New p-menthanetriols and their glucosides from the fruit of caraway

Article abstract of DOI:10.1016/S0040-4020(01)00776-1

Ten new p-menthanetriols, including eight stereoisomers of p-menthane-2,8,9-triol, and five new glucosides were isolated from the water-soluble portion of the methanol extract of the fruit of caraway (Carum carvi L.), which has been used as a spice and medicine. Their structures were clarified by spectral investigation.

Full text of DOI:10.1016/S0040-4020(01)00776-1

TETRAHEDRON
Pergamon
Tetrahedron 57 +2001) 8067±8074
New p-menthanetriols and their glucosides from the
fruit of carawayq
Tetsuko Matsumura, Toru Ishikawa and Junichi Kitajimap
Showa Pharmaceutical University, Higashi-Tamagawagakuen 3, Machida, Tokyo 194-8543, Japan
Received 16 May 2001; accepted 25July 2001
AbstractÐTen new p-menthanetriols, including eight stereoisomers of p-menthane-2,8,9-triol, and ®ve new glucosides were isolated from
the water-soluble portion of the methanol extract of the fruit of caraway +Carum carvi L.), which has been used as a spice and medicine. Their
structures were clari®ed by spectral investigation. q 2001 Elsevier Science Ltd. All rights reserved.
1. Introduction
eluate fraction was chromatographed on Sephadex LH-20,
and subjected to a combination of silica gel, Lobar RP-8
column chromatography and HPLC to isolate mono-
terpenoid triols +1±10) and their glucosides +11±15). All
glucosides described in this paper were b-d-glucopyrano-
sides as shown by their 13C NMR data +Table 2), and this
was con®rmed by hydrolysis to yield d-glucose or by a
comparison of the [a]D or [M]D values with those of their
aglycones except 15.5 Their molecular formulae were
suggested from the accurate mass number of [M1H]1 or
[M1Na]1 or [M1K]1 ion peaks in the high-resolution
positive FAB-MS.
Caraway [Carum carvi L.; Umbelliferae] has been used as a
popular aromatic herb and spice since antiquity and has
been cultivated in Europe since the Middle Ages.1,2 Its
fruit has been used for medicine and in cooking, and is listed
in British, German and European pharmacopoeia.2,3 For
medicinal purpose, it is used to relieve ¯atulent indigestion,
colic and bronchitis.1,2 Studies on the fruit were made on the
essential oil, and many monoterpenoids [d-carvone +main;
50±60%), l-limonene, carvacrol, trans-carveol, d-dihydro-
carveol, l-dihydrocarveol, etc.] were reported as the con-
stituents.4 However, no report has been published on the
water-soluble portion of this fruit. In continuation of our
studies on the water-soluble constituents of spices, and to
reveal the relationship between the essential oil and the
water-soluble constituent, we undertook a detailed investi-
gation on the constituents of this fruit. In this paper, we
discuss the isolation and the characterization of 10 new
monoterpenoid triols and ®ve new monoterpenoid triol
glucosides.
Triol 1 +C10H20O3, an amorphous powder, [a]21D1148) and
2 +C10H20O3, an amorphous powder, [a]21D188) showed
[M1H]1 ion peaks at m/z 189 in the positive FAB-MS.
They revealed quite similar 1H and 13C NMR spectral
features +Tables 1 and 2), and have one tert-methyl, one
sec-methyl, four methylenes +one of them was oxygenated),
three methines +one of them was oxygenated) and one
oxygenated quaternary carbon. From the analysis
of HMBC spectral data of 1, they were suggested to be
p-menthane-2,8,9-triol. Hirai et al. isolated two mono-
terpenoidtriols, +1R,2R,4R,8S)- and +1R,2R,4R,8R)-p-
menthane-2,8,9-triol +1a and 2a) from the fruiting body of
Flammulina velutipes +Tricholomataceae),6 and 1 and 2
2. Results and discussion
Commercial caraway was extracted with 70% methanol,
and the methanolic extract was suspended in water and
successively extracted with ether and ethyl acetate. The
aqueous layer was chromatographed on Amberlite XAD-II
to give water and methanol eluate fractions. The methanol
1
showed the identical H and 13C NMR spectral data with
those of 1a and 2a. Furthermore, 1a and 2a had the opposite
optical rotation values to that of 1 and 2 +1a; [a]24D217.08,
2a; [a]24D25.38). Therefore, 1 and 2 were concluded to be
+1S,2S,4S,8R)- and +1S,2S,4S,8S)-p-menthane-2,8,9-triol,
respectively.
q This is the ®rst report of the isolation of all eight stereoisomeric terpe-
noids having four asymmeric carbons in the molecule, from the plant.
Triol 3 +C10H20O3, mp 119±1228C, [a]25D2318) and 4
+C10H20O3, mp 115±1178C, [a]21D2358) were indicated
to be p-menthane-2,8,9-triol by the 1H and 13C NMR
spectral data +Tables 1 and 2) and the result of HMBC
experiment of 3. The observed NOE interactions between
Keywords: caraway; Carum carvi fruit; p-menthanetriol; p-menthanetriol
glucoside; p-menthane-2,8,9-triol; stereoisomers.
Corresponding author. Tel.: 181-427-21-1577; fax: 181-427-21-1576;
p
e-mail: kitajima@ac.shoyaku.ac.jp
0040±4020/01/$ - see front matter q 2001 Elsevier Science Ltd. All rights reserved.
PII: S0040-4020+01)00776-1

8068
T. Matsumura et al. / Tetrahedron 57 ,2001) 8067±8074
Table 1. 1H NMR chemical shifts of 1±15 +in pyridine-d5, 500 MHz)
1
2
3
4
H-1ax
eq
1.56 m
±
1.56 m
±
±
2.28 m
±
2.28 m
H-2ax
H-3ax
eq
H-4ax
H-5ax
eq
H-6ax
eq
H3-7
H2-9
3.44 ddd +3.0, 10.0, 11.0)
1.64 ddd +11.0, 12.0, 12.0)
2.58 dddd +3.0, 3.0, 3.0, 12.0)
2.09 dddd +3.0, 3.0, 12.0, 12.0)
1.43 dddd +3.0, 12.0, 12.0, 12.0)
2.20 dddd +3.0, 3.0, 3.0, 12.0)
1.09 dddd +3.0, 12.0, 12.0, 12.0)
1.81 dddd +3.0, 3.0, 3.0, 12.0)
1.24 d +6.5)
3.45 ddd +3.5, 10.0, 11.0)
1.73 ddd +11.0, 12.0, 12.0)
2.83 dddd +3.5, 3.5, 3.5, 12.0)
2.10 dddd +3.5, 3.5, 12.5, 12.0)
1.30 dddd +3.5, 12.0, 12.0, 12.0)
1.92 dddd +3.5, 3.5, 3.5, 12.0)
1.09 dddd +3.5, 12.0, 12.0, 12.0)
1.79 dddd +3.5, 3.5, 3.5, 12.0)
1.25 d +6.5)
4.10 ddd +3.5, 3.5, 11.0)
1.84 ddd +11.0, 12.0, 12.0)
2.24 ddd +3.5, 3.5, 12.0)
2.05 dddd +3.5, 3.5, 12.0, 12.0)
1.64 dddd +3.5, 12.0, 12.0, 12.0)
1.92 dddd +3.5, 3.5, 3.5, 12.0)
1.58 dddd +3.5, 3.5, 12.0, 12.0)
1.72 dddd +3.5, 3.5, 3.5, 12.0)
1.22 d +7.0)
4.10 ddd +4.0, 4.0, 11.0)
1.94 ddd +11.0, 12.0, 12.0)
2.48 ddd +3.5, 4.0, 12.0)
2.06 dddd +3.5, 3.5, 12.0, 12.0)
1.58 dddd +4.0, 12.0, 12.0, 12.0)
1.65 m
1.58 dddd +4.0, 4.0, 12.0, 12.0)
1.67 dddd +4.0, 6.0, 12.0, 12.0)
1.22 d +7.0)
3.91 d +10.5)
3.96 d +10.5)
1.42 s
3.91 d +11.0)
3.94 d +11.0)
1.41 s
3.92 d +11.0)
3.96 d +11.0)
1.45s
3.92 d +11.0)
3.95 d +11.0)
1.44 s
H3-10
5
6
7
8
H-1ax
eq
1.55 m
±
1.55 m
±
±
2.13 m
±
2.11 m
H-2eq
H-3ax
eq
H-4ax
H-5ax
eq
H-6ax
eq
H3-7
H2-9
4.13 br dd +3.0, 3.0)
1.60 ddd +3.0, 13.0, 13.0)
2.37 ddd +3.0, 3.0, 13.0)
2.64 dddd +3.0, 3.0, 13.0, 13.0)
1.49 dddd +3.0, 13.0, 13.0, 13.0)
2.30 dddd +3.0, 3.0, 3.0, 13.0)
1.85dddd +3.0,13.0, 13.0, 13.0)
1.54 dddd +3.0, 3.0, 3.0, 13.0)
1.17 d +7.0)
4.16 br dd +3.0, 3.0)
1.68 ddd +3.0, 13.0, 13.0)
2.62 ddd +3.0, 3.0, 13.0)
2.66 dddd +3.0, 3.0, 13.0, 13.0)
1.35dddd +3.0, 13.0, 13.0, 13.0)
2.03 dddd +3.0, 3.0, 3.0, 13.0)
1.84 dddd +3.0, 13.0, 13.0, 13.0)
1.51 dddd +3.0, 3.0, 3.0, 13.0)
1.17 d +7.0)
4.11 br dd +3.0, 3.0)
1.83 ddd +3.0, 13.0, 13.0)
2.13 ddd +3.0, 3.0, 13.0)
2.67 dddd +3.0, 3.0, 13.0, 13.0)
1.71 dddd +3.0, 13.0, 13.0, 13.0)
2.01 dddd +3.0, 3.0, 3.0, 13.0)
2.35dddd +3.0, 3.0, 13.0, 13.0)
1.49 br ddd +3.0, 3.0, 13.0)
1.02 d +7.0)
4.12 br dd +3.0, 3.0)
1.88 ddd +3.0, 13.0, 13.0)
2.34 ddd +3.0, 3.0, 13.0)
2.66 dddd +3.0, 3.0, 13.0, 13.0)
1.54 dddd +3.0, 13.0, 13.0, 13.0)
1.73 dddd +3.0, 3.0, 3.0, 13.0)
2.31 dddd +3.0, 3.0, 13.0, 13.0)
1.43 br ddd +3.0, 3.0, 13.0)
0.99 d +7.0)
3.91 d +10.5)
3.95 d +10.5)
1.44 s
3.89 d +10.5)
3.96 d +10.5)
1.42 s
3.92 d +10.5)
3.96 d +10.5)
1.46 s
3.88 d +10.5)
3.93 d +10.5)
1.42 s
H3-10
9
10
11
12
H-1eq
H-2ax
eq
H-3ax
eq
H-4ax
H-5ax
eq
H-6ax
eq
±
±
±
2.46 m
4.30 br d +12.0)
±
1.81 ddd +12.0, 13.0, 13.0)
2.29 br d +13.0)
1.93 dddd +3.0, 3.0, 13.0, 13.0)
1.57 dddd +3.0, 13.0, 13.0, 13.0)
1.85 m
1.43 dddd +3.0, 3.0, 13.0, 13.0)
1.6 br dd +3.0, 13.0)3
1.20 d +7.0)
±
±
3.87 d +11.0)
3.90 d +11.0)
1.38 s
2.25m
4.03 ddd +4.5, 4.5, 12.0)
±
±
4.26 dd +3.0, 3.0)
2.43 ddd +3.0, 12.5, 12.5)
2.36 ddd +3.0, 3.0, 12.5)
2.48 dddd +3.0, 3.0, 12.5, 12.5)
2.13 dddd +3.0, 12.5, 12.5, 12.5)
2.02 ddd +3.0, 3.0, 12.5)
2.27 ddd +3.0, 12.5, 12.5)
1.93 ddd +3.0, 3.0, 12.5)
1.69 s
±
1.44 s
4.18 dd +3.5, 3.5)
2.33 ddd +3.5, 13.0, 13.0)
2.04 ddd +3.5, 3.5, 13.0)
1.86 m
2.05 dddd +3.5, 13.0, 13.0, 13.0)
1.71 dddd +3.5, 3.5, 3.5, 13.0)
2.22 ddd +3.5, 13.0, 13.0)
1.85 ddd +3.5, 3.5, 13.0)
1.69 s
2.42 m
1.46 s
3.78 dd +7.0, 10.5)
3.99 dd +5.5, 10.5)
1.17 d +7.0)
1.73 ddd +12.0, 12.0, 12.0)
2.24 br d +12.0)
2.03 br dd +12.0, 12.0)
1.56 dddd +3.0, 12.0, 12.0, 12.0)
1.84 br d +12.0)
1.53 dddd +3.0, 3.0, 12.0, 12.0)
1.67 br d +12.0)
1.18 d +7.0)
H3-7
H-8
H3-9
H2-9
±
±
3.83 d +10.0)
4.30 d +10.0)
1.35s
4.95 d +7.5)
±
±
1.44 s
±
13
H3-10
Glc-1
±
14
5.03 d +7.5)
15
H-2eq
H-3ax
eq
H-4ax
H-5ax
eq
H-6ax
eq
H3-7
H-8
H3-9
H-9a
b
H3-10
H2-10
Glc-1
4.18 dd +3.0, 3.0)
2.36 ddd +3.0, 12.0, 12.0)
2.42 ddd +3.0, 3.0, 12.0)
2.54 dddd +3.0, 3.0, 12.0, 12.0)
2.00 dddd +3.0, 12.0, 12.0, 12.0)
1.95ddd +3.0, 3.0, 12.0)
2.20 ddd +3.0, 12.0, 12.0)
1.85ddd +3.0, 3.0, 12.0)
1.69 s
±
1.46 s
±
±
1.46 s
±
5.10 d +7.5)
4.16 dd +3.0, 3.0)
2.22 ddd +3.0, 13.0, 13.0)
2.33 ddd +3.0, 3.0, 13.0)
1.80 m
2.00 dddd +3.0, 13.0, 13.0, 13.0)
1.54 dddd +3.0, 3.0, 3.0, 13.0)
2.06 ddd +3.0, 13.0, 13.0)
1.79 ddd +3.0, 3.0, 13.0)
1.81 s
2.41 m
±
3.73 dd +7.0, 10.0)
3.93 dd +6.0, 10.0)
1.07 d +7.0)
4.20 dd +3.0, 3.0)
2.49 ddd +3.0, 13.0, 13.0)
2.50 m
3.00 dddd +3.0, 3.0, 13.0, 13.0)
2.14 dddd +3.0, 13.0, 13.0, 13.0)
1.75dddd +3.0, 3.0, 3.0, 13.0)
2.07 ddd +3.0, 13.0, 13.0)
1.78 ddd +3.0, 3.0, 13.0)
1.82 s
±
±
5.10 br s
5.41 br s
±
4.48 br s
5.03 d +7.5)
±
5.03 d +8.0)
d in ppm from TMS [coupling constants +J) in Hertz are given in parentheses].

T. Matsumura et al. / Tetrahedron 57 ,2001) 8067±8074
Table 2. 13C NMR chemical shifts of 1±15 and 18 +in pyridine-d5, 125MHz)
8069
1
2
3
4
5
6
7
8
9
10
C-1
C-2
C-3
C-4
41.11
41.11
34.90
72.53
31.22
44.13
34.91
72.58
30.29
44.01
37.46
70.16
36.01
38.04
37.41
35.06
71.44
29.95
38.55
35.07
71.48
29.10
38.49
70.82
74.47
31.48
42.31
71.09
74.24
32.73
41.40
76.28
38.18
44.31
76.37
37.30
44.20
70.20
35.21
37.94
C-526.74
C-6
C-7
C-8
C-9
27.64
20.14
21.12
31.27
27.21
29.09
28.05
20.94
27.06
19.31
74.10
69.00
21.50
21.88
23.26
34.97
17.07
74.2574.18
26.45
34.01
19.37
74.01
69.0569.13
21.75
34.0531.20
19.34
73.92
29.07
19.29
74.16
68.91
21.86
27.06
17.04
34.91
28.87
71.63
11.55
74.08
11.58
74.02
28.59
32.65
a
69.00
21.55
69.15 69.03
69.14
21.74
27.77
65.75
14.52
C-10
21.96
21.93
21.67
27.93a
11
12
13
14
15
18
C-1
C-2
C-3
C-4
C-519.99
C-6
C-7
C-8
C-9
C-10
Glc-1
Glc-2
Glc-3
Glc-4
Glc-5
Glc-6
30.32 +24.6)
78.61 +16.1)
29.15+ 22.1)
44.02
34.73
72.43
31.08
43.99
70.78
70.73 +20.4)
110.5)
70.41
84.14
33.94
30.29 +24.6)
78.44 +15.9)
28.21 +22.1)
74.4584.72 +
31.10
40.94 +21.4)
30.38 +22.3)
40.91
34.3544.09
27.61 21.00
35.30
19.91
23.17
26.13
30.78
11.41
74.06
68.90
21.7521.63
101.35106.00
75.42
31.08
11.41
73.55 +20.5)
77.43 +18.4)
24.41
98.63
75.30
78.6578.95 78.73
71.88
78.55
63.02
35.03
28.71
79.77 +18.1)
35.40
28.26
32.43
65.60
13.95
30.88
11.41
73.93
69.00
21.63
28.27
156.13
107.01
64.75
101.28
75.70
24.28a +23.5)
+23.5)
a
106.47
75.52
106.31
75.73
75.45
78.52
78.63
78.74
78.71
71.74
71.7571.64
78.39
62.7562.73
71.61
62.86
71.73
78.34
62.75
78.10
78.30
62.90
d in ppm from TMS. Dd +d glucoside±d aglycone) are given in parentheses.
Assignments may be interchanged in each column.
a
Figure 1. Structures of 1±15, and NOE interactions between H2-9 and H2-3 or H2-5observed in the NOESY spectra of 1±8, 11 and 12.

8070
T. Matsumura et al. / Tetrahedron 57 ,2001) 8067±8074
H3-7/H-3ax, H-5ax and between H-2/H-4 of both triols
suggested that the con®guration of C-1 methyls was axial
and that of C-2 hydroxyls was equatorial, and the conforma-
tion of the cyclohexane rings were chair-form with 7±8 cis
relation +Fig. 1). So, 3 and 4 were concluded to be stereo-
isomers of cis-p-menthane-2eq,8,9-triol at C-8. The stereo-
chemical relationship between 3 and 4 was established by
comparison of their 13C NMR spectra with those of epimeric
pairs of 1 and 2, and +4R,8R)- and +4R,8S)-p-menth-8,9-diol
+16 and 17),7 where C-3 signals in +4Rp,8Sp)-forms +1; d
38.18, 16; d 26.93) appeared signi®cantly down®eld to
those in the +4Rp,8Rp)-forms +2; d 37.30, 17; d 25.81),
and C-5in +4 Rp,8Sp)-forms +1; d 26.74, 16; d 23.05)
appeared signi®cantly up®eld to those in the +4Rp,8Rp)-
forms +2; d 27.64, 17; d 24.30). For 3 and 4, the 13C chemi-
cal shift at C-3 of 3 +d 31.22) was down®eld to that of 4 +d
30.29), whereas C-5of 3 +d 20.14) was up®eld to that of 4 +d
21.12). Thus, the stereochemical relationship between C-4
and C-8 was considered to be 4Rp,8Sp in 3 and 4Rp,8Rp in 4
as in the pair of 1 and 2. This was also supported by the
results of the NOESY spectrum which showed interactions
between H3-10/H2-3, H2-5, and between H2-9/H-3eq in 1 and
3, and interactions between H3-10/H2-3, H2-5, and between
H2-9/H-5eq in 2 and 4. Glucoside 11 +C16H30O8, an
amorphous powder, [a]25D2468) and 12 +C16H30O8, an
amorphous powder, [a]21D2358) showed [M1H]1 ion
peaks at m/z 351 in the positive FAB-MS. Both glucosides
were hydrolyzed with hesperidinase and, from the hydro-
lyzed mixtures, 3 and d-glucose were obtained. Conse-
quently, 11 and 12 were monoglucosides of 3,
respectively. The position of the b-glucosyl unit of 11 was
proved to be C-2 from the HMBC correlation of glucosyl
H-1/C-2, and from the observed NOE interaction between
the glucosyl H-1/H-2 in the NOESY spectrum. The absolute
con®guration at C-2 of 3 was indicated as R by the values of
the glycosylation shift of the a- and b-carbon and the
chemical shift of the glucosyl anomeric carbon as shown
in Table 2.8 Thus, 3 and 11 were characterized as
+1S,2R,4R,8S)-p-menthane-2,8,9-triol and +1S,2R,4R,8S)-p-
menthane-2,8,9-triol 2-O-b-d-glucopyranoside, respec-
tively. The position of the b-glucosyl unit of 12 was proved
to be C-9 by the down®eld shift of the C-9 +by 8.4 ppm)
signal and up®eld shift of C-8 +by 0.5ppm) signal, and from
the observed NOE interaction between the glucosyl H-1/H2-
9 in the NOESY spectrum. So, 12 was characterized as
+1S,2R,4R,8S)-p-menthane-2,8,9-triol 9-O-b-d-glucopyrano-
side. On the other hand, the absolute con®guration of 4 was
indicated to be 1S,2R,4R,8R as +1S,2R,4R,8R)-p-menthane-
2,8,9-triol 2-O-b-d-glucopyranoside +18), which was
isolated from the fruit of Anethum gravealens +Dill) by
us,9 gave an aglycone identical to 4. Therefore, 4 was
suggested to be +1S,2R,4R,8R)-p-menthane-2,8,9-triol.
between H3-7/H-2eq, H2-6, and between H3-10/H-3ax,
H-5ax in their NOESY spectra. So, 5 and 6 were revealed
to be stereoisomers of trans-p-menthane-2ax,8,9-triol at C-8,
respectively. Further, the 13C chemical shift of C-3 of 5 +d
36.01) showed down®eld to that of 6 +d 35.21) and C-5 of 5
+d 27.21) appeared up®eld to that of 6 +d 28.05); the
relationship between 5 and 6 was indicated to be the same
as that between 3 and 4. This conclusion was supported by
the results of the NOESY spectra of 5 and 6, which showed
the same interactions as 3 +between H3-10/H2-3, H2-5, and
between H2-9/H-3eq) and 4 +between H3-10/H2-3, H2-5, and
between H2-9/H-5eq). Then, 5 and 6 were concluded to be
rel-+1R,2S,4R,8S)-p-menthane-2,8,9-triol and rel-+1R,2S,
4R,8R)-p-menthane-2,8,9-triol, respectively.
Triol 7 +C10H20O3, mp 130±1338C, [a]21D278) and 8
+C10H20O3, an amorphous powder, [a]21D2138) were indi-
cated to be p-menthane-2,8,9-triol by the same way
described for 1±6, and the con®guration of C-2 hydroxyls
was concluded to be axial by the equatorial H-2 signal
1
patterns of their H NMR spectra +Table 1). The stereo-
chemical relationship between C-7 and C-8 was suggested
to be cis from the observed NOE interactions between H3-7/
H-2eq, H-3ax, H-5ax, H-6eq, and between H3-10/H-3ax, H-5ax
in their NOESY spectra. So, 7 and 8 were concluded to be
stereoisomers of cis-p-menthane-2ax,8,9-triol at C-8, respec-
tively. The 13C chemical shifts of C-3 +7; d 29.95, 8; d
29.10) and C-5+ 7; d 20.94, 8; d 21.88) indicated the relative
con®guration at C-4 and C-8 was 4Rp,8Sp for 7 and 4Rp,8Rp
for 8. In addition, the NOESY spectrum of 7 showed NOE
interactions between H3-10/H2-3, H2-5and between H 2-9/H-
3eq which were also shown in the NOESY spectra of 1, 3 and
5, and the NOESY or 1D-NOE spectra of 8 showed the NOE
interactions between H3-10/H2-3, H2-5and between H 2-9/H-
5eq which were also observed in the NOESY spectra of 2, 4
and 6. Then, 7 and 8 were concluded to be rel-+1S,2S,
4R,8S)-p-menthane-2,8,9-triol and rel-+1S,2S,4R,8R)-p-
menthane-2,8,9-triol, respectively.
Triol 9 +C10H20O3, mp 134±1358C, [a]23D2258) showed
[M1H]1 ion peaks at m/z 189 in the positive FAB-MS, and
the 1H and 13C NMR spectral data +Tables 1 and 2) revealed
the presence of three tert-methyls, three methylenes, two
methines +one of them was oxygenated) and two oxygenated
quaternary carbons. From the analysis of the HMBC spec-
tral data, 9 was suggested to be p-menthane-1,2,8-triol. By
comparison of NMR data with those of +4R)-p-menthane-
1,2,8-triols, which were synthesized by Carman et al.,10
9
was indicated to be +1Sp,2Sp,4Rp)-form. Furthermore,
+1S,2S,4R)-p-menthane-1,2,8-triol showed a positive optical
rotation +[a]24D1448+CHCl3)) contrary to that of 9
+[a]21D2478+CHCl3)), and 9 was characterized as +1R,
2R,4S)-p-menthane-1,2,8-triol. Glucoside 13 +C16H30O8, an
amorphous powder, [a]23D2318) showed [M1H]1 ion
peaks at m/z 351 and [M2C6H12O61H]1 ion peaks at m/z
171 in the positive FAB-MS, and gave an aglycone, which
was identical to 9, and d-glucose by enzymatic hydrolysis.
The position of attachment of the glucosyl unit was revealed
to be C-8 of 9 from the H-C long-range correlation between
the glucosyl anomeric proton signal and the C-8 carbon in
the HMBC spectrum. Therefore, 13 was determined to be
+1R,2R,4S)-p-menthane-1,2,8-triol 8-O-b-d-glucopyrano-
side.
Triol 5 +C10H20O3, mp 123±1268C, [a]21D2228) and 6
+C10H20O3, an amorphous powder, [a]21D2308) showed
[M1K]1 ion peaks at m/z 227 and were indicated to be
1
p-menthane-2,8,9-triol by the H and 13C NMR spectral
data +Tables 1 and 2) and the result of HMBC experiment
of 5. The con®guration of C-2 hydroxyls was suggested to
be axial by the H-2 signals which found double doublets
with half bandwidths of 3 Hz in their 1H NMR spectra. The
conformation of 5 and 6 was found to be 7,8-trans form
from the observed NOE interactions between H-1/H-3ax,

T. Matsumura et al. / Tetrahedron 57 ,2001) 8067±8074
8071
Triol 10 +C10H20O3, an amorphous powder, [a]24D1258)
3. Experimental
3.1. General procedures
1
was also indicated to be monoterpenoidtriol by H and 13C
NMR spectral data +Tables 1 and 2), and its planar
structure was revealed to be p-menthane-1,2,9-triol by the
HMBC experiment. The con®guration of C-1 methyl, C-4
isopropyl was suggested to be equatorial, and the conforma-
tion of the cyclohexane ring was found to be chair-form
from the NOE interactions between H3-7/H-2, H2-6, and
between H-4ax/H-6ax observed in the NOESY spectrum.
The con®guration of C-2 hydroxyl was indicated to be
Melting points were determined on a Yanagimoto micro-
melting point apparatus and are uncorrected. Optical rota-
tions were measured on a JASCO DIP-370 digital
polarimeter. FAB-MS were recorded with a JEOL HX-
110 spectrometer using glycerol as matrix. 1H and 13C
NMR spectra were taken on JEOL JNM GX-270 and
A-500 spectrometers with tetramethylsilane as an internal
standard, and chemical shifts were recorded in d value.
1H±13C COSY, HMBC and NOESY spectra were obtained
with the usual pulse sequence, and data processing was
performed with standard JEOL software. Column chromato-
graphy +C. C.) was carried out under TLC monitoring using
Kieselgel 60 +70±230 mesh, Merck), Sephadex LH-20 +25±
100 mm, Pharmacia), Lobar RP-8 column +Merck) and
Amberlite XAD-II +Organo). TLC was performed on silica
gel +Merck 5721) and spots were detected with p-anis-
aldehyde±H2SO4 reagent. HPLC separation was carried
out on a JASCO chromatograph +980-system) with a
JASCO RI-930 detector, and Symmetryprep C18 7 mm
[Waters; column size, 7.8£300 mm; ODS], Carbohydrate
Analysis [Waters; column size, 3.9£300 mm; CHA] were
used as columns. Acetylation was done in the usual way
using Ac2O and pyridine, and no acetoxyl group was
detected by NMR spectral data for those acetylated
fractions.
1
axial by the narrow equatorial H-2 signal of its H NMR
spectrum +Table 1). The stereochemical relationship
between C-4 and C-8 was revealed to be 4Rp,8Rp from the
observed NOE interactions between H3-10/H2-3, and
between H-3ax/H-8 in the NOESY spectrum. Then, the
relative con®guration was concluded to be 1Sp,2Sp,4Rp,8Rp,
respectively. Glucoside 14 +C16H30O8, mp 174±1788C,
[a]24D1218) showed [M1H]1 ion peaks at m/z 351 and
[M2C6H12O61H]1 ion peaks at m/z 171 in the positive
FAB-MS. 14 was hydrolyzed with b-glucosidase and,
from the hydrolyzed mixtures, 10 and d-glucose were
obtained. So, 14 was a monoglucoside of 10. The position
of the b-glucosyl unit was suggested to be C-2 from the
HMBC correlation of glucosyl H-1/C-2, and from the
observed NOE interaction between the glucosyl H-1/H-2
in the NOESY spectrum. The absolute con®guration at
C-2 of 10 was indicated as S by the values of the glycosyl-
ation shift of the a- and b-carbon and the chemical shift of
the glucosyl anomeric carbon as shown in Table 2.8 Thus, 10
and 14 were characterized as +1S,2S,4R,8R)-p-mentane-
1,2,9-triol and +1S,2S,4R,8R)-p-menthane-1,2,9-triol 2-O-
b-d-glucopyranoside, respectively.
3.2. Plant material, extraction and isolation
Commercial caraway +the fruit of Carum carvi L.;
purchased from Asaoka Spices Ltd., Lot. No. 93010;
2.0 kg) was extracted with 70% methanol +4**L£2) at
room temperature for two weeks. After evaporation of the
solvent, the residue +402.3 g) was partitioned into ether±
water, ethyl acetate±water. Removal of the solvent from
each phase gave the ether +200.9 g), ethyl acetate +3.6 g)
and aqueous +40.6 g) extracts, The aqueous extract was
chromatographed over Amberlite XAD-II +H2O±MeOH).
The methanol eluate +27.6 g) was subjected to Sephadex
LH-20 +MeOH) to give eight fractions +frs. A±H). Fraction
B +18.9 g) was chromatographed over silica gel [CHCl3±
MeOH±H2O +17:3:0.2-4:1:0.1-7:3:0.5)-MeOH] to give 14
fractions +frs. B1±B14). Fraction B3 +1.70 g) was passed
through a Lobar RP-8 column [MeCN±H2O +3:17)] to
give nine fractions +frs. B3-1±B3-9), and fr. B3-5 was
subjected to HPLC [ODS, MeCN±H2O +3:37)]. The main
fraction was acetylated with Ac2O and pyridine, and the
acetylated fraction was subjected to HPLC [ODS, MeCN±
H2O +2:3)] to give two fractions. These two fractions were
deacetylated by heating in a water bath with 5% NH4OH±
MeOH for 2 h, and passed through Sephadex LH-20
+MeOH) to give 9 +6 mg) and 10 +2 mg). Fraction B3-7
was subjected to HPLC [ODS, MeCN±H2O +1:9)] to give
3 +4 mg), 1 +10 mg), 4 +43 mg) and 2 +9 mg). Fraction B3-7
was subjected to HPLC [ODS, MeCN±H2O +1:9)] to give 7
+8 mg), 8 +1 mg), 5 +9 mg) and a mixture of 6 and an alkyl
glucoside. From this mixture, 6 +3 mg) was isolated by silica
gel column chromatography [CHCl3±MeOH±H2O
+9:1:0.1)]. Fraction B9 +0.83 g) was subjected to a Lobar
RP-8 column [MeCN±H2O +3:17)] and HPLC [CHA,
Glucoside 15 +C16H28O8, an amorphous powder, [a]23D178)
showed [M1H]1 ion peaks at m/z 349 and
[M2C6H12O61H]1 ion peaks at m/z 169 in the
positive FAB-MS. The 1H, 13C and 13C±1H COSY
NMR spectral data +Tables 1 and 2) showed the
presence of one b-glucopyranosyl, one tert-methyl,
four methylenes +one of them was oxygenated), two
methines +one of them was oxygenated), one oxygenated
quaternary carbon and one terminal-methylene group. By
comparison of its 13C NMR data with that of 14, 15
was suggested to be a dehydro-derivative of 14 having a
double bond at C-8. The NOE interactions between H-2eq/
glucosyl H-1, between H3-7/H-2eq, H2-6, glucosyl H-1,
between H-9b/H-5ax, and between H2-10/H2-3 also
supported the suggested structure, and the con®guration
of the C-2 hydroxyl group was con®rmed to be axial
by the coupling constant +dd, Jˆ3.0, 3.0 Hz) of the
H-2 signal proton. Since this glucose was considered to be
d-form the same as other glucosides, the absolute con-
®guration at C-2 should be S. So, 15 was concluded to
be +1S,2S,4R)-p-menth-8-ene-1,2,10-triol 2-O-b-d-gluco-
pyranoside.
This is the ®rst report of the isolation of all eight stereo-
isomeric monoterpenoids, except the optical isomer, having
four asymmetric carbons in the molecule +1±8). Further-
more, the relationship between the essential oil and the
water-soluble constituent was con®rmed by the isolation
of these monoterpenoids and glucosides which showed a
biosynthetic relation to d-carbone.

8072
T. Matsumura et al. / Tetrahedron 57 ,2001) 8067±8074
MeCN±H2O +9:1)] to give 12 +5mg). Fraction B 11 +0.71 g)
was also subjected to a Lobar RP-8 column [MeCN±H2O
+3:17)] and HPLC [CHA, MeCN±H2O +9:1)] to give 13
+6 mg). Fraction B10 +1.38 g) was passed through a Lobar
RP-8 column [MeCN±H2O +3:17)] to give eight fractions
+frs. B10-1±B10-8). Fraction B10-4, fr. B10-5 and B10-7 were
subjected to HPLC [CHA, MeCN±H2O +9:1)]. to give 15
+2 mg), 14 +11 mg) and 11 +610 mg), respectively.
C-1, C-3, C-4, C-6; H-5eq/C-3, C-4, C-6; H-6ax/C-2, C-4,
C-5, C-7; H-6eq/C-2, C-4, C-5, C-7; H3-7/C-1, C-2, C-6;
H2-9/ C-4, C-8, C-10; H3-10/C-4, C-8, C-9.
3.2.6. Rel-,1R,2S,4R,8R)-p-menthane-2,8,9-triol ,6). An
amorphous powder, [a]21D2308 +cˆ0.4, MeOH). Positive
FAB-MS m/z: 377 [2M1H]1, 227.1051 [M1K]1 +Calcd for
C10H20KO3; 227.1050), 171 [M2H2O1H]1, 15
3
1
[M22H2O1H]1 +base), 135[M 23H2O1H]1. H NMR
3.2.1. ,1S,2S,4S,8R)-p-Menthane-2,8,9-triol ,1). An
amorphous powder, [a]21D1148 +cˆ0.2, MeOH).
Positive FAB-MS m/z: 377 [2M1H]1, 211 [M1Na]1,
189.1486 [M1H]1 +base, Calcd for C10H21O3; 189.1490),
+pyridine-d5, 500 MHz) d: Table 1. 13C NMR +pyridine-
d5, 125MHz) d: Table 2.
3.2.7. Rel-,1S,2S,4R,8S)-p-menthane-2,8,9-triol ,7).
Colorless needles +MeOH), mp 130±1338C, [a]21D278
+cˆ0.3, MeOH). Positive FAB-MS m/z: 399 [2M1Na]1,
377 [2M1H]1, 227.1032 [M1K]1 +Calcd for C10H20KO3;
227.1050), 189 [M1H]1, 171 [M2H2O1H]1, 15
171 [M2H2O1H]1, 15
3
2[M2H2O1H]1, 135
[M23H2O1H]1. 1H NMR +pyridine-d5, 500 MHz) d:
Table 1. 13C NMR +pyridine-d5, 125MHz) d: Table 2.
HMBC correlations: H-1ax/C-2, C-3, C-5, 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-
4ax/C-2, C-3, C-5, C-6, C-8, C-9, C-10; H-5ax/C-1, C-3, C-4,
C-6, C-8; 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; H2-9/ C-4, C-8, C-10; H3-10/C-4, C-8, C-9.
3
1
[M22H2O1H]1 +base), 135[M 23H2O1H]1. H NMR
+pyridine-d5 500 MHz) d: Table 1. 13C NMR +pyridine-d5,
125MHz) d: Table 2. HMBC correlations: H-2eq/C-3;
H-3ax/C-4, C-8; H-4ax/C-3, C-8, C-10; H-5ax/C-4, C-8;
H-6ax/C-1, C-5, C-7; H-6eq/C-1; H3-7/C-1, C-2, C-6; H2-9/
C-4, C-8, C-10; H3-10/C-4, C-8, C-9.
3.2.2. ,1S,2S,4S,8S)-p-Menthane-2,8,9-triol ,2). An amor-
phous powder, [a]21D188 +cˆ0.2, MeOH). Positive FAB-
MS m/z: 377 [2M1H]1, 211 [M1Na]1, 189.1476 [M1H]1
+base, Calcd for C10H21O3; 189.1490), 171 [M2H2O1H]1,
153 [M22H2O1H]1, 135[M 23H2O1H]1. 1H NMR
+pyridine-d5, 500 MHz) d: Table 1. 13C NMR +pyridine-
d5, 125MHz) d: Table 2.
3.2.8. Rel-,1S,2S,4R,8R)-p-menthane-2,8,9-triol ,8). An
amorphous powder, [a]21D2138 +cˆ0.1, MeOH). Positive
FAB-MS m/z: 399 [2M1Na]1, 377 [2M1H]1, 227.1047
[M1K]1 +Calcd for C10H20KO3; 227.1050), 189 [M1H]1,
171 [M2H2O1H]1, 15 3 [M22H2O1H]1 +base), 135
[M23H2O1H]1. 1H NMR +pyridine-d5, 500 MHz) d:
Table 1. 13C NMR +pyridine-d5, 125MHz) d: Table 2.
3.2.3. ,1S,2R,4R,8S)-p-Menthane-2,8,9-triol ,3). Colorless
needles +MeOH), mp 119±1228C, [a]25D2318 +cˆ3.1,
MeOH). Positive FAB-MS m/z: 377 [2M1H]1, 227
[M1K]1, 211 [M1Na]1, 189.1502 [M1H]1 +base, Calcd
for C10H21O3; 189.1490), 171 [M2H2O1H]1, 15
[M22H2O1H]1, 135[M 23H2O1H]1. 1H NMR +pyridine-
d5, 500 MHz) d: Table 1. 13C NMR +pyridine-d5, 125MHz)
d: Table 2. HMBC correlations: H-1eq/C-2, C-3, C-5, C-6,
C-7; 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; H-4ax/C-2, C-3, C-5,
C-6, C-8, C-9, C-10; H-5ax/C-1, C-3, C-4, C-6, C-8; H-5eq/
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; H2-9/
C-4, C-8, C-10; H3-10/C-4, C-8, C-9.
3.2.9. ,1R,2R,4S)-p-Menthane-1,2,8-triol ,9). Colorless
needles +MeOH), mp 134±1358C, [a]23D2258 +cˆ0.2,
MeOH), [a]21D2478 +cˆ0.1, CHCl3). Positive FAB-MS
m/z: 377 [2M1H]1, 211.1305[M 1Na]1 +Calcd for
3
C10H20NaO3;
211.1310),
189
[M1H]1,
171
[M2H2O1H]1, 15
3
2[M2H2O1H]1 +base), 135
[M23H2O1H]1. 1H NMR +pyridine-d5, 500 MHz) d:
Table 1. 13C NMR +pyridine-d5, 125MHz) d: Table 2.
HMBC correlations: H-3ax/C-1, C-2, C-5; H-3eq/C-1, C-2,
C-5; H-5ax/C-3, C-4, C-6; H-6ax/C-4, C-5; H-6eq/C-1, C-2,
C-4, C-5; H3-7/C-1, C-2, C-6; H3-9/ C-4, C-8, C-10; H3-10/
C-4, C-8, C-9.
3.2.10. ,1S,2S,4R,8R)-p-Menthane-1,2,8-triol ,10). An
amorphous powder, [a]24D1258 +cˆ0.2, MeOH). Positive
FAB-MS m/z: 377 [2M1H]1, 211.1323 [M1Na]1 +Calcd
for C10H20NaO; 211.1310), 189 [M1H]1, 171
3.2.4. ,1S,2R,4R,8R)-p-Menthane-2,8,9-triol ,4). Color-
less needles +MeOH), mp 115±1178C, [a]21D2358 +cˆ0.4,
MeOH). Positive FAB-MS m/z: 377 [2M1H]1, 211
[M1Na]1, 189.1493 [M1H]1 +base, Calcd for C10H21O3;
189.1490), 171 [M2H2O1H]1, 15 3 [M22H2O1H]1, 135
[M23H2O1H]1. 1H NMR +pyridine-d5, 500 MHz) d:
Table 1. 13C NMR +pyridine-d5, 125MHz) d: Table 2.
[M2H2O1H]1, 15
3
2[M2H2O1H]1 +base), 135
[M23H2O1H]1. 1H NMR +pyridine-d5, 500 MHz) d:
Table 1. 13C NMR +pyridine-d5, 125MHz) d: Table 2.
HMBC correlations: H-2eq/C-1, C-3, C-6; H-3ax/C-2, C-4,
C-5, C-8; H-3eq/C-1, C-2, C-4, C-5, C-8; H-4ax/C-2, C-3,
C-5, C-6, C-8, C-9, C-10; H-5ax/C-1, C-4, C-6; H-5eq/C-1,
C-3, C-6, C-8; H-6ax/C-1, C-5, C-7; H-6eq/C-1, C-2, C-5,
C-7; H3-7/C-1, C-2, C-6; H2-9/ C-4, C-8, C-10; H3-10/
C-4, C-8, C-9.
3.2.5. Rel-,1R,2S,4R,8S)-p-menthane-2,8,9-triol ,5).
Colorless needles +MeOH), mp 123±1268C, [a]21D2228
+cˆ0.2, MeOH). Positive FAB-MS m/z: 377 [2M1H]1,
227.1040 [M1K]1 +Calcd for C10H20KO3; 227.1050), 171
[M2H2O1H]1, 15
3
2[M2H2O1H]1 +base), 135
[M23H2O1H]1. 1H NMR +pyridine-d5, 500 MHz) d:
Table 1. 13C NMR +pyridine-d5, 125MHz) d: Table 2.
HMBC correlations: H-1ax/C-2, C-5, C-7; H-3ax/C-4, C-5;
H-3eq/C-2, C-4, C-5; H-4ax/C-3, C-5, C-6, C-8, C-10; H-5ax/
3.2.11. ,1S,2R,4R,8S)-p-Menthane-2,8,9-triol 4-O-b-d-
glucopyranoside
,11).
An
amorphous
powder,
[a]25D2468 +cˆ1.8, MeOH). Positive FAB-MS m/z: 373
[M1Na]1, 351.2048 [M1H]1 +base, Calcd for C16H31O8;

T. Matsumura et al. / Tetrahedron 57 ,2001) 8067±8074
8073
351.2019), 189 [M2C6H10O51H]1. 1H NMR +pyridine-d5,
500 MHz) d: Table 1. 13C NMR +pyridine-d5, 125MHz) d:
Table 2. HMBC correlations: H-1eq/C-2; H-2ax/C-3, C-4,
C-7, Glc C-1; H-3ax/C-1, C-2, C-4, C-5; H-3eq/C-1, C-2,
C-4, C-5; H-4ax/C-3, C-8, C-10; H-5ax/C-4, C-6; H-5eq/
C-1, C-3, C-4, C-6; H-6ax/C-1, C-4, C-5, C-7; H-6eq/C-2,
C-4, C-5; H3-7/C-1, C-2, C-6; H2-9/C-4, C-8, C-10;
H3-10/C-4, C-8, C-9; Clc H-1/C-2.
3.5.1. ,1S,2S,4R,8R)-p-Menthane-1,2,9-triol 2-O-b-d-
glucopyranoside ,14). Colorless needles +MeOH), mp
174±1788C, [a]24D1218 +cˆ0.2, MeOH). Positive FAB-
MS m/z: 723 [2M1Na]1, 701 [2M1H]1, 389 [M1K]1,
373 [M1Na]1, 351.2018 [M1H]1 +Calcd for C16H31O8;
351.2019), 171 [M2C6H12O61H]1 +base). 1H NMR
+pyridine-d5, 500 MHz) d: Table 1. 13C NMR +pyridine-
d5, 125MHz) d: Table 2. HMBC correlations: H-2eq/C-1,
C-6, C-7, Glc C-1; 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-4ax/C-8, C-9, C-10; H-5ax/C-1,
C-4, C-6; H-5eq/C-1, C-3, C-4, C-8; H-6ax/C-1, C-5, C-7;
H-6eq/C-1, C-3, C-5; H3-7/C-1, C-2, C-6; H2-9/ C-4, C-8,
C-10; H3-10/C-4, C-8, C-9; Clc H-1/C-2.
3.3. Enzymatic hydrolysis of 11
A mixture of 11 +12 mg) and hesperidinase +5mg, ICN
Biomedicals Inc., Lot. 72635) in water +5 ml) was shaken
in a water bath at 378C for 20 days. The mixture was evapo-
rated in vacuo to dryness and the residue was chromato-
graphed over silica gel [CHCl3±MeOH±H2O +4:1:0.1 and
1:1:0.1)] to afford 3 +6 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 min21; t R
4.50 min +same location as that of d-glucose)] show the
presence of d-glucose.
3.6. Enzymatic hydrolysis of 14
A mixture of 14 +8 mg) and b-glucosidase +5mg) in water
+5ml) was shaken in a water bath at 37 8C for 7 days. The
mixture was treated in the same way described for 11 to
afford 10 +4 mg) and a sugar fraction. From the sugar frac-
tion, the presence of d-glucose was revealed as 11.
3.6.1. ,1S,2S,4R)-p-Menthane-1,2,10-triol 2-O-b-d-gluco-
pyranoside ,15). An amorphous powder, [a]23D178
+cˆ0.1, MeOH). Positive FAB-MS m/z: 371.1681
[M1Na]1 +Calcd for C16H30NaO8; 371.1681), 349
[M1H]1, 169 [M2C6H12O61H]1 +base). 1H NMR
+pyridine-d5, 500 MHz) d: Table 1. 13C NMR +pyridine-
d5, 125MHz) d: Table 2.
3.3.1. ,1S,2R,4R,8S)-p-Menthane-2,8,9-triol 9-O-b-d-
glucopyranoside
,12).
An
amorphous
powder,
[a]21D2358 +cˆ0.2, MeOH). Positive FAB-MS m/z: 723
[2M1Na]1, 701 [2M1H]1, 389 [M1K]1, 373 [M1Na]1,
351.2018 [M1H]1 +Calcd for C16H31O8; 351.2019), 171
[M2C6H12O61H]1 +base). 1H NMR +pyridine-d5,
500 MHz) d: Table 1. 13C NMR +pyridine-d5, 125MHz)
d: Table 2.
Acknowledgements
The authors thank Messrs Y. Takase and H. Suzuki of the
Analytical Center of this University for NMR and MS
measurements.
3.4. Enzymatic hydrolysis of 12
A mixture of 12 +9 mg) and hesperidinase +5mg) in water
+5ml) was shaken in a water bath at 37 8C for 20 days. The
mixture was treated in the same way described for 11 to
afford 3 +2 mg) and a sugar fraction. From the sugar frac-
tion, the presence of d-glucose was revealed as 11.
References
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3.4.1. ,1S,2R,4S)-p-Menthane-1,2,8-triol 8-O-b-d-gluco-
pyranoside ,13). An amorphous powder, [a]23D2318
+cˆ0.4, MeOH). Positive FAB-MS m/z: 701 [2M1H]1,
389 [M1K]1, 373.1847 [M1Na]1 +base, Calcd for
2. Herbal Drugs and Phytopharmaceuticals; Wichtl, M., Ed.;
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C16H30NaO8;
373.1838),
351
[M1H]1,
171
1
[M2C6H12O61H]1. H NMR +pyridine-d5, 500 MHz) d:
Table 1. 13C NMR +pyridine-d5, 125MHz) d: Table 2.
HMBC correlations: H-2eq/C-4; H-3ax/C-4, C-5; H-3eq/C-1,
C-2, C-4, C-5; H-4ax/C-5, C-8, C-9, C-10; H-5ax/C-1, C-3,
C-4, C-6; H-5eq/C-1, C-6; H-6ax/C-1, C-4, C-5; H-6eq/C-1,
C-2, C-4; H3-7/C-1, C-2, C-6; H2-9/ C-4, C-8, C-10; H3-10/
C-4, C-8, C-9; Clc H-1/C-8.
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3.5. Enzymatic hydrolysis of 13
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A mixture of 13 +5mg) and b-glucosidase +5mg, TOYOBO
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bath at 378C for 7 days. The mixture was treated in the same
way described for 11 to afford 9 +2 mg) and a sugar fraction.
From the sugar fraction, the presence of d-glucose was
revealed as 11.
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+b). The absolute con®guration at C-2 of 4 was indicated as R
by the values of the glycosylation shift of the a-carbon +d18±
d4) and the chemical shift of the glucosyl anomeric carbon of
18 as shown in Table 2.
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9. +a) Ishikawa, T.; Kudo, M.; Kitajima, J. Abstract papers II,