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Y. Hori et al. / Phytochemistry 62 (2003) 613–617
3.2. Extraction and isolation
(OH), 1714 (C¼O), 1527 (benzene ring), 1194, 1177,
1
1050 (SO3H). H NMR (CD3OD) d: 0.88 (3H, t, J=7.3
Commercial Shokyo (Zingiberis rhizome) powder (5.0
kg, Uchida Wakanyaku Co., imported from China; Lot.
No. 193012) was percolated with MeOH–H2O (4:1, 28 l)
at rt for 3 days; with the resulting for 80% MeOH
extract concentrated in vacuo at 40 ꢀC. The residual
syrup was suspended in H2O (1 l) and successfully
extracted with diethyl ether (3 Â 500 ml) to afford an
ether extract (155 g) after concentration. The ether
extract was repeatedly subjected to silica gel chromato-
graphy to give six known compounds, 6-, 8-, 10-ginger-
ols, 6-shogaol, 6-paradol and 6-gingeacetate,
identification of which was performed by comparison of
spectral data with those reported. The aqueous layer
(546 g) was, after concentration, subjected to an ODS
column with a gradient mixture of H2O and MeOH
providing the following six fractions: Frs. A (H2O, 498
g), B (H2O, 12.2 g), C (50% MeOH, 1.2 g), D (50%
MeOH, 19.0 g), E (50% MeOH, 8.2 g) and F (MeOH,
7.2 g). Fr. E was separated into three fractions by
Sephadex LH-20 CC (MeOH): Frs. E-1 (0.09 g), E-2
(7.86 g), and E-3 (0.33 g). Fr. E-2 (1.0 g) was purified by
silica gel CC [CHCl3–MeOH–AcOEt–H2O (2:2:4:1),
lower phase] and then Sephadex LH-20 CC (MeOH) to
give 1 (98 mg). Fr. D was applied to a silica gel column
eluted with CHCl3–MeOH–H2O (7:3:0.4) to give six
fractions: Frs. D-1 (0.8 g), D-2 (2.3 g), D-3 (5.8 g), D-4
(2.9 g), D-5 (0.9 g), and D-6 (4.2 g). Fr. D-2 was suc-
cessively applied to ODS (10% MeOH), silica gel
[CHCl3–MeOH–AcOEt–H2O (2:2:4:1), lower phase],
and Sephadex LH-20 (MeOH) columns to afford 2 (31
mg) and 3 (62 mg). A similar separation protocol was
applied for frs. D-3 and D-4, giving five compounds: 2
(22 mg) and 3 (203 mg) from the former and 4 (24 mg),
5 (4 mg) and 6 (14 mg) from the latter, respectively.
6-Gingesulfonic acid (1) White amorphous powder,
mp. 177–181 ꢀC (dec.). [ꢁ]2D1 +0.7ꢀ (MeOH, c 1.00).
Hz, H3-8), 1.36, 1.87 (1H each, m, H2-6), 1.36 (2H, m,
H-7), 2.79 (4H, m, H2-1 and H2-2), 2.50 (1H, dd,
J=17.4, 6.4 Hz, H-4), 3.03 (1H, dd, J=17.4, 6.1 Hz, H-
4), 3.31 (1H, m, H-5), 3.82 (3H, s, O-CH3), 6.61 (1H, dd,
J=8.2, 1.8 Hz, H-60), 60.68 (1H, d, J=8.2 Hz, H-50), 6.75
(1H, d, J=1.8 Hz, H-2 ). 13C NMR: Table 1.
Shogasulfonic acid A (3) Pale yellowish amorphous
21
ꢀ
powder, mp. 205 C (dec.), [ꢁ]D À0.5ꢀ (MeOH, c 2.00).
Positive FAB-MS (NBA) m/z: 439.1417 ([M+H]+,
C21H27O8S: 439.1427). EI-MS, m/z: 356 ([M-H2SO3]+).
KBr
max
IR n
cmÀ1: 3379 (OH), 1698 (C¼O), 1523 (benzene
ring), 1222, 1179, 1154, 1054 (SO3H). 1H NMR
(CD3OD) ꢀ: 1.71 (1H, m, H-6), 2.19 (1H, m, H-6), 2.61
(2H, t, J=7.6 Hz, H2-7), 2.76 (4H, m, H2-1 and H2-2),
2.57 (1H, dd, J=17.4, 6.4 Hz, H-4), 3.05 (1H, dd,
J=17.4, 6.1 Hz, H-4), 3.34 (1H, m, H-5), 3.79, 3.81 (3H
each, s, O-CH3 Â 2), 6.58, 6.60 (1H each, dd, J=8.2, 2.4
Hz, H-60 and H-600), 6.67, 6.68 (1H each, d, J=8.2 Hz,
H-50 and H-500), 6.75, 6.76 (1H each, d, J=2.4 Hz, H-20
and H-200). 13C NMR: Table 1.
Shogasulfonic acid B (4) Pale green oil, [ꢁ]2D1 À1.0ꢀ
(MeOH,
c 1.60). Positive FAB-MS (NBA) m/z:
425.1247 ([M+H]+, C20H25O8S: 425.1270). IR nKmBaxr
cmÀ1: 3421 (OH), 1708 (C¼O), 1519 (benzene ring),
1
1196, 1153, 1039 (SO3H). H NMR (CD3OD) ꢀ: 1.70
(1H, m, H-6), 2.16 (1H, m, H-6), 2.55 (2H, m, H2-7), 2.57
(1H, m, H-4), 2.74 (4H, m, H2-1 and H2-2), 3.04 (1H, dd,
J=17.4, 5.8 Hz, H-4), 3.36 (1H, m, H-5), 3.80 (3H, s, O-
CH3), 6.47 (1H, dd, J=7.9, 1.8 Hz, H-60), 6.60 (1H, dd,
J=7.9, 1.8 Hz, H-600), 6.62 (1H, d, J=1.8 Hz, H-20), 6.64
(1H, d, J=7.9 Hz, H-50), 6.67 (1H, d, J=7.9 Hz, H-500),
6.75 (1H, d, J=1.8 Hz, H-200). 13C NMR: Table 1.
Shogasulfonic acid C (5) Pale yellowish oily sub-
stance, [ꢁ]2D1 À5.6ꢀ (MeOH, c 0.25). Positive FAB-MS
(NBA) m/z: 411.1084 ([M+H]+, C19H23O8S: 411.1114).
KBr
max
IR n
cmÀ1: 3445 (OH), 1707 (C¼O), 1527 (benzene
1
Positive FAB-MS (NBA), m/z: 359.1457 ([M+H]+,
ring), 1198, 1042 (SO3H) H NMR (CD3OD) ꢀ: 2.70
(4H, m, H2-1 and H2-2), 2.54 (1H, dd, J=17.1, 6.4 Hz,
H-4), 3.04 (1H, dd, J=17.1, 5.8 Hz, H-4), 3.33 (1H, m,
H-5), 1.68 (1H, m, H-6), 2.16 (1H, m, H-6), 2.54 (2H, m,
H2-7), 6.48 (1H, dd, J=8.0, 2.2 Hz, H-60), 6.49 (1H, dd,
J=8.2, 2.2 Hz, H-600), 6.615, 6.619 (1H each, d, J=2.2
Hz, H-20 and H-200), 6.638 (1H, d, J=8.0 Hz, H-50),
6.641 (1H, d, J=8.2 Hz, H-500). 13C NMR: Table 1.
Shogasulfonic acid D (6) Pale yellowish crystaline
KBr
max
C17H27O6S: 359.1528). IR n
cmÀ1: 3182 (OH), 1711
(C¼O), 1525 (benzene ring), 1219, 1175, 1056 (SO3H).
1H NMR (CD3OD) ꢀ: 0.88 (3H, t, J=7.3 Hz, H3-10),
1.30- 1.35 (6H, m, H2-7, H2-8 and H2-9), 1.42 (1H, m, H-
6), 1.90 (1H, m, H-6), 2.80 (4H,m, H2-1 and H2-2), 2.50
(1H, dd, J=17.4, 6.4 Hz, H-4a), 3.03 (1H, dd, J =17.4,
6.4 Hz, H-4b), 3.30 (1H, m, H-5), 3.82 (3H,s, O-CH3),
6.60 (1H, dd, J=8.0, 1.9 Hz, H-60), 6.67 (1H, d, J=8.0
Hz, H-50), 6.77 (1H, d, J=1.9 Hz, H-20). 13C NMR:
Table 1. HMBC correlations were observed between
H-10/C-8, H-4a/C-6 and H-4b/C-6. The spectral data of 1
were coincident with those reported for 6-gingesulfonic
acid (Yoshikawa et al., 1992; Yoshikawa et al., 1994)
21
ꢀ
powder, mp. 154–158 C, [ꢁ]D À0.3ꢀ (MeOH, c 1.00).
Positive FAB-MS (NBA), m/z: 471.1370 ([M+H]+,
KBr
max
C21H27O10S: 471.1325). IR n
cmÀ1: 3421 (OH), 1714
(C¼O), 1527 (benzene ring), 1194, 1177, 1050 (SO3H).
1H NMR (CD3OD) ꢀ: 1.72 (1H, m, H-6), 2.17 (1H, m,
H-6), 2.56 (2H, m, H2-7), 2.71 (4H, m, H2-1 and H2-2),
2.55 (1H, dd, J=17.4, 6.4 Hz, H-4), 3.05 (1H, dd,
J=17.4, 6.1 Hz, H-4), 3.34 (1H, m, H-5), 3.78, 3.79 (3H
each, s, O-CH3 Â 2), 6.30, 6.31 (1H each, d, J=1.8 Hz,
4-Gingesulfonic acid (2) Pale brownish amorphous
21
ꢀ
powder, mp. 180–190 C (dec.), [ꢁ]D +1.0ꢀ (MeOH, c
2.00). Positive FAB-MS (NBA), m/z: 331.1173
([M+H]+, C15H23O6S: 331.1215). IR nmKaBxr cmÀ1: 3447