J Nat Med
20
based on TLC analysis. FIV.1 (1.5 g) was subjected to
Sephadex LH-20 column chromatography ([ 2.5 9 60 cm)
eluted with MeOH to give three subfractions (FIV.3.1–FIV.3.3).
FIV.3.1 (876.0 mg) was subjected to further silica gel CC
(200–300mesh, [ 2.0 9 30 cm, 25 g)elutedwithpetroleum
ether–EtOAc (10:1, 5:1), and FIV.3.1.1 (170 mg) was purified
by preparative semi-HPLC [Waters SymmetryPrepTM C-18
column (7 lm, [ 19.0 9 300 mm), 8 mL/min, 55 % MeOH
in H2O] to afford compounds 8 (30.0 mg), and 10 (1.5 mg).
Compound 2: Colorless needles (MeOH); ½aꢁD -20.8
(c 0.125, MeOH); UV (MeOH) kmax (log e), 242 (2.96) nm;
CD (MeOH, c 6.31 9 10-4 M): 205 (?11.5), 240 (-20.1)
nm (De). IR (KBr) mmax 3408, 2926, 2860, 1660, 1621,
1445, 1384, 1114, 1101, 1075, 1041, 641, 591 cm-1
;
1H- and 13C-NMR data see Table 1. HR-ESI-MS at m/z
419.2049 [M?Na]? (calcd. for C21H32O7Na, 419.2046).
20
Compound 4: Colorless needles (MeOH); ½aꢁD ?6.5
(c 0.750, MeOH); UV (MeOH) kmax (log e), 252 (2.96) nm;
CD (MeOH, c 3.61 9 10-4 M): 202 (?21.5), 244 (?23.3),
280 (-12.3) nm (De). IR (KBr) mmax 3395, 2915, 2872,
2855, 1657, 1617, 1461, 1430, 1372, 1306, 1229, 1162,
FIV.3.1.2 (108.0 mg) was further purified with the same HPLC
system using 58 % MeOH in H2O resulting in the isolation of
compounds 10 (70.0 mg) and 11 (3.0 mg). FIV.3.1.3 (38 mg)
was subjected to the same HPLC procedures (50 % MeOH in
H2O) to afford compounds 6 (4.3 mg), 2 (5.0 mg) and 5
(4.3 mg). FIV.3.1.4 (184.0 mg) was purified with HPLC to
afford compounds 1 (11.0 mg), 4 (25.4 mg), and 3 (37.1 mg)
by the above HPLC system (29 % CH3CN in H2O). FIV.3.2
(205.0 mg) was separated by RP-18 silica gel column CC
([ 1.5 9 30 cm, 8 g) using 35 % MeOH in H2O, followed
by HPLC (30 % CH3CN in H2O) to afford compound 12
(7.0 mg).
1085, 1048, 998, 885, 644, 580 cm-1; H- and 13C-NMR
1
data see Table 2. HR-ESI-MS at m/z 419.2037 [M?Na]?
(calcd. for C21H32O7Na, 419.2046).
20
Compound 5: Amorphous solid; ½aꢁD ?75.8 (c 0.215,
MeOH); UV (MeOH) kmax (log e), 251 (2.94) nm; CD
(MeOH, c 4.32 9 10-4 M): 201 (?11.7), 245 (?21.6), 284
(-7.1) nm (De). IR (KBr) mmax 3396, 2929, 2859, 1658, 1622,
1441, 1372, 1319, 1302, 1227, 1196, 1101, 1075,1037,
900,884, 634 cm-1; 1H- and 13C-NMR data see Table 2. HR-
ESI-MS at m/z 419.2027 [M?Na]? (calcd. for C21H32O7Na,
419.2046).
Petroleum ether extract (150.0 g) was subjected to CC
on silica gel (100–200 mesh, [ 8.0 9 60 cm, 600 g eluted
with petroleum ether:EtOAc in a gradient manner (50:1,
20
Compound 6: Amorphous solid; ½aꢁD ?52.1 (c 0.215,
20:1, 10:1, 5:1, 1:1, 0:1) to yield seven fractions (F2.1–F2.7
)
MeOH); UV (MeOH) kmax (log e), 264 (2.68) nm; CD (MeOH,
c 1.03 9 10-3 M): 216 (?12.4), 255 (?16.4), 282 (-7.2) nm
(De). IR (KBr) mmax 3386, 2928, 2859, 1761, 1743, 1662, 1619,
1384, 1306, 1260, 1241, 1199, 1101, 1074, 1033, 641,
564 cm-1;1H- and 13C-NMR data see Table 2. HR-ESI-MS at
m/z 433.1830 [M?Na]? (calcd. for C21H30O8Na, 433.1838).
based on TLC analysis. F2.1 (3.9 g) was also subjected to
silica gel CC (200–300 mesh, [ 4.0 9 40 cm, 80 g) eluted
with petroleum ether to obtain compounds 13 (38.0 mg)
and 14 (15.0 mg). F2.2 (5.0 g) was treated exactly like F2.1
,
eluted with petroleum ether:EtOAc (80:1) to afford com-
pounds 15 (29.0 mg), 7 (3.0 mg) and 22 (13.0 mg). F2.3
(4.6 g) was separated by RP-18 silica gel column CC ([
3.5 9 60 cm, 40 g) using 90 % MeOH in H2O, and F2.3.2
was purified by silica gel column CC eluted with petroleum
ether:EtOAc (30:1) to afford compounds 16 (4.0 mg) and
17 (119.0 mg), F2.3.3 was purified by silica gel column CC
eluted with petroleum ether:EtOAc (35:1) to afford com-
pound 9 (19.0 mg). F2.5 (6.6 g) was separated by silica gel
column CC ([ 4.5 9 60 cm, 70 g) eluted with petroleum
ether:EtOAc (20:1). F2.5.3 was purified by RP-18 silica gel
column CC ([ 1.5 9 60 cm, 8 g) using 42 % CH3CN in
H2O to afford compounds 18 (81.0 mg), 19 (5 mg), and 20
(20 mg). F2.5.3 was separated by Sephadex LH-20 ([
1.5 9 60 cm, MeOH) to get compound 21 (25.6 mg).
Compound 7: Colorless oil; ½aꢁ2D0 -48.7 (c 0.150, CHCl3);
UV (MeOH) kmax (log e), 233 (3.01) nm; IR (KBr) mmax 2972,
2927, 2858, 1761, 1701, 1649, 1441, 1386, 1307, 1284, 1204,
1112, 1100, 1045, 1022, 976, 945, 915, 749 cm-1; 1H- and
13C-NMR data see Table 1. HR-ESI-MS at m/z 299.1626
[M?Na]? (calcd. for C17H24O3Na, 299.1623).
Crystal data of 2 (ref 22): C21H32O7, Mr 396.47, ortho-
˚
rhombic, space group P212121, a = 7.1264(5) A, a = 90°,
˚
˚
b = 7.6320(9) A, b = 90°, c = 37.822(3) A, c = 90°,
3
3
˚
V = 2057.1(3) A , Z = 4, Dc = 1.280 Mg/m , F(000) =
856; crystal dimensions/shape/color 0.35 9 0.30 9
0.01 mm/block/colorless. Reflections collected 4694, inde-
pendent reflections 3277. Number of parameters refined 265;
final R indices [I [ 2r(I)] R1 = 0.0409, wR2 = 0.0929;
R indices (all data) R = 0.0484, wR2 = 0.0982.
20
Compound 1: Amorphous solid; ½aꢁD ?31.3 (c 0.550,
MeOH); UV (MeOH) kmax (log e), 240 (3.04) nm; CD
(MeOH, c 5.50 9 10-4 M): 200 (?25.0), 239 (?8.7), 271
(-3.6) nm (De). IR (KBr) mmax 3391, 2929, 1651, 1621,
1462, 1434, 1371, 1323, 1262, 1235, 1201, 1170, 1074,
1043, 892, 845, 654, 596 cm-1; 1H- and 13C-NMR data see
Table 1. HR-ESI-MS at m/z 419.2053 [M?Na]? (calcd. for
C21H32O7Na, 419.2046).
Crystaldata of 4 (ref22): C21H32O7,Mr 224.75,monoclinic,
˚
˚
space group P21, a = 7.412(2) A, a = 90°, b = 7.943(3) A,
o
3
˚
˚
b = 92.45(3) , c = 20.031(7) A; c = 90°, V = 1178.2(7) A ,
Z = 4, Dc = 1.267 Mg/m3, F(000) = 486, crystal dimen-
sions/shape/color 0.03 9 0.03 9 0.02 mm/block/colorless.
Reflections collected 7289, independent reflections 3989.
123