Journal of Asian Natural Products Research
305
(4.4 g) were re-chromatographed over
LiChroprep RP-18 (ODS silica gel; 40–
63 mm: 200 g; each fraction 100 ml). The
elution was sequentially performed with
methanol and water to yield 20 fractions
(50 ml each): fractions 1–4 with H2O–
MeOH (8:2, v/v), fractions 5–8 with
H2O–MeOH (6:4, v/v), fractions 9–12
with H2O–MeOH (4:6, v/v), fractions 13–
16 with H2O–MeOH (2:8, v/v), and
fractions 17–20 with methanol. Fractions
9–12 (0.56 g) were re-chromatographed
over Lichroprep RP18 ODS (80 g, each
fraction of 50 ml). The elution was
sequentially performed with methanol
containing 80%, 60%, 40%, 20%, 10%,
and 0% of water to yield new compound 1
(20 mg).
C6H14–EtOAc to yield one new aliphatic
acid (2, 19 mg) and known compound (3,
22 mg).
3.3.1 Stigmast-5-en-3b-ol-3-O-b-D-(20-
n-triacontanoyl) glucopyranoside
White solid; 20 mg; Rf 0.60 (CHCl3:
20
MeOH: 8.5:1.5); ½aꢀD þ 47.1 (c ¼ 1,
MeOH); IR nmax (KBr): 3430, 3350,
2919, 2850, 1736, 1645, 1463, 1377,
1
1242, and 1020 cm21; H and 13C NMR
spectral data see Table 1; þ ve FAB-MS
m/z (rel. int.): 1011 [M þ H]þ(1.5), 452
(11.3), 413 (12.6), 183 (9.1), 163 (10.2);
HR-FAB-MS: m/z 1011.8954 [M]þ (calcd
for C65H119O7, 1011.8960).
The entire ethyl acetate extract was
subjected to normal phase column chro-
matography over silica gel (500 g) to yield
30 fractions (each of 500 ml) with the
following eluants: fractions 1 and 2 with
C6H14, fractions 3 and 4 with C6H14–
EtOAc (9:1, v/v), fractions 5 and 6 with
C6H14–EtOAc (8:2, v/v), fractions 7 and 8
with C6H14–EtOAc (7:3, v/v), fractions 9
and 10 with C6H14–EtOAc (6:4, v/v),
fractions 11 and 12 with C6H14–EtOAc
(1:1, v/v), fractions 13 and 14 with
C6H14–EtOAc (4:6, v/v), fractions 15
and 16 with C6H14–EtOAc (3:7, v/v),
fractions 17 and 18 with C6H14–EtOAc
(2:8, v/v), fractions 19 and 20 with
C6H14–EtOAc (1:9, v/v), fractions 21
and 22 fractions with EtOAc, fractions
23–26 with EtOAc–MeOH (9.5:0.5), and
fractions 27 and 30 with EtOAc–MeOH
(9:1). All fractions were examined by
TLC. Fractions 5 and 6 (0.9 g) were
crystallized after purification by column
chromatography, yielding b-sitosterol
whose identity was confirmed through
the comparison of TLC and spectroscopic
data with those of an authentic sample.
Fractions 17 and 18 (4.4 g) were
re-chromatographed over silica gel
(100 g, each fraction of 50 ml). The
elution was sequentially performed with
3.3.2 19,21-Dimethyl triacont-
17,22,24,26,28-pentaene-1-oic acid (2)
Semi-solid; IR (KBr): nmax 3010
(COOH), 2925, 2854, 1711 (COOH),
1463, 1282, and 723 cm21
;
1H NMR
(CDCl3): d 5.38 (2H, m, H-23, H-24),
5.36 (3H, m, H-25, H-26, H-27), 5.34
(2H, m, H-28, H-29), 5.33 (2H, m, H-22,
H-17), 5.32 (1H, m, H-18), 2.80 (2H, m,
H2-2), 2.75 (1H, m, H-19), 2.33 (2H, m,
H2-16), 2.07 (1H, m, H-21), 2.04 (2H, m,
H2-30), 1.62 (2H, m, CH2), 1.60 (2H, m,
CH2), 1.34 (4H, m, 2 £ CH2), 1.32 (8H,
br s, 4 £ CH2), 1.29 (5H, br s, 3 £ CH2),
1.26 (10H, br s, 5 £ CH2), 0.97 (3H, t,
J ¼ 6.1 Hz, Me-32), 0.89 (3H, d,
J ¼ 6.5 Hz, Me-33), 0.86 (3H, d,
J ¼ 6.3 Hz, Me-34); 13C NMR (CDCl3):
d 181.3 (C-1), 133.2 (C-23), 131.5 (C-24),
131.4 (C-25), 131.3 (C-26), 129.6 (C-27),
129.5 (C-28), 129.4 (C-22), 129.3 (C-18),
129.1 (C-17), 128.5 (C-29), 35.4 (C-2),
32.9 (C-19), 31.1 (C-21), 31.0 (C-20),
30.9 (CH2), 30.8 (CH2), 30.8 (CH2), 30.7
(CH2), 30.7 (CH2), 30.5 (CH2), 30.4
(CH2), 30.4 (CH2), 28.5 (CH2), 27.0
(CH2), 26.9 (CH2), 26.9 (CH2), 26.0
(CH2), 24.1 (CH2), 24.0 (CH2), 21.9
(CH2), 15.6 (Me-33),15.5 (Me-34), 15.4
(Me-32); þ ve FAB-MS m/z (rel. int.):