1310
H. Kikuzaki et al. / Phytochemistry 52 (1999) 1307±1312
aq. residue was partitioned with EtOAc to give the
EtOAc soluble and H2O soluble parts. The n-hexane
extract (5.0 g) was dissolved with n-hexane (200 ml)
and the soln. was extracted two times with 0.5 N
NaOH aq. (200 ml). Acidi®cation of the alkaline soln.
with 4 N HCl, followed by extraction with EtOAc,
gave the acidic fraction (2.6 g), which was subjected to
CC on silica gel eluted with CH2Cl2 to aord eugenol
(3, 1.6 g). The CH2Cl2 extract (5.3 g) was chromato-
graphed on silica gel and eluted with CH2Cl2 and
MeOH mixture of increasing polarity to give 12 frs.
Fr. 2 which was eluted with CH2Cl2, was identi®ed as
eugenol (1.6 g). Fr. 3, eluted with CH2Cl2, was recrys-
tallized with water to aord vanillin (6, 230 mg). Fr. 4,
eluted with CH2Cl2, was rechromatographed on silica
gel (C6H6±Me2CO) to give 4 (20 mg) and 5 (7 mg). Fr.
7, eluted with CH2Cl2±MeOH (99:1), was rechromato-
graphed on Sephadex LH-20 (iso-PrOH), followed by
repeated CC on silica gel (CH2Cl2±MeOH, C6H6±
Me2CO, isopropylether) and recrystallization with
CHCl3, to give 1 (29 mg). Fr. 8, eluted with CH2Cl2±
MeOH (97:3), was rechromatographed on Sephadex
LH-20 (iso-PrOH) and silica gel (CH2Cl2±MeOH) to
give 2 (37 mg).
5.38 (1H, ddd, J = 4.0, 4.9, 7.9 Hz, H-2), 6.04 (1H, d,
J = 7.9 Hz, H-1), 6.99 (1H, dd, J = 2.0, 8.5 Hz, H-
6'), 7.00 (1H, d, J = 2.0 Hz, H-2'), 7.04 (1H, d, J =
8.5 Hz, H-5'). EIMS 20 eV m/z (rel. int.): 360
[M+2]+ (3.1, [M+2]/[M] = 0.39), 358 [M]+ (7.9),
318 (23), 316 (68), 256 (99), 214 (100), 195 (99), 179
(99), 153 (89).
3.5. Compound 2
25
D
film
Colorless oil, a
128 (EtOH; c 0.99). IR nmax
1
cm
:
3600±3200, 1605, 1516, 1274. 1H-NMR
((CD3)2CO): d 2.59 (1H, dd, J = 4.4, 13.2 Hz, H-3a),
2.70 (1H, dd, J = 3.4, 13.2 Hz, H-3b), 3.43 (1H, ddd,
J = 5.8, 5.8, 10.8 Hz, H-1a), 3.51 (1H, ddd, J = 4.4,
6.0, 10.8 Hz, H-1b), 3.80 (1H, m, H-2), 3.83 (3H, s,
OCH3), 6.67 (1H, dd, J = 2.0, 8.0 Hz, H-6'), 6.72
(1H, d, J = 8.0 Hz, H-5'), 6.85 (1H, d, J = 2.0 Hz,
H-2'), 7.31 (1H, s, 4'-OH). 13C-NMR ((CD3)2CO): d
40.4 (C-3), 56.1 (OCH3), 66.5 (C-1), 74.2 (C-2), 113.7
(C-2'), 115.5 (C-5'), 122.6 (C-6'), 131.4 (C-1'), 145.7
(C-4'), 148.0 (C-3'). EIMS 70 eV m/z (rel. int.): 198
[M]+ (20), 180 (9), 167 (17), 137 (100). HR-EIMS 70
eV: found m/z 198.0914 [M]+, C10H14O4, requires
198.0892.
3.3. Compound 1
25
D
3.6. Acetylation of 2
Colorless needles, mp 1218 (CHCl3). a
28
(EtOH; c 0.52). IR nmax cm 1: 3400±3200, 1609, 1522.
1H-NMR ((CD3)2CO): d 3.37 (1H, dd, J = 6.5, 11.5
Hz, H-3a), 3.62 (1H, dd, J = 4.5, 11.5 Hz, H-3b), 3.81
(1H, m, H-2), 3.84 (3H, s, OCH3), 4.32 (1H, d, J =
5.0 Hz, 2-OH), 4.45 (1H, d, J = 4.5 Hz, 1-OH), 4.64
(1H, dd, J = 4.5, 5.0 Hz, H-1), 6.79 (1H, d, J = 8.0
Hz, H-5'), 6.85 (1H, dd, J = 2.0, 8.0 Hz, H-6'), 7.04
(1H, d, J = 2.0 Hz, H-2'), 7.55 (1H, s, 4'-OH). 13C-
NMR ((CD3)2CO): d 47.4 (C-3), 56.2 (OCH3), 74.9 (C-
1), 76.7 (C-2), 111.0 (C-2'), 115.3 (C-5'), 120.2 (C-6'),
134.1 (C-1'), 146.9 (C-4'), 148.1 (C-3'). EIMS 20 eV
m/z (rel. int.): 234 [M+2]+ (0.9, [M+2]/[M] = 0.41),
232 [M]+ (2.2), 216 (1), 214 (4), 196(4), 178 (11), 151
(100), 137 (47). HR-EIMS 20 eV: found: m/z 232.0547
[M]+, C10H13O4Cl requires: 232.0501.
nujol
2 (5.0 mg) was treated in the same manner as
1
described above to give 2a (8.5 mg), IR nmax cm
film
:
1
1735, 1606, 1510, 1231. H-NMR (CDCl3): d 2.04 (3H,
s, OAc), 2.08 (3H, s, OAc), 2.30 (3H, s, OAc), 2.86
(1H, dd, J = 6.7, 14.0 Hz, H-3a), 2.93 (1H, dd, J =
6.7, 14.0 Hz, H-3b), 3.82 (3H, s, OCH3), 4.05 (1H, dd,
J = 6.1, 11.6 Hz, H-1a), 4.25 (1H, dd, J = 3.4, 11.6
Hz, H-1b), 5.27 (1H, dddd, J = 3.4, 6.1, 6.7, 6.7 Hz,
H-2), 6.78 (1H, dd, J = 1.8, 7.9 Hz, H-6'), 6.82 (1H,
d, J = 1.8 Hz, H-2'), 6.95 (1H, d, J = 7.9 Hz, H-5').
EIMS 70 eV m/z: (rel. int.): 324 (4), 282 (6), 264 (3),
222 (100), 180 (22), 179 (32), 137 (48).
3.7. Methylation of 2
3.4. Acetylation of 1
To a soln. of 2 (10.5 mg) in dried Me2CO (5 ml)
were added dry K2CO3 (0.50 g) and CH3I (0.94 g).
After stirring overnight at room temp., CH2Cl2 (20 ml)
was added and the mixture was ®ltered. After remov-
ing the solvent from the ®ltrate, the residue was sub-
mitted to CC on silica gel (CH2Cl2:MeOH = 97:3) to
A soln. of 1 (3.3 mg) in pyridine (0.5 ml) and Ac2O
(0.5 ml) was allowed to stand overnight at room temp.
The reaction mixture was poured into cold 2 N HCl,
and then extracted with EtOAc. The organic layer was
washed with water, dried over anhydrous MgSO4 and
25
1
give 2b (8.4 mg), a
148 (EtOH; c 0.66). H-NMR
D
film
evaporated to dryness to give 1a (5.7 mg), IR nmax
(CDCl3): d 2.69 (1H, dd, J = 8.0, 12.0 Hz, H-3a), 2.76
(1H, dd, J = 5.0, 12.0 Hz, H-3b), 3.53 (1H, dd, J =
7.0, 11.0 Hz, H-1a), 3.71 (1H, dd, J = 3.0, 11.0 Hz,
H-1b), 3.86 (3H, s, OCH3), 3.88 (3H, s, OCH3), 3.92
(1H, m, H-2), 6.75 (1H, br s, H-2'), 6.76 (1H, dd, J =
cm 1: 1735, 1609, 1510, 1219. 1H-NMR (CDCl3): d
2.08 (3H, s, OAc), 2.11 (3H, s, OAc), 2.31 (3H, s,
OAc), 3.33 (1H, dd, J = 4.9, 12.2 Hz, H-3a), 3.60
(1H, dd, J = 4.0, 12.2 Hz, H-3b), 3.85 (3H, s, OCH3),