International Journal of Food Science and Technology 2014, 49, 787–796 787

Original article
Headspace solid-phase micro extraction coupled to
comprehensive two-dimensional with time-of-flight mass
spectrometry applied to the evaluation of Nebbiolo-based wine
volatile aroma during ageing

Matteo Bordiga,* Gianluca Piana, Jean Daniel Co€ısson, Fabiano Travaglia & Marco Arlorio
Universita’ degli Studi del Piemonte Orientale ‘A. Avogadro’, Dipartimento di Scienze del Farmaco, Largo Donegani 2, 28100 Novara, Italy
(Received 26 March 2013; Accepted in revised form 28 August 2013)

Summary This study presents the application of a headspace solid-phase microextraction (HS-SPME) method on
the analysis of Nebbiolo-based wine volatiles by comprehensive two-dimensional gas chromatography
(GC9GC) and time-of-flight mass spectrometry (TOF-MS). The aroma patterns were established for dif-
ferent samples of Nebbiolo-based wines, aged in oak wood barrels for 18 months at constant temperature.
The methodology allowed for the simultaneous analysis of over 130 different volatile compounds detected
in the headspace. The odour activity values (OAVs) were assessed to identify potentially important odor-
ants of Nebbiolo-based wine during ageing. The highest OAVs were obtained for several compounds such
as ethyl octanoate, ethyl hexanoate, isoamyl acetate, b-damascenone, ethyl cinnamate, ethyl 3-methylbu-
tanate and acetaldehyde. Ethyl butanoate, octanoic acid, ethyl dihydrocinnamate and c-nonalactone were
also at concentrations higher than their corresponding threshold. Also vanillin, (E)-whisky lactone,
(Z)-whisky lactone, guaiacol and 4-ethyl guaiacol seem to be important odorants after oak wood ageing
period.
Keywords Comprehensive two-dimensional gas chromatography, headspace solid-phase micro extraction, Nebbiolo, oak wood, wine
aroma.

Extraction of volatile compounds from oak barrels

Introduction
The flavour of wine results from a series of different
biochemical and technological processes. Aroma for-
mation is influenced by several factors: environment,
grape variety, ripeness, fermentation conditions and
biological factors (Antonelli et al., 1999; Estevez et al.,
2004). The flavour of wine is due to the presence of an
extremely complex pattern, in both quantitative and
qualitative terms. The chemical composition of wine
aroma continues to change during ageing especially if
the technological procedure is performed in wood bar-
rel (primarily oak wood) (Bloem et al., 2008). In wine-
making, this common practice helps refine the
stability, spontaneous clarification and the formation
of a complex aroma, and thus improving the quality
of the wine (Puech et al., 1999; Tominaga et al., 2000).
Contact with the barrel modifies the wine composi-
tion due to compounds extracted from the wood (e.g.
tannins, phenolic acids and volatile compounds).
*Correspondent: Fax: +39 0321 375751;
e-mail: matteo.bordiga@pharm.unipmn.it
depends on several factors, including the botanical and
geographical origin of the wood (Garde-Cerdan et al.,
2002) as well as the degree of oak toasting (Chatonnet
et al., 1999). The thermal treatment causes thermal deg-
radation of some components of oak wood, which pro-
duces several volatile compounds. Furanic compounds
are formed through thermal degradation of carbohy-
drates, volatile phenols from the thermal degradation
of lignin, and oak lactones are products of the dehydra-
tion of the acids present in wood (Sanz et al., 2011).
Oak wood contains a high level of volatile com-
pounds that have a great impact on wood-matured wine
aroma. The main volatile compounds susceptible to
migration from oak wood to wine are the two whisky
lactone isomers [(Z)-Whisky lactone and (E)-Whisky
lactone], furfural and its derivatives, phenolic aldehyde
such as vanillin, and volatile phenols such as guaiacol
and its derivatives (Ribereau-Gayon et al., 2006).
The sample preparation is a fundamental step for a
representative and significant analysis of complex
matrices such as wine, and commonly the applied
methods are time-consuming, expensive and often offer

doi:10.1111/ijfs.12366
© 2013 Institute of Food Science and Technology
788 2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al.
specific advantages or disadvantages for certain spe-
cific chemical class extraction.
Headspace solid-phase micro extraction (HS-SPME)
is simple, fast, inexpensive, sensitive, easy-to-automate,
solvent-free and can be considered an excellent alterna-
tive to other methods (Zalacain et al., 2004). An
important analytical problem is related to the presence
of several different chemical classes in wine volatile
aroma. Then, a specific process of extraction must be
considered, and SPME may be regarded as a good
device for this specific target (Bonino et al., 2003).
Comprehensive multidimensional gas chromatogra-
phy (GC9GC) coupled with time-of-flight (TOF) mass
detection combined with HS-SPME represents a rela-
tively new technique for the characterisation of vola-
tiles complex profiles. This method has provided a
major advantage in analysing complex samples where
the number of analytes may be both large and/or pres-
ent at trace levels as is the case with wine (Ryan et al.,
2004; Ryona et al., 2010).
The principal aim of this work was the evaluation
of HS-SPME techniques coupled with GC9GC/TOF-
MS for the headspace analyses of different Nebbiolo-
based wines from Piedmont, Italy. With this cultivar,
vinified as a single variety, high quality and interna-
tionally renowned wines are produced, namely Barolo
D.O.C.G. (Controlled and Guaranteed Designation of
Origin) characterised by long oak wood ageing period.
Oak barrels have long been used in fine winemaking,
especially for wood-positive effects, namely the ceding
of pleasant aromas and the regulation of red wine col-
our. This study proposes to assess the changes in the
volatile profile of Nebbiolo-based wine aged in oak
wood over 18 months.
Materials and methods
Samples
In this work, the influence of oak wood storage/ageing
on the aromatic profile of Nebbiolo-based wines was
investigated. For this purpose, typical wine samples
(100% Nebbiolo cultivar) were used. Wine samples
were checked at four time points over 18 months. The
first point, designated ‘starting wine’, was analysed
immediately after the wines were considered ready to
undergo barrel ageing period. Then, the other time
points were defined at ‘6’, ‘12’ and ‘18 months’, respec-
tively, at constant storage temperatures in oak wood
barrels. Each time point was the result of three repli-
cates of four different barrels (n = 12). The model wines
were produced following the standard protocols used
for the ‘Barolo D.O.C.G.’ wine appellation that requires
a period of refining in wood for at least 18 months;
otherwise, the wine cannot be denominated ‘Barolo’ but
only ‘Nebbiolo’. This is the reason why this study has
International Journal of Food Science and Technology 2014
focused on this period of time. The barrels used in this
study were new and constituted from French Quercus
robur oak species, medium type of toasting.
All standard chemicals (Table S1) used were pur-
chased from Sigma–Aldrich (Milan, Italy). Water was
obtained from Milli-Q instrument (Millipore Corp.,
Bedford, MA, USA).
Headspace SPME extraction
A Supelco fibre holder (Bellefonte, PA, USA) and a
50/30-lm divinylbenzene-carboxenpoly(dimethylsilox-
ane) (DVB-CAR-PDMS) fibre were used. This triphasic
fibre has been previously described as the most suitable
for adsorbing a large range of volatile compounds from
wines and other alcoholic beverages (Rodrigues et al.,
2008; Bordiga et al., 2013). 5 mL of each wine was
added with 3 g of NaCl (salt saturation of the sample
was reached to obtain maximum extraction perfor-
mance) and maintained for 10 min at 35 °C under
magnetic stirring. The fibre was exposed for 20 min for
the extraction of all samples. Subsequently, the fibre
was removed from the sample, and the analytes were
thermally desorbed in the injector port of the GC.
GC3GC/TOF-MS
All chromatographic analyses were performed using a
GC9GC/TOF-MS from Thermo Electron S.p.A.
(Rodano, Milan, Italy). The apparatus used was
equipped with a split/splitless injector. The first col-
umn was a DB-5 (30 m 9 0.25 mm i.d. 9 0.25 lm
film thickness); the second column a DB-WAX
(1.20 m 9 0.1 i.d. mm 9 0.1 lm film thickness). Both
columns were from J&W Scientific (Folsom, CA,
USA). The columns, both contained in a single oven,
were connected using a glass press-fit connector. The
cryogenic modulation was obtained with a CO2 dual-
jet system, installed at the head of the second dimen-
sion column. This device was controlled by a 2D-GC
on-off Wave Modulator (Thermo Electron). Separa-
tion was performed starting at 50 °C, held for 5 min,
temperature increased at a rate of 5 °C/min to 230 °C
and finally held for 15 min. Analysis was performed at
1.0 mL min 1 constant flow; helium was used as car-
rier gas. Due to the signal enhancement obtained by
modulation of flow eluting from the first column into
the second, with a smaller diameter, analysis was
performed with 10:1 split ratio, to not saturate column
2. Desorption of compounds from fibres was per-
formed with split-injection at a temperature of 250 °C.
A modulation of 2.5 s was applied on the dual-jet
modulator. The transfer line temperature was 230 °C,
and the MS source temperature was 220 °C. The mass
spectral data acquisition rate was 25 Hz for all analy-
ses. The data were collected over a mass range of
© 2013 Institute of Food Science and Technology

35–500 lm. Total ion current chromatograms (TIC)
were finally processed by automated data processing
software Xcalibur 1.4 SR1.
Peak identifications were based on comparison of
their mass spectra with pure standard from Sigma-
Aldrich and/or with those reported by the MainLib
NIST05 commercial library (Table S1). Furthermore,
more careful attention was provided by the manual
inspection of the mass spectra and/or by the use of
additional data, such as the experimentally determined
retention index (RI) values and values reported in the
bibliography for chromatographic columns similar to
that used as the first dimension column in the present
work. For the determination of the RI, calculated on
the first dimension (DB5 column), a C6–C24 n-alkane
series was used. Table S1 lists the compounds
identified in the volatile fraction of the wine samples.
Semi-quantitative data of the aroma compounds were
calculated by relating the peak areas of volatiles to the
peak area of butylated hydroxytoluene (BHT) used as
internal standard. The concentration of volatiles was
expressed as lg L 1 (Table 1).
Odour activity values
The contribution of each volatile compound to wine
aroma was evaluated qualitatively via its associated
descriptor and quantitatively via its OAVs. OAVs were
calculated using the equation OAV = c/t, where c is
the total concentration (lg L 1) of each compound
detected in the samples and t is the odour threshold
value (lg L 1) of the compound in water/ethanol solu-
tion (references are shown in Table 2).
Statistical analysis
Results were expressed as mean  standard deviation
(SD) of at least three independent experiments. Differ-
ences among samples were estimated by analysis of
variance (ANOVA) followed by Tukey’s ‘honest signifi-
cant difference’; the statistical significance level was set
to 0.05. All statistical analyses were performed using
the free statistical software R 2.10.0 version (R Devel-
opment Core Team, 2008).
Results and discussion
Evolution of Nebbiolo-based wine aroma during ageing
A HS-SPME-GC9GC/TOF-MS method was applied
for the comprehensive separation of the volatile aroma
of Nebbiolo-based wines matured in oak barrel for
18 months at constant temperature. The SPME tech-
nique represents a good ‘trap’ for major and minor
volatiles of wine, both polar and nonpolar organic
compounds. Comprehensive 2D-GC coupled with
© 2013 Institute of Food Science and Technology
2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al. 789
time-of-flight mass detection may now be regarded as
an emerging technology that provides considerably
more chemical selectivity than GC/MS, and thus
should be ideally suited to the analysis of complex
matrices like wine.
GC9GC/TOF-MS provides multidimensional data
(retention times on two columns, peak intensity) and a
complete mass spectral plot at each retention time
coordinate in the 2D separation space. Separation on
the two columns and MS analysis are performed in
real time by means of a modulation system. The differ-
ence of polarity between two columns leads to the
optimal separation of complex mixtures of volatiles.
Clearly, the effect of saturation in a GC9GC system is
more critical on second column, which is generally a
very short column. A balance between a very low satu-
ration effect of principal compounds and the resolu-
tion of ‘in trace’ compounds is required. This aspect
could be particularly important regarding the identifi-
cation of these compounds often characterised by a
highly odoriferous impact.
Table 1 reports the qualitative/quantitative charac-
terisation results of aromatic profile from the wine
samples. There were valuable changes in the wine vola-
tile composition during the oak barrel ageing period.
Esters quantitatively represented the largest group
of volatile compounds in Nebbiolo-based wines. Sig-
nificant quantitative differences in the behaviour of
esters during storage in oak barrels were detected.
Esters are important components of wine aroma, espe-
cially for their fruity and floral notes, and their levels
revealed the importance of wood contact time even to
distinguish wines in the same commercial category.
Fatty acid ethyl esters such as ethyl hexanoate, ethyl
octanoate or ethyl decanoate together with acetates
like isoamyl acetate, 1-hexyl acetate or 2-phenylethyl
acetate gradually reduced their concentrations from 6
to 18 months (Fig. 1a). On the contrary, levels of
diethyl succinate and isobutyl acetate increased pro-
gressively as a consequence of hydrolysis-esterification
balances previously reported (Ferreira et al., 2000).
Alcohols contents were generally affected during
storage. Average alcohol content, mainly 1-hexanol
(Fig. 1b) and 1-octanol (Fig. 1c), decreased in major
proportion in wines aged over 6 months. Their con-
centration in wines matured for 12 months was around
25% and 30% lower compared with that wines
matured 6 months. C6 compounds such as 1-hexanol,
(E)-3-hexen-1-ol and (Z)-3-hexen-1-ol, which supply
‘vegetal’ and ‘herbaceous’ tones to the wine, usually
have a negative effect on wine quality when their con-
centration values are above their odour thresholds.
However, in this study, only 1-hexanol, in starting
wine and 6 months aged wine, respectively, showed
values slightly higher than its threshold. Concerning
1-heptanol, a major proportion decreased over
International Journal of Food Science and Technology 2014
790 2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al.

Table 1 Mean quantitative value (lg L 1) and standard deviation (SD) of volatile compounds in wine samples during oak wood ageing at
constant temperature

Starting wine 6 months 12 months 18 months

Wine lg/L (n = 12) (n = 12) (n = 12) (n = 12)

Esters
4 Ethyl acetate 27 620A  353 27 823A  35.1 28 536B  68.6 28 998C  180
7 Ethyl propionate 2967A  41.0 2725B  38.8 2530C  45.8 2440C  65.5
12 Ethyl isobutyrate 2020A  158 2176A  39.5 2286AB  15.2 2456B  145
15 Isobutyl acetate 1512A  28.9 1908B  45.1 2526C  73.7 3060D  60.0
16 Ethyl butanoate 163.0A  10.0 152.0A  8.00 134.0B  4.00 88.0C  13.1
19 Ethyl 2-Butenoate 447.3A  12.2 482.2AB  10.1 513.0B  16.0 599.8C  12.7
20 Ethyl 2-methyl-butanoate 11.0A  3.02 13.4A  3.01 25.6B  5.06 30.6B  3.05
22 Ethyl 3-methyl-butanoate 18.0A  2.08 20.1A  2.00 24.0AB  6.02 27.0B  3.00
28 Isoamyl acetate 651.0A  9.16 592.2B  12.3 343.3C  14.5 258.5D  10.1
32 Ethyl pentanoate 240.3A  12.3 243.3A  40.4 258.3AB  7.64 272.3B  6.81
34 Methyl hexanoate 622.6A  7.37 620.4A  2.34 610.6B  10.0 600.8B  11.2
37 Ethyl 2-hydroxy-3-methyl-butanoate 102.3A  9.50 140.0B  20.0 155.3B  5.03 210.2C  8.38
38 Isoamyl propionate 487.3A  20.0 411.6B  10.4 376.6C  15.2 351.6C  11.5
44 Ethyl hexanoate 739.4A  3.14 710.9B  10.0 631.3C  15.0 505.0D  2.11
47 1-Hexyl acetate 2096A  96.0 1790B  105 1420C  131 1042D  11.3
53 Ethyl 2-hexenoate 1216A  19.2 1226A  30.5 1290B  10.0 1310B  10.1
55 Ethyl 2-furancarboxylate 920.3A  10.0 930.0A  26.4 976.6B  25.1 1043C  6.21
56 Isoamyl butanoate 401.6A  9.07 400.5A  0.96 386.6B  15.2 388.6B  6.11
58 Ethyl 2-hydroxy-4-methyl-pentanoate 148.3A  4.73 201.3B  6.11 320.0C  20.0 346.6C  14.5
59 Isoamyl lactate 1212A  9.85 1213A  37.8 1279B  17.9 1315B  14.6
67 Ethyl heptanoate 1256A  9.07 1060B  52.9 896.6C  65.0 836.3C  4.73
69 Ethyl 3-(methylthio)propanoate 717.6A  6.81 699.2A  18.8 623.3B  25.1 614.0B  5.29
70 Isoamyl 2-methyl butyrate 248.0A  7.21 256.0A  6.93 295.3B  17.4 374.6C  9.61
72 Ethyl methyl succinate 948.6A  6.11 972.8AB  25.7 1093B  100 1228C  2.65
73 Heptyl acetate 241.0A  11.0 203.3B  15.2 38.0C  10.5 0.00  0.00
75 Methyl octanoate 446.3A  12.9 472.1A  26.9 506.6B  40.4 531.3B  7.77
80 Benzyl ethanoate 1221A  11.5 1137B  72.9 753.3C  55.0 614.3D  6.03
82 Ethyl benzoate 447.3A  6.43 333.3B  41.6 256.6C  45.0 226.3C  6.03
83 Diethyl succinate 6506A  40.4 7113B  80.3 8353C  233 8774D  24.2
85 Ethyl 7-octenoate 10.0A  2.00 9.67AB  0.58 9.33B  0.78 9.31B  0.80
87 Ethyl octanoate 875.0A  58.2 771.6B  44.5 698.6C  43.0 621.4D  51.8
89 Methyl salicylate 32.6A  3.06 25.5B  6.48 19.3B  2.08 16.6C  1.53
90 Diethylmethylsuccinate 306.3A  5.69 314.4A  7.18 316.6A  15.2 353.3B  4.16
95 Ethyl benzene acetate 206.0A  5.29 197.6A  9.71 148.3B  17.5 145.3B  5.51
96 2-Methylbutyl hexanoate 89.0A  4.58 78.6B  9.61 58.3C  7.64 42.6D  2.52
97 2-Phenylethyl acetate 438.6A  8.08 416.2A  20.0 336.6B  35.1 280.3C  3.51
99 Ethyl salicylate 31.3A  1.53 28.8A  4.23 20.3B  2.52 18.6B  1.53
101 Diethyl glutarate 53.0A  3.61 55.3A  4.04 65.6B  5.13 67.6B  5.69
108 4-Terpineol acetate 10.3A  0.58 9.89A  0.77 9.85AB  0.92 8.67B  0.58
111 Methyl decanoate 49.0A  4.58 46.0A  5.20 44.0B  4.58 42.6B  2.52
112 Methyl geranate 40.0A  3.00 33.3B  11.5 27.6C  4.04 28.0C  2.00
114 Isobutyl octanoate 21.3A  3.21 22.5A  2.12 23.1A  1.84 23.6A  3.21
115 Ethyl dihydrocinnamate 10.2A  1.03 12.4AB  2.02 14.3AB  5.03 17.3B  5.01
118 Butyl benzoate 20.6A  1.15 22.7B  3.66 23.5B  3.20 25.0C  1.73
119 Ethyl 9-decenoate 237.6A  6.81 215.8B  19.5 183.3C  15.2 166.6D  3.06
122 Ethyl decanoate 354.0A  6.00 284.6B  60.8 223.3BC  20.8 208.6C  9.02
126 Isoamyl octanoate 878.0A  7.21 832.6A  40.5 633.3B  35.1 651.6B  12.5
129 Ethyl cinnamate 11.3A  1.53 14.2A  5.00 18.0B  1.73 17.6B  0.67
130 Methyl dodecanoate 9.00A  1.73 9.30A  0.50 9.14A  0.78 8.67A  0.58
131 Ethyl vanillate 58.6A  8.08 96.6B  25.1 273.3C  25.1 289.6C  7.51
133 Ethyl dodecanoate 130.3A  3.51 125.7A  5.27 116.6B  20.8 107.3B  4.93
136 2-Phenylethyl hexanoate 19.3A  3.06 17.4A  3.86 17.3A  3.79 17.0A  0.85
138 Ethyl tetradecanoate 11.6A  1.53 9.78AB  3.27 8.22B  3.02 8.33B  1.53
139 Ethyl hexadecanoate 30.3A  2.52 24.7AB  5.21 21.6B  2.08 16.6C  1.34

International Journal of Food Science and Technology 2014 © 2013 Institute of Food Science and Technology
 2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al. 791

Table 1 (Continued)

Starting wine 6 months 12 months 18 months

Wine lg/L (n = 12) (n = 12) (n = 12) (n = 12)

Alcohols
3 1-Propanol 155.0A  7.00 149.3A  2.08 132.0B  3.00 122.6C  7.09
5 Isobutanol 4474A  55.0 4829B  71.7 5192C  65.8 5372D  121
10 3-Methyl-1-butanol 48 895A  510 48 086AB  74.5 46 730B  98.4 46 203C  83.7
11 2-Methyl-1-butanol 31 717A  235 31 121AB  94.7 30 669B  35.8 30 411B  207
13 1-Pentanol 3763A  58.5 3830A  35.1 3863AB  30.5 3945B  43.6
21 (E)-3-Hexen-1-ol 249.0A  10.5 204.3B  4.51 142.3C  19.6 82.1D  5.80
23 (Z)-3-Hexen-1-ol 350.0A  3.61 326.6AB  30.5 270.0B  30.0 182.6C  7.51
25 Furfuryl alcohol 1562A  20.4 1605A  22.0 2193B  179 2748C  42.3
27 1-Hexanol 9424A  83.0 9093B  60.8 7137C  99.0 6803C  76.3
39 1-Heptanol 2306A  33.4 2026B  83.2 1823BC  107 1302C  46.0
49 1-Hexanol, 2-ethyl- 713.6A  5.69 723.3A  15.8 816.6B  15.8 866.3C  10.9
52 Benzyl alcohol 2.20A  1.01 3.05A  2.03 6.01B  1.51 7.62B  2.14
62 1-Octanol 866.0A  6.56 760.0B  40.0 526.6C  30.5 489.6C  5.51
74 2-Phenylethyl alcohol 23 663A  77.6 23 003AB  105 21 696B  195 20 133C  10.4
84 Nonanol 29.3A  2.08 25.2A  7.12 13.6B  5.51 8.67C  0.58
Aldehydes
2 Acetaldehyde 3160A  92.0 3247A  65.0 4087B  140 4497C  185
17 Hexanal 161.0A  7.00 156.3A  3.46 116.0B  29.4 49.1C  3.82
18 Furfural 935.0A  15.1 1356B  81.4 2246C  40.8 2635D  60.6
24 Isobutyraldehyde, diethyl acetal 152.0A  7.00 205.6B  5.13 223.3BC  25.1 245.3C  12.8
35 Benzaldehyde 3.02A  2.14 4.20A  1.05 7.08B  1.09 9.10C  1.14
36 5-Methyl-furfural 83.6A  7.51 82.2A  1.95 154.0B  5.29 244.2C  3.04
46 Octanal 1173A  66.5 1006A  41.6 583.3B  85.0 423.6C  12.9
54 Benzeneacetaldehyde 467.3A  6.43 410.0B  26.4 297.0C  20.6 179.5D  4.39
71 Nonanal 217.3A  11.3 207.7AB  8.80 185.3B  12.8 158.6C  8.08
78 (E)-2-Nonenal 21.6A  2.08 18.8B  2.83 0.00  0.00 0.00  0.00
79 3-Phenylpropanal 29.0A  2.65 27.0A  3.46 25.0AB  1.00 23.6B  2.08
91 Decanal 130.0A  4.00 116.6B  15.2 96.6B  15.2 49.3C  3.06
100 Cinnamylaldehyde 244.0A  5.29 231.3A  12.0 220.0AB  20.0 210.0B  0.00
109 Undecanal 19.3A  2.08 16.2B  5.39 10.8C  0.96 8.67D  0.52
123 Vanillin 31.3A  1.53 38.4A  7.93 92.3B  6.81 108.6C  10.9
134 Tetradecanal 355.6A  5.13 317.5B  33.0 230.0C  26.4 137.0D  6.08
Ketones
6 3-Pentanone 102.3A  2.52 122.6B  9.02 186.3C  6.51 245.0D  27.8
41 2-Methyl-2-hepten-6-one 472.0A  9.17 435.6B  12.5 320.0C  20.0 309.0C  6.56
42 2-Octanone 457.6A  10.7 411.0B  10.1 336.6C  35.1 258.3D  6.66
60 Acetophenone 1.03A  0.94 2.06AB  1.02 3.06B  0.91 3.10B  1.05
65 2-Nonanone 660.6A  7.02 560.0B  40.0 436.6C  47.2 480.6C  6.51
86 4-Methylacetophenone 41.6A  1.53 35.8B  5.83 19.3C  2.08 19.3C  2.52
106 2-Undecanone 20.3A  2.52 19.2A  1.92 17.6B  2.08 17.3B  0.58
125 2-T-butyl-5-propyl-[1,3]dioxolan-4-one 9.67A  0.58 13.2AB  3.67 14.3B  1.15 16.6B  0.58
127 Trans-Geranylacetone 10.6A  1.15 10.8A  2.01 9.56A  4.11 8.67B  0.58
Terpenes
43 b-Myrcene 551.3A  9.07 453.3B  25.1 140.0C  52.9 85.3C  3.79
50 Limonene 11.3A  1.53 9.67A  0.58 9.33AB  0.58 8.92B  1.01
51 Eucalyptol 10.0A  1.00 12.0A  2.00 15.3B  1.53 18.1C  1.61
61 (E)-Linalool oxide (trans furan) 29.3A  .04 106.6B  20.8 280.3C  20.0 312.0C  9.85
64 (Z)-Linalool oxide (cis furan) 52.6A  3.06 123.3B  15.2 192.0C  23.0 216.0C  7.94
66 p-Cymenene 19.6A  1.53 18.4A  2.12 13.6B  2.08 11.5B  2.65
68 Linalool 40.0A  3.00 38.0A  3.46 18.0B  7.21 0.00  0.00
76 ( )-Camphor 324.0A  14.0 296.6A  15.2 83.3B  15.2 0.00  0.00
77 Nerol oxide 10.6A  2.08 11.7A  1.92 13.6B  1.53 16.3B  2.08
88 a-Terpineol 20.0A  3.00 18.0A  3.46 15.0AB  4.36 8.33B  0.58
92 b-Cyclocitral 10.3A  1.53 10.8A  0.96 13.3B  1.53 14.3B  2.52
121 b-Damascenone 2.16A  0.38 1.89A  0.27 1.32B  0.32 0.73C  0.38

© 2013 Institute of Food Science and Technology International Journal of Food Science and Technology 2014
792 2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al.

Table 1 (Continued)

Starting wine 6 months 12 months 18 months

Wine lg/L (n = 12) (n = 12) (n = 12) (n = 12)

124 Thujopsene 0.00  0.00 3.33A  1.53 5.00AB  1.00 8.67B  1.15
Aromatic Hydrocarbons
14 Toluene 98.3A  8.02 93.1A  2.25 89.0AB  2.65 83.1B  3.88
26 Ethylbenzene 734.6A  15.0 1223B  60.2 2376C  75.0 2586C  41.6
29 Styrene 61.0A  6.56 57.0A  3.00 49.6B  1.53 48.5B  4.30
30 Benzene, 1,3-dimethyl- 84.0A  4.00 115.6B  16.2 212.6C  15.5 232.1C  10.6
31 Benzene, 1,2-dimethyl- 32.0A  2.65 46.3B  6.03 89.6C  2.52 99.6D  6.81
57 Benzene, (ethoxymethyl)- 612.6A  10.2 569.3B  26.1 396.6C  15.2 348.6D  4.16
94 Benzene, 2-methoxy-4-methyl-1-(1-methylethyl)- 151.0A  3.61 158.0A  6.24 175.3B  18.5 177.3B  5.13
98 4-Ethoxystyrene 310.6A  3.06 283.5B  25.5 199.0C  11.5 130.0D  6.00
120 Biphenyl 228.3A  6.66 208.7B  16.9 190.0C  10.0 154.3D  5.13
Lactones
33 c-Butyrolactone 3821A  89.4 3546B  61.1 2309C  27.6 2095D  46.3
107 (Z)-Whisky lactone 23.6A  3.51 55.3B  18.5 120.0C  20.0 194.6D  3.79
113 (E)-Whisky lactone 51.6A  2.08 83.3B  20.8 203.3C  25.1 277.0D  6.56
117 c-Nonalactone 218.6A  6.11 212.4A  10.7 159.0B  2.65 155.0B  4.36
Volatile Phenols
63 Guaiacol 8.86A  2.13 21.6B  2.08 29.2C  2.25 31.3C  1.74
102 4-Ethyl guaiacol 10.3A  1.53 23.4B  15.1 83.3C  15.2 96.0C  4.00
Others
1 Sulphur dioxide 384.6A  11.2 391.0A  3.37 416.3B  4.51 424.5B  12.3
8 Acetal 104.0A  9.17 203.3B  7.09 299.3C  10.0 422.0D  10.4
9 1,3-Dioxolane, 2,4,5-trimethyl- 147.3A  7.37 215.6B  8.73 315.3C  13.6 354.6D  23.3
40 2H-pyran, 2-ethenyltetrahydro-2,6,6-trimethyl- 506.6A  16.2 663.3B  70.9 900.0C  40.0 1031D  21.2
45 Benzofuran 101.0A  8.54 123.3AB  5.77 155.3B  5.03 174.0C  3.61
48 1,4-Cineol 609.6A  10.0 573.3A  40.4 520.0B  20.0 515.0B  13.2
81 Octanoic acid 2139A  10.1 1953B  50.3 1483C  104 1345D  5.13
93 Cyclohexane, isothiocyanato- 11.3A  1.53 14.2B  5.00 14.0B  5.29 18.0C  1.73
103 Geranylethyl ether 30.0A  3.00 27.0A  5.20 16.6B  5.77 16.0B  1.73
104 Anethol 22.3A  2.52 20.2A  3.66 14.0B  5.29 8.67B  0.58
105 Vitispirane 11.3A  1.15 14.2AB  5.00 14.6AB  5.03 16.6B  1.53
110 Azulene 29.6A  2.52 26.7A  5.00 19.3B  1.15 18.6B  1.15
116 Naphthalene, 1,2-dihydro-1,5,8-trimethyl- 30.0A  3.61 27.6A  4.04 24.6B  1.53 25.0B  3.61
128 2-Phenylpyridine 9.67A  1.53 9.11A  0.96 7.89B  1.84 8.33B  0.51
132 1,6,10-Dodecatrien-3-ol, 3,7,11-trimethyl- 11.3A  1.53 10.4A  4.07 8.22B  2.91 9.33AB  1.15
135 2-(Methylthio)phenyl isothiocyanate 11.6A  2.08 10.5A  3.15 9.33AB  0.58 7.67B  0.32
137 Naphthalene, 1,6-dimethyl-4-(1-methylethyl)- 12.3A  2.08 10.8A  2.50 9.78AB  2.27 7.67B  0.51

Values with different letters in the same row are significantly different (estimated by analysis of variance (ANOVA) followed by Tukey’s ‘honest signifi-
cant difference’ test, P < 0.05). Bold type (Compounds that increase their content during ageing).

12 months, and its concentration in wines matured
18 months was 30% lower (Fig. 1c). The tendency
observed in 1-propanol, 3-methyl-1-butanol and
2-methyl-1-butanol was quite similar, although a smal-
ler amount decreased. On the contrary, isobutanol and
1-pentanol showed a positive trend during ageing, the
latter slightly less pronounced than the former. The
content of 2-phenylethyl alcohol, aromatic compound
characterised by ‘flowery’ and ‘sweet’ notes, appeared
higher in the starting wine than in the 18 months aged
wine, but with values always above its perception
thresholds (Fig. 1b). Octanoic acid contents showed
significant decreases mainly in the 6–12 months period,
while hexanoic acid methyl ester and decanoic acid
methyl ester also behaved similarly, but with a more
regular trend. According to Jarauta et al. (2005), this
fact may be related to the disappearance of com-
pounds in the wood-aged wines as a consequence of
sorption processes.
Similarly, levels of terpenic alcohols, compounds
typically characterised by fruity aromas, showed a sig-
nificant reduction during ageing (e.g. a-terpineol, linal-
ool). The storage in barrels can result in natural
changes in the content of these aromatic compounds,
leading to the conversion of their respective oxides.
For example, linalool was not detected after
18 months of ageing because its molecule quickly gen-
erates furanic and pyranic derivates (linalool oxides)

International Journal of Food Science and Technology 2014 © 2013 Institute of Food Science and Technology
 2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al. 793

Table 2 Odour activity values of free aroma compounds in Nebbiolo-based wines during oak wood ageing

OAVs

1
Odour description Odour threshold (lg L ) Starting 6 months 12 months 18 months

E
(E)-3-Hexen-1-ol Green 400 0.62 0.51 0.36 0.21
(E)-Linalool oxide (trans furan) Sweet, pine, floral 500A 0.06 0.21 0.56 0.62
(E)-Whisky lactone Coconut 200D 0.26 0.42 1.02 1.39
(Z)-3-Hexen-1-ol Cut grass 400E 0.88 0.82 0.68 0.46
(Z)-Linalool oxide (cis furan) Sweet, pine, floral 500A 0.11 0.25 0.38 0.43
(Z)-Whisky lactone Coconut 200D 0.12 0.28 0.60 0.97
1-Hexanol Green, cut grass 8000D 1.18 1.14 0.89 0.85
1-Hexyl acetate Green, floral 1500D 1.40 1.19 0.95 0.70
2-Methyl-1-butanol Burnt, alcohol 60 000B 0.53 0.52 0.51 0.51
2-Phenylethyl alcohol Floral, roses 14 000D 1.69 1.64 1.55 1.44
3-Methyl-1-butanol Burnt, alcohol 60 000B 0.81 0.80 0.78 0.77
4-Ethyl guaiacol Leather, phenolic 33B 0.31 0.71 2.53 2.91
5-Methyl-furfural Spicy, caramel 20 000D 0.00 0.00 0.01 0.01
Acetaldehyde Pungent, ripe apple 500A 6.32 6.49 8.17 8.99
Benzaldehyde Sweet, fruity 350B 0.01 0.01 0.02 0.03
Diethyl succinate Vinous 200 000D 0.03 0.04 0.04 0.04
Ethyl 2-methyl-butanoate Green-fruity, pineapple 18C 0.61 0.72 1.39 1.67
Ethyl 3-methyl-butanoate Green-fruity, pineapple 3C 6.00 6.67 8.00 9.00
Ethyl 9-decenoate Light fruity, fatty 150E 1.58 1.44 1.22 1.11
Ethyl butanoate Fruity, pineapple 20C 8.15 7.60 6.70 4.40
Ethyl cinnamate Honey, cinnamon 1.5B 7.56 9.48 12.00 11.78
Ethyl decanoate Sweet, fruity 200B 1.77 1.42 1.12 1.04
Ethyl dihydrocinnamate Floral, pollen 2B 5.00 6.00 7.00 8.50
Ethyl hexanoate Green, apple 14B 52.79 50.71 45.07 36.07
Ethyl octanoate Sweet, fruity 6D 145.83 128.50 116.33 103.50
Ethyl vanillate Honey, vanillin 990A 0.06 0.10 0.28 0.29
Furfural Sweet, woody, almond 14 000D 0.07 0.10 0.16 0.19
Furfuryl alcohol Faint burnt 2000D 0.78 0.80 1.10 1.37
Guaiacol Smoky, hospital 10C 0.80 2.10 2.90 3.10
Isoamyl acetate Banana 30A 21.70 19.73 11.43 8.63
Linalool Floral 25A 1.60 1.52 0.72 0.00
Octanoic acid Sweet, cheese 500B 4.28 3.91 2.97 2.69
Vanillin Vanillin 60A 0.52 0.64 1.54 1.81
b-Damascenone Sweet, fruity 0.05A 42.00 36.00 26.00 14.00
b-Phenylethyl acetate Pleasant, floral 250D 1.75 1.66 1.35 1.12
c-Butyrolactone Sweet, caramel 35 000E 0.11 0.10 0.07 0.06
c-Nonalactone Sweet, coconut 30C 7.29 7.08 5.30 5.17

A
 et al. (2004); EGo
Guth (1997); BFerreira et al. (2000); CEscudero et al. (2007); DCullere  mez-Mıguez et al. (2007).

(Fig. 2a), characterised by lower fruity notes (Bordiga
et al., 2013). These results suggest that terpene levels
in Nebbiolo-aged wines are hardly affected by the
casks storage.
Benzenic compound levels underwent considerable
changes during ageing as concentrations of benzalde-
hyde and benzyl alcohol were lower in young wines
than in aged wines, although their levels increased
regularly up to 18 months.
Another group of aromatic molecules detected were
the furanic compounds, formed by degradation of car-
bohydrates. The major components of this group were
furfural, furfuryl alcohol and 5-methyl-furfural, whose
concentrations showed a positive trend during ageing
period (Fig. 2a, b). Generally, in this study, the furan-
ic compound levels were lower than their perception
thresholds; only in 12 and 18 months samples, furfuryl
alcohol showed values slightly higher.
Several lactones were identified in the samples.
These compounds are extremely important to the sen-
sory characteristics of wines namely when aged in oak
wood. Their odour description depends on the chemi-
cal structure, functional groups and side chain length
obtained following cyclisation of the corresponding
c-hydroxycarboxylic acid. Two of the most affected
compounds by the storage in oak casks were whisky
lactone isomers, according to previous studies (Garde-
Cerdan et al., 2002; Cadahia et al., 2009).

© 2013 Institute of Food Science and Technology International Journal of Food Science and Technology 2014
794 2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al.
(a)
(b)
(c)
Figure 1 Change in different aromatic compounds content
(expressed as lg L 1 wine) in Nebbiolo-based wine during oak wood
storage (four time points from left to right: starting wine, 6, 12 and
18 months). (a) esters, (b) and (c) alcohols.
Both levels underwent positive exponential increase
from 6 to 18 months providing a direct relation with
the length of ageing period of wines (Fig. 2c). On the
contrary, c-butyrolactone and c-nonalactone showed a
decreasing trend during time.
Volatile phenols such as guaiacol and derivatives
(e.g. 4-ethyl guaiacol) showed a direct relationship
between the period of wood ageing and the wines’ vol-
atile composition. Guaiacol was extracted constantly
and almost linearly in wines matured from 6 to
International Journal of Food Science and Technology 2014
18 months according to other studies (Jarauta et al.,
2005; Fernandez de Sim on et al., 2008). However, the
accumulation of these compounds was much greater
after 12–18 months of ageing, indicating the impor-
tance of wood contact time (Fig. 2c).
The most important compounds of this group in
terms of its contribution to wine aroma are vanillin
and, even if in a secondary way, ethyl vanillate, wood-
extractable compounds associated with ‘vanilla’ and
‘chocolate’ odour descriptors, which were mainly
extracted in the first 12 months of ageing.
Lastly, among the chemical groups of norisopre-
noids, b-damascenone was detected in sample wines.
This molecule, normally considered a positive contribu-
tor to wine aroma, is responsible for the sweet, baked
apple and fruity notes. Moreover, due to its low odour
threshold, it can have a great sensorial impact on wine.
Odour activity values
As a preliminary step to achieve the identification of
the potentially most important odorants of Nebbiolo-
based wine during ageing, the OAVs (the ratio
between the concentration of each volatile compound
with the corresponding odour threshold) were assessed.
On the basis of their threshold and odour description,
the most representative odorants in samples were ten-
tatively established. As shown in Table 2, at least
twenty four components were present at concentra-
tions higher (OAVs > 1) than their corresponding
odour thresholds, nine components with OAVs com-
prised between 0.1–1 and 4 with values <0.1. Thus,
these twenty four compounds exhibit an aroma index
value higher than the unity considered to contribute
individually to the Nebbiolo-based wine aroma. Obvi-
ously, during ageing, it was possible to highlight both
positive and negative trends among the intensities of
these odorants in the different wine samples.
According to the results presented in Table 2, the
seven most potent aromas of each wine are substan-
tially the same. The highest OAVs were obtained for
several well-known by-products of yeast metabolism
such as ethyl octanoate, ethyl hexanoate, isoamyl
acetate, b-damascenone, ethyl cinnamate, ethyl 3-meth-
ylbutanate and acetaldehyde. Ethyl butanoate, octanoic
acid, ethyl dihydrocinnamate and c-nonalactone were
also at concentrations higher than their corresponding
thresholds and seem to be important odorants of this
wine.
The relevant content of vanillin, associated with
‘vanilla’ and ‘chocolate’ odour descriptors, with OAVs
higher than 1 for 12 and 18 months of ageing samples,
should be considered to have a sensorial contribution
for these red wines.
Similarly, the two whisky lactone isomers showed a
substantially linear increasing trend during oak wood
© 2013 Institute of Food Science and Technology

(a)
(c)
Figure 2 Change in different aromatic com-
pounds content (expressed as lg L 1 wine)
in Nebbiolo-based wine during oak wood
storage (four time points from left to right:
starting wine, 6, 12 and 18 months). Terp-
enes, aldehydes, lactones and volatile phe-
nols evenly distributed (a, b and c)
considering their content in wine samples.
storage period. Particularly, (E)-whisky lactone
achieved OAVs higher than 1 for 12 and 18 months,
while (Z)-whisky lactone reached a value of 0.97 only
in 18 months samples.
Guaiacol and 4-ethyl guaiacol, responsible for
‘smoky’ and ‘leather’ tones, respectively, generally
showed OAVs higher than 1 for 12 and 18 months of
ageing samples (guaiacol even starting from the 6th
month with a value of 2.10). Their contents showed
positive trends during ageing so proving that their sen-
sorial contribute becomes increasingly important with
time.
Conclusions
The results of this work allowed us to conclude that
the combination of a GC9GC/TOF-MS system with
HS-SPME technique is a useful tool for the analysis of
volatile compounds in wines. A wide range of organic
compounds from wine aroma were easily extracted
and identified by overcoming problems of co-elution
from the presence of a different range of molecule clas-
ses in untreated wine samples thanks to considerably
more chemical selectivity than GC/MS, and thus ide-
ally suited for the analysis of complex matrices.
Moreover, it was possible to highlight the odour
activity values (OAVs) of the potentially important
odorants of Nebbiolo-based wine during ageing and
also to describe the changes (increasing and decreas-
ing) of these compounds. Vanillin, (E)-whisky lactone,
(Z)-whisky lactone, guaiacol and 4-ethyl guaiacol
showed to be important odorants especially after oak
wood ageing period, proving that these compounds
© 2013 Institute of Food Science and Technology
2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al. 795
(b)
are responsible for the ‘vanilla’, ‘smoky’ and ‘toasty’
tones of this wine.
Acknowledgments
This work was funded by a grant from Regione Pie-
monte (Ricerca Agricola 2008). Dr. Matteo Bordiga
was the recipient of a fellowship from the Italian
Politiche Agricole, Alimentari e Forestali Ministry
(Food-Link project). The authors thank Shelby Punu
for editorial assistance.
References
Antonelli, A., Castellari, L., Zambonelli, C. & Carnacini, A. (1999).
Yeast influence on volatile composition of wines. Journal of Agri-
cultural and Food Chemistry, 47, 1139–1144.
Bloem, A., Lonvaud-Funel, A. & de Revel, G. (2008). Hydrolysis of
glycosidically bound flavour compounds from oak wood by Oeno-
coccus oeni. Food Microbiology, 25, 99–104.
Bonino, M., Schellino, R., Rizzi, C., Aigotti, R., Delfini, C. & Baioc-
chi, C. (2003). Aroma compounds of an Italian wine (Ruche′) by
HS–SPME analysis coupled with GC–ITMS. Food Chemistry, 80,
125–133.
Bordiga, M., Rinaldi, M., Locatelli, M. et al. (2013). Characteriza-
tion of Muscat wines aroma evolution using comprehensive gas
chromatography followed by a post-analytic approach to 2D con-
tour plots comparison. Food Chemistry, 140, 57–67.
Cadahia, E., Fernandez de Sim on, B., Sanz, M., Poveda, P. &
Colio, J. (2009). Chemical and chromatic characteristics of
Tempranillo, Cabernet Sauvignon and Merlot wines from DO
Navarra aged in Spanish and French oak barrels. Food Chemis-
try, 115, 639–649.
Chatonnet, P., Cutzach, I., Pons, M. & Dubourdieu, D. (1999).
Monitoring toasting intensity of barrels by chromatographic analy-
sis of volatile compounds from toasted oak wood. Journal of Agri-
cultural and Food Chemistry, 47, 4310–4318.
International Journal of Food Science and Technology 2014
796 2D-GC evaluation of Nebbiolo-based wine ageing M. Bordiga et al.
Cullere, L., Escudero, A., Cacho, J. & Ferreira, V. (2004). Gas chro-
matography–olfactometry and chemical quantitative study of the
aroma of six premium quality Spanish aged red wines. Journal of
Agricultural and Food Chemistry, 52, 1653–1660.
van den Dool, H. & Kratz, P.D. (1963). A generalization of the
retention index system including linear temperature programmed
gas–liquid partition chromatography. Journal of Chromatography
A, 11, 463–471.
Escudero, A., Campo, E., Fari~ na, L., Cacho, J. & Ferreira, V.
(2007). Analytical characterization of the aroma of five premium
red wines. insights into the role of odor families and the concept of
fruitiness of wines. Journal of Agricultural and Food Chemistry, 55,
4501–4510.
Estevez, P., Gil, M. & Falque, E. (2004). Effects of seven yeast
strains on the volatile composition of Palomino wines. Interna-
tional Journal of Food Science and Technology, 39, 61–69.
Fernandez de Sim on, B., Cadahia, E., Sanz, M. et al. (2008). Vola-
tile compounds and sensorial characterization of wines from four
Spanish denominations of origin, aged in Spanish Rebollo (Quer-
cus pyrenaica willd.) oak wood barrels. Journal of Agricultural and
Food Chemistry, 56, 9046–9055.
Ferreira, V., Lopez, R. & Cacho, J.F. (2000). Quantitative determi-
nation of the odorants of young red wines from different grape
varieties. Journal of the Science of Food and Agriculture, 80, 1659–
1667.
Garde-Cerdan, T., Rodriguez-Mozaz, S. & Ancin-Azpilicueta, C.
(2002). Volatile composition of aged wine in used barrels of French
oak and of American oak. Food Research International, 35, 603–610.
Gauvin, A., Lecomte, H. & Smadja, J. (2004). Comparative investi-
gations of the essential oils of two scented geranium (Pelargonium
spp.) cultivars grown on Reunion Island. Flavour and Fragrance
Journal, 19, 455–460.
Gomez-Mıguez, M.J., Cacho, J.F., Ferreira, V., Vicario, I.M. &
Heredia, F.J. (2007). Volatile components of Zalema white wines.
Food Chemistry, 100, 1464–1473.
Guth, H. (1997). Identification of character impact odorants of dif-
ferent white wine varieties. Journal of Agricultural and Food Chem-
istry, 45, 3027–3032.
Hognadottir, A. & Rouseff, R.L. (2003). Identification of aroma
active compounds in orange essence oil using gas chromatogra-
phy–olfactometry and gas chromatography–mass spectrometry.
Journal of Chromatography A, 998, 201–211.
Jarauta, I., Cacho, J.F. & Ferreira, V. (2005). Concurrent phenom-
ena contributing to the formation of the aroma of wine during
aging in oak wood: an analytical study. Journal of Agricultural and
Food Chemistry, 53, 4166–4177.
Ozel, M.Z., Gogus, F. & Lewis, A.C. (2006). Comparison of direct
thermal desorption with water distillation and superheated water
extraction for the analysis of volatile components of Rosa damasc-
ena Mill. using GC9GC-TOF/MS. Analytica Chimica Acta, 566,
172–177.
Puech, J.L., Feuillat, F. & Mosedale, J.R. (1999). The tannins of
oak heartwood: structure, properties, and their influence on wine
flavor. American Journal of Enology and Viticulture, 50, 469–478.
International Journal of Food Science and Technology 2014
R Development Core Team. (2008). R: A Language and Environment
for Statistical Computing. Vienna, Austria: R Foundation for Sta-
tistical Computing.
Ribereau-Gayon, P., Glories, Y., Maujean, A. & Durboridieu, D.
(2006). Handbook of enology. Vol. 2. In: The Chemistry of Wine,
Stabilization and Treatments. (edited by John Wiley and Sons). Pp.
91–108. Chichester, UK: Ellis Horwood.
Rodrigues, F., Caldeira, M. & C^amara, J.S. (2008). Development of
a dynamic headspace solid phase microextraction procedure cou-
pled to GC–qMSD for evaluation the chemical profile in alcoholic
beverages. Analytica Chimica Acta, 609, 82–104.
Ryan, D., Shellie, R., Tranchida, P., Casilli, A., Mondello, L. &
Marriott, P. (2004). Analysis of roasted coffee bean volatiles by
using comprehensive two-dimensional gas chromatography–time-
of-flight mass spectrometry. Journal of Chromatography A, 1054,
57–65.
Ryona, I., Leclerc, S. & Sacks, G.L. (2010). Correlation of 3-Isobu-
tyl-2-methoxypyrazine to 3-Isobutyl-2-hydroxypyrazine during mat-
uration of bell pepper (Capsicum annuum) and wine grapes (Vitis
vinifera). Journal of Agricultural and Food Chemistry, 58, 9723–
9730.
Sanz, M., Fernandez de Sim on, B., Esteruelas, E. et al. (2011). Effect
of toasting intensity at cooperage on phenolic compounds in acacia
(Robinia pseudoacacia) heartwood. Journal of Agricultural and Food
Chemistry, 59, 3135–3145.
Tominaga, T., Blanchard, L., Darriet, P. & Dubourdieu, D. (2000).
A powerful aromatic volatile thiol, 2-furanmethanethiol, exhibiting
roast coffee aroma in wines made from several Vitis vinifera grape
varieties. Journal of Agricultural and Food Chemistry, 48, 1799–
1802.
Zalacain, A., Alonso, G.L., Lorenzo, C., I~ niguez, M. & Salinas,
M.R. (2004). Stir bar sorptive extraction for the analysis of wine
cork taint. Journal of Chromatography A, 1033, 173–178.
Supporting Information
Additional Supporting Information may be found in
the online version of this article:
Table S1. Identification of compounds detected in
the volatile fraction of wine samples. aRI lit: retention
index [van den Dool & Kratz (1963)] reported in the
literature DB-5 [(5%-phenyl)-methylpolysiloxane] GC
column or equivalents [Gauvin et al. (2004); Hogna-
dottir & Rouseff (2003); Ozel et al. (2006)]. bThe reli-
ability of the identification or structural proposal is
indicated by the following: (M) mass spectrum and
retention time consistent with those of an authentic
standard; (N) structural proposals given on the basis
of mass spectral data (NIST05); (O) mass spectrum
consistent with spectra found in literature.
© 2013 Institute of Food Science and Technology