0 Syiringic acid 2.3±0.0 3.0±0.0 2.7±0.0 2.0±0.0 2.9±0.0 4.0±0.2 1.5±0.1 1.8±0.0 1.4±0.0 1.4±0.0 2.7±0.1 2.4±0.0 p-Coumaric acid 8.5±0.2 8.9±0.4 5.1±0.2 6.7±0.1 8.9±0.1 7.8±0.2 9.9±0.3 7.8±0.0 4.9±0.0 8.4±0.1 9.9±0.5 4.7±0.1 Gallic acid 67.2±1.0 43.0±0.4 38.1±0.0 43.7±2.0 34.3±0.6 54.8±3.0 41.4±1.0 32.2±0.6 11.4±0.0 37.8±0.4 35.3±0.0 38.0±0.3 Ferulic acid 3.6±0.1 3.7±0.0 3.1±0.1 3.4±0.0 3.1±0.0 3.6±0.0 3.6±0.0 3.5±0.0 6.6±0.0 4.3±0.1 3.5±0.0 2.8±0.0 Caffeic acid 9.3±0.1 10.0±0.3 6.4±0.2 11.2±0.2 8.8±0.1 14.4±1.0 7.2±0.1 17.5±0.0 10.5±0.1 23.3±0.4 8.5±0.1 10.5±0.1 Sinapinic acid 3.9±0.0 4.2±0.1 4.2±0.1 4.4±0.2 4.4±0.1 4.2±0.1 4.1±0.2 5.9±0.1 2.9±0.0 4.1±0.1 4.3±0.2 3.5±0.0 cis-Resveratrol 1.7±0.0 2.6±0.0 0.4±0.0 1.0±0.0 3.0±0.0 0.6±0.0 0.2±0.0 0.3±0.0 0.3±0.0 0.1±0.0 0.4±0.1 0.3±0.0 trans-Resveratrol 2.9±0.0 2.8±0.0 1.2±0.2 1.0±0.0 3.5±0.0 3.2±0.0 1.7±0.1 0.9±0.0 1.5±0.2 0.6±0.0 2.0±0.0 1.5±0.0 Epicatechin 28.7±0.3 31.0±1.3 28.5±0.0 30.3±0.2 33.3±0.4 35.0±1.1 35.0±1.0 21.2±1.0 31.9±0.5 27.3±0.7 32.8±0.1 29.9±0.1 Catechin 25.7±0.4 28.7±0.0 22.9±0.2 42.3±0.0 35.5±0.3 33.4±0.7 26.2±0.7 20.7±0.9 23.9±0.4 27.6±0.8 34.4±2.6 26.0±0.1 Fisetin 12.5±0.1 12.0±0.0 11.3±0.0 11.8±0.1 12.6±0.2 12.0±0.2 11.9±0.0 11.5±0.1 11.2±0.0 11.2±0.0 12.3±3.0 11.5±0.1 § mean values ± SD, n=3 3,4-DIHY acid, 3,4-dihydroxyphenilacetic acid Antioxidant properties of Italian red wines 203 coming from the same region. It is known that the quali- and quanti- tative phenol composition of grape and wine shows a great variability in relation to environmental, viticultural, enological or storage prac- tices (IVANOVA-PETROPULOS et al., 2012). SIMONETTI et al. (1997) analyzed ten red Italian wines and reported the myricetin content ranging from 0.6 mg/L (Squinzano) to 9.6 mg/L (Cabernet Sauvi- gnon), while in this study the value changed between 3.2 and 7.2 for CDO wines and from 2.3 to 5.1 mg/L for TGI wines, analyzed in 2010. With regard to rutin, it was detected only in some samples as also reported by SIMONETTI et al. (1997). Rhamnetin was never detected in Umbrian wine samples. After one-year storage, a slight decrease of single phenol content was observed, even if only the concentration of quercetin changed significantly (P<0.05) in DOC wine category. In conclusion, this is the first study of phenol content, qualitative composition and antioxidant capacity in wines produced in central Italy. The results confirmed the good quality of CDO and TGI Umbrian red wines, in fact there were no significant differences between wine categories, both as regards TP content, ORAC value and phenol composition. Generally, one-year storage in bottle did not affect significantly the quality of the analyzed wines, in particular for CDO category. The relevance of these results is evident when con- sidering that some Umbrian red wines, as Montefalco Sagrantino, are appreciated all around the word. A broader examination of Umbrian red wine is actually in progress. Acknowledgments This work was funded by the Fondazione Cassa di Risparmio di Peru- gia (Project 2009.020.0004- “I vini umbri: speciazione della frazione polifenolica per via HPLC-MS e/o HRGC-MS ed aspetti salutistici correlati”). References ARNOUS, A., MAKRIS, D.P., KEFALAS, P., 2002: Correlation of pigment and flavanol content wine antioxidant properties in selected aged regional wines from Greece. J. Food Comp. Anal. 15, 655-665. DOI: 10.1006/jfca.2002.1070 ARRANZ, S., CHIVA-BLANCH, G., VALDERAS-MARTÍNEZ, P., MEDINA- REMÓN, A., LAMUELA-RAVENTÓS, R.M., ESTRUCH, R., 2012: Wine, beer, Tab. 5: Content§ of phenol compounds analyzed by high-performance liquid chromatography in 2010 Phenols CDO1 CDO2 CDO3 CDO4 CDO5 CDO6 TGI1 TGI2 TGI3 TGI4 TGI5 TGI6 (mg/L) Rutin 3.1±0.4 13.3±0.6 3.0±0.1 3.5±0.3 0.1±0.0 5.7±0.3 1.8±0.2 7.0±0.2 0.1±0.0 1.1±0.0 0.1±0.0 3.8±0.1 Ethyl gallate 14.2±0.7 16.3±0.3 16.7±0.8 20.9±1.1 21.6±1.0 19.6±1.0 7.0±0.3 7.5±0.1 3.8±0.0 8.7±0.9 8.0±0.2 7.9±0.1 Quercetin-3-β-D- 12.4±0.6 13.5±0.2 10.8±0.7 16.5±0.8 15.9±0.6 14.5±0.5 16.8±0.3 12.3±0.3 6.4±0.2 20.7±1.3 19.0±1.0 15.3±0.1 glucoside Kaempferol-3-O- 0.5±0.1 0.8±0.1 2.0±0.1 0.7±0.1 1.8±0.0 0.9±0.0 0.1±0.0 0.3±0.0 0.1±0.0 0.3±0.0 1.0±0.0 0.6±0.2 glucoside Isorhamnetin-3-O- 2.1±0.4 3.1±0.1 2.6±0.1 1.1±0.0 1.3±0.0 2.5±0.1 1.4±0.1 0.1±0.0 0.2±0.0 0.1±0.0 2.4±0.1 2.0±0.3 glucoside Myricetin 3.2±0.2 7.2±0.1 5.1±0.1 6.2±0.4 4.8±0.4 3.0±0.2 3.6±0.1 3.1±0.1 3.3±0.3 2.3±0.1 5.1±0.1 2.7±0.1 Quercetin 6.0±0.7 10.9±0.1 6.8±0.1 8.5±0.6 5.7±0.6 2.3±0.1 6.2±0.1 6.5±0.0 3.5±0.2 1.8±0.1 4.4±0.0 3.8±0.2 Kaempferol 0.2±0.1 0.3±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 Isorhamnetin 0.3±0.1 2.3±0.0 1.1±0.0 0.7±0.1 1.4±0.3 0.1±0.0 0.2±0.0 1.1±0.0 0.6±0.0 0.1±0.0 0.8±0.0 0.6±0.1 § mean values ± SD, n=3 Tab. 6: Content§ of phenol compounds analyzed by high-performance liquid chromatography in 2011 Phenols CDO1 CDO2 CDO3 CDO4 CDO5 CDO6 TGI1 TGI2 TGI3 TGI4 TGI5 TGI6 (mg/L) Rutin 2.0±0.0 6.8±0.4 2.2±0.1 3.4±0.3 0.1±0.0 4.8±0.1 0.1±0.0 5.3±0.0 0.1±0.0 1.1±0.0 0.1±0.0 3.8±0.1 Ethyl gallate 11.2±0.1 14.1±0.1 15.6±0.7 19.1±0.8 6.4±0.2 18.0±0.3 8.7±0.1 2.7±0.1 3.1±0.0 7.8±0.1 5.8±0.2 7.8±0.2 Quercetin-3-β-D- 11.9±0.2 14.8±0.2 9.0±1.2 15.5±0.6 5.6±0.2 15.5±0.5 15.5±0.2 11.5±0.1 6.3±0.0 14.0±0.6 10.4±0.5 13.9±0.6 glucoside Kaempferol-3-O- 0.4±0.0 0.7±0.0 1.4±0.1 0.6±0.0 1.2±0.2 0.8±0.0 0.1±0.0 0.2±0.0 0.5±0.0 0.3±0.0 1.0±0.0 0.2±0.0 glucoside Isorhamnetin-3-O- 1.5±0.1 3.0±0.1 2.5±0.0 0.7±0.0 1.2±0.0 2.0±0.0 1.3±0.0 0.1±0.0 0.1±0.0 0.1±0.0 2.4±0.1 2.4±0.3 glucoside Myricetin 2.1±0.3 6.5±0.6 0.5±0.0 2.2±0.2 3.9±0.0 1.4±0.1 2.4±0.0 2.9±0.1 2.8±0.0 0.7±0.0 5.1±0.1 1.4±0.2 Quercetin 2.6±0.0 7.2±0.3 0.4±0.0 1.7±0.3 4.1±0.0 0.7±0.0 4.3±0.1 3.6±0.1 2.6±0.0 0.3±0.0 4.4±0.0 0.5±0.0 Kaempferol 0.1±0.0 0.1 ±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 0.1±0.0 Isorhamnetin 0.1±0.0 1.7±0.2 1.4±0.1 0.1±0.0 1.1±0.0 0.1±0.0 0.1±0.0 0.6±0.0 1.3±0.5 0.1±0.0 0.8±0.0 0.1±0.0 § mean values ± SD, n=3 204 G. 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