August 2007
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spectra, Shimadzu FTIR-8100 spectrometer; EI-MS and high-resolution MS,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C24H44O12Na
JEOL JMS-GCMATE mass spectrometer; FAB-MS and high-resolution MS, (MꢁNa)ꢁ: 547.2730; Found: 547.2728. IR (KBr, cmꢀ1): 3410, 2940, 2918,
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JEOL JMS-SX 102A mass spectrometer; H-NMR spectra, JEOL EX-270
(270 MHz) and JNM-LA500 (500 MHz) spectrometers; 13C-NMR spectra,
JEOL EX-270 (68 MHz) and JNM-LA500 (125 MHz) spectrometers with
1541, 1474, 1171, 1081, 1047. 1H-NMR (500 MHz, pyridine-d5) d: 0.51
(1H, ddd, Jꢂ2.8, 4.6, 10.7 Hz, 6-H), 0.77, 0.92 (3H each, both s, 11, 12-H3),
0.90 (3H, d, Jꢂ6.1 Hz, 13-H3), 1.16, 1.92 (1H each, both m, 7-H2), 1.19
tetramethylsilane as an internal standard; and HPLC detector, Shimadzu (1H, ddd, Jꢂ11.3, 11.3, 11.3 Hz, 4a-H), 1.27 (1H, dd, Jꢂ11.9, 11.9 Hz, 2a-
RID-6A refractive index and SPD-10Avp UV–VIS detectors. HPLC col-
umn, Cosmosil 5C18-MS-II (Nacalai Tesque Inc., 250ꢇ4.6 mm i.d.) and
(250ꢇ20 mm i.d.) columns were used for analytical and preparative pur- 11.3 Hz), 4.35 (1H, d, Jꢂ5.2, 11.3 Hz), 5ꢆ-H2], 3.96 (2H, m, 10-H2), 3.98
H), 1.27 (1H, m, 5-H), 1.66, 1.80 (1H each, both m, 8-H2), 2.02 (1H, ddd,
Jꢂ2.0, 3.1, 11.9 Hz, 2b-H), 2.21 (1H, m, 4b-H), [3.70 (1H, dd, Jꢂ10.4,
poses, respectively.
(1H, m, 5ꢃ-H), 4.04 (2H, m, 9, 2ꢆ-H), 4.05 (1H, m, 2ꢃ-H), 4.12 (1H, m, 3-H),
The following experimental conditions were used for chromatography: or- 4.15 (1H, dd, Jꢂ8.8, 8.8 Hz, 3ꢆ-H), 4.21 (1H, m, 4ꢆ-H), 4.29 (2H, m, 3ꢃ, 4ꢃ-
dinary-phase silica gel column chromatography, Silica gel BW-200 (Fuji H), [4.43 (1H, dd, Jꢂ5.2, 11.9 Hz), 4.58 (1H, dd, Jꢂ2.5, 11.9 Hz), 6ꢃ-H2],
Silysia Chemical, Ltd., Aichi, Japan, 150—350 mesh); reverse-phase silica 5.02 (1H, d, Jꢂ8.0 Hz, 1ꢃ-H), 5.04 (1H, d, Jꢂ8.2 Hz, 1ꢆ-H). 1H-NMR
gel column chromatography, Chromatorex ODS DM1020T (Fuji Silysia (500 MHz, CD3OD) d: 0.53 (1H, ddd, Jꢂ2.2, 4.6, 10.7 Hz, 6-H), 0.82, 0.95
Chemical, Ltd., Aichi, Japan, 100—200 mesh); TLC, precoated TLC plates
with Silica gel 60F254 (Merck, 0.25 mm) (ordinary phase) and Silica gel RP-
18 F254S (Merck, 0.25 mm) (reverse phase); reverse-phase HPTLC, precoated dd, Jꢂ12.2, 12.2 Hz, 2a-H), 1.44 (1H, m, 5-H), 1.49, 1.65 (1H each, both
TLC plates with Silica gel RP-18 WF254S (Merck, 0.25 mm); and detection
was achieved by spraying with 1% Ce(SO4)2–10% aqueous H2SO4, followed 3.11 (1H, dd, Jꢂ7.6, 9.2 Hz, 2ꢃ-H), [3.17 (1H, dd, Jꢂ10.4, 11.0 Hz), 3.83
by heating. (1H, m), 5ꢆ-H2], 3.18 (1H, dd, Jꢂ7.6, 9.4 Hz, 2ꢆ-H), 3.25 (2H, m, 4ꢃ, 5ꢃ-H),
(3H each, both s, 11, 12-H3), 0.97 (3H, d, Jꢂ6.5 Hz, 13-H3), 1.02 (1H, ddd,
Jꢂ11.9, 11.9, 11.9 Hz, 4a-H), 1.06, 1.65 (1H each, both m, 7-H2), 1.13 (1H,
m, 8-H2), 1.79 (1H, ddd, Jꢂ2.2, 4.3, 12.2 Hz, 2b-H), 2.01 (1H, m, 4b-H),
Plant Material S. sarmentosum was cultivated in Huangshan, Anhui 3.31 (1H, m, 3ꢆ-H), 3.33 (1H, m, 3ꢃ-H), 3.47 (1H, m, 4ꢆ-H), [3.51 (1H, dd,
province, China, and plant material was identified by one of authors (M. Y.). Jꢂ7.0, 12.8 Hz), 3.60 (1H, m), 10-H2], 3.61 (1H, m, 9-H), [3.64 (1H, dd,
A voucher specimen (2005.01. Eishin-02) of this plant is on file in our labo-
Jꢂ5.2, 11.9 Hz), 3.85 (1H, m), 6ꢃ-H2], 3.84 (1H, m, 3-H), 4.33 (1H, d,
Jꢂ7.7 Hz, 1ꢃ-H), 4.33 (1H, d, Jꢂ7.7 Hz, 1ꢆ-H). 13C-NMR (125 MHz, pyri-
dine-d5 and CD3OD) dC: see Table 2. Positive-ion FAB-MS: m/z 547
(MꢁNa)ꢁ.
ratory.1,16)
Extraction and Isolation Fractions 1-5 (1510 mg), 2-5 (3300 mg), 2-8
(1800 mg), 2-10 (1360 mg), 3-7 (230 mg), and 5-6 (665 mg) were obtained
from the methanol-eluted fraction of the hot water extract from the fresh
whole plant of S. sarmentosum as reported previously.1,16) Fraction 1-5
Sedumoside H (18): An amorphous powder, [a]D27 ꢁ71.4° (cꢂ0.21,
MeOH). High-resolution positive-ion FAB-MS: Calcd for C19H32O8Na
(1510 mg) was purified on Sephadex LH-20 column chromatography [150 g, (MꢁNa)ꢁ: 411.1995; Found: 411.1989. CD [MeOH, nm (De)]: 211
CHCl3–MeOH (1 : 1, v/v)] and finally HPLC [MeOH–H2O (35 : 65, v/v)] to
furnish sedumoside I (19, 107.2 mg, 0.00020%). Fraction 2-5 (3300 mg) was
further separated by HPLC [CH3CN–H2O (15 : 85, v/v)] to furnish sedumo-
(ꢁ4.40), 237 (ꢁ3.42), 335 (ꢁ0.75). UV [MeOH, nm (log e)]: 240 (4.08). IR
(KBr, cmꢀ1): 3389, 3011, 2961, 2876, 1669, 1471, 1076, 1038, 752. 1H-
NMR (500 MHz, CD3OD) d: 1.01, 1.09, 2.04 (3H each, all s, 12, 11, 13-H3),
side H (18, 83.7 mg, 0.00016%) and 19 (34.1 mg, 0.00006%). Fraction 1.49, 1.98 (1H each, both m, 7-H2), 1.51, 1.61 (1H each, both m, 8-H2), 1.96
2-8 (1800 mg) was purified on Sephadex LH-20 column chromatography (1H, m, 6-H), 2.00 (1H, d, Jꢂ17.1 Hz, 2b-H), 2.46 (1H, d, Jꢂ17.1 Hz, 2a-
[150 g, CHCl3–MeOH (1 : 1, v/v)] and finally HPLC [CH3CN–MeOH–H2O H), 3.21 (1H, dd, Jꢂ7.7, 8.6 Hz, 2ꢃ-H), 3.28 (1H, m, 5ꢃ-H), 3.30 (1H, m, 4ꢃ-
(20 : 8 : 72, v/v/v) and MeOH–H2O (40 : 60, v/v)] to furnish sedumoside
A4 (15, 4.9 mg, 0.00001%). Fraction 2-10 (1360 mg) was further separated
by HPLC [CH3CN–MeOH–H2O (20 : 8 : 72, v/v/v) and MeOH–H2O (40 :
60, v/v)] to furnish 18 (37.5 mg, 0.00007%). Fraction 3-7 (230 mg) was
purified by HPLC [MeOH–H2O (29 : 71, v/v)] to furnish sedumoside
A6 (17, 12.0 mg, 0.00002%). Fraction 5-6 (665 mg) was purified by
HPLC [CH3CN–MeOH–H2O (10 : 8 : 82, v/v/v)] to give sedumoside A5
(16, 18.4 mg, 0.00003%).
H), 3.36 (1H, m, 3ꢃ-H), [3.43 (1H, dd, Jꢂ6.4, 10.1 Hz), 4.74 (1H, dd, Jꢂ3.4,
10.1 Hz), 10-H2], [3.64 (1H, dd, Jꢂ4.9, 11.6 Hz), 3.84 (1H, br d, Jꢂca.
12 Hz), 6ꢃ-H2], 3.75 (1H, m, 9-H), 4.28 (1H, d, Jꢂ7.7 Hz, 1ꢃ-H), 5.81 (1H, s,
4-H). 13C-NMR (125 MHz, CD3OD) dC: see Table 2. Positive-ion FAB-MS:
m/z 411 (MꢁNa)ꢁ.
Sedumoside I (19): An amorphous powder, [a]D27 ꢀ0.2° (cꢂ1.41, MeOH).
High-resolution EI-MS: Calcd for C19H32O8 (Mꢁ): 388.2097; Found:
388.2095. CD [MeOH, nm (De)]: 285 (ꢁ0.09). IR (KBr, cmꢀ1): 3431, 2961,
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Sedumoside A4 (15): An amorphous powder, [a]D26 ꢀ11.1° (cꢂ0.25, 1723, 1715, 1559, 1472, 1076, 1044, 753. H-NMR (500 MHz, CD3OD) d:
MeOH). High-resolution positive-ion FAB-MS: Calcd for C18H34O7Na 0.77, 1.07 (3H each, both s, 11, 12-H3), 1.08 (3H, d, Jꢂ6.7 Hz, 13-H3), 1.21
(MꢁNa)ꢁ 385.2202; Found 385.2209. IR (KBr, cmꢀ1): 3405, 2924, 2870, (1H, ddd, Jꢂ3.1, 6.1, 10.7 Hz, 6-H), 1.46, 1.81 (1H each, both m, 7-H2),
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1473, 1072, 1039. H-NMR (500 MHz, CD3OD) d: 0.53 (1H, ddd, Jꢂ2.0,
1.81 (1H, m, 5-H), 1.96 (1H, dd, Jꢂ2.2, 13.2 Hz, 2b-H), 2.22 (1H, ddd,
4.6, 10.7 Hz, 6-H), 0.84, 0.95 (3H each, both s, 11, 12-H3), 0.98 (3H, d, Jꢂ2.2, 4.6, 14.1 Hz, 4b-H), 2.16 (1H, dd, Jꢂ14.1, 14.1 Hz, 4a-H), 2.39
Jꢂ6.8 Hz, 13-H3), 0.90 (1H, ddd, Jꢂ11.9, 11.9, 11.9 Hz, 4a-H), 1.07, 1.66 (1H, d, Jꢂ13.2 Hz, 2a-H), 2.69 (2H, m, 8-H2), 3.25 (1H, m, 2ꢃ-H), 3.26
(1H each, both m, 7-H2), 1.09 (1H, dd, Jꢂ11.9, 11.9 Hz, 2a-H), 1.45 (1H,
m, 5-H), 1.49, 1.65 (1H each, both m, 8-H2), 1.64 (1H, ddd, Jꢂ2.4, 4.0,
(1H, m, 4ꢃ-H), 3.27 (1H, m, 5ꢃ-H), 3.36 (1H, m, 3ꢃ-H), 3.64, 3.87 (1H each,
both m, 6ꢃ-H2), 4.31 (1H, d, Jꢂ7.7 Hz, 1ꢃ-H), 4.33, 4.52 (1H each, both d,
11.9 Hz, 2b-H), 1.89 (1H, m, 4b-H), [3.18 (1H, dd, Jꢂ10.4, 11.3 Hz), 3.84 Jꢂ17.4 Hz, 10-H2). 13C-NMR (125 MHz, CD3OD) dC: see Table 2. EI-MS
(1H, dd, Jꢂ5.2, 11.3 Hz), 5ꢃ-H2], 3.20 (1H, dd, Jꢂ7.7, 9.2 Hz, 2ꢃ-H), 3.31
(1H, m, 3ꢃ-H), 3.47 (1H, ddd, Jꢂ5.2, 8.9, 10.4 Hz, 4ꢃ-H), [3.53 (1H, dd,
Jꢂ6.7, 12.5 Hz), 3.62 (1H, m), 10-H2], 3.62 (1H, m, 9-H), 3.70 (1H, m, 3-
(%): m/z 388 (Mꢁ, 1), 370 (MꢁꢀH2O, 1), 255 (100), 227 (66), 208 (42).
Acid Hydrolysis of 15—19 A solution of sedumosides A4 (15, 3.1 mg),
A5 (16, 2.0 mg), or A6 (17, 2.5 mg) in 1 M HCl (1.0 ml) was heated under re-
H), 4.34 (1H, d, Jꢂ7.7 Hz, 1ꢃ-H). 13C-NMR (125 MHz, CD3OD) dC: see flux for 1 h. After cooling, the reaction mixture was extracted with EtOAc.
Table 2. Positive-ion FAB-MS m/z: 385 (MꢁNa)ꢁ.
The EtOAc extract was neutralized with Amberlite IRA-400 (OHꢀ form),
Sedumoside A5 (16): An amorphous powder, [a]D19 ꢀ16.7° (cꢂ0.93, and the resin was removed by filtration. Removal of the solvent from the fil-
MeOH). High-resolution positive-ion FAB-MS: Calcd for C25H46O13Na trate under reduced pressure furnished a residue, which was purified by
(MꢁNa)ꢁ 577.2836; Found 577.2831. IR (KBr, cmꢀ1): 3410, 2941, 2898, HPLC [MeOH–H2O (35 : 65, v/v)] to give sarmentol A16) (2, 1.2 mg, 91%
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1474, 1171, 1076, 1030. H-NMR (500 MHz, CD3OD) d: 0.55 (1H, ddd, from 15; 1.3 mg, 79% from 16; or 0.8 mg, 73% from 17). Through the simi-
Jꢂ1.9, 5.2, 11.3 Hz, 6-H), 0.83, 0.97 (3H each, both s, 11, 12-H3), 0.98 (3H, lar procedure, a solution of sedumosides H (18) or I (19) (each 1.0 mg) in
d, Jꢂ6.5 Hz, 13-H3), 1.02 (1H, ddd, Jꢂ12.2, 12.2, 12.2 Hz, 4a-H), 1.08,
1.65 (1H each, both m, 7-H2), 1.13 (1H, dd, Jꢂ12.2, 12.2 Hz, 2a-H), 1.45
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 15—18 were sub-
(1H, m, 5-H), 1.57, 1.64 (1H each, both m, 8-H2), 1.79 (1H, ddd, Jꢂ2.2, 3.7, jected to HPLC analysis under the following conditions, respectively: HPLC
12.2 Hz, 2b-H), 2.01 (1H, m, 4b-H), 3.17 (1H, dd, Jꢂ7.7, 9.2 Hz, 2ꢃ-H), column, Kaseisorb LC NH2-60-5, 4.6 mm i.d.ꢇ250 mm (Tokyo Kasei Co.,
3.20 (1H, dd, Jꢂ7.7, 9.2 Hz, 2ꢆ-H), 3.25—3.29 (4H, m, 4ꢃ, 5ꢃ, 4ꢆ, 5ꢆ-H), Ltd., Tokyo, Japan); detection, optical rotation [Shodex OR-2 (Showa Denko
3.33 (1H, dd, Jꢂ9.2, 9.2 Hz, 3ꢆ-H), 3.35 (1H, dd, Jꢂ9.2, 9.2 Hz, 3ꢃ-H), [3.52 Co., Ltd., Tokyo, Japan); mobile phase, CH3CN–H2O (85 : 15, v/v); flow rate
(1H, dd, Jꢂ5.8, 11.9 Hz), 3.65 (1H, dd, Jꢂ3.4, 11.9 Hz), 10-H2], [3.64 (2H,
m), 3.85 (2H, dd, Jꢂ2.0, 12.0 Hz), 6ꢃ, 6ꢆ-H2], 3.69 (1H, m, 9-H), 3.84 (1H,
0.8 ml/min]. Identification of D-xylose (i) from 15 and 17 and D-glucose (ii)
from 15—19 present in the aqueous layer was carried out by comparison of
m, 3-H), 4.33 (1H, d, Jꢂ7.7 Hz, 1ꢃ-H), 4.42 (1H, d, Jꢂ7.7 Hz, 1ꢆ-H). 13C- the retention time and optical rotation with those of authentic sample, tR: (i)
NMR (125 MHz, CD3OD) dC: see Table 2. Positive-ion FAB-MS m/z: 577
9.5 min (D-xylose, positive optical rotation), and (ii) 13.9 min (D-glucose,
positive optical rotation), respectively.
Enzymatic Hydrolysis of 18 and 19 with b-Glucosidase A solution of
(MꢁNa)ꢁ.
Sedumoside A6 (17): An amorphous powder, [a]D17 ꢀ26.8° (cꢂ0.60,