Comparative Study of Aromatic Compounds in Young Red Wines from Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet Varieties in China

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1 JFS C: Food Chemistry and Toxicology Comparative Study of Aromatic Compounds in Young Red Wines from Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet Varieties in China M. ZHANG,Q.XU,C.DUAN, W.QU, AND Y. WU ABSTRACT: The aromatic composition and key odorants of young red wines produced from Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet wines were compared and the reasons for the difference in their aromatic compounds were discussed. Forty-three odorants were detected in Cabernet Sauvignon and Cabernet Franc wines compared to 50 in Cabernet Gernischet wine. Quantitatively, acids formed the most abundant group in the aromatic components of the 3 wines, followed by alcohols and esters. Compared to Cabernet Sauvignon and Cabernet Franc wines, the profiles of alcohols and esters for Cabernet Gernischet wine were more diverse. Monoterpenes, namely, 4- terpinenol, citronellol, and nerol, were found solely in Cabernet Gernischet wine. Only 10 compounds, namely, ethyl octanoate, ethyl hexanoate, isoamyl acetate, ethyl butyrate, β-damascenone, ethyl decanoate, isoamyl alcohol, acetic acid, octanoic acid, and phenylethyl acetate, were always present in the 3 wines at concentrations higher than their threshold values. However, ethyl octanoate, ethyl hexanoate, and isoamyl acetate were found to jointly contribute to 97%, 98.9%, and 99% of the global aroma of Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet wines, respectively. This result showed that the aroma indistinguishableness of the 3 wines was mainly due to the dominance of the fruity notes exerted by the ethyl esters and, to a lesser extent, to the contribution of varietal aromatic compounds to the global aroma of the wines. Keywords: aromatic components, Cabernet Franc, Cabernet Gernischet, Cabernet Sauvignon, headspace/solidphase microextraction Introduction Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet (also called as Shelongzhu) are known as The Three Pearls of wine grapes in China; these grape varieties are used by the Chinese wineries to prepared premium quality wines. Compared to Cabernet Sauvignon and Cabernet Franc, Cabernet Gernischet is special in the sense that this grape variety can only be found in China. Different conclusions have been drawn on the origin and authenticity of Cabernet Gernischet. Based on RAPD (Random Amplification Polymorphism DNA) analysis, Cabernet Sauvignon, Cabernet Gernischet, and Cabernet Franc were found to belong to the same group (Yao and others 2005). The SSR (Simple Sequence Repeat) analysis, however, indicated that they were 3 independent cultivars (Yao and others 2005). Headspace/solid-phase microextraction (HS-SPME) is a suitable technique to obtain representative extracts of wine aromatic compounds by partitioning them from a liquid or gaseous sample into an immobilized poly-coated fiber (Wang and others 2004). This technique provides information about the composition of volatile fraction that would be perceived by the consumer MS Submitted 12/12/2006, Accepted 3/19/2007. Authors Zhang, Xu, Duan, Qu, and Wu are with Center for Viticulture and Enology, College of Food Science & Nutritional Engineering, China Agriculture Univ., P.O. Box 301, Qinghua Donglu 17, Beijing , China. Author Zhang is with Henan Inst. of Science and Technology, Xinxiang , Henan Province, China. Direct inquiries to author Duan ( chqduan@yahoo.com.cn). when smelling wine (Martí and others 2003). It is the most sensitive and powerful method for the identification and determination of trace organic constituents in complex matrices (Francioli and others 1999). Simplicity, speediness, solvent-free extraction, and a little sample manipulation are among the advantages offered by this extraction technique (Whiton and Zoecklein 2000; Begala and others 2002; Martí and others 2003; Howard and others 2005). It is possible to assess and differentiate wine quality according to the aromatic composition as the volatile components of wine represent a group of compounds with high discriminating power that can be determined by objective methods (Begala and others 2002). Grape variety is known to exert a significant influence on the aromatic composition of wines as 3 important families of aromatic compounds derived from yeast amino acid metabolism (isoacids and fusel alcohols, ethyl esters of isoacids, fusel alcohol acetates) are strongly linked to the variety of grape (Ferreira and others 2000). In China, the aromatic characteristics of monovarietal wine from Cabernet Gernischet are found to closely resemble those of the monovarietal wines from Cabernet Sauvignon and Cabernet Franc. These grape varieties, particularly Cabernet Gernischet and Cabernet Franc, when cultivated in the same region, are perceived to produce red wines with similar color and flavor. However, there are no systematic studies to confirm this observation. Hence, in this article, we compare the aromatic compounds of 3 monovarietal wines from Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet and we also attempt to elucidate the reasons behind the aroma indistinguishableness between the wines. C248 JOURNAL OF FOOD SCIENCE Vol. 72, Nr. 5, 2007 C 2007 Institute of Food Technologists doi: /j x Further reproduction without permission is prohibited 转载

2 Materials and Methods Winemaking Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet grapes were harvested from Huailai, Hebei Province, China and vinificated in the same winery. All 3 wines were fermented in 20 m 3 stainless steel tanks. Each monovarietal wine was vinificated in duplicate using the traditional process and the same vinification process was performed on the 3 grapes. Briefly, the grapes were crushed, destemmed, and placed separately in stainless steel tanks. Sulfur dioxide (50 mg/l) was added to the musts and the contents were mixed with pumping. After maceration of the musts at 20 C for 24 h, 200 mg/l activated dry yeast BM45 (Lallemend Company, Toulouse, France) was added to the musts. Alcoholic fermentation was carried out at 22 to 25 Ctodryness (reducing sugar < 4 g/l) followed by malolactic fermentation by MBR B1 (Lallemend Company, France) at 15 to 18 C for 1 mo. After the wines were stored in stainless steel tanks at 15 C for 3 mo, three 50-mL wine samples were collected from each tank and analyzed for aromatic compounds. Solid-phase microextraction Aromatic compounds of the wine samples were extracted by HS- SPME and analyzed using gas chromatography/mass spectrometry as described by Begala and others (2002) with minor modifications. Five milliliters of wine sample and 1 g NaCl were placed in a 15-mL sample vial. The vial was tightly capped with a PTFE-silicon septum and heated at 40 C for 30 min on a heating platform agitation at 400 rpm. The SPME (50/30-µm DVB/Carboxen/PDMS, Supelco, Bellefonte, Pa., U.S.A.), preconditioned according to manufacturer s instruction, was then inserted into the headspace, where extraction was allowed to occur for 30 min with continued heating and agitation by a magnetic stirrer. The fiber was subsequently desorbed in the GC injector for 25 min. Gas chromatography-mass spectrometry (GC-MS) analysis The GC-MS system used was an Agilent 6890 GC equipped with an Agilent 5975 mass spectrometry. The column used was a 60 m 0.25 mm HP-INNOWAX capillary with 0.25-µm film thickness (J & W Scientific, Folsom, Calif., U.S.A.). The carrier gas was helium at a flow rate of 1 ml/min. Samples were injected by placing the SPME fiber at the GC inlet for 25 min with the splitless mode. The oven s starting temperature was 50 C, which was held for 1 min, then raised to 220 Catarate of 3 C/min and held at 220 C for 5 min. The mass spectrometry in the electron impact mode (MS/EI) at 70 ev was recorded in the range m/z 20 to 450 U. The mass spectrophotometer was operated in the selective ion mode under autotune conditions and the area of each peak was determined by ChemStation software (Agilent Technologies). Analyses were carried out in triplicate. All standards were purchased from Aldrich (Milwaukee, Wis., U.S.A.) and Fluka (Buchs, Switzerland). Purity of all standards was above 99%. Model solutions were prepared using the methods reported by Howard and others (2005). 4-Methyl-2-pentanol was used as the internal standard. For quantification, 5-point calibration curves for each compound were prepared using the method described by Ferreira and others (2000), which was also used as a reference to determine the concentration range of standard solutions. The regression coefficients of calibration curves were above 96%. The standard deviation for the SPME method was below 10%. Titratable acidity (expressed as tartaric acid), volatile acidity (expressed as acetic acid), ethanol, free and total SO 2, total sugar, and reducing sugar of the 3 musts and wines were determined by the methods described by Ough and Amerine (1988). ph was measured with a Beckman model 250-pH meter (Beckman Coulter Inc., Fullerton, Calif., U.S.A.). All determinations were performed in triplicate. Odor activity values (OAVs) were used to estimate the sensory contribution of the aromatic compounds to the overall flavor of wine (Guth 1997). The OAVs of aromatic compounds in the 3 wines were calculated by using corresponding odor threshold values reported previously (Guth 1997; Tominaga and others 1998; Ferrier and others 2000; Peinado and others 2004). OAVs were calculated by dividing the mean concentration of an aromatic compound by its odor threshold value. Statistical analysis One-way ANOVA was used to evaluate differences in the aromatic composition among the 3 wines studied. Significant difference was calculated at 0.05 level. SPSS version 11.5 Statistical Package for Windows was used for all statistical analysis. Results and Discussion Physicochemical characteristics of musts and wines Table 1 shows some of the physicochemical characteristics of musts and wines from Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet. Results showed that the sugar, titratable acidity, and ph of musts were quite similar in all the 3 varieties. Furthermore, after being stored in stainless steel tanks at 15 C for 3 mo, young red wines made from the 3 grapes did not show marked differences in titratable acidity, volatile acidity, ph, total and free SO 2, reducing sugar, or ethanol concentration. Composition of aroma Three typical total ion chromatograms were generated for Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet wines using HS-SPME coupled with GC-MS. The key aromatic compounds of the 3 wines were identified and grouped into alcohols, esters, acids, aldehydes, and ketones (Table 2). Table General composition of 3 musts and wines Cabernet Sauvignon Cabernet Franc Cabernet Gernischet Must Wine Must Wine Must Wine Total sugar (g/l) Total acidity (g/l) PH Total SO 2 (mg/l) Free SO 2 (mg/l) Volatile acidity (g/l) Reducing sugar (g/l) Ethanol (%, v/v) The data were mean values of triplicate samples (maximum SD: ± 10%). Vol. 72, Nr. 5, 2007 JOURNAL OF FOOD SCIENCE C249

3 Table The threshold values and concentration of volatile components found in the Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet red wines Concentration (µg/l) Compounds Threshold (µg/l) Cabernet Sauvignon Cabernet Franc Cabernet Gernischet Significance Alcohols 1-Propanol d Isobutyl alcohol a 30857a 22465c 27823b 1-Butanol c 819b 927b 1262a Isoamyl alcohol a a b c 1-Pentanol 16b 12b 25a 3-Buten-1-ol, 3-methyl Pentanol, 4-methyl- 34a 27b 18c 1-Hexanol 8000 a 987a 734b 1075a 3-Hexen-1-ol, (E)- 24a 16b 18b 3-Hexen-1-ol, (Z)- 400 a 23a 17a 19a ns 2-Hexen-1-ol, (Z) Heptanol 35a 44a 33a ns 2-Ethyl-1-hexanol 14 (S)-3-Ethyl-4-methylpentanol - 40b 56b 188a 2-Nonanol ,3-Butanediol, [R-(R,R )]- 52a 64a 26b 1-Octanol 15b 24b 166a 1-Decanol 400 a 4a 5a 5a ns 1-Dodeanol 3a 3a 3a ns 3-(Methylthio)-1-propanol 500 a 7 5 Benzyl alcohol 294b 238b 497a Phenylethanol a 24155a 16514b 5276c 3-Ethoxy-1-propanol 12 Subtotal (µg/l) a b c Subtotal (%) Esters Ethyl acetate 7500 a 35781b 68347a 42386b Ethyl butyrate 20 a 4992 Isoamyl acetate 30 a 18327b 11163c 57271a Ethyl hexanoate 5 a 15141a 12610a 14805a ns Ethyl lactate c 24407a 19847b 25661a Hexyl acetate 670 d Methyl octanoate 65b 75b 153a Ethyl octanoate 2 a 14330b 15358b 26290a Isoamyl hexanoate 97a 73b 96a Ethyl 2,4-hexadienoate 140 Isoamyl lactate Metyl decanoate 65 Ethyl decanoate 200 a 3327c 5429b 10025a Ethyl benzoate 575 a 168 Diethyl succinate c 18688c 24645b 31125a Methyl benzoate, 2-hydroxy Phenylethyl acetate 250 a 301b 243b 746a Ethyl laurate 312c 621b 891a Ethyl myristate 113 Subtotal (µg/l) b b a Subtotal (%) Acids Acetic acid a a a a ns Isobutyric acid a Hexanoic acid 3000 a 1143a 858b 1325a Octanoic acid 500 a 775b 759b 1968a n Decanoic acid a 6b 7b 17a Subtotal (µg/l) b a c Subtotal (%) Aldehydes and ketones Nonanal 15 Decanal 13b 19a 23a Benzaldehyde 2000 d 3528 β-damascenone 0.05 a 3b 4b 6a Acetoin a 272c 474b 630a Subtotal (µg/l) 288c 512b 4187a Subtotal (%) <0.1 < Terpenyl compounds α-terpinene Terpinenol 250 b 27 Citronellol 100 a 5 Nerol 2 Continued C250 JOURNAL OF FOOD SCIENCE Vol. 72, Nr. 5, 2007

4 Table Continued Concentration (µg/l) Compounds Threshold (µg/l) Cabernet Sauvignon Cabernet Franc Cabernet Gernischet Significance Subtotal (µg/l) 21b 23b 34a Subtotal (%) <0.1 <0.1 <0.1 Others Furfural a 78b 120a 23c Butyrolactone d 1033 Methoxy-phenyl-oxime 591a 452a 515a ns Subtotal (µg/l) 1702a 572b 538b Subtotal (%) 0.1 <0.1 <0.1 Total b a c ns = no significance; = indicates significance (P < 0.05); = indicates significance (P < 0.01). a Guth (1997). The matrix was a 10% water/ethanol solution. b Ferrier and others (2000). The matrix was a 11% water/ethanol solution containing 7 g/l glycerol and 5 g/l tartartic acid, with the ph adjusted to 3.4 with 1 M NaOH. c Tominaga and others (1998). Thresholds were calculated in a 12% water/ethanol solution. d Peinado and others (2004). Thresholds were determined in 10% ethanol solution adjusted to ph 3.5 with tartaric acid. e The data were mean values of triplicate samples (maximum SD: ± 10%). The different letter following values represents significant difference at P < 0.05 in the same compound. Quantitatively, acids formed the most abundant group in the aromatic components of the 3 wines, followed by alcohols and esters. The production of fatty acids has been reported to be dependent on the composition of the must and fermentation conditions (Schreirer 1979). Acetic acid was the major fatty acid found, constituting 99.6% to 99.8% of the total fatty acid content of the wines. Acetic acid is produced during alcoholic and malolactic fermentation. At low levels this compound lifts wine flavors; however, at high levels, it is detrimental to the taste of wine by leaving the wine tasting sour and thin (Joyeux and others 1984). Except isobutyric acid, which was absent in Cabernet Gernischet wine, hexanoic acid, octanoic acid, and decanoic acid were found in all the 3 wines. These C6 to C10 fatty acids at concentrations of 4 to 10 mg/l impart mild and pleasant aroma to wine; however, at levels beyond 20 mg/l, their impact on wine becomes negative (Shinohara 1985). The C6 to C10 fatty acids might not have a significant impact on the aroma of the 3 wines examined in the current study since their levels were all far below 4 mg/l. Alcohols, with 23 compounds identified, represented the largest group in terms of the numbers of aromatic compounds identified. Alcohols are formed from the degradation of amino acid, carbohydrates, and lipids (Antonelli and others 1999). The composition of alcohols differed both qualitatively and quantitatively among the 3 wines. Isoamyl alcohol was the most abundant alcohol accounting for > 80% of the total higher alcohols in all the 3 wines studied, and it was significantly higher in the Cabernet Sauvignon wine (P < 0.05). Compared to Cabernet Sauvignon and Cabernet Franc wines, the alcohol profile of Cabernet Gernischet wine was more diverse, containing 21 types of alcohols compared to only 17 and 19 in Cabernet Sauvignon and Cabernet Franc, respectively. 3-Methyl-3-buten- 1-ol, 2-ethyl-1-hexanol, 2-nonanol, 4-terpinenol, and 3-ethoxy-1- propanol were absent in the wine made from Cabernet Sauvignon and Cabernet Franc. Other alcohols that were missing in Cabernet Sauvignon wine were 1-propanol and (Z)-2-hexen-1-ol. On the other hand, (Z)-2-hexen-1-ol and 3-(methylthio)-1-propanol, which were found to be present in very small amounts in Cabernet Franc wine, were absent in Cabernet Gernischet wine. There were also significant differences in the type and amount of esters present in the 3 wines. In general, the number and quantity of esters in Cabernet Gernischet wine (15.7%) were higher than those of Cabernet Sauvignon (9.5%) and Cabernet Franc (10.5%). Although their amount varied between the 3 wines, ethyl acetate, isoamyl acetate, ethyl lactate, ethyl octanoate, and diethyl succinate were the major esters found in the aromatic components of the 3 wines. Hexyl acetate, ethyl-2,4-hexadienoate, methyl decanoate, and methyl myristate were esters found only in Cabernet Gernischet wine, while ethyl butyrate was unique for Cabernet Sauvignon wine. Ethyl esters of the fatty acid are formed from ethanolysis of acyl-coa during fatty acid synthesis or degradation (Lee and others 2004); these compounds appear mainly during the phase of alcoholic fermentation (Gil and others 2006). On the other hand, the formation of acetate esters is the result of the reaction between acetyl- CoA and alcohols (Lee and others 2004). Ethyl lactate, a product of malolactic fermentation during wine vinification (Gil and others 2006), was lower in Cabernet Franc wine than in Cabernet Sauvignon and Cabernet Gernischet wines (P < 0.05). Monoterpenes, an important component of varietal aroma, are not affected by yeast metabolism during fermentation (Rapp 1988); they are hence a good indicator for the variety and quality of grape (Begala and others 2002). In the present study, the monoterpenes, namely, 4-terpinenol citronellol and nerol, were found solely in Cabernet Gernischet wine. On the other hand, α- terpinene could only be detected in Cabernet Sauvinon and Cabernet Franc wines. Hence, these terpenyl compounds could serve as potential indicators to distinguish wine derived from Cabernet Gernischet from those from Cabernet Sauvignon and Cabernet Franc. The composition of aldehydes and ketones varied greatly between the 3 wines. Decanal, β-damascenone, and acetoin were found in all the 3 wines. On the other hand, nonanal and benzaldehyde existed only in the aromatic components of Cabernet Franc and Cabernet Gernischet wines, respectively. Other compounds isolated from the 3 wines included furfural, butyrolactone, and methoxy-phenyl-oxime. Odor activity values (OAVs) The flavor of wine depends on many varietal and fermentative compounds, which are present in highly diverse amounts, and constitute mainly of alcohols, esters, terpenes, sulfur compounds, acids, and lactones (Francioli and others 1999). Table 3 shows the OAVs for compounds that exceeded their thresholds in the 3 wines. Only 10 compounds, namely, ethyl octanoate, ethyl hexanoate, isoamyl acetate, ethyl butyrate, β-damascenone, ethyl decanoate, isoamyl alcohol, acetic acid, octanoic acid, and phenylethyl acetate, were always present in the 3 wines at concentrations higher than their threshold values. The OAVs for isoamyl acetate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, and β-damascenone in Cabernet Gernishcet wine were at least several-fold higher than those of Cabernet Sauvignon and Cabernet Franc. The computation of relative odor contribution (ROC) proposed by Ohloff (1994) is a useful index for determining the important Vol. 72, Nr. 5, 2007 JOURNAL OF FOOD SCIENCE C251

5 Table Odor activity values (OAVs) and relative odor contribution a (ROC) for the aroma compounds in Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet young red wines Compounds Odor descriptor b Cabernet Sauvignon Cabernet Franc Cabernet Gernischet Ethyl octanoate Fruity, fresh b 7170b (64.3%) 7680b (71.8%) 13100a (72.2%) Ethyl hexanoate Fruity c 3030a (27.2) 2520a (23.6%) 2961a (16.3%) Isoamyl acetate Banana c 611b (5.5%) 372c (3.5%) 1910a (10.5) Ethyl butyrate Fruity c 250 (2.2%) β-damascenone Sweet, apple c 54.9b (0.5%) 78.2b (0.7%) 109a (0.6%) Ethyl decanoate Brandy, fruity, grape d 17.4b (0.2%) 27.1b (0.3%) 50.1a (0.3%) Isoamyl alcohol Cheese c 10.2a (0.1%) 8.4b (0.1%) 7.1b (0.35%) Ethyl acetate Pineapple, fruity, solvent, balsamic d Acetic acid Acid, fatty c 4.7a 5.3a 4.3a Octanoic acid Fatty, unpleasant b 1.6b 1.5b 3.9a Phenylethyl acetate Flowery c 1.2b 1.0b 3.0a Benzaldehyde Almond d 1.8 Phentylethanol Roses c 1.7a 1.2b 0.4b a Relative odor contribution (ROC) of each aroma compound is shown in parentheses and was calculated as the ratio of the OAV of the respective compound to the total OAV of each wine. b Odor descriptor as reported by Escudero and others (2004). c Odor descriptor as reported by Ferreira and others (2002). d Odor descriptor as reported by Peinado and others (2004). The different letter following values represents significant difference at P < 0.05 in the same compound. aromatic components in a complex system. Based on the ROC values of individual compounds, we found that the global aroma of all the wines was dominated by fermentative aromas, namely, the ethyl esters of fatty acids (ethyl octanoate and ethyl hexanoate) that conferred fruity notes to all the wines. Specifically, ethyl octanoate, ethyl hexanoate, and isoamyl acetated jointly accounted for 97%, 98.9%, and 99%, of the global aroma of Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet wines, respectively (Table 3). In contrast, the contribution of the varietal aromatic compounds derived from grapes, namely, β-damascenone, phenylethyl acetate, and phenylethanol, was minimal. Based on their ROC, β- damascenone, phenylethyl acetate, and phenylethanol each accounted for less than 1% of the global aroma perceptions of all the 3 wines. According to Escudero and others (2004), even if they were present at a concentration higher than their threshold values, compounds such as fusel alcohols, acids, esters, β-damascenone, and volatile phenols were not able to affect individually the flavor of the wines. This is because these compounds (except β-damascenone) are common in any kind of fermented alcoholic beverages that share similar aromatic properties. Moreover, the aromatic buffer in wine as a result of the presence of a large amount of ethanol, ethyl esters, fusel alcohols, fatty acids, and β-damascenone could only be broken down by the presence of compounds with distinctive aromatic properties, such as 4-methyl-4-mercapto-pentan-2-one (Escudero and others 2004). Hence, based on the result of ROC, the aroma indistinguishableness of the 3 wines was mainly due to the dominance of the fruity notes exerted by the ethyl esters and, to a lesser extent, to the contribution of varietal aromatic compounds to the global aroma of the wines. Conclusion The present study demonstrated that there were significant differences in the volatile components of young red wines made from Cabernet Sauvignon, Cabernet Franc, and Cabernet Gernischet (P < 0.05). The aroma indistinguishableness of the 3 wines was probably due to the dominance of ethyl esters of fatty acids and their contributions to the global aroma of the 3 wines. Acknowledgments This article was edited by Dr. Tan Sze Sze and Dr. Jian Zhao. This research was supported by 948 Research Programs of China Ministry of Agriculture (grant nr 2006-G26 to C.-Q.D). References Antonelli A, Castellari L, Zambonelli C, Carnacini A Yeast influence on volatile composition of wines. J Agric Food Chem 47: Begala M, Corda L, Podda G, Fedrigo MA, Traldi P Headspace solid-phase microextraction gas chromatography/mass spectrometry in the analysis of the aroma constituents of Cannonau of Jerzu wine. Rapid Comm Mass Spectrom 16: Escudero A, Gogorza B, Melús MA, Ortín N, Cacho J, Ferreira V Characterization of the aroma of a wine from Maccabeo. Key role played by compounds with low odor activity values. J Agric Food Chem 52: Ferreira V, Lápez R, Cacho JF Quantitative determination of the odorants of young red wines from different grape varieties. J Sci Food Agric 80: Ferreira V, Ortín N, Escudero A, López R, Cacho J Chemical characterization of the aroma of Grenache rose wines: aroma extract dilution analysis, quantitative determination, and sensory reconstitution studies. J Agric Food Chem 50: Francioli S, Guerra M, López-Tamanes E, Guadayoi JM, Caixach J Aroma of sparkling wines by headspace/solid phase microextraction and gas chromatography/mass spectrometry. Am J Enol Vitic 50: GilM,Cabellos JM, Arroyo T, Prodanov M Characterization of the volatile fraction of young wines from the denomination of origin Vinos de Madrid (Spain). Anal Chim Acta 563: Guth H Quantitation and sensory studies of character impact odorants of different white varieties. J Agric Food Chem 45: Howard KL, Mike JH, Riesen R Validation of a solid-phase microextraction method for headspace analysis of wine aroma components. Am J Enol Vitic 56: Joyeux A, Lafon-Lafourcade S, Ribéreau-Gayon P Evolution of acetic acid bacteria during fermentation and storage of wine. Appl Environ Microbiol 48: Lee SJ, Rathbone D, Asimont S, Adden R, Ebeler SE Dynamic changes in ester formation during chardonnay juice fermentations with different yeast inoculation and initial Brix conditions. Am J Enol Vitic 55: Martí MP, Mestres M, Sala C, Busto O, Guasch J Solid-phase microextraction and gas chromatography olfactometry analysis of successively diluted samples. A new approach of the aroma extract dilution analysis applied to the characterization of wine aroma. J Agri Food Chem 51: Ohloff G Scent and fragrances: the fashion of odors and their chemical perspectives. Berlin: Springer-Verlag, 57 p. Ough CS, Amerine MA Methods for musts and wines. 2nd ed. New York: Wiley- Interscience. 377 p. Peinado RA, Moreno J, Bueno JE, Moreno JA, Mauricio JC Comparative study of aromatic compounds in two young white wines subjected to pre-fermentative cryomaceration. Food Chem 84: Rapp A Micro-element analysis in wine and grapes. In: Linskens HF, Jackson HF, Conte LS, editors. Wine analysis, Vol. 6. Berlin: Springer-Verlag. p Schreirer P Flavor composition of wines a review. Rev Food Sci Nutri 12: Shinohara T Gas chromatographic analysis of volatile fatty acids in wines. Agric Biol Chem 49: Tominaga T, Murat ML, Dubourdieu D Development of a method for analyzing the volatile thiols involved in the characteristic aroma of wines made from Vitis vinifera L. cv Sauvignon Blanc. J Agric Food Chem 46: Yao Y,Zuo F, Zhai H, Liu L The RAPD and SSR analysis of Cabernet Shelongzhu and other wine grape cultivars. Acta Hortic Sinica 32: Wang L, Xu Y, Zhao G, Li J Rapid analysis of flavor volatiles in apple wine using headspace solid-phase microextraction. J Inst Brew 110: Whiton RS, Zoecklein BW Optimization of headspace solid-phase microextraction for analysis of wine aroma compounds. Am J Enol Vitic 51: C252 JOURNAL OF FOOD SCIENCE Vol. 72, Nr. 5, 2007