Glycosides and phenylpropanoid glycerol in Vitis vinifera Cv. Gewurztraminer wine

Article abstract of DOI:10.1021/jf0002600

Eight glycosides and a phenylpropanoid glycerol were isolated from Vitis vinifera cv. Gewurztraminer wine, and their structures were elucidated by MS and NMR spectroscopies, cis-l-(5-Ethenyl-5methyltetrahydrofuran-2-yl)-l-methylethyl O-β-D-apiofuranosyl-(l→6)-Oβ-D-glucopyranoside, (E)3,6,9-trihydroxymegastigm-7-ene 9-O-β-D-glucopyranoside, 2-phenylethyl O-ss-D-apiofuranosyl-(1→6-O-β-D-glucopyranoside, and 2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]propane-1,3-diol are reported for the first time as wine components.

Full text of DOI:10.1021/jf0002600

6178
J. Agric. Food Chem. 2000, 48, 6178−6182
Glycosid es a n d P h en ylp r op a n oid Glycer ol in Vitis vin ifer a Cv.
Gew u r ztr a m in er Win e
Raymonde Baltenweck-Guyot,† J ean-Michel Trendel,† Pierre Albrecht,*,† and Alex Schaeffer‡
Laboratoire de Chimie Organique des Substances Naturelles, UMR 7509 CNRS/Universite´ Louis Pasteur,
Institut de Chimie, 1 rue Blaise Pascal, 67000 Strasbourg, France, and Station de Recherches Vigne et Vin,
INRA, 8 rue Kle´ber, 68000 Colmar, France
Eight glycosides and a phenylpropanoid glycerol were isolated from Vitis vinifera cv. Gewurztraminer
wine, and their structures were elucidated by MS and NMR spectroscopies. cis-1-(5-Ethenyl-5-
methyltetrahydrofuran-2-yl)-1-methylethyl O-â-D-apiofuranosyl-(1f6)-O-â-D-glucopyranoside, (E)-
3,6,9-trihydroxymegastigm-7-ene 9-O-â-D-glucopyranoside, 2-phenylethyl O-â-D-apiofuranosyl-(1f6)-
O-â-D-glucopyranoside, and 2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]propane-1,3-diol are reported
for the first time as wine components.
Keyw or d s: Vitis vinifera; Gewurztraminer wine; mono- and diglycosides; cis-furan linalool oxide;
(E)-3,6,9-trihydroxymegastigm-7-ene; benzyl alcohol; phenylethanol; isolariciresinol; phenylpropanoid
glycerol
INTRODUCTION
Increasing interest has been devoted in the past years
to the study of glycosidically bound volatile compounds
occurring in Vitis vinifera grape and wine, in relation
with their potential organoleptic role. Sugars involved
in these glycoconjugates are mono- and disaccharides,
mainly â-D-glucopyranose, R-L-arabinofuranosyl-(1f6)-
O-â-D-glucopyranose, R-L-rhamnnopyranosyl-(1f6)-O-
â-D-glucopyranose (Williams et al., 1982, 1983), and â-D-
apiofuranosyl-(1f6)-O-â-D-glucopyranose (Voirin et al.,
1992b). Many aglycons have been identified as mono-
terpenoids with various oxidation states, norisoprenoids
with 13 carbon atoms, and shikimate-derived com-
pounds (Winterhalter et al., 1990). As most of these
products are volatile and show interesting sensory
properties, their flavorless glycosidic counterparts, often
by far more abundant than the free aglycons, may
represent an important potential source of aroma.
Glycosides are especially abundant in aromatic grape
varieties, in particular in those growing in Alsace
(France) (Gu¨nata et al., 1985; Voirin et al., 1992b).
In our previous publications on the nonvolatile aroma
precursors aiming to investigate the relationships be-
tween wine molecular markers and grape varieties, we
reported on the isolation and characterization of new
megastigmane (1-4) (Baltenweck-Guyot et al., 1996)
and hemiterpene (5 and 6) (Baltenweck-Guyot et al.,
1997) glycosides (Figure 1) occurring in an Alsatian
Gewurztraminer white wine. We have now obtained,
from the same source, eight additional glycosides and
a phenylpropanoid glycerol. This paper deals with their
structural elucidation and gives detailed or revised
F igu r e 1. Structures of compounds 1-15. Configurations
shown for compounds 7-9 are relative.
NMR data. Among these products, four are reported for
the first time as wine components.
MATERIALS AND METHODS
Sp ectr oscop y. NMR. 1H, 1H-1H COSY, NOESY, HSQC,
1
and HMBC spectra were obtained at 500 and 125 MHz for H
and 13C, respectively, on a Bruker ARX 500 spectrometer using
the standard Bruker software package. The 13C signal and the
residual 1H signal of the solvent (CDCl3) were used as
secondary references (δ 77.0 and 7.26 from TMS, respectively).
13C data were taken from HMBC and HSQC spectra.
LRMS. Spectra were recorded on a Finnigan MAT TSQ 700
spectrometer.
* Author to whom correspondence should be addressed
[telephone 33 (0)3 88 41 68 41; fax 33 (0)3 88 61 00 04; e-mail
albrecht@chimie.u-strasbg.fr].
† Laboratoire de Chimie Organique des Substances Naturel-
les.
‡ Station de Recherches Vigne et Vin.
10.1021/jf0002600 CCC: $19.00 © 2000 American Chemical Society
Published on Web 10/07/2000

Glycosides in Gewurztraminer Wine
J. Agric. Food Chem., Vol. 48, No. 12, 2000 6179
CIMS. NH3 was used as reagent gas, source pressure ) 40
mbar, source temperature ) 150 °C, 70 eV.
1.7, 17.0 Hz, Hb-7), 5.21 (1H, dd, J ) 9.5, 9.5 Hz, Glc-H-3),
5.83 (1H, dd, J ) 10.4, 17.0 Hz, H-6); GC-EIMS 70 eV, in
agreement with literature data (Winterhalter et al., 1990);
HRMS, m/z 523.2159 (calcd for C24H36O11Na, 523.2155).
3,6,9-Trihydroxymegastigm-7-ene 9-O-â-D-Glucopyranoside
(9), Pentaacetate: 1H NMR (500 MHz, CDCl3) δ 0.77 (3H, d, J
) 6.7 Hz, H-13), 0.88 (3H, s, H-12), 1.02 (3H, s, H-11), 1.25
(H-10, multiplicity obscured due to partial overlapping with
signals of impurities), 1.48 (3H, m, Ha-2 and H-4), 1.77 (1H,
m, Hb-2), 1.96 (1H, m, H-5), 2.00-2.15 (15H, 5s, H of acetate
CH3), 3.61 (1H, ddd, J ) 2.5, 4.5, 9.6 Hz, Glc-H-5), 4.13 (1H,
dd, J ) 4.5, 12.4 Hz, Glc-H-6), 4.18 (1H, dd, J ) 2.5, 12.4 Hz,
Glc-H-6), 4.25 (1H, m, H-9), 4.55 (1H, d, J ) 8.0 Hz, Glc-H-1),
4.94 (1H, m, H-3), 4.98 (1H, dd, J ) 8.0, 9.6 Hz, Glc-H-2), 5.08
(1H, dd, J ) 9.6, 9.6 Hz, Glc-H-4), 5.18 (1H, dd, J ) 9.6, 9.6
Hz, Glc-H-3), 5.58 (1H, d, J ) 15.8 Hz, H-7), 5.73 (1H, dd, J )
6.6, 15.8 Hz, H-8); 13C NMR (125 MHz) δ 15.8 (C-13), 20.4-
21.2 (acetate CH3), 24.5 (C-11), 25.1 (C-12), 34.0 (C-5), 34.8
(C-4), 39.5 (C-1), 40.7 (C-2), 62.0 (Glc-C-6), 68.3 (Glc-C-4), 69.9
(C-3), 71.5 (Glc-C-2), 71.8 (Glc-C-5), 73.0 (Glc-C-3), 77.0 (C-6),
77.8 (C-9), 99.5 (Glc-C-1), 131.9 (C-8), 134.7 (C-7), 169.5-171.0
(acetate CdO), C-10 not determined; GC-EIMS 70 eV, m/z (rel
int) 540 (2), 331 (30), 271 (13), 253 (13), 252 (9), 211 (6), 193
(23), 192 (78), 169 (100), 149 (21), 135 (18), 127 (15), 109 (54),
97 (13), 82 (15); HRMS, m/z 623.2678 (calcd for C29H44O13Na,
623.2680).
GC-EIMS. The column used was a DB5 J &W (60 m × 0.25
mm i.d. × 0.1 µm film thickness), He at 2 mL min-1; 38 °C (1
min), 38-220 °C at 10 °C min-1, 220-300 °C at 3 °C min-1
300 °C (30 min); source temperature ) 200 °C, 70 eV.
,
HRMS. Measurements on (M + Na)+ ions of the peracetyl-
ated compounds were performed on a Micromass Autospec
spectrometer in positive LSIMS mode using a mixture of
3-nitrobenzyl alcohol, TFA (1%), and NaI as the matrix.
Win e. The Gewurztraminer wine from year 1993 was
obtained from the cave vinicole of Ribeauville´ (Alsace, France).
Solven ts a n d Rea gen ts. Usual solvents (CH2Cl2, MeOH,
Me2CO, and EtOAc) were distilled twice. Acetic anhydride
(Prolabo), iso-PrOH (Carlo Erba), and pyridine (Prolabo,
distilled over CaH2 and stored over 4 Å molecular sieves) were
of analytical grade. MeCN (Aldrich) was of HPLC grade. H2O
was distilled and deionized.
Isola tion of Com p ou n d s. The crude glycosidic extract (500
mg) obtained from the wine (5 L) was subjected to HPLC gross
separation, affording 10 fractions A1-A10 (Baltenweck-Guyot
et al., 1996, 1997). Further fractionation of A5 (60 mg) and
A6 (40 mg) by analytical HPLC (Du Pont Zorbax ODS, 250 ×
4.6 mm, H2O/MeCN 9:1 for A5, H2O/Me2CO 9:1 for A6, 1 mL
min-1) furnished in each case 10 subfractions B1-B10 and
C1-C10, respectively. Final purification of compounds from
B6 (5 and 6), B7 (10, 1 mg; and 11, 0.8 mg), B9 (1 and 2), B10
(3, 4, and 14, 0.5 mg), C3 (9, 0.6 mg; and 15, 1.6 mg), C6 (13,
1 mg), and C10 (7, 0.7 mg; and 8, 0.3 mg) was achieved by
analytical HPLC [Astec cyclobond I acetylated â-cyclodextrin,
250 × 4.6 mm, H2O (H2O/MeCN 95:5 for C10), 0.8-1 mL
min-1]. Glycoside 12 (3.5 mg) was purified by TLC (silica gel
60 F254 Merck, EtOAc/iso-PrOH/H2O 60:30:10) from B8. Owing
to possible loss of product during the isolation procedure,
weights indicated for the investigated compounds may be
underestimated with regard to their true concentrations in the
wine.
Phenylmethyl O-â-D-Glucopyranoside (10), Tetraacetate: 1H
NMR (500 MHz, CDCl3) δ 1.99-2.12 (12H, 4s, H of acetate
CH3), 3.67 (1H, ddd, J ) 2.3, 4.5, 9.4 Hz, Glc-H-5), 4.16 (1H,
dd, J ) 2.3, 12.2 Hz, Glc-H-6), 4.28 (1H, dd, J ) 4.5, 12.2 Hz,
Glc-H-6), 4.54 (1H, d, J ) 7.9 Hz, Glc-H-1), 4.62 (1H, d, J )
12.4 Hz, H-1), 4.90 (1H, d, J ) 12.4 Hz, H-1), 5.07 (1H, dd, J
) 7.9, 9.4 Hz, Glc-H-2), 5.11 (1H, dd, J ) 9.4, 9.4 Hz, Glc-H-
4), 5.16 (1H, dd, J ) 9.4, 9.4 Hz, Glc-H-3), 7.28 (2H, m, H-2aro
and H-6aro), 7.28-7.35 (1H, m, H-4aro), 7.31-7.37 (2H, m,
H-3aro and H-5aro); 13C NMR (125 MHz) δ 20.5-21.2 (acetate
CH3), 62.0 (Glc-C-6), 68.0 (Glc-C-4), 70.5 (C-1), 71.1 (Glc-C-2),
71.8 (Glc-C-5), 72.8 (Glc-C-3), 99.0 (Glc-C-1), 127.8 (C-2aro and
C-6aro), 128.2 (C-4aro), 128.5 (C-3aro and C-5aro), 136.5 (C-
1aro), 169.0-171.0 (acetate CdO); GC-EIMS 70 eV, in agree-
ment with literature data (Williams et al., 1983); HRMS, m/z
461.1426 (calcd for C21H26O10Na, 461.1424).
Phenylmethyl O-R-L-Arabinofuranosyl-(1f6)-O-â-D-glucopy-
ranoside (11), Hexaacetate: 1H NMR (500 MHz, CDCl3) δ 1.99-
2.13 (18H, 6s, H of acetate CH3), 3.66 (1H, dd, J ) 1.0, 10.2
Hz, Glc-H-6), 3.68 (1H, m, Glc-H-5), 3.78 (1H, m, Glc-H-6), 4.22
(1H, dd, J ) 5.8, 11.9 Hz, Ara-Hb-5), 4.30 (1H, ddd, J ) 3.3,
5.2, 5.8 Hz, Ara-H-4), 4.41 (1H, dd, J ) 3.3, 11.9 Hz, Ara-Ha-
5), 4.53 (1H, d, J ) 8.0 Hz, Glc-H-1), 4.62 (1H, d, J ) 12.4 Hz,
H-1), 4.87 (1H, d, J ) 12.4 Hz, H-1), 4.99 (1H, dd, J ) 1.5, 5.2
Hz, Ara-H-3), 5.02 (1H, dd, J ) 8.0, 9.5 Hz, Glc-H-2), 5.04 (1H,
dd, J ) 9.5, 9.5 Hz, Glc-H-4), 5.08 (1H, d, J ) 1.5 Hz, Ara-H-
2), 5.10 (1H, s, Ara-H-1), 5.14 (1H, dd, J ) 9.5, 9.5 Hz, Glc-
H-3), 7.26-7.32 (1H, m, H-4aro), 7.28 (2H, m, H-2aro and
H-6aro), 7.34 (2H, m, H-3aro and H-5aro); 13C NMR (125 MHz)
δ 20.0-21.0 (acetate CH3), 63.3 (Ara-C-5), 65.8 (Glc-C-6), 69.0
(Glc-C-4), 70.2 (C-1), 71.3 (Glc-C-2), 72.9 (Glc-C-3), 73.1 (Glc-
C-5), 77.0 (Ara-C-3), 80.3 (Ara-C-4), 81.2 (Ara-C-2), 99.0 (Glc-
C-1), 105.9 (Ara-C-1), 127.8 (C-2aro and C-6aro), 128.0 (C-
4aro), 128.2 (C-3aro and C-5aro), 136.8 (C-1aro), 169.6-170.2
(acetate CdO); GC-EIMS 70 eV, in agreement with literature
data (Williams et al., 1983); HRMS, m/z 677.2067 (calcd for
Acetyla tion of Com p ou n d s. Compounds were acetylated
in a mixture of acetic anhydride and anhydrous pyridine for 3
days at room temperature.
(Z)-1-(5-Ethenyl-5-methyltetrahydrofuran-2-yl)-1-methyleth-
yl O-â-D-Apiofuranosyl-(1f6)-O-â-D-glucopyranoside (7), Hexaac-
etate: 1H NMR (500 MHz, CDCl3) δ 1.16 (3H, s, H-10), 1.21
(3H, s, H-9), 1.26 (3H, s, H-8), 1.71 (m, Hb-4), 1.76 (1H, m,
Hb-3), 1.79 (1H, m, Ha-4), 1.92 (1H, m, Ha-3), 1.99, 2.01, 2.02,
2.03, 2.08, 2.11 (18H, 6s, H of acetate CH3), 3.54 (1H, dd, J )
6.9, 11.1 Hz, Glc-H-6), 3.64 (1H, ddd, J ) 2.3, 6.9, 9.5 Hz, Glc-
H-5), 3.68 (1H, dd, J ) 2.3, 11.1 Hz, Glc-H-6), 3.92 (1H, t, J )
6.8 Hz, H-2), 4.14 (1H, d, J ) 10.6 Hz, Api-Ha-4), 4.21 (1H, d,
J ) 10.6 Hz, Api-Hb-4), 4.49 (1H, d, J ) 12.3 Hz, Api-H-5),
4.80 (1H, d, J ) 12.3 Hz, Api-H-5), 4.81 (1H, d, J ) 8.0 Hz,
Glc-H-1), 4.90 (1H, dd, J ) 8.0, 9.5 Hz, Glc-H-2), 4.90 (1H, dd,
J ) 9.5, 9.5 Hz, Glc-H-4), 4.96 (1H, dd, J ) 1.5, 10.8 Hz, Ha-
7), 5.01 (1H, d, J ) 0.4 Hz, Api-H-1), 5.15 (1H, dd, J ) 1.5,
17.4 Hz, Hb-7), 5.19 (1H, dd, J ) 9.5, 9.5 Hz, Glc-H-3), 5.32
(1H, d, J ) 0.4 Hz, Api-H-2), 5.92 (1H, dd, J ) 10.8, 17.4 Hz,
H-6); 13C NMR (125 MHz) δ 20.6-21.9 (acetate CH3), 21.8 (C-
10), 24.2 (C-9), 25.5 (C-8), 26.9 (C-3), 37.5 (C-4), 63.0 (Api-C-
5), 66.7 (Glc-C-6), 69.3 (Glc-C-4), 71.6 (Glc-C-2), 72.4 (Api-C-
4), 72.8 (Glc-C-5), 73.2 (Glc-C-3), 76.5 (Api-C-2), 79.6 (C-1), 83.0
(C-5), 84.0 (Api-C-3), 84.2 (C-2), 95.3 (Glc-C-1), 105.9 (Api-C-
1), 111.3 (C-7), 144.3 (C-6), 169.5-171.0 (acetate CdO); GC-
EIMS 70 eV, m/z (rel int) 547 (1), 317 (4), 260 (10), 259 (100),
169 (6), 154 (5), 153 (86), 139 (58), 135 (11), 129 (8), 127 (6),
111 (7), 109 (9), 97 (10), 93 (7), 71 (13); HRMS, m/z 739.2794
(calcd for C33H48O17Na, 739.2789).
C
30H38O16Na, 677.2058).
Phenylmethyl O-â-D-Apiofuranosyl-(1f6)-O-â-D-glucopyra-
noside (12), Hexaacetate: 1H NMR (500 MHz, CDCl3) δ 2.00-
2.15 (18H, 6s, H of acetate CH3), 3.57 (1H, dd, J ) 5.5, 11.0
Hz, Glc-H-6), 3.62 (1H, m, Glc-H-5), 3.68 (1H, dd, J ) 3.0, 11.0
Hz, Glc-H-6), 4.11 (1H, d, J ) 10.5 Hz, Api-H-4), 4.20 (1H, d,
J ) 10.5 Hz, Api-H-4), 4.51 (1H, d, J ) 7.9 Hz, Glc-H-1), 4.56
(1H, d, J ) 12.2 Hz, Api-H-5), 4.57 (1H, d, J ) 12.3 Hz, H-1),
4.72 (1H, d, J ) 12.2 Hz, Api-H-5), 4.82 (1H, d, J ) 12.3 Hz,
H-1), 4.90 (1H, dd, J ) 9.5, 9.5 Hz, Glc-H-4), 4.97 (1H, dd, J
) 7.9, 9.5 Hz, Glc-H-2), 5.02 (1H, s, Api-H-1), 5.12 (1H, dd, J
) 9.5, 9.5 Hz, Glc-H-3), 5.32 (1H, s, Api-H-2), 7.28 (2H, m,
H-2aro and H-6aro), 7.28-7.34 (1H, m, H-4aro), 7.34 (2H, m,
(Z)-1-(5-Methyl-5-vinyltetrahydrofuran-2-yl)-1-methylethyl O-â-
D-Glucopyranoside (8), Tetraacetate: 1H NMR (500 MHz,
CDCl3) δ 1.17 (3H, s, H-10), 1.22 (3H, s, H-9), 1.27 (3H, s, H-8),
1.79 (2H, m, H-4), 1.85 (2H, m, H-3), 2.00, 2.02, 2.04, 2.06 (12H,
4s, H of acetate CH3), 3.67 (1H, m, Glc-H-5), 3.74 (1H, t, J )
6.4 Hz, H-2), 4.10 (1H, m, Glc-H-6), 4.21 (1H, dd, J ) 10.6,
12.4 Hz, H-6), 4.85 (1H, d, J ) 7.7 Hz, Glc-H-1), 4.95 (1H, dd,
J ) 7.7, 9.5 Hz, Glc-H-2), 4.98 (1H, dd, J ) 1.7, 10.4 Hz, Ha-
7), 5.03 (1H, dd, J ) 9.5, 9.5 Hz, Glc-H-4), 5.18 (1H, dd, J )

6180 J. Agric. Food Chem., Vol. 48, No. 12, 2000
Baltenweck-Guyot et al.
H-3aro and H-5aro); 13C NMR (125 MHz) δ 20.5-21.2 (acetate
CH3), 63.2 (Api-C-5), 66.7 (Glc-C-6), 69.3 (Glc-C-4), 70.7 (C-1),
71.4 (Glc-C-2), 72.6 (Api-C-4), 72.8 (Glc-C-3), 73.3 (Glc-C-5),
76.1 (Api-C-2), 84.0 (Api-C-3), 99.3 (Glc-C-1), 106.2 (Api-C-1),
127.8 (C-2aro and C-6aro), 128.0 (C-4aro), 128.2 (C-3aro and
C-5aro), 137.0 (C-1aro), 169.1-170.3 (acetate CdO); GC-EIMS
70 eV, m/z (rel int) 317 (2), 289 (7), 260 (5), 259 (48), 245 (7),
217 (9), 189 (4), 169 (4), 145 (9), 140 (8), 139 (82), 109 (13), 97
(18), 91 (100); HRMS, m/z 677.2070 (calcd for C30H38O16Na,
677.2058).
2-Phenylethyl O-â-D-Apiofuranosyl-(1f6)-O-â-D-glucopyra-
noside (13), Hexaacetate: 1H NMR (500 MHz, CDCl3) δ 2.00-
2.15 (18H, 6s, H of acetate CH3), 2.80 (2H, m, H-2), 3.59 (1H,
dd, J ) 6.5, 11.1 Hz, Glc-H-6), 3.65 (1H, m, Ha-1), 3.67 (1H,
m, Glc-H-5), 3.70 (1H, m, Glc-H-6), 4.11 (1H, m, Hb-1), 4.14
(1H, d, J ) 10.7 Hz, Api-Ha-4), 4.21 (1H, d, J ) 10.7 Hz, Api-
Hb-4), 4.46 (1H, d, J ) 8.0 Hz, Glc-H-1), 4.56 (1H, d, J ) 12.3
Hz, Api-Ha-5), 4.74 (1H, d, J ) 12.3 Hz, Api-Hb-5), 4.92 (1H,
dd, J ) 9.5, 9.5 Hz, Glc-H-4), 4.95 (1H, dd, J ) 8.0, 9.5 Hz,
Glc-H-2), 5.03 (1H, s, Api-H-1), 5.17 (1H, dd, J ) 9.5, 9.5 Hz,
Glc-H-3), 5.33 (1H, s, Api-H-2), 7.19 (2H, m, H-2aro and
H-6aro), 7.26-7.30 (signals obscured due to overlapping with
residual protons of the solvent, 3H, m, H-3aro, H-4aro and
H-5aro); 13C NMR (125 MHz) δ 20.5-21.2 (acetate CH3), 36.0
(C-2), 63.3 (Api-C-5), 66.5 (Glc-C-6), 69.3 (Glc-C-4), 70.5 (C-1),
72.7 (Glc-C-2), 72.7 (Api-C-4), 73.0 (Glc-C-3), 73.3 (Glc-C-5),
76.1 (Api-C-2), 100.7 (Glc-C-1), 106.0 (Api-C-1), 128.2-128.8
(C-3aro, C-4aro and C-5aro), 129.0 (C-2aro and C-6aro), 169.3-
170.2 (acetate CdO); GC-EIMS 70 eV, m/z (rel int) 317 (2),
289 (2), 260 (7), 259 (100), 217 (12), 169 (5), 145 (4), 139 (77),
127 (7), 109 (4), 105 (81), 104 (31), 97 (7), 91 (7), 65 (2); HRMS,
m/z 691.2225 (calcd for C31H40O16Na, 691.2214).
Isolariciresinol 4-O-â-D-Glucopyranoside (14), Heptaacetate:
1H NMR (500 MHz, CDCl3) δ 2.00, 2.01, 2.04, 2.07, 2.29, 2.62,
2.62 (21H, 7s, H of acetate CH3), 2.04 (1H, m overlapping with
acetate signals, H-8), 2.29 (1H, m overlapping with acetate
signals, H-8′), 2.84 (2H, m, H-7′), 3.44 (1H, ddd, J ) 2.4, 2.7,
9.4 Hz, Glc-H-5), 3.78 (3H, s, CH3-O-C-3′), 3.79 (3H, s, CH3-
O-C-3), 3.81 (1H, dd, J ) 2.4, 12.6 Hz, Glc-H-6), 3.91 (1H, d
broad J ) 10.8 Hz, H-7), 3.98 (1H, dd, J ) 3.4, 11.6 Hz, H-9′),
4.09 (1H, m, H-9), 4.10 (1H, m, H-9′), 4.16 (1H, dd, J ) 2.7,
12.6 Hz, Glc-H-6), 4.23 (1H, dd, J ) 4.2, 11.1 Hz, H-9), 4.52
(1H, m, Glc-H-1), 5.12-5.20 (3H, m, Glc-H-2, Glc-H-3 and Glc-
H-4), 6.47 (1H, s broad, H-5′), 6.61 (1H, s broad, H-2′), 6.66
(1H, dd, J ) 1.7, 8.0 Hz, H-6), 6.71 (1H, d, J ) 1.7 Hz, H-2),
6.96 (1H, d, J ) 8.0 Hz, H-5); GC-EIMS 70 eV, m/z (rel int)
486 (1), 426 (11), 395 (1), 366 (6), 331 (17), 289 (4), 271 (9),
224 (10), 211 (5), 170 (6), 169 (100), 164 (10), 149 (6), 137 (11),
127 (9), 109 (37), 103 (11), 97 (7); HRMS, m/z 839.2776 (calcd
for C40H48O18Na, 839.2738).
F igu r e 2. Most relevant NOESY connectivities (A) and
HMBC connectivities (B) observed for the aglycon moiety of
the peracetylated compound 7. Configurations shown for the
aglycon residue are relative.
cis-F u r a n Lin a lool Oxid e Glycosid es 7 a n d 8. The
CI(NH3) mass spectrum of the nonderivatized diglyco-
side 7 displays a pseudomolecular [M + NH4]+ ion at
m/z 482, whereas the GC-EI mass spectrum of peracetyl-
ated 7 shows the typical fragments of a tri-O-acetyl-
pentafuranose (m/z 259 and 139) and a tri-O-acetyl-
hexapyranose (m/z 169 and 109) together with ions at
m/z 153, 111, and 93 arising from the aglycon moiety,
all data consistent with the proposed structure. NMR
data, including NOESY and HMBC spectra, allowed the
nature and the assemblage of the sugars to be deter-
mined as a â-D-apiofuranosyl-(1f6)-O-â-D-glucopyra-
noside, as previously described for diglycosides 3-6
(Baltenweck-Guyot et al., 1996, 1997). Concerning the
1
aglycon residue, H and 1H-1H COSY spectra reveal
the presence of three olefinic protons H-6 and H-7
belonging to a vinyl group, of three methyl singlets, and
one proton H-2 coupling with two geminal protons H-3,
which in turn are coupled with the protons H-4. These
data support the structure of a linalyl furanic oxide
moiety. This proposal was fully confirmed by analysis
of the connectivity networks of NOESY and HMBC
spectra (Figure 2), allowing in particular the glycosidic
linkage and the cis configuration of the aglycon to be
determined. Consequently, the structure of 7 was
established as being cis-1-(5-ethenyl-5-methyltetrahy-
drofuran-2-yl)-1-methylethyl O-â-D-apiofuranosyl-(1f6)-
O-â-D-glucopyranoside. Chemical shift values of the
aglycon are in agreement with those obtained by
Strauss et al. (1987) for an arabinosyl-glucoside of cis-
furan linalool oxide isolated from a Riesling grape juice.
Although Baumes et al. (1993) have reported the isola-
tion from grape of a mixture of the cis and trans isomers
bound to the apiosyl-glucosyl residue, we nevertheless
present here the first unambiguous characterization of
compound 7 in a wine. In a similar manner, from both
MS and NMR (1H-1H COSY and NOESY) studies, the
structure of 8 was elucidated as being the â-D-glucopy-
ranoside of cis-furan linalool oxide. This compound was
previously identified by Winterhalter et al. (1990) in a
Riesling wine. We report here its detailed 1H NMR data.
Nor isop r en oid glycosid e 9. Mass spectral, 1H
NMR, and 2D NMR data for the derivatized component
2-[4-(3-Hydroxypropyl)-2-methoxyphenoxy]propane-1,3-di-
ol (15), Triacetate: 1H NMR (500 MHz, CDCl3) δ 1.93 (2H, m,
H-8′), 1.96 (3H, s, CH3 of acetate at C-9′), 1.98 (6H, 2s, CH3 of
acetates at C-1 and C-3), 2.63 (2H, t broad, J ) 8.1 Hz, H-7′),
3.82 (3H, s, H3C-O-C-2′), 4.08 (2H, t broad, J ) 6.6 Hz, H-9′),
4.43 (4H, d, J ) 5.1 Hz, H-1 and H-3), 4.49 (1H, q, J ) 5.1 Hz,
H-2), 6.69 (1H, dd, J ) 1.5, 8.1 Hz, H-5′), 6.71 (1H, d, J ) 1.5
Hz, H-3′), 6.93 (1H, d, J ) 8.1 Hz, H-6′); 13C NMR (125 MHz)
δ 21.8 (2 × acetate CH3 at C-1 and C-3), 22.2 (acetate CH3 at
C-9′), 30.4 (C-8′), 33.1 (C-7′), 57.4 (H3C-O-C-2′), 63.1 (C-1 and
C-3), 63.9 (C-9′), 76.1 (C-2), 112.6 (C-3′), 120.5 (C-5′), 120.9
(C-6′), 137.0 (C-4′), 144.8 (C-1′), 150.9 (C-2′), 170.8 (2 × acetate
CdO at C-1 and C-3), 171.2 (acetate CdO at C-9′); GC-EIMS
70 eV, m/z (rel int) 382 (7), 224 (1), 164 (7), 160 (8), 159 (100),
139 (5), 137 (7), 117 (2), 103 (3), 99 (10), 57 (4); HRMS, m/z
405.1526 (calcd for C21H25O8Na, 405.1549).
RESULTS AND DISCUSSION
The investigated compounds 7-15 were obtained
from the glycosidic extract of the wine according to the
procedure described under Materials and Methods. The
products were acetylated before their examination by
GC-MS and by NMR.

Glycosides in Gewurztraminer Wine
J. Agric. Food Chem., Vol. 48, No. 12, 2000 6181
F igu r e 3. Most relevant NOESY connectivities observed for
the aglycon moiety of the peracetylated compound 13.
F igu r e 4. Most relevant NOESY connectivities observed for
the aglycon moiety of the peracetylated compound 14.
9 allowed its structure to be established as the (E)-3,6,9-
trihydroxymegastigm-7-ene 9-O-â-D-glucopyranoside, al-
ready characterized by Lutz et al. (1993) from starfruit
and by Otsuka et al. (1994) from leaves of Alangium
premnifolium. The relative configurations at C-3, C-5,
and C-6 shown in Figure 1 follow from NOE effects
observed between H-11 and protons H-3, H-5, and H-7.
As for megastigmane derivatives 1-4, the configuration
at C-9 remains undetermined. This glucoside has not
been previously reported as a constituent of grape or
wine, although the free aglycon was characterized in
glycoside hydrolysates of various grape cultivars (Sefton
et al., 1996).
F igu r e 5. Most relevant NOESY connectivities (A) and
HMBC connectivities (B) observed for the peracetylated com-
pound 15.
Ben zyl Alcoh ol a n d P h en yleth a n ol Glycosid es
10-13. The peracetylated components 10-12, identified
by mass spectral and NMR studies as benzyl alcohol
glycosides, were previously characterized in various
wines and grapes. Thus, the benzyl O-â-D-glucopyrano-
side 10 was isolated from Riesling and Muscat of
Alexandria grapes by Williams et al. (1983), the benzyl
O-R-L-arabinofuranosyl-(1f6)-O-â-D-glucopyranoside 11
was found by Williams et al. (1983) in the same grape
varieties and by Voirin et al. (1992a,b) in Muscat of
Alexandria, Muscat Frontignan, Muscat Hambourg,
Muscat Ottonel, and Gewurztraminer grapes, and the
apiosyl glucoside 12 was identified in a Muscat Fron-
tignan grape by Baumes et al. (1993). Except for H-1
and H-2 of the apiose moiety, the assignments of which
should be reversed on the basis of the NOESY, HMBC,
and HSQC correlations observed, NMR data of 12 are
very similar to that of the compound obtained by
synthesis by Suzuki et al. (1988) and Mba¨ıraroua et al.
(1994). The GC-EI mass spectrum of the diglycoside 13
comprises, in addition to the usual fragments corre-
sponding to an apiosyl- or arabinosyl-glucoside, ions at
m/z 91 and 105, which may arise from a phenylethyl
unit. The pseudomolecular [M + NH4]+ ion at m/z 434
obtained by CIMS further supports the hypothesis of
such a diglycoside of phenylethanol. NMR spectra of the
peracetylated compound 13 reveal the presence of the
same sugar moiety as for the diglycoside 12, that is, a
peracetyl â-D-apiofuranosyl-â-D-glucopyranoside, and for
the aglycon part, a 2-phenylethyl unit linked at C-1 to
the sugars, as shown in particular by NOE effects
observed (Figure 3). Hence, the structure of 13 is that
of the 2-phenylethyl O-â-D-apiofuranosyl-(1f6)-O-â-D-
glucopyranoside. Again except for H-1 and H-2 of the
apiose moiety, our NMR data are similar to those of
Mba¨ırouara et al. (1994) for the same compound ob-
tained by a synthetic way. However, it is the first time
that the glycoside 13 has been identified in a wine,
although several authors have attempted to characterize
it, especially by MS (Voirin et al., 1992b).
H-5′ chemical shifts, obtained in our case on the basis
of NOE effects, should, however, be reversed.
P h en ylp r op a n oid Glycer ol 15. The CIMS (NH3) of
the nonderivatized compound 15 shows a pseudomo-
lecular [M + NH4]+ ion at m/z 274, shifted to m/z 400
for its peracetylated counterpart, which indicates the
presence of three derivatizable groups. The GC-EI mass
spectrum of acetylated 15 displays, besides the molec-
ular ion M+ ) 382, a major fragment at m/z 159
[(AcOCH2)2C]+ and ions at m/z 175 [AcOCH2)2CH - O]+,
117 [159 - CH2CO]+, and 99 [159 - CH3COOH]+, which
can be issued from a diacetyl glyceryl unit. Additional
ions at m/z 224 [M - (AcOCH2)2C]+ and 164 [224 - CH3-
COOH]+ clearly arise from the second part of the
molecule, which bears one acetylated hydroxyl group,
as confirmed by MS/MS studies showing that fragments
at m/z 224 and 159 belong to two different subunits of
1
1
15. H and H-1H COSY NMR spectra exhibit down-
field signals of three aromatic protons, two of which are
vicinal, at δ 6.69 (J ) 1.5, 8.1 Hz, H-5′), 6.71 (J ) 1.5
Hz, H-3′), and 6.93 (J ) 8.1 Hz, H-6′), thus indicating
the presence of a trisubstituted aromatic ring. These
spectra also include a methyl singlet at δ 3.82 ascribable
to a methoxy group, a proton quintuplet at δ 4.49 (H-2)
coupled with four protons appearing as a doublet at δ
4.33 (H-1 and H-3), these five protons characteristic of
an acetylated glycerol subunit linked by its C-2 hydroxy
group, and a sequence of three methylene groups at δ
4.08 (H-9′), 2.83 (H-7′), and 1.93 (H-8′) typical of a
phenylpropanol side chain. The arrangement of these
three substituent groups on the aromatic ring easily
follows from the NOESY and HMBC spectra correla-
tions as pictured in Figure 5. Thus, the structure of
compound 15 was identified as 2-[4-(3-hydroxypropyl)-
2-methoxyphenoxy]propane-1,3-diol. Our NMR data are
in good agreement with those obtained by Marinos et
al. (1992) for the glucosidic derivative of 15 isolated by
these authors from a Riesling wine. However, the free
aglycon 15, which has a marked butter and caramel
smell as also noted by Marinos et al. (1992), has not
been previously reported as a wine component.
1
Isola r icir esin ol 4-O-â-D-Glu cop yr a n osid e 14. H
NMR data of the peracetylated compound 14 (Figure
4) are similar to those of peracetylated isolariciresinol
4-O-â-D-glucopyranoside already identified by Marinos
et al. (1992) in a Riesling wine. Assignment of H-2′ and
Besides glycosides 1-6 previously characterized in an
Alsatian Gewurztraminer wine, we have reported here

6182 J. Agric. Food Chem., Vol. 48, No. 12, 2000
Baltenweck-Guyot et al.
Otsuka, H.; Kamada, K.; Ogimi, C.; Hirata, E.; Takushi, A.;
Takeda, Y. Alangionosides A and B, ionol glycosides from
leaves of Alangium premnifolium. Phytochemistry 1994, 35,
1331-1334.
Sefton, M. A.; Francis, I. L.; Williams, P. J . The free and bound
volatile secondary metabolites of Vitis vinifera grape cv.
Semillon. Austr. J . Grape Wine Res. 1996, 2, 179-183.
Strauss, C. R.; Gooley, P. R.; Wilson, B.; Williams, P. J .
Application of droplet countercurrent chromatography to the
analysis of conjugated forms of terpenoids, phenols, and
other constituents of grape juice. J . Agric. Food Chem. 1987,
35, 519-524.
the unambiguous identification, based on MS and 2D-
NMR data, of nine additional compounds isolated from
the same wine. Four of them are reported for the first
time as wine components, and most of the others, the
NMR data of which have been revised or made more
precise, have not been recorded before from the Gewurz-
traminer cultivar. A more extensive study of glycosides
of Gewurztraminer and other Alsatian white wines,
with regard to their role as aroma precursors and their
use as markers of wine variety, is under way and will
be reported later.
Suzuki, Y.; Yamaguchi, I.; Murofushi, N.; Takahashi, N.;
Sugawara, F.; Yoshida, S.; Nukada, T.; Ogawa, T. Synthesis
of benzyl 6-O-â-D-apiofuranosyl-â-D-glucopyranoside, a me-
tabolite of benzoic acid in Lemna paucicostata. Agric. Biol.
Chem. 1988, 52, 1261-1266.
Voirin, S. G.; Baumes, R. L.; Gu¨nata, Z. Y.; Bitteur, S. M.;
Bayonove, C. L. Analytical methods for monoterpene gly-
cosides in grapes and wine. I. XAD-2 extraction and gas
chromatographic-mass spectrometric determination of syn-
thetic glycosides. J . Chromatogr. 1992a , 590, 313-328.
Voirin, S. G.; Baumes, R. L.; Sapis, J .-C.; Bayonove, C. L.
Analytical methods for monoterpene glycosides in grapes
and wine. II. Qualitative and quantitative determination
of monoterpene glycosides in grape. J . Chromatogr. 1992b,
595, 269-281.
Williams, P. J .; Strauss, C. R.; Wilson, B.; Massy-Westropp,
R. A. Novel monoterpene disaccharide glycosides of Vitis
vinifera grapes and wines. Phytochemistry 1982, 21, 2013-
2020.
Williams, P. J .; Strauss, C. R.; Wilson, B.; Massy-Westropp,
R. A. Glycosides of 2-phenylethanol and benzyl alcohol in
Vitis vinifera grapes. Phytochemistry 1983, 22, 2039-2041.
Winterhalter, P.; Sefton, M. A.; Williams, P. J . Two-dimen-
sional GC-DCCC analysis of the glycoconjugates of mono-
terpenes, norisoprenoids, and shikimate-derived metabolites
from Riesling wine. J . Agric. Food Chem. 1990, 38, 1041-
1048.
ACKNOWLEDGMENT
We thank Roland Graff, Estelle Mastio, and Patrick
Wehrung for assistance in NMR and mass spectral
measurements and the referees for helpful comments.
LITERATURE CITED
Baltenweck-Guyot, R.; Trendel, J .-M.; Albrecht, P.; Schaeffer,
A. Mono- and diglycosides of (E)-6,9-dihydroxymegastigma-
4,7-dien-3-one in Vitis vinifera wine. Phytochemistry 1996,
43, 621-624.
Baltenweck-Guyot, R.; Trendel, J .-M.; Albrecht, P.; Schaeffer,
A. New hemiterpene glycosides in Vitis vinifera wine. J . Nat.
Prod. 1997, 60, 1326-1327.
Baumes, R.; Dugelay, I.; Gu¨nata, Y. Z.; Tapie´ro, C.; Bitteur,
S.; Bayonove, C. Analyse structurale de quelques glycosides
de raisin. In Connaissance Aromatique des Ce´pages et
Qualite´ des Vins; Bayonove, C., Crouzet, J ., Flanzy, C.,
Martin, J . C., Sapis, J . C., Eds.; Revue Franc¸aise
d’Œnologie: Be´ziers, France, 1993; pp 90-98.
Gu¨nata, Y. Z.; Bayonove, C. L.; Baumes, R. L.; Cordonnier, R.
E. The aromas of grapes. I. Extraction and determination
of free and glycosidically bound fractions of some grape
aroma components. J . Chromatogr. 1985, 331, 83-90.
Lutz, A.; Winterhalter, P.; Schreier, P. The structure of a new
ionone glucoside from starfruit (Averrhoa carambola L.).
Nat. Prod. Lett. 1993, 3, 95-99.
Marinos, V. A.; Tate, M. E.; Williams, P. J . Lignan and
phenylpropanoid glycerol glucosides in wine. Phytochemistry
1992, 31, 4307-4312.
Mba¨ıraroua, O.; Ton-That, T.; Tapie´ro, C. Synthe`ses de 6-O-
â-D-apiofuranosyl-â-D-glucopyranosides de monoterpe´nyle.
Carbohydr. Res. 1994, 253, 79-99.
Received for review March 1, 2000. Revised manuscript
received August 24, 2000. Accepted August 25, 2000. We thank
the Re´gion Alsace and the Comite´ Interprofessionnel des Vins
d’Alsace for financial support.
J F0002600