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J Nat Med (2011) 65:617–622
Plant material
The residue (28.5 g) in fractions 9–15 obtained on
Diaion HP-20 CC was subjected to silica gel (600 g) CC
with increasing amounts of MeOH in CHCl3 [CHCl3 (3 l),
and CHCl3–MeOH (49:1, 3 l), (24:1, 3 l), (23:2, 3 l), (9:1,
3 l), (7:1, 3 l), (17:3, 3 l), (4:1, 3 l), (3:1, 3 l), and (7:3,
3 l)], 500 ml fractions being collected. The residue
(933 mg) in fractions 31–35 was separated by ODS open
CC, followed by DCCC (117 mg in fractions 157–171) to
give 58.5 mg of 11 in fractions 73–97. The residue (5.23 g)
in fractions 45–51 was separated by ODS open CC to give
153 mg of 4 in fractions 118–125 and 75.4 mg of 6 in
fractions 170–177. The residue (7.04 g) in fractions 52–59
was separated by ODS open CC to give 4.1 mg of 16 in
fractions 65–81. ODS fractions 45–60 (30.1 mg) were
combined and purified by HPLC (MeOH–H2O, 1:9) to give
8.1 mg of 15 from the peak at 7 min.
Leaves of S. jasminodora were collected in Kunigami-son,
Kunigami-gun, Okinawa, Japan in June 2005, and a vou-
cher specimen was deposited in the Herbarium of Phar-
maceutical Sciences, Graduate School of Biomedical
Sciences, Hiroshima University (05-SJ-Okinawa-0627).
Extraction and isolation
Air-dried leaves of S. jasminodora (5.66 kg) were extrac-
ted three times with MeOH (30 l 9 3) at room temperature
for one week and then concentrated to 3 l in vacuo. The
concentrated extract was washed with n-hexane (3 l,
72.0 g) and then the MeOH layer was concentrated to a
gummy mass. The latter was suspended in water (3 l) and
then extracted with EtOAc (3 l) to give 138 g of an EtOAc-
soluble fraction. The aqueous layer was extracted with
1-BuOH (3 l) to give a 1-BuOH-soluble fraction (106 g),
and the remaining water-layer was concentrated to furnish
358 g of a water-soluble fraction. The 1-BuOH-soluble
fraction (106 g) was subjected to Diaion HP-20 CC
(U = 55 mm, L = 45 cm), using H2O–MeOH (4:1, 3 l),
(3:2, 3 l), (2:3, 3 l), and (1:4, 3 l), and MeOH (3 l), 500 ml
fractions being collected. The residue (20.5 g) in fractions
4–6 from the 20% MeOH eluate was subjected to silica gel
(450 g) CC with increasing amounts of MeOH in CHCl3
[CHCl3 (3 l), and CHCl3–MeOH (49:1, 3 l), (24:1, 3 l),
(23:2, 3 l), (9:1, 3 l), (7:1, 3 l), (17:3, 3 l), (4:1, 3 l), (3:1,
3 l), (7:3, 3 l), and (3:2, 3 l)], 500 ml fractions being col-
lected. The residue (1.44 g) in fractions 44–49 was sepa-
rated by ODS open CC, followed by DCCC (114 mg in
fractions 47–54) to give 73.6 mg of 13 in fractions 8–14.
The residue (91.3 mg) in fractions 63–72 was purified by
HPLC (MeOH–H2O, 1:4) to yield 12.1 mg of 2 from the
peak at 12 min.
The residue (7.22 g) in fraction 16–20 obtained on
Diaion HP-20 CC was subjected to silica gel (180 g) CC
with increasing amounts of MeOH in CHCl3 [CHCl3
(1.5 l), and CHCl3–MeOH (97:3, 1.5 l), (19:1, 1.5 l), (23:2,
1.5 l), (9:1, 1.5 l), (7:1, 1.5 l), (17:3, 1.5 l), (4:1, 1.5 l),
(3:1, 1.5 l), (7:3, 1.5 l), and (3:2, 1.5 l)], 250 ml fractions
being collected. The residue (650 mg) in fractions 41–46
was separated by ODS open CC, followed by DCCC
(160 mg in fraction 175–188) to give 26.2 mg of 7 in
fractions 36–46 and 66.0 mg of 9 in fractions 47–55.
Compound 5 (30.6 mg) was obtained from fractions
194–205 on ODS open CC as a precipitate.
The residue (20.9 g) in fractions 21–27 obtained on
Diaion HP-20 CC was subjected to silica gel (500 g) CC
with increasing amounts of MeOH in CHCl3 [CHCl3 (3 l),
and CHCl3–MeOH (49:1, 3 l), (24:1, 3 l), (19:1, 3 l), (23:2,
3 l), (9:1, 3 l), (7:1, 3 l), (17:3, 3 l), (4:1, 3 l), (3:1, 3 l),
and (7:3, 3 l)], 500 ml fractions being collected. The res-
idue (5.81 g) in fractions 25–31 was separated by ODS
open CC to give 95.6 mg of 8 in fractions 211–223.
Isoorientin 400-O-gallate (1). Amorphous powder, [a]2D3
?8.4 (c 0.95, MeOH). IR (film) cm-1: 3340, 1695, 1650,
1616, 1491, 1446, 1348, 1234, 1115, 1039. UV (MeOH)
The residue (8.1 g) in fractions 7–8 obtained on Diaion
HP-20 CC was subjected to silica gel (350 g) CC with
increasing amounts of MeOH in CHCl3 [CHCl3 (1.5 l), and
CHCl3–MeOH (97:3, 1.5 l), (19:1, 1.5 l), (23:2, 1.5 l), (9:1,
1.5 l), (7:1, 1.5 l), (17:3, 1.5 l), (4:1, 1.5 l), (3:1, 1.5 l), (7:3,
1.5 l), and (3:2, 1.5 l)], 250 ml fractions being collected. The
residue (799 mg) in fractions 28–35 was separated by ODS
open CC, followed by DCCC (102 mg in fractions 87–102)
to give 26.6 mg of 10 in fractions 70–84. The residue
(710 mg) in fractions 36–41 was separated by ODS open CC,
followed by DCCC (142 mg in fractions 90–108) to afford
53.0 mg of 14 in fractions 50–59 and 9.7 mg of 12 in frac-
tions 73–83. The residue (880 mg) in fractions 52–69 was
separated by ODS open CC to give 38.4 mg of 3 in fractions
147–164 and 28.1 mg of 1 in fractions 194–204. The residue
(820 mg) in fractions 70–78 was separated by ODS open CC
to give 195 mg of 17 in fractions 60–88.
1
nm (log e): 348 (4.32), 271 (4.36), 217 (4.47). H-NMR
(400 MHz, DMSO-d6) d: 13.63 (1H, s, 5-OH), 7.42 (1H,
dd, J = 8, 2 Hz, H-60), 7.40 (1H, d, J = 2 Hz, H-20), 7.00
(2H, s, H-2000 and 6000), 6.90 (1H, d, J = 8 Hz, H-50), 6.67
(1H, s, H-3), 6.51 (1H, s, H-8), 4.80 (1H, dd, J = 10,
10 Hz, H-400), 4.69 (1H, d, J = 10 Hz, H-100), 4.24 (1H, dd,
J = 10, 10 Hz, H-200), 3.51 (1H, dd, J = 10, 10 Hz, H-300),
3.47 (1H, ddd, J = 12, 6, 2 Hz, H-500), 3.41–3.28 (2H, m,
H-600a and 600b). 1H-NMR (400 MHz, CD3OD) d: 7.35 (1H,
d, J = 2 Hz, H-20), 7.34 (1H, dd, J = 9, 2 Hz, H-60), 7.14
(2H, s, H-2000 and 6000), 6.89 (1H, d, J = 9 Hz, H-50), 6.52
(1H, s, H-3), 6.48 (1H, s, H-8), 5.11 (1H, dd, J = 10,
10 Hz, H-400), 4.99 (1H, d, J = 10 Hz, H-100), 4.38 (1H, dd,
J = 10, 10 Hz, H-200), 3.79 (1H, dd, J = 10, 10 Hz, H-300),
123