Monoterpenoids and their glycosides from the leaf of thyme

Article abstract of DOI:10.1016/j.phytochem.2004.09.010

Seven monoterpenoid glycosides, including thymuside A, were isolated from the methanol extract of thyme leaf. From the polar portion of the methanol extract of thyme (leaf of Thymus vulgaris; Labiatae), which has been used as an important stomachic, carminative, a component of prepared cough tea, and a spice, seven monoterpenoid glycosides were isolated together with two known monoterpenoids and three known monoterpenoid glucosides. Structures of the seven monoterpenoid glycosides were determined by spectral analysis.

Full text of DOI:10.1016/j.phytochem.2004.09.010

PHYTOCHEMISTRY
Phytochemistry 65 (2004) 3279–3287
www.elsevier.com/locate/phytochem
Monoterpenoids and their glycosides from the leaf of thyme
*
Junichi Kitajima , Toru Ishikawa, Atushi Urabe, Mitsuru Satoh
Showa Pharmaceutical University, Higashi-Tamagawagakuen 3, Machida, Tokyo 194-8543, Japan
Received 26 March 2004; received in revised form 3 September 2004
Abstract
From the polar portion of the methanol extract of thyme (leaf of Thymus vulgaris; Labiatae), which has been used as an impor-
tant stomachic, carminative, a component of prepared cough tea, and a spice, seven monoterpenoid glycosides were isolated
together with two known monoterpenoids and three known monoterpenoid glucosides. Structures of the seven monoterpenoid gly-
cosides were determined by spectral analysis.
ꢀ 2004 Elsevier Ltd. All rights reserved.
Keywords: Thyme; Thymus vulgaris; Labiatae; Monoterpenoid; p-Menthane; Monoterpenoid glucoside; Thymuside A
1. Introduction
leaves, and this paper discuss the isolation and charac-
terization of compounds 1–12, of which six were new.
Thyme [Thymus vulgaris; Labiatae] is indigenous to
central and southern Europe, and is now widely culti-
vated as a tea, spice, and herbal medicine. Its leaf is
listed in the German and British Herbal Pharmacopoeia,
and has been used as a stomachic, carminative, diuretic,
urinary disinfectant, and vermifuge. Thyme leaf tea is
said to promote rest and sleep (Wichtl, 1994). As the
constituents of this leaf, an essential oil (1.0–2.5%) con-
taining mainly the isomeric monoterpenoid thymol (30–
70%) and carvacrol (3–15%), and other monoterpenoids
such as methylcarvacrol, p-cymene, a-phellandrene,
limonene, a-terpineol, terpinen-4-ol, 1,8-cineole, bor-
neol, bornyl acetate, etc., are known (Reddy et al.,
1998; Rustaiyan et al., 2000). However, no terpenoid
glycosides have been reported from the polar portion
of this leaf. In continuation of our studies on the polar
constituents of spices and herbal medicines (Kitajima
et al., 2003), we undertook an investigation of thyme
2. Results and discussion
The commercial thyme leaf tissue was extracted
with methanol, and the methanolic extract was sus-
pended in water and successively extracted with ether
and ethyl acetate. The aqueous layer was applied to
an Amberlite XAD-II column to give water and
methanol eluate fractions, respectively. The ethyl ace-
tate extract was subjected to Sephadex LH-20 chroma-
tography, and then to a combination of silica gel,
Lobar RP-8 column chromatography and HPLC to
isolate four monoterpenoid glycosides (2, 3, 4 and
10). The methanol eluate fraction was applied to a
Sephadex LH-20 column, and then subjected to a
combination of silica gel, Lobar RP-8 column chroma-
tography and HPLC to isolate two monoterpenoids (1
and 9), and eight monoterpenoid glycosides (2, 5–8,
and 10–12). Among them, 2–6 and 11 are new, and
10 is a newly isolated compound. All new glucosides
*
Corresponding author. Tel.: +81 427 21 1577; fax: +81 427 21
described in this paper are b-
D
-glucopyranosides as
1588.
1
shown by their H and 13C NMR spectral data (Tables
E-mail address: kitajima@ac.shoyaku.ac.jp (J. Kitajima).
0031-9422/$ - see front matter ꢀ 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.phytochem.2004.09.010

3280
J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
1–3), and this was confirmed by hydrolysis to yield
D
-
of the [M + H]+, [M + Na]+ or [M + K]+ ion peak in
the high-resolution positive FAB-MS.
Monoterpenoid 1 (C10H18O2, an amorphous powder,
glucose and comparison of the [M]D values with those
of their aglycones (Klyne, 1950). Their molecular for-
mulae were suggested from the accurate mass number
24
½aꢀ ¼ ꢁ22ꢂ) was suggested to be p-menth-4(5)-ene-1,2-
D
Table 1
H NMR chemical shifts of 1-6 and 6a (in pyridine-d5, 500 MHz)a
1
1
2
3
4
H-2ax
H-3ax
4.21 dd (5.5, 9.0)
2.31 dd (9.0, 17.0)
2.69 dd (5.5, 17.0)
5.38 dd (2.5, 4.5)
2.52 br d (17.5)
2.48 dd (4.5, 17.5)
1.54 s
4.32 dd (5.0, 10.0)
2.25 dd (10.0, 17.0)
2.58 dd (5.0, 17.0)
5.23 dd (2.5, 5.0)
2.74 br d (17.5)
2.35 dd (5.0, 17.5)
1.54 s
4.32 dd (5.0, 10.0)
2.25 dd (10.0, 17.0)
2.62 dd (5.0, 17.0)
5.32 dd (2.5, 5.0)
2.70 br d (17.0)
2.42 dd (5.0, 17.0)
1.52 s
4.13 dd (5.0, 9.0)
2.19 dd (9.0, 17.0)
2.64 dd (5.0, 17.0)
5.29 dd (2.5, 4.0)
2.44 br d (17.0)
2.39 dd (4.0, 17.0)
1.57 s
eq
H-5
H-6ax
eq
H3-7
H-8
2.19 septet (7.0)
0.99 d (7.0)
0.99 d (7.0)
2.13 septet (6.5)
0.95 d (6.5)
0.95 d (6.5)
2.16 septet (7.0)
0.97 d (7.0)
0.97 d (7.0)
2.16 septet (7.0)
0.98 d (7.0)
0.98 d (7.0)
H3-9
H3-10
Glc H-1
Glc H-2
Glc H-6a
b
5.19 d (8.0)
5.15 d (7.5)
5.24 d (8.0)
4.06 dd (8.0, 8.0)
4.36 dd (6.0, 12.0)
4.56 dd (2.5, 12.0)
2.02 s
4.03 dd (7.5, 8.0)
4.75 dd (6.5, 11.0)
4.95 dd (2.0, 11.0)
2.13 s
5.54 dd (8.0, 8.0)
4.28 dd (6.0, 11.5)
4.55 dd (2.0, 11.5)
OAc
5
6
6a
H-2ax
H-3ax
eq
4.93 dd (7.5, 10.0)
2.06 m
2.06 m
H-2ax
H-3
4.83 br s
5.60 br s
2.03 m
4.59 br s
5.77 br s
H-5ax
eq
2.10 ddd (3.0, 8.0, 17.0)
2.23 ddd (5.0, 5.5, 17.0)
1.98 ddd (5.5, 8.0, 13.0)
2.05 ddd (3.0, 5.0, 13.0)
1.62 s
H-5
5.16 br d (5.0)
2.45 br d (16.5)
2.21 dd (5.0, 16.5)
1.64 s
2.03 m
2.02 m
H-6ax
eq
H3-7
H-6ax
eq
2.02 m
1.58 s
H3-7
H-8
H-8
H3-9
2.08 septet (6.5)
0.87 d (6.5)
0.85 d (6.5)
2.11 septet (7.0)
0.93 d (7.0)
0.93 d (7.0)
5.29 d (7.5)
2.18 septet (7.0)
0.99 d (7.0)
0.99 d (7.0)
H3-9
H3-10
Glc H-1
H3-10
sug H-1
sug H-3
sug H-4
sug H-5
sug H-6a
b
5.32 s
4.27 d (9.5)
–
4.46 dd (9.5, 9.5)
3.89 ddd (2.5, 6.0, 9.5)
4.42 dd (6.0, 11.5)
4.60 dd (2.5, 11.5)
a
d in ppm from TMS [coupling constants (J) in Hz are given in parentheses].
Table 2
1H NMR chemical shifts of 10 and 11 (in pyridine-d5, 500 MHz)a
10
11
H-2exo
H-3endo
exo
4.41ddd (2.0, 3.0, 9.0)
1.48 br dd (3.0, 13.0)
2.25 ddd (4.5, 9.0, 13.0)
1.58 br dd (4.5, 4.5)
1.29 br ddd (4.0, 9.0, 13.0)
1.65 dddd (4.5, 9.0, 13.0, 13.0)
2.40 ddd (4.5, 9.0, 13.0)
1.20 dddd (2.0, 4.0, 13.0, 13.0)
0.79 s
4.63 ddd (2.0, 3.0, 9.0)
1.63 br dd (3.0, 13.0)
2.52 ddd (4.5, 9.0, 13.0)
2.20 br dd (4.5, 4.5)
1.40 br ddd (4.5, 9.0, 13.0)
1.74 dddd (4.5, 9.0, 13.0, 13.0)
2.49 ddd (4.5, 9.0, 13.0)
1.31 dddd (2.0, 4.5, 13.0, 13.0)
1.21 s
H-4
H-5endo
exo
H-6endo
exo
H3-8
H3-9
H2-9
0.77 s
–
–
3.66 d (11.0)
–
1.01 s
4.88 d (7.5)
3.92 d (11.0)
1.17 s
4.90 d (8.0)
H3-10
Glc H-1
a
d in ppm from TMS [coupling constants (J) in Hz are given in parentheses].

J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
3281
Table 3
13C NMR chemical shifts of 1–6, 6a, 10 and 11 (in pyridine-d5, 125 MHz)a
1
2 (d 2–1)
3 (d 3–2)
4 (d 4–2)
5
6 (d 6–6a)
6a
71.83
10
49.44
11
C-1
C-2
71.61
73.86
34.33
80.02 (+8.4)
69.68 (ꢁ4.2)
33.71
80.14
69.82
33.61
79.66
71.91
33.36
73.18
70.68
30.46
79.96 (+8.1)
72.95 (ꢁ1.8)
123.64
54.47
83.97
36.33
42.27
28.67
27.99
49.96
15.11
64.38
14.80
103.71
75.30
78.66
71.74
78.31
62.84
74.78
123.55
144.25
25.45
83.79
36.42
45.25
28.54
27.14
48.23
19.32
18.97
13.96
103.70
75.32
78.68
71.79
78.37
62.91
C-3
C-4
141.58
117.35
39.99
141.44
117.30
141.51
117.17
36.71
141.30
116.06
36.59
142.22
117.16
39.77
20.42
34.92
21.05
21.62
143.45
25.68
C-5
C-6
36.83 (ꢁ3.2)
18.62 (ꢁ2.9)
34.65
21.23b
21.66b
97.06
33.23 (ꢁ1.6)
17.02 (ꢁ5.6)
34.28
21.35b
21.53b
98.12
34.87
22.65
C-7
C-8
21.50
34.84
21.38b
21.66b
18.49
18.60
34.75
34.67
34.60
21.54b
21.65b
C-9
C-10
21.23b
21.63b
97.15
21.28
21.28
Glc C-1
Glc C-2
Glc C-3
Glc C-4
Glc C-5
Glc C-6
OAc
96.11 (ꢁ1.0)
75.46 (ꢁ0.1)
76.42 (ꢁ2.4)
71.91
75.52
78.84
75.31
78.57
71.61
75.56
78.85
71.71
78.61
62.73
72.13
78.42
62.96
75.08 (ꢁ3.5)
64.81 (+2.1)
20.75
78.47
62.47
21.32
169.93
170.81
Sug C-1
Sug C-2
Sug C-3
Sug C-4
Sug C-5
Sug C-6
a
97.85
94.08
80.31
69.74
78.51
63.13
d in ppm TMS.
b
Assignments may be interchanged in each column.
1
diol from its H, 13C NMR spectral data (Tables 1 and
3), and the results of heteronuclear multiple bond con-
nectivity (HMBC) spectral data (see Section 3). The ob-
served NOE interactions between H-2/H-6ax, and
between H3-7/H-3ax in its nuclear Overhauser and ex-
change spectroscopy (NOESY) suggested that the con-
figuration of C-1 methyl was axial, and C-1, C-2
hydroxyls were equatorial (Fig. 1).
Consequently, 1 was found to be p-menth-4(5)-ene-
1b,2a-diol, which was reported as a constituent of the
Fig. 1. Structures of 1–12, and NOE interactions observed in the NOESY spectra of 1, 5, 6 and 10.

3282
J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
carbonyl carbon of the acetoxyl group. So, 3 was deter-
mined to be (1R,2R)-p-menth-4(5)-ene-1,2-diol 1-O-b-
D
-(6-O-acetyl)-glucopyranoside.
Glucoside 4 (C18H30O8, an amorphous powder,
22
½aꢀ ¼ ꢁ32ꢂ) showed [M + Na]+ and [M + H]+ ion
D
peaks at m/z 397 and 375, and an [M ꢁ C8H14O7 + H]+
ion peak at m/z 153 in the positive FAB-MS. Since 4 was
deacetylated to 2 by alkaline hydrolysis, it was also a
monoacetate of 2. The position of the acetoxyl group
was revealed to be C-2 of the glucose moiety by the
downfield shift of H-2 (by 1.48 ppm) and the upfield
shift of C-1 (by 1.0 ppm) and C-3 (by 2.4 ppm) signals
of the glucosyl moiety (Yamazaki et al., 1977; Konishi
et al., 1978), and the HMBC correlation between the
glucosyl H-2 and the carbonyl carbon of the acetoxyl
group. Then, 4 was characterized as (1R,2R)-p-menth-
Fig. 2. Dd Values in Hz [=dS-(ꢁ) ꢁ dR-(+)] of 1-MTPA and 6a-
MTPA ester.
steam distilled oil of Ferula jaeschkeana rhizome (Garg
and Agarwal, 1988). Since the absolute structure of 1
was not previously defined, its absolute configuration
at C-2 was investigated by means of the modified
MosherÕs method (Ohtani et al., 1991; Kitajima et al.,
2002). Treatment of 1 with S-(ꢁ)- and R-(+)-2-meth-
oxy-2-trifluoromethyl acetic acid (MTPA) chloride gave
the 1-O-ester of S-(ꢁ)- MTPA (1-S) and R-(+)-MTPA
(1-R), respectively. Signals due to protons on C-3, C-8,
C-9 and C-10 of 1-S appeared at lower field than those
of 1-R, while the proton signals due to C-6 and C-7 of 1-
S were observed at higher field than those of 1-R (Fig.
2). Therefore, the absolute configuration at C-2 of 1 is
assigned as R, and the absolute structure of 1 was de-
scribed as shown in Fig. 1 [(1R,2R)-p-menth-4(5)-ene-
4(5)-ene-1,2-diol 1-O-b-D-(2-O-acetyl)-glucopyranoside.
Thymuside A (5, C16H26O7, an amorphous powder,
22
½aꢀ ¼ þ2ꢂ) showed [M + Na]+, [M ꢁ H2O ꢁ H]+ and
D
[M ꢁ C6H12O7 + H]+ ion peaks at m/z 353, 313 and
135 in the positive FAB-MS, and [M ꢁ H]ꢁ and
[M ꢁ H2O + H]ꢁ ion peaks at m/z 329 and 311 in the
negative FAB-MS. The 1H, 13C and 13C–1H COSY
NMR spectral data of 5 (Tables 1 and 3) showed the
presence of a monoterpenoid which was characterized
to be p-menth-4(5)-ene-1,2-diol, and a sugar moiety
which corresponded to hydroxyhexose. From compari-
son of the NMR spectral data with those of 1 and 2,
and analysis of its HMBC spectral data (see Section
3), 5 was concluded to be composed of p-menth-4(5)-
ene-1,2-diol and 2-hydroxyhexose, which were linked
with two ether linkages. As the HMBC correlations be-
tween H0-1 of the sugar moiety and C-1 of the mono-
terpenoid, and between H-2 of the monoterpenoid and
C0-2 of the sugar moiety were observed, the monoterpe-
noid and 2-hydroxyhexose should be linked between C-1
and C0-1, and between C-2 and C0-2, respectively. The
NOE interaction between H3-7/H-3ax, and between H-
2/H-6ax confirmed that the configuration of C-1 and
C-2 hydroxyl was equatorial, the same as that of 1,
and the NOE interactions between H0-1/H0-3, H0-5 of
the sugar moiety in its NOESY spectrum suggested that
the configuration of H0-1, H0-3 and H0-5 was axial, the
1,2-diol].
24
D
Glucoside 2 (C16H28O7, m.p. 137–139 ꢁC, ½aꢀ
¼
ꢁ42ꢂ) showed [M + Na]+ and [M + H]+ ion peaks at
m/z 355 and 333, and an [M ꢁ C6H12O6 + H]+ ion peak
at m/z 153 in the positive FAB-MS. Enzymatic hydroly-
sis of 2 with b-glucosidase gave 1 and
D-glucose, and 2
was suggested to be a monoglucoside of 1. The position
of the glucosyl unit was found to be C-1 by the observed
cross-peak between Glc H-1/C-1 in the HMBC spec-
trum. Therefore, 2 was characterized as (1R,2R)-p-
menth-4(5)-ene-1,2-diol 1-O-b-D-glucopyranoside. The
glycosylation shift values of the a- and the b-pro-S-side
carbons and the chemical shift of glucosyl C-1 are
shown in Table 3.
Glucoside 3 (C18H30O8, an amorphous powder,
24
½aꢀ ¼ ꢁ38ꢂ) showed [M + Na]+, [M + H]+ and
same as that of methyl b-D-glucopyranoside. Further-
D
[M ꢁ C8H14O7 + H]+ ion peaks at m/z 397, 375 and
153 in the positive FAB-MS. It displayed similar NMR
spectral features to those of 2 except for the presence
of an acetoxyl group (Tables 1 and 3), and the IR spec-
trum of 2 showed an absorption due to an ester carbonyl
functionaly (1720 cmꢁ1). Glycoside 3 was deacylated to 2
by heating in a water bath with 5% NH4OH–MeOH for 2
h. From comparison of the NMR spectral data with
those of 2, 3 was the monoacetate of 2, and the position
of the acetoxyl group was revealed to be C-6 of the glu-
cose from the analysis of the HMBC spectrum, which
showed a cross-peak between the glucosyl H2-6 and the
more, the downfield shift of the H-2ax (by 0.60 ppm) sig-
nal to that in 2 suggested that the H-2ax and the free
hydroxyl group of C0-2 were in 1,3-diaxial relationship.
Therefore, 5 was concluded to be as shown in Fig. 1.
Glucoside 6 (C16H28O7, an amorphous powder,
22
½aꢀ ¼ ꢁ23ꢂ) showed [M + K]+, [M + Na]+ and
D
[M ꢁ C6H12O6 + H]+ ion peaks at m/z 371, 355 and
153 in the positive FAB-MS. Enzymatic hydrolysis of
6 with b-glucosidase gave an aglycone (6a; C10H18O2,
D-glucose. It
22
an amorphous powder, ½aꢀ ¼ ꢁ38ꢂ) and
D
1
showed similar H and 13C NMR spectral data (Tables
1 and 3) to those of 2, except for the chemical shifts

J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
3283
and coupling constants of the H-2ax and methylene pro-
tons. From the results of HMBC spectral data (see Sec-
tion 3), the structure of the aglycone moiety was
concluded to be p-menth-3-ene-1,2-diol, and the posi-
tion of the glucosyl unit was C-1. As the NOE interac-
tions between H-2/H-6ax, and between H3-7/H-6eq were
observed in its NOESY spectrum in CD3OD, the config-
uration of C-1 methyl and C-2 hydroxyl was suggested
to be axial and equatorial, respectively (Fig. 1). The
absolute structure of 6a was defined by the same way de-
scribed for 1. Treatment of 6a with S-(ꢁ)- and R-(+)-
MTPA chloride gave the 1-O-ester of S-(ꢁ)-MTPA
(6a-S) and R-(+)-MTPA (6a-R), respectively, and the
signals due to protons on C-3, C-9 and C-10 of 6a-S ap-
peared at lower field than those of 6a-R, while the pro-
ton signals due to C-6 and C-7 of 6a-S were observed at
higher field than those of 6a-R (Fig. 2). Therefore, the
absolute configuration at C-2 of 6a is assigned as R,
and the absolute structure of 6a was characterized as
(1R,2R)-p-menth-3-ene-1,2-diol. Therefore, 6 was char-
physical and NMR spectral data with those reported
(Orihara and Furuya, 1993).
Glucoside 11 (C16H28O7, an amorphous powder,
23
½aꢀ ¼ ꢁ48ꢂ) showed [M + K]+, [M + Na]+, [M + H]+
D
and [M ꢁ C6H12O6 + H]+ ion peaks at m/z 371, 355,
333 and 153 in the positive FAB-MS. It showed similar
NMR spectral features with those of 10 except the ab-
sence of one tert-methyl and the presence of a hydroxyl-
ated methylene group (Tables 2 and 3). Enzymatic
hydrolysis of 11 gave an aglycone 11a (C10H18O2, an
D-glucose, and
23
D
amorphous powder, ½aꢀ ¼ ꢁ16ꢂ) and
the NMR spectra of 11a were identical with those of
vicodiol first isolated from Vicoa indica (Vasanth
et al., 1990). The position of the glucosyl unit was con-
firmed to be C-2 by the HMBC analysis (see Section 3).
As the vicodiol, which has the 1S, 2R, 4S, 7R configura-
tion, showed a minus [a]D value (ꢁ16ꢁ) the same as 11a,
11 was concluded to have the 1S, 2R, 4S, 7R configura-
tion. It was also supported by comparison of the 13C
NMR glycosylation shift values [Dd(d glucoside ꢁ d
aglycone)] of the a- and b-carbon and the chemical shift
of Glc C-1 [C-1 (b-pro-R), Dd +0.2; C-2 (a-carbon), Dd
+7.7; C-40 (b-pro-S), Dd-3.2, glucusyl C-1, d 103.71] with
those of 10. So, 11 was characterized as (1S, 2R, 4S,
acterized as (1R,2R)-p-menth-3-ene-1,2-diol 1-O-b-D-
glucopyranoside.
25
D
Glucosides 7 (C16H24O7, m.p. 178–180 ꢁC, ½aꢀ
25
¼
ꢁ45ꢂ), and 8 (C16H24O7, an amorphous powder,
½aꢀ ¼ ꢁ39ꢂ) were identified as 6-hydroxythymol 6-O-
7R)-vicodiol 2-O-b-D-glucopyranoside.
Glucoside 12 (C16H28O7, an amorphous powder,
D
b-
glucopyranoside, respectively, which were isolated from
D
-glucopyranoside and 6-hydroxythymol 3-O-b-
D
-
22
D
½aꢀ ¼ ꢁ32ꢂ) was identified as (1S, 2R, 4S, 5R)-angelico-
the fruit of ajowan (Ishikawa et al., 2001b). Monoterp-
idenol 2-O-b-D-glucopyranoside which was isolated
from the root and rhizoma of Glehenia littoralis (Kitaj-
ima et al., 1998).
23
D
enoid 9 (C10H20O3, m.p. 134–135 ꢁC, ½aꢀ ¼ ꢁ31ꢂ) was
identified as (1S,2S,4R)-p-menthane-1,2,8-triol which
was isolated from the fruit of Glehenia littoralis (Ish-
ikawa et al., 2001a).
The ingredient relationship between the essential oil
and the polar portion was confirmed by the isolation
of these glycosides which showed the biosynthetic rela-
tion to thymol, carvacrol, a-terpineol and borneol. It
is worthy of note that, a novel glycoside thymuside A,
which is linked a monoterpenoid and a hydroxyhexose
with two ether linkages, was isolated from the leaf of
thyme.
Glucoside 10 (C16H28O6, m.p. 126–130 ꢁC,
24
½aꢀ ¼ ꢁ53ꢂ) showed [M + Na]+, [M + H]+ and
D
[M ꢁ C6H12O6 + H]+ ion peaks at m/z 339, 317 and
137 in the positive FAB-MS. Its 1H and 13C NMR spec-
tral data (Tables 2 and 3) revealed the presence of three
tert-methyls, three methylenes, one methine, two quater-
nary carbons, and one hydroxylated methine, in addi-
tion to the b-D-glucopyranosyl moiety. From analysis
of the HMBC spectrum (see Section 3), the aglycone
of 10 was clarified as borneol and the location of the
glucosyl group was C-2. The observed NOE interactions
between H-2/H3-9 suggested the configuration of C-2
hydroxyl was endo (Fig. 1). Enzymatic hydrolysis of
3. Experimental
3.1. General
Melting points were determined on a Yanagimoto
micromelting point apparatus and are uncorrected.
Optical rotations were measured on a JASCO DIP-370
digital polarimeter. IR spectra were obtained with a
JASCO A-103 IR spectrophotometer. FAB-MS were re-
corded with a JEOL HX-110 spectrometer using glyc-
10 gave an aglycone 10a (C10H18O2, m.p. 205–209 ꢁC,
24
D
½aꢀ ¼ ꢁ33ꢂ) and
D-glucose, and the absolute configura-
tion at C-2 of 10a was indicated to be R by the empirical
rule of 13C NMR glycosylation shift (Kasai et al., 1977;
Tori et al., 1977), [C-1 (b-pro-R), Dd ꢁ0.5; C-2 (a-car-
bon), Dd +7.7; C-40 (b-pro-S), Dd ꢁ3.1, glucosyl C-1,
d 103.70]. Further, glucoside 10 was identified as
erol as matrix. H and 13C NMR spectra were taken
1
on JEOL A-500 spectrometers with tetramethylsilane
as an internal standard, and chemical shifts were re-
corded in d value. CC was carried out under TLC mon-
itoring using Kieselgel 60 (70–230 mesh, Merck),
Sephadex LH-20 (25–100 lm, Pharmacia), Lobar RP-8
(1S,2R,4S)-borneol b-D-glucopyranoside, which was re-
ported as a biotransformation product from a cell sus-
pension culture of Eucalyptus perriniana following
administration of (ꢁ)-borneol, by comparison of its

3284
J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
column (Merck) and Amberlite XAD-II (Organo),
respectively. TLC was performed on silica gel (Merck
5721) and spots were detected with p-anisaldehyde–
H2SO4 reagent. HPLC separation was carried out with
tions (frs. I5–1–I5–20). Fraction I5–12 was subjected to
HPLC [ODS, MeCN–H2O (1:1)] to give 6 (20 mg),
and fr. I5–15 was further purified by HPLC [CHA,
MeCN–H2O (19:1)] to give 2 (440 mg). Fraction I6
(5.79 g) was passed through a Lobar RP-8 column
[MeCN–H2O (3:17)] to give 15 fractions (frs. I6–1–I6–
15), and fr. I6–5 was subjected to HPLC [CHA,
MeCN–H2O (19:1)] to give 11 (117 mg) and 12 (7 mg).
The following compounds were identified by compar-
ison with authentic compounds: 6-hydroxythymol
Symmetryprep C18
7 lm [Waters; column size,
7.8 · 300 mm; ODS] and carbohydrate analysis [Waters;
column size, 3.9 · 300 mm; CHA] columns.
3.2. Extraction and separation
Commercial thyme leaves (from Thymus vulgaris L.;
purchased from Asaoka Spices Ltd., Lot. No.
00011801; 2.0 kg) were extracted with MeOH (5 L · 2)
for two weeks at room temperature, and the extract
(419.1 g) was partitioned into Et2O–H2O and EtOAc–
H2O extracts, respectively. Removal of the solvents
from each phase gave Et2O (140.3 g), EtOAc (20.9 g)
and aqueous (257.9 g) fractions.
The EtOAc fraction was subjected to Sephadex LH-
20 (MeOH) chromatography to give seven fractions
(frs. A–G). Fraction C (2.48 g) was applied to a silica
gel column [CHCl3–MeOH–H2O (17:3:0.2) ! MeOH]
to give nine fractions (frs. C1–C9). Fraction C4 (0.40 g)
was passed through a Lobar RP-8 column [MeCN–
H2O (3:2)] to give eight fractions (frs. C4–1–C4–8). Frac-
tion C4–2 was subjected to silica gel CC [CHCl3–MeOH
(19:1)] to give four fractions (frs. C4–2–1–C4–2–4), and fr.
6-O-b-
D
D
-glucopyranoside (7), 6-hydroxythymol 3-O-b-
-glucopyranoside (8), (1S,2S,4R)-p-menthane-1,2,8-
triol (9) and (1S,2R,4S,5R)-angelicoidenol 2-O-b-
glucopyranoside (12).
D-
3.3. (1R,2R)-p-menth-4(5)-ene-1,2-diol (1)
24
Amorphous powder, ½aꢀ ¼ ꢁ22ꢂ (c = 0.6, MeOH).
D
Positive FAB-MS m/z: 209.0935 [M + K]+ (Calcd for
C10H18KO2; 209.0944), 153 [M ꢁ H2O + H]+ (base),
135 [M ꢁ 2H2O + H]+ (base). For 1H NMR and 13C
NMR spectral data, see Tables 1 and 3. HMBC correla-
tions: H-2ax/C-1, C-3, C-4, C-6, C-7; H-3ax/C-1, C-2,
C-4, C-5, C-8; H-3eq/C-1, C-2, C-4, C-5, C-8; H-5/C-1,
C-3, C-4, C-6, C-8; H-6ax/C-1, C-2, C-4, C-5, C-7; H-
6eq/C-1, C-2, C-4, C-5, C-7; H3-7/C-1, C-2, C-6; H-8/
C-3, C-4, C-5, C-9, C-10; H3-9/C-4, C-8, C-10; H3-10/
C-4, C-8, C-9. NOE correlations: H-2ax/H-6ax; H-3ax/
H3-7, H3-10; H-3eq/H3-10; H-5/H-8, H3-9; H-6eq/H3-7.
C
4–2–1 was applied to HPLC [CHA, MeCN–H2O (49:1)]
to give 3 (40 mg) and 4 (11 mg). Fraction C6 (0.30 g) was
passed through a Lobar RP-8 column [MeCN–H2O
(3:2)] to give 23 fractions (frs. C6–1–C6–23), and fr.
C6–20 was subjected to HPLC [ODS, MeOH–H2O (3:2)
to give 10 (120 mg). Fraction C7 (0.11 g) was passed
through a Lobar RP-8 column [MeCN–H2O (3:7)] to
give 2 (75 mg).
3.4. 2-O-MTPA ester of 1
A solution of 1 (2 mg) in a mixture of CHCl3 and
pyridine (each 0.5 ml) was treated with R-(ꢁ)-MTPA-
chloride (5 drops) and the mixture was stirred for 2 h
at room temperature. The reaction mixture was
extracted with EtOAc, and the EtOAc extract was sub-
jected to silica gel column chromatography (hexane–
EtOAc = 4:1) to give 1-O-S-(ꢁ)-MTPA ester of 1-S
(1a-S-(ꢁ)-MTPA) In a similar procedure, 1-R (1a-R-
(+)-MTPA) was obtained with S-(+)-MTPA-chloride.
The aqueous fraction was applied to an Amberlite
XAD-II column (H2O ! MeOH) to give a water
(106.6 g) and MeOH eluate (143.4 g) fractions. The
MeOH eluate fraction was subjected to Sephadex LH-
20 (MeOH) chromatography to give five fractions
(frs. H–L). Fraction I (124.31 g) was applied to silica
gel CC [CHCl3–MeOH–H2O (17:3:0.2 ! 4:1:0.1 !
7:3:0.5) ! MeOH] to give 13 fractions (frs. I1–I13). Frac-
tion I3 (6.74 g) was passed through a Lobar RP-8 col-
umn [MeCN–H2O (3:17 ! 3:7)] to give 25 fractions
(frs. I3–1–I3–25), and fr. I3–4 was subjected to HPLC
[ODS, MeOH–H2O (3:37)] to give 9 (9 mg). Fraction
I3–20 was applied to HPLC [CHA, MeCN–H2O (19:1)]
to give 5 (20 mg), and fr.I3–22 was subjected to HPLC
[ODS, MeOH–H2O (3:2)] to give 1 (9 mg) and 10 (105
mg). Fraction I4 (5.51 g) was passed through a Lobar
RP-8 column [MeCN–H2O (3:17 ! 3:7)] to give 21 frac-
tions (frs. I4–1–I4–21), and fr. I4–6 was subjected to HPLC
[ODS, MeCN–H2O (3:17)] to give 8 (2 mg) and 7 (123
mg). Fraction I5 (4.30 g) was passed through a Lobar
RP-8 column [MeCN–H2O (3:17 ! 3:7)] to give 20 frac-
3.5. (1R,2R)-p-menth-4(5)-ene-1,2-diol 1-O-b-
glucopyranoside (2)
D-
Colorless needles (MeOH), m.p. 137–139 ꢁC,
24
½aꢀ ¼ ꢁ42ꢂ (c = 2.0, MeOH). Positive FAB-MS m/z:
D
355 [M + Na]+, 333.1918 [M + H]+ (Calcd for
C16H29O7; 333.1914), 315 [M ꢁ H2O + H]+, 153
[M ꢁ C6H12O6 + H]+ (base). For 1H NMR and 13C
NMR spectral data, see Tables 1 and 3. HMBC correla-
tions: H-2ax/C-1, C-3, C-4, C-6, C-7; H-3ax/ C-1, C-2,
C-4, C-5, C-8; H-3eq/C-1, C-2, C-4, C-5, C-8; H-5/C-1,
C-3, C-4, C-6, C-8; H-6ax/C-1, C-2, C-4, C-5, C-7; H-
6eq/C-1, C-2, C-4, C-5, C-7; H3-7/C-1, C-2, C-6;

J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
3285
H-8/C-3, C-4, C-5, C-9, C-10; H3-9/C-4, C-8, C-10; H3-
10/C-4, C-8, C-9; Glc H-1/C-1.
C-8, C-10; H3-10/C-4, C-8, C-9; Glc H-1/C-1; Glc H-2/
OCOCH3; OCOCH3/OCOCH3.
3.10. Alkaline hydrolysis of 4
3.6. Enzymatic hydrolysis of 2
Glucoside 4 (2 mg) was deacetylated in the same way
as described for 3 to afford 2 (1 mg).
A mixture of 2 (100 mg) and b-glucosidase (5 mg,
Toyobo Co. Ltd., Lot 52275) in water (5 ml) was shaken
in a water bath at 37 ꢁC for 15 d. The mixture was con-
centrated in vacuo to dryness and the residue was sub-
jected to silica gel CC [CHCl3–MeOH (9:1 ! 1:1)] to
afford 1 (45 mg) and a sugar fraction. The sugar fraction
was passed through Sephadex LH-20 (MeOH) to give a
syrup, and HPLC [carbohydrate analysis (Waters),
detector; JASCO RI-930 detector and JASCO OR-990
chiral detector, solv.; MeCN–H2O (17:3), 2 ml/min; t
3.11. Thymuside A (5)
22
Amorphous powder, ½aꢀ ¼ þ2ꢂ (c = 0.5, MeOH).
D
Positive FAB-MS m/z: 353.1565 [M + Na]+ (Calcd for
C16H26NaO7; 353.1577), 313 [M ꢁ H2O + H]+ (base),
135 [M ꢁ C6H12O6 + H]+. Negative FABMS m/z: 329
[M ꢁ H]+
(base),
311
[M ꢁ H2O ꢁ H]+,
293
[M ꢁ 2H2OH]+. For H NMR and 13C NMR spectral
data, see Tables 1 and 3. HMBC correlations: H-2ax/
C-1, C-3, C-6, C-7, Sug C-2; H-3ax/C-1, C-2, C-4, C-5;
H-3eq/C-1, C-2, C-4, C-5; H-5/C-1, C-3, C-6, C-8;
H-6ax/C-1, C-4, C-5; H-6eq/C-1, C-2, C-4, C-5, C-7;
H3-7/C-1, C-2, C-6; H-8/C-3, C-4, C-5, C-9, C-10; H3-
9/C-4, C-8, C-10; H3-10/C-4, C-8, C-9; Sug H-1/C-1,
Sug C-2, Sug C-5; Sug H-3/Sug C-2, Sug C-4, Sug C-
5; Sug H-4/Sug C-3, Sug C-5, Sug C-6; Sug H-5/Sug
C-1, Sug C-3, Sug C-4, Sug C-6; Sug H2-6/Sug C-4,
Sug C-5. NOE correlations: H-2ax/H-6ax; H-3ax/H3-7,
H3-10; H-3eq/H3-10; H-5/H-8, H3-9; H-6eq/H3-7; Sug
H-1/Sug H-3, Sug H-5; Sug H-3/Sug H-5.
1
R 4.50 min (same location as that of
lished the presence of -glucose.
D-glucose)] estab-
D
3.7. (1R,2R)-p-menth-4(5)-ene-1,2-diol 1-O-b-
acetyl) glucopyranoside (3)
D-(6-O-
24
Amorphous powder, ½aꢀ ¼ ꢁ38ꢂ (c = 0.4, MeOH).
D
IRmmax nujol cmꢁ1: 3350 (OH), 1720 (acetoxyl). Positive
FAB-MS m/z: 397 [M + Na]+, 375.2029 [M + H]+
(Calcd for C18H31O8; 375.2019), 153 [M ꢁ C8H14O7 +
1
H]+ (base). For H NMR and 13C NMR spectral data,
see Tables 1 and 3. HMBC correlations: H-2ax/C-1, C-
6, C-7; H-3ax/C-1, C-2, C-4, C-5, C-8; H-3eq/C-1, C-2,
C-4, C-5, C-8; H-5/C-1, C-3, C-4, C-6, C-8; H-6ax/C-1,
C-2, C-4, C-5, C-7; H-6eq/C-1, C-2, C-4, C-5, C-7; H3-
7/C-1, C-2; H-8/C-3, C-4, C-5, C-9, C-10; H3-9/C-4, C-
8, C-10; H3-10/C-4, C-8, C-9; Glc H-1/C-1; Glc H-6a/
OCOCH3; Glc H-6b/OCOCH3; OCOCH3/OCOCH3.
3.12. (1R,2R)-p-menth-3-ene-1,2-diol 2-O-b-
glucopyranoside (6)
D-
22
Amorphous powder, ½aꢀ ¼ ꢁ23ꢂ (c = 1.7, MeOH).
D
Positive FAB-MS m/z: 371.1484 [M + K]+ (Calcd for
C16H28KO7; 371.1472), 355.1746 [M + Na]+ (Calcd for
C16H28NaO7; 355.1733), 153 [M ꢁ C6H12O6 + H]+
3.8. Alkaline hydrolysis of 3
1
(base). For H NMR (in pyridine-d5), see Table 1; (in
Glucoside 3 (5 mg) was deacetylated by heating in a
water bath with 5% NH4OH–MeOH for 6 hr. Glucoside
2 (3 mg) was obtained after Sephadex LH-20 (MeOH)
column chromatography.
CD3OD): 1.02 (6H, s, H3-9 and H3-10), 1.19 (3H, s,
H3-7), 1.73 (1H, ddd, J = 5.0, 10.5, 13.0 Hz, H-6ax),
1.88 (1H, ddd, J = 3.0, 5.0, 13.0 Hz, H-6eq), 2.09 (1H,
ddd, J = 3.0, 10.5, 17.0 Hz, H-5ax), 2.11 (1H, ddd,
J = 5.0, 5.0, 17.0 Hz, H-5eq), 2.21 (1H, septet. J = 7.0,
H-8), 4.28 (1H, br s, H-2ax), 5.22 (1H, br s, H-3), 4.58
(1H, d, J = 7.5 Hz, Glc H-1). For 13C NMR (in pyri-
dine-d5), see Table 3; (in CD3OD): 80.85 (C-1), 74.18
(C-2), 122.56 (C-3), 145.92 (C-4), 25.98 (C-5), 33.73
(C-6), 16.62 (C-7), 35.35 (C-8), 21.70, 21.87 (C-9, C-
10), 98.03 (Glc C-1), 75.29 (Glc C-2), 78.15 (Glc C-3),
71.94 (Glc C-4), 77.71 (Glc C-5), 62.85 (Glc C-6).
HMBC correlations: H-2ax/ C-1, C-3, C-4, C-7; H-3/
C-1, C-5, C-8; H-5ax/C-1, C-3, C-4, C-6; H-5eq/C-1,
C-3, C-4, C-6; H-6ax/C-1, C-2, C-4, C-5, C-7; H-6eq/
C-1, C-2, C-4, C-5, C-7; H3-7/C-1, C-2, C-6; H-8/C-3,
C-4, C-5, C-9, C-10; H3-9/C-4, C-8, C-10; H3-10/C-4,
C-8, C-9; Glc H-1/C-1. NOE correlations:
3.9. (1R,2R)-p-Menth-4(5)-ene-1,2-diol 1-O-b-
acetyl) glucopyranoside (4)
D-(2-O-
22
Amorphous powder, ½aꢀ ¼ ꢁ32ꢂ (c = 0.4, MeOH).
D
IRmmax cmꢁ1: 3350 (OH), 1720 (acetoxyl). Positive
FAB-MS m/z: 397 [M + Na]+, 375.2002 [M + H]+
(Calcd for C18H31O8; 375.2019), 153 [M ꢁ C8H14O7 +
1
H]+ (base). For H NMR and 13C NMR spectral data,
see Tables 1 and 3. HMBC correlations: H-2ax/C-1, C-
3, C-4, C-6, C-7; H-3ax/C-2, C-4, C-5, C-8; H-3eq/C-1,
C-2, C-4, C-5; H-5/C-1, C-3, C-6, C-8; H-6ax/C-1, C-2,
C-4, C-5, C-7; H-6eq/C-1, C-2, C-4, C-5, C-7; H3-7/C-
1, C-2, C-6; H-8/C-3, C-4, C-5, C-9, C-10; H3-9/C-4,

3286
J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
H-2ax/H-6ax; H-3/H-8, H3-10; H-5ax/H3-7, H3-9; H-
5eq/H3-9; H-6eq/H3-7; H3-7/Glc H-1.
filtrate was subjected to silica gel CC [CHCl3–MeOH
D-
(6:4)] to give a sugar fraction which was detected as
glucose as described for 2.
3.13. Enzymatic hydrolysis of 6
3.18. (1S,2R,4S,7R)-vicodiol 2-O-b-
(11)
D-glucopyranoside
A mixture of 6 (10 mg) and b-glucosidase (3 mg) in
water (5 ml) was shaken in a water bath at 37 ꢁC for 7
d. The mixture was applied to a silica gel column
[CHCl3–MeOH (9:1 ! 1:1)] to afford 6a (5 mg) and a
23
Amorphous powder, ½aꢀ ¼ ꢁ48ꢂ (c = 1.7, MeOH).
D
Positive FAB-MS m/z: 371.1485 [M + K]+ (Calcd for
sugar fraction. From the sugar fraction,
D
-glucose was
C16H28KO7; 371.1472), 355.1724 [M + Na]+ (Calcd for
detected as described for 2.
C16H28NaO7;
355.1733),
333
[M + H]+,
315[M ꢁ H2O + H]+, 153 [M ꢁ C6H12O6 + H]+ (base).
1
3.14. (1R,2R)-p-menth-3-ene-1,2-diol (6a)
For H NMR and13C NMR spectral data, see Tables
2 and 3. HMBC correlations: H-2exo/C-4, C-6, C-7,
C-10; H-3endo/C-1, C-2, C-4, C-5; H-3exo/C-1, C-2,
C-4, C-5, C-7; H-4/C-1, C-2, C-5, C-6, C-7, C-8, C-9;
H-5endo/C-1, C-3, C-4, C-6, C-7; H-5exo/C-3, C-4, C-
6; H-6endo/C-1, C-2, C-4, C-5, C-7; H-6exo/C-2, C-5,
C-7, C-10; H3-8/C-1, C-4, C-7, C-9; H2-9/C-1, C-4, C-
7, C-8; H3-10/C-1, C-2, C-6, C-7; Glc H-1/C-2. NOESY
correlations: H-2exo/H-9a, H-9b, H3-10, Glc H-1; H-
3endo/H-5endo; H-3exo/H2-9; H-5exo/H3-8; H-6endo/
H3-8.
22
Amorphous powder, ½aꢀ ¼ ꢁ38ꢂ (c = 0.3, MeOH).
D
1
For H NMR and 13C NMR spectral data, see Tables
1 and 3.
3.15. 2-O-MTPA ester of 6a
A solution of 6a (2 mg) in a mixture of CHCl3 and
pyridine (each 0.5 ml) was treated with R-(ꢁ)-MTPA-
chloride (5 drops) and the mixture was stirred for 2 h
at room temperature. The reaction mixture was treated
as described for 1 to give 1-O-S-(ꢁ)-MTPA ester of
6a-S (6a-S-(ꢁ)-MTPA). In the similar procedure, 6a-R
(6a-R-(+)-MTPA) was obtained withS-(+)-MTPA-
chloride.
3.19. Enzymatic hydrolysis of 11
A mixture of 11 (10 mg) and b-glucosidase (5 mg) in
water (5 ml) was shaken in a water bath at 37 ꢁC for 30
d. The mixture was applied to a silica gel [CHCl3–
3.16. (1S,2R,4S)-borneol b-
D
-glucopyranoside (10)
MeOH (9:1 ! 1:1)] column to afford 11a [(1S,2R,4S,
23
23
5R)-vicodiol, amorphous powder, ½aꢀ ¼ ꢁ16ꢂ (c = 0.3,
D
Colorless needles (MeOH), m.p. 126–130 ꢁC,
MeOH), ½aꢀ ¼ ꢁ19ꢂ (c = 0.3, CHCl3); 3 mg] and a su-
D
24
½aꢀ ¼ ꢁ53ꢂ (c = 3.2, MeOH). Positive FAB-MS m/z:
gar fraction. From the sugar fraction, D-glucose was de-
tected as described for 2.
D
339 [M + Na]+, 317.1978 [M + H]+ (Calcd for
C16H29O6; 317.1964), 137 [M ꢁ C6H12O6 + H]+ (base).
1
For H NMR and 13C NMR spectral data, see Tables
2 and 3. HMBC correlations: H-2exo/C-1, C-3, C-4,
C-6, C-7, C-10; H-3endo/C-1, C-2, C-4, C-5, C-7; H-
3exo/C-1, C-2, C-4, C-5, C-7; H-4/C-1, C-2, C-3, C-5,
C-6, C-7, C-8, C-9; H-5endo/C-1, C-3, C-4, C-6, C-7;
H-5exo/C-1, C-3, C-4, C-6, C-7; H-6endo/C-1, C-2, C-
4, C-5, C-7, C-8, C-10; H-6exo/C-1, C-2, C-4, C-5, C-
7, C-10; H3-8/C-1, C-4, C-7, C-9; H3-9/C-1, C-4, C-7,
C-8; H3-10/C-1, C-2, C-6, C-7; Glc H-1/C-2.NOESY
correlations: H-2exo/H3-9, H3-10, Glc H-1; H-3endo/
H-5endo; H-3exo/H3-9; H-5exo/H3-8; H-6endo/H3-8.
Acknowledgements
The authors thank Y. Takase, H. Suzuki and T.
Kiyotani of the Analytical Center of this University
for NMR and MS measurements.
References
Garg, S.N., Agarwal, S.K., 1988. New monoterpene diols from
essential oil of Ferula jaeschkeana. Phytochemistry 27, 936–937.
Ishikawa, T., Sega, Y., Kitajima, J., 2001a. Water-soluble constituents
of Glehnia littoralis fruit. Chem. Pharm. Bull. 49, 583–584.
Ishikawa, T., Sega, Y., Kitajima, J., 2001b. Water-soluble constituents
of ajowan. Chem. Pharm. Bull. 49, 840–844.
3.17. Enzymatic hydrolysis of 10
A mixture of 10 (20 mg) and naringinase (10 mg, ICN
Biomedicals Inc., Lot 2421c) in water (5 ml) was placed
in a glass test tube closed with a glass plug, and shaken
in a water bath at 37 ꢁC for 14 d. The precipitated
prisms were filtered out and washed with water to give
Kasai, R., Suzuo, M., Asakawa, J., Tanaka, O., 1977. Carbon-13
chemical shifts of isoprenoid-b- -glucopyranosides and b-D-man-
nopyranosides. Stereochemical influences of aglycone alcohols.
D
Tetrahedron Lett., 175–178.
Kitajima, J., Okamura, C., Ishikawa, T., Tanaka, Y., 1998. Mono-
terpenoid glycosides of Glehnia littoralis root and rhizoma. Chem.
Pharm. Bull. 46, 1595–1598.
10a [(1S,2R,4S)-borneol, colorless prisms (H2O), m.p.
23
D
205–209 ꢁC, ½aꢀ ¼ ꢁ33ꢂ (c = 0.3, MeOH); 8 mg]. The

J. Kitajima et al. / Phytochemistry 65 (2004) 3279–3287
3287
Kitajima, J., Suzuki, N., Satoh, M., Watanabe, M., 2002.
Sesquiterpenoids of Torilis japonica fruit. Phytochemistry 59,
811–815.
of three Thymus species grown wild in Iran. Planta Med. 66,
197–198.
Reddy, M.V.B., Angers, P., Gosselin, A., Arul, J., 1998. Character-
ization and use of essential oil from Thymus vulgaris against
Botrytis cinerea and Rhizopus stolonifer in strawberry fruits.
Phytochemistry 47, 1515–1520.
Kitajima, J., Ishikawa, T., Satoh, M., 2003. Polar constituents of celery
seed. Phytochemistry 64, 1003–1011.
Klyne, W., 1950. The configuration of the anomeric carbon atoms in
some cardiac glycoside. Biochem. J. 47, XIi–XIii.
Konishi, T., Tada, A., Shoji, J., Kasai, R., Tanaka, O., 1978. The
structures of platycodin A and C, monoacetylated saponins of the
roots of Platycodon grandiflorum A. DC Chem. Pharm. Bull. 26,
668–670.
Tori, K., Seo, S., Yoshimura, Y., Atita, H., Tomita, Y., 1977.
Glycosylation shifts from aglycone and glucose to glucoside.
Tetrahedron Lett., 179–182.
Vasanth, S., Kundu, A.B., Purushothaman, K.K., Patra, A., Pattabhi,
V., Connolly, J.D., 1990. Isolation and characterization of vicodiol,
a new monoterpenediol from Vicoa indica. J. Nat. Prod. 53, 354–
358.
Ohtani, I., Kusumi, T., Kashman, Y., Kakisawa, H., 1991. High-
Field FT-NMR application of MosherÕs method. Absolute
configurations of marine terpenoids. J. Am. Chem. Soc. 113,
4092–4096.
Wichtl, M. (Ed.), 1994. Herbal Drugs and Phytopharmaceuticals.
CRC Press, Stuttgart, pp. 493–495.
Orihara, Y., Furuya, T., 1993. Biotransformation of (ꢁ)-borneol by
cultured cells of Eucalyptus perriniana. Phytochemistry 34, 1045–
1048.
Yamazaki, K., Kasai, R., Masaki, Y., Okihara, M., Tanaka, O.,
Ohshiro, H., Takagi, S., Yamaki, M., Masuda, K., Nonaka, G.,
Tsuboi, M., Nishioka, I., 1977. Application of C-13 NMR to the
structural elucidation of acylated plant glycosides. Tetrahedron
Lett., 1231–1234.
Rustaiyan, A., Masoudi, S., Monfared, A., Kamalinejad, M.,
Lajevardi, T., Sedaghat, S., Yari, M., 2000. Volatile constituents