440
Vol. 55, No. 3
both s, 11, 12-H3), 0.90 (1H, ddd, Jꢃ12.2, 12.2, 12.2 Hz, 4a-H), 1.11 (1H,
dd, Jꢃ12.2, 12.2 Hz, 2a-H), 1.32 (1H, dd, Jꢃ9.8, 10.4 Hz, 6-H), 1.28 (3H,
d, Jꢃ6.4 Hz, 10-H3), 1.53 (1H, m, 5-H), 1.69 (1H, ddd, Jꢃ2.2, 4.0, 12.2 Hz,
2b-H), 1.96 (1H, m, 4b-H), 3.17 (1H, m, 2ꢁ-H), 3.29 (1H, m, 4ꢁ-H), 3.30
(2H, m, 3ꢁ, 5ꢁ-H), [3.56 (1H, dd, Jꢃ4.6, 11.0 Hz), 3.79 (1H, dd, Jꢃ2.4,
11.0 Hz), 6ꢁ-H2], 3.73 (1H, m, 3-H), 4.35 (1H, m, 9-H), 4.35 (1H, d,
Jꢃ7.9 Hz, 1ꢁ-H), 5.35 (1H, dd, Jꢃ9.8, 15.6 Hz, 7-H), 5.53 (1H, dd, Jꢃ7.0,
15.6 Hz, 8-H). 13C-NMR (125 MHz, CD3OD) dC: given in Table 1. Positive-
ion FAB-MS: m/z 397 (MꢂNa)ꢂ.
(1161 mg)]. Fr. 5-6 (664.5 mg) was purified by HPLC [CH3CN–MeOH–H2O
(10 : 8 : 82, v/v/v)] to give (ꢂ)-larisiresinol 4,4ꢁ-bis-O-b-D-glucopyranoside
(14, 167.7 mg, 0.00030%). Fr. 5-8 (229.8 mg) was crystallized in MeOH
to give limocitrin 3,7-di-O-b-D-glucopyranoside (29, 295.3 mg, 0.00055%)
and the mother liquid was purified by HPLC [CH3CN–MeOH–H2O
(28 : 24 : 246, v/v/v)] to give herhacetin 8-methyl ester 3,7-di-O-b-D-glu-
copyranoside (27, 14.9 mg, 0.00003%). Fr. 5-9 (589.3 mg) was separated by
Sephadex LH-20 column chromatography [150 g, CHCl3–MeOH (1 : 1, v/v)]
and finally HPLC [MeOH–H2O (35 : 65, v/v)] to give isorhamnetin 3,7-di-O-
b-D-glucopyranoside (25, 44.3 mg, 0.00008%) and limocitrin 3,7-di-O-b-D-
glucopyranoside (29, 10.1 mg, 0.00002%). Fr. 5-10 (1818 mg) was separated
by Sephadex LH-20 column chromatography [150 g, CHCl3–MeOH (1 : 1,
v/v)] and finally HPLC [MeOH–H2O (35 : 65 or 40 : 60, v/v) or
CH3CN–MeOH–H2O (15 : 8 : 77, v/v/v)] to give 19 (79.9 mg, 0.00015%),
grosvenorine (20, 53.3 mg, 0.00010%), quercetin 3,7-di-O-b-D-glucopyra-
noside (21, 37.6 mg, 0.00007%), 22 (23.9 mg, 0.00004%), 25 (32.8 mg,
0.00006%), and 26 (26.8 mg, 0.00005%).
Sedumoside F2 (5): An amorphous powder, [a]D24 ꢀ26.0° (cꢃ1.08,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C25H44O11Na
(MꢂNa)ꢂ: 543.2781. Found: 543.2776. IR (KBr, cmꢀ1): 3410, 2967, 2940,
1
1669, 1474, 1341, 1140, 1055, 968. H-NMR (500 MHz, CD3OD) d: 0.83
(3H, d, Jꢃ6.4 Hz, 13-H3), 0.87, 0.91 (3H each, both s, 11, 12-H3), 0.91 (1H,
ddd, Jꢃ12.2, 12.2, 12.2 Hz, 4a-H), 1.12 (1H, dd, Jꢃ12.2, 12.2 Hz, 2a-H),
1.32 (1H, dd, Jꢃ9.8, 10.4 Hz, 6-H), 1.27 (3H, d, Jꢃ6.1 Hz, 6ꢄ-H3), 1.28 (3H,
d, Jꢃ6.1 Hz, 10-H3), 1.54 (1H, m, 5-H), 1.69 (1H, ddd, Jꢃ1.9, 4.3, 12.2 Hz,
2b-H), 1.96 (1H, m, 4b-H), 3.16 (1H, dd, Jꢃ7.9, 8.6 Hz, 2ꢁ-H), 3.29 (1H, m,
4ꢁ-H), 3.30 (2H, m, 3ꢁ, 5ꢁ-H), 3.35 (1H, m, 4ꢄ-H), [3.56 (1H, dd, Jꢃ4.6,
11.0 Hz), 3.92 (1H, br d, Jꢃca. 11 Hz), 6ꢁ-H2], 3.65 (1H, m, 5ꢄ-H), 3.68 (1H,
dd, Jꢃ3.4, 9.8 Hz, 3ꢄ-H), 3.72 (1H, m, 3-H), 3.84 (1H, dd, Jꢃ1.5, 3.4 Hz,
2ꢄ-H), 4.31 (1H, m, 9-H), 4.32 (1H, d, Jꢃ7.9 Hz, 1ꢁ-H), 4.71 (1H, d,
Jꢃ1.5 Hz, 1ꢄ-H), 5.35 (1H, dd, Jꢃ9.8, 15.6 Hz, 7-H), 5.52 (1H, dd, Jꢃ7.0,
15.6 Hz, 8-H). 13C-NMR (125 MHz, CD3OD) dC: given in Table 1. Positive-
ion FAB-MS: m/z 543 (MꢂNa)ꢂ.
The known compounds were identified by comparison of their physical
1
data ([a]D, IR, H-, 13C-NMR, MS) with reported values16—22,24—32,34—42) or
authentic samples.23)
Sedumoside E1 (1): An amorphous powder, [a]D24 ꢀ33.9° (cꢃ1.08,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C25H46O11Na
(MꢂNa)ꢂ 545.2938; Found 545.2933. IR (KBr, cmꢀ1): 3431, 2967, 2932,
1
1509, 1473, 1458, 1081, 1046. H-NMR (500 MHz, CD3OD) d: 0.54 (1H,
ddd, Jꢃ2.4, 5.4, 11.3 Hz, 6-H), 0.84, 0.96 (3H each, both s, 11, 12-H3), 0.98,
1.14 (3H each, both d, Jꢃ6.4 Hz, 13, 10-H3), 1.02 (1H, ddd, Jꢃ11.6, 11.6,
11.6 Hz, 4a-H), 1.04, 1.43 (1H each, both m, 7-H2), 1.14 (1H, dd, Jꢃ11.9,
11.9 Hz, 2a-H), 1.26 (3H, d, Jꢃ6.4 Hz, 6ꢄ-H3), 1.46, 1.53 (1H each, both m,
8-H2), 1.48 (1H, m, 5-H), 1.79 (1H, ddd, Jꢃ1.9, 3.7, 11.9 Hz, 2b-H), 2.04
(1H, m, 4b-H), 3.11 (1H, dd, Jꢃ8.0, 9.2 Hz, 2ꢁ-H), 3.26 (1H, dd, Jꢃ9.2,
9.2 Hz, 4ꢁ-H), 3.32 (1H, m, 3ꢁ-H), 3.35 (1H, m, 4ꢄ-H), 3.37 (1H, m, 5ꢁ-H),
[3.59 (1H, dd, Jꢃ6.1, 11.0 Hz), 3.96 (1H, dd, Jꢃ1.8, 11.0 Hz), 6ꢁ-H2], 3.63
(1H, m, 9-H), 3.64 (1H, m, 3ꢄ-H), 3.65 (1H, m, 5ꢄ-H), 3.78 (1H, m, 3-H),
3.82 (1H, dd, Jꢃ1.5, 3.7 Hz, 2ꢄ-H), 4.32 (1H, d, Jꢃ8.0 Hz, 1ꢁ-H), 4.73 (1H,
d, Jꢃ1.5 Hz, 1ꢄ-H). 13C-NMR (125 MHz, CD3OD) dC: given in Table 1. Pos-
itive-ion FAB-MS m/z: 545 (MꢂNa)ꢂ.
Sedumoside G (6): An amorphous powder, [a]D19 ꢀ35.7° (cꢃ0.17,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C25H44O11Na
(MꢂNa)ꢂ: 543.2781. Found: 543.2787. IR (KBr, cmꢀ1): 3406, 2932, 1716,
1
1456, 1368, 1066, 1047. H-NMR (500 MHz, CD3OD) d: 0.59 (1H, ddd,
Jꢃ2.3, 5.3, 10.8 Hz, 6-H), 0.85, 0.95 (3H each, both s, 11, 12-H3), 0.97 (3H,
d, Jꢃ6.5 Hz, 13-H3), 1.03 (1H, ddd, Jꢃ12.1, 12.1, 12.1 Hz, 4a-H), 1.15 (1H,
dd, Jꢃ12.1, 12.1 Hz, 2a-H), 1.26 (3H, d, Jꢃ6.3 Hz, 6ꢄ-H3), 1.31, 1.70 (1H
each, both m, 7-H2), 1.48 (1H, m, 5-H), 1.79 (1H, ddd, Jꢃ2.5, 4.4, 12.1 Hz,
2b-H), 2.05 (1H, m, 4b-H), 2.12 (3H, s, 10-H3), [2.46 (1H, ddd, Jꢃ5.8,
11.0, 16.9 Hz), 2.59 (1H, ddd, Jꢃ5.2, 10.7, 16.9 Hz), 8-H2], 3.11 (1H, dd,
Jꢃ7.9, 9.2 Hz, 2ꢁ-H), 3.78 (1H, m, 3-H), 3.25 (1H, dd-like, 4ꢁ-H), 3.32 (1H,
m, 3ꢁ-H), 3.35 (1H, m, 4ꢄ-H), 3.37 (1H, m, 5ꢁ-H), [3.59 (1H, dd, Jꢃ6.2,
11.1 Hz), 3.95 (1H, dd, Jꢃ1.7, 11.1 Hz), 6ꢁ-H2], 3.65 (1H, m, 5ꢄ-H), 3.64
(1H, m, 3ꢄ-H), 3.82 (1H, dd, Jꢃ1.6, 3.5 Hz, 2ꢄ-H), 4.32 (1H, d, Jꢃ7.9 Hz,
1ꢁ-H), 4.73 (1H, d, Jꢃ1.8 Hz, 1ꢄ-H). 13C-NMR (125 MHz, CD3OD) dC:
given in Table 1. Positive-ion FAB-MS: m/z 543 (MꢂNa)ꢂ.
Sedumoside E2 (2): An amorphous powder, [a]D22 ꢀ38.6° (cꢃ0.27,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C25H46O11Na
(MꢂNa)ꢂ 545.2938; Found 545.2932. IR (KBr, cmꢀ1): 3432, 2967, 2934,
1
1541, 1509, 1474, 1458, 1069, 1046. H-NMR (500 MHz, CD3OD) d: 0.51
(1H, ddd, Jꢃ2.5, 4.9, 11.1 Hz, 6-H), 0.83, 0.94 (3H each, both s, 11, 12-H3),
0.90 (1H, ddd, Jꢃ11.9, 11.9, 11.9 Hz, 4a-H), 0.97, 1.17 (3H each, both d,
Jꢃ6.4 Hz, 13, 10-H3), 1.08, 1.55 (1H each, both m, 7-H2), 1.08 (1H, dd,
Jꢃ12.0, 12.0 Hz, 2a-H), 1.26 (3H, d, Jꢃ6.1 Hz, 6ꢄ-H3), 1.55, 1.59 (1H each,
both m, 8-H2), 1.45 (1H, m, 5-H), 1.63 (1H, ddd, Jꢃ1.9, 3.7, 12.0 Hz, 2b-
H), 1.87 (1H, m, 4b-H), 3.14 (1H, dd, Jꢃ7.7, 9.2 Hz, 2ꢁ-H), 3.25 (1H, dd,
Jꢃ9.2, 9.2 Hz, 4ꢁ-H), 3.33 (1H, m, 3ꢁ-H), 3.37 (2H, m, 4ꢄ, 5ꢁ-H), [3.57 (1H,
dd, Jꢃ6.4, 11.3 Hz), 3.97 (1H, dd, Jꢃ1.5, 11.3 Hz), 6ꢁ-H2], 3.65 (1H, m, 5ꢄ-
H), 3.67 (1H, m, 3ꢄ-H), 3.69 (1H, m, 3-H), 3.77 (1H, m, 9-H), 3.83 (1H, dd,
Jꢃ1.6, 3.4 Hz, 2ꢄ-H), 4.29 (1H, d, Jꢃ7.7 Hz, 1ꢁ-H), 4.73 (1H, d, Jꢃ1.6 Hz,
1ꢄ-H). 13C-NMR (125 MHz, CD3OD) dC: given in Table 1. Positive-ion
FAB-MS m/z: 545 (MꢂNa)ꢂ.
Acid Hydrolysis of 1—6 A solution of 1—6 (each 1.0 mg) in 1 M HCl
(1.0 ml) was heated under reflux for 3 h. After cooling, the reaction mixture
was extracted with EtOAc. The aqueous layers of 1—6 were subjected to
HPLC analysis under the following conditions, respectively: HPLC column,
Kaseisorb LC NH2-60-5, 4.6 mm i.d.ꢅ250 mm (Tokyo Kasei Co., Ltd.,
Tokyo, Japan); detection, optical rotation [Shodex OR-2 (Showa Denko Co.,
Ltd., Tokyo, Japan); mobile phase, CH3CN–H2O (85 : 15, v/v); flow rate
0.8 ml/min]. Identification of D-apiose45ꢀ47) (i) from 3, L-rhamnose (ii) from
1, 2, 5, or 6, D-glucose (iii) from 1—6 present in the aqueous layer was car-
ried out by comparison of its retention time and optical rotation with those
of authentic samples, tR: (i) 6.6 min (D-apiose, positive optical rotation), (ii)
7.8 min (L-rhamnose, negative optical rotation), and (iii) 13.9 min (D-glu-
cose, positive optical rotation), respectively.
Sedumoside E3 (3): An amorphous powder, [a]D21 ꢀ41.5° (cꢃ0.35,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C24H44O11Na
(MꢂNa)ꢂ: 531.2781. Found: 531.2774. IR (KBr, cmꢀ1): 3339, 2922, 1471,
Enzymatic Hydrolysis of 1—3, 5, and 6 with Hesperidinase A solu-
tion of 1 (4.7 mg) in H2O (1.0 ml) was treated with hesperidinase (11.6 mg,
from Aspergillus nigar, Sigma) and the solution was stirred at 37 °C for
12 h. After EtOH was added to the reaction mixture, the solvent was re-
moved under reduced pressure and the residue was purified by HPLC
[MeOH–H2O (55 : 45, v/v)] to furnish (3S,5R,6S,9R)-megastigman-3,9-diol
(1a,44) 1.3 mg, 64%). Through the similar procedure, a solution of 2
(17.3 mg), 3 (3.5 mg), 5 (21.0 mg), and 6 (3.2 mg) in H2O (1.0 ml) was
treated with hesperidinase (49.0, 45.0, 61.6, and 14.5 mg, respectively) and
the solution was stirred at 37 °C for 12 h gave 1a (5.0 mg, 70% from 2,
0.6 mg, 42% from 3), sarmentol F (4a, 5.2 mg, 61% from 5), and sarmentol
G (6a, 0.9 mg, 67% from 6), respectively. Compound 1a was identified by
1
1387, 1039, 1028. H-NMR (500 MHz, CD3OD) d: 0.51 (1H, ddd, Jꢃ2.4,
5.8, 11.0 Hz, 6-H), 0.83, 0.95 (3H each, both s, 11, 12-H3), 0.90 (1H, ddd,
Jꢃ12.2, 12.2, 12.2 Hz, 4a-H), 0.98 (3H, d, Jꢃ6.5 Hz, 13-H3), 1.08, 1.54
(1H each, both m, 7-H2), 1.08 (1H, dd, Jꢃ12.2, 12.2 Hz, 2a-H), 1.17 (3H, d,
Jꢃ6.5 Hz, 10-H3), 1.45 (1H, m, 5-H), 1.55, 1.59 (1H each, both m, 8-H2),
1.63 (1H, ddd, Jꢃ2.4, 4.0, 12.2 Hz, 2b-H), 1.87 (1H, m, 4b-H), 3.14 (1H,
dd, Jꢃ8.0, 9.2 Hz, 2ꢁ-H), 3.26 (1H, dd, Jꢃ9.2, 9.2 Hz, 4ꢁ-H), 3.33 (1H, m,
3ꢁ-H), 3.37 (1H, m, 5ꢁ-H), 3.57 (2H, s, 5ꢄ-H), [3.57 (1H, dd, Jꢃ6.1,
11.3 Hz), 3.97 (1H, dd, Jꢃ1.8, 11.3 Hz), 6ꢁ-H2], 3.69 (1H, m, 3-H), 3.75,
3.94 (1H each, both d, Jꢃ9.8 Hz, 4ꢄ-H), 3.77 (1H, m, 9-H), 3.88 (1H, d,
Jꢃ2.5 Hz, 2ꢄ-H), 4.29 (1H, d, Jꢃ8.0 Hz, 1ꢁ-H), 5.00 (1H, d, Jꢃ2.5 Hz, 1ꢄ-
H). 13C-NMR (125 MHz, CD3OD) dC: given in Table 1. Positive-ion FAB-
MS: m/z 531 (MꢂNa)ꢂ.
1
comparison of its physical data ([a]D and H- and 13C-NMR) with reported
values.44)
Sarmentol F (4a): Colorless oil, [a]D24 ꢀ12.1° (cꢃ0.27, MeOH). High-
resolution EI-MS: Calcd for C13H24O2 (Mꢂ): 212.1776. Found: 212.1769. IR
(film, cmꢀ1): 3303, 2967, 2924, 1669, 1470, 1385, 1296, 1273, 1130, 1061,
976, 943, 916, 878, 756. 1H-NMR (500 MHz, CDCl3) d: 0.82 (3H, d,
Jꢃ6.4 Hz, 13-H3), 0.84, 0.88 (3H each, both s, 11, 12-H3), 0.92 (1H, ddd,
Sedumoside F1 (4): An amorphous powder, [a]D24 ꢀ11.2° (cꢃ1.14,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C19H34O7Na
(MꢂNa)ꢀ: 397.2202. Found: 397.2206. IR (KBr, cmꢀ1): 3389, 2960, 2919,
1734, 1684, 1671, 1559, 1541, 1509, 1474, 1341, 1078, 1034. 1H-NMR
(500 MHz, CD3OD) d: 0.82 (3H, d, Jꢃ6.4 Hz, 13-H3), 0.88, 0.90 (3H each,