W. Wei et al. / Phytochemistry xxx (2016) xxx–xxx
9
andafocoumarins C (5, 25 mg, t
R
10 min), G (9, 25 mg, t
R
25 min),
4.3.10. Andafocoumarin H (10)
2
0
and H (10, 21 mg, t
R
28 min).
Pale yellow oil; [
log
2925, 2854, 1734, 1623, 1456; H NMR (CDCl
3
a
]
D
+ 40.3 (c 0.1, CHCl
3
); UV (MeOH) kmax nm
ꢀ1
(
e
): 248 (4.14), 258 (4.09), 307 (4.00); IR (KBr)
m
max cm : 3466,
, 400 MHz) data, see
3
Table 1; C NMR (CDCl , 100 MHz) data, see Table 2; HRESIMS m/z
1
0
0
00
4
.3.1. 5-(3 -Hydroxy-3 -methylbutyl)-8-hydroxyfurocoumarin (1)
2
0
13
White amorphous solid; [
max nm (log ): 268 (3.88), 311 (3.72); IR (KBr)
a]
D
+ 30.0 (c 0.1, CHCl
max cm : 3377,
OD, 400 MHz) data, see
OD, 100 MHz) data, see Table 2; HRESIMS
3
); UV (MeOH)
ꢀ1
+
k
2
e
m
51 6
555.3688 [M+H] (calcd for C34H O , 555.3680).
1
972, 1697, 1599, 1480; H NMR (CD
3
1
3
Table 1; C NMR (CD
3
4.3.11. Andafocoumarin I (11)
ꢀ
20
m/z 287.0920 [MꢀH] (calcd for C16
H
15
O
5
, 287.0925).
Pale yellow oil; [
log
2854, 1737, 1479; H NMR (CDCl , 400 MHz) data, see Table 1;
D 3
a] + 23.9 (c 0.1, CHCl ); UV (MeOH) kmax nm
ꢀ1
(
e): 267 (4.19), 312 (4.00); IR (KBr) mmax cm : 3460, 2925,
1
0
0
4
.3.2. Isobyakangelicin hydrate-3 -ethyl ether (2)
3
1
3
2
0
C NMR (CDCl , 100 MHz) data, see Table 2; HRESIMS m/z
Pale yellow oil; [
a]
D
+ 41.2 (c 0.1, CHCl
3
); UV (MeOH) kmax nm
3
+
ꢀ1
602.4060 [M+NH ] (calcd for C35H NO , 602.4051).
(
2
log ): 265 (3.49), 307 (3.34); IR (KBr)
e
m
max cm : 3466, 2975,
930, 1736, 1593, 1479; H NMR (CDCl , 400 MHz) data, see
, 100 MHz) data, see Table 2; HRESIMS
, 361.1293).
4
56
7
1
3
1
3
4.3.12. Andafocoumarin J (12)
Table 1; C NMR (CDCl
m/z 361.1281 [MꢀH] (calcd for C19
3
2
0
ꢀ
Pale yellow oil; [
log ): 248 (3.69), 266 (3.61), 309 (3.51); IR (KBr)
926, 2854, 1736, 1623, 1457; H NMR (CDCl
a
]
+ 34.5 (c 0.1, CHCl ); UV (MeOH) k
max
nm
H
21
O
7
D
3
ꢀ
1
(
2
e
m
max cm : 3433,
, 400 MHz) data, see
, 100 MHz) data, see Table 2; HRESIMS m/z
567.3316 [M+H] (calcd for C34 , 567.3316).
1
3
4
.3.3. Andafocoumarin A (3)
1
3
2
0
Table 1; C NMR (CDCl
3
D 3
Pale yellow oil; [a] + 38.4 (c 0.1, CHCl ); UV (MeOH) kmax nm
+
ꢀ1
47 7
H O
(
log ): 250 (3.66), 265 (3.51), 301 (3.45); IR (KBr) mmax cm : 2925,
e
1
13
2
853, 1730, 1464; H NMR (CDCl
, 100 MHz) data, see Table 2; HRESIMS m/z 495.2729,
M+Na] (calcd for C28 40NaO , 495.2717).
3
, 400 MHz) data, see Table 1;
C
4.4. Determination of absolute configuration of the secondary alcohol
NMR (CDCl
3
+
units in 2ꢀ11
[
H
6
According to the published procedure (Frelek and Szczepek,
1999), each tested compound (1.0 mg) and [Rh (OCOCF ] (3 mg)
was mixed and dissolved in CH Cl (600 L) to form a 1.6 mg/mL
of solution. Then, the solution was subjected to CD measurements.
The first CD spectrum was recorded immediately and its time evo-
lution was monitored until stationary (about 15 min after mixing).
The inherent CD was subtracted. The observed sign of the E band at
around 350 nm in the induced CD spectrum was correlated to the
absolute configuration of the secondary alcohol moiety.
4
.3.4. Andafocoumarin B (4)
2
0
2
3 4
)
Pale yellow oil; [
D 3
a] + 20.1 (c 0.2, CHCl ); UV (MeOH) kmax nm
ꢀ
1
2
2
l
(
log ): 269 (3.41), 307 (3.36); IR (KBr)
e
m
max cm : 3466, 2925,
853, 1732, 1623, 1579, 1455; H NMR (CDCl , 400 MHz) data,
, 100 MHz) data, see Table 2; HRESIMS
1
2
3
1
3
see Table 1; C NMR (CDCl
m/z 517.2804 [M+HCOO] (calcd for C29
3
ꢀ
H
41
O
8
, 517.2807).
4.3.5. Andafocoumarin C (5)
2
0
Pale yellow oil; [
D 3
a] + 26.1 (c 0.1, CHCl ); UV (MeOH) kmax nm
ꢀ1
(
log ): 268 (5.00), 311 (4.84); IR (KBr)
e
m
max cm : 3462, 2925,
854, 1737, 1595, 1479; H NMR (CDCl , 400 MHz) data, see
, 100 MHz) data, see Table 2; HRESIMS
4.5. Oxidations of 9a and 9ꢀ11
1
2
3
1
3
Table 1; C NMR (CDCl
m/z 520.3276 [M+NH
3
Following the reported procedure (Pappo et al., 1956; Wyche
+
4 7
] (calcd for C29H46NO , 520.3269).
et al., 2012), each tested compound (1.0 mg) was dissolved in
THF (1 mL), and then H O (1 mL) was added. While stirring, NaIO
30 mg) and OsO (1 mg) were slowly added to the solution,
respectively. The mixture was stirred at room temperature for
.5 h. THF was removed under reduced pressure at 40 °C, and the
remaining aqueous phase was extracted with CH Cl
2
4
4
.3.6. Andafocoumarin D (6)
(
4
2
0
Pale yellow oil; [
D 3
a] + 39.6 (c 0.1, CHCl ); UV (MeOH) nm kmax
(
log ): 241 (3.71), 248 (3.70), 266 (3.55), 304 (3.46); IR (KBr) m
e
1
max
1
ꢀ
1
cm : 3465, 2914, 2837, 1766, 1476; H NMR (CDCl
data, see Table 1; C NMR (CDCl
HRESIMS m/z 518.3473 [M+NH
3
, 400 MHz)
, 100 MHz) data, see Table 2;
] (calcd for C30 , 518.3476).
2
2
(50 mL ꢁ 3).
1
3
3
The organic layers were dried using rotary evaporation and ana-
lyzed by HRESIMS and GC–MS. Specially, as to the oxidation of
+
4
H48NO
6
9
a (8 mg), 5 mg of the crude product was obtained. The latter
was subjected to silica gel CC eluting with CH Cl –MeOH (30:1,
v/v) to give 6-formyl-7-hydroxy-6,8-dimethoxycoumarin (9c,
.3 mg). The structure of 9c was established by analysis of NMR
4
.3.7. Andafocoumarin E (7)
2
2
2
0
D 3
Pale yellow oil; [a] + 17.2 (c 0.3, CHCl ); UV (MeOH) kmax nm
(
log ): 248 (3.96), 258 (3.91), 266 (3.89), 308 (3.82); IR (KBr) m
e
1
max
3
ꢀ
1
cm : 3467, 2925, 2853, 1726, 1623, 1579, 1456; H NMR (CDCl
4
Table 2; HRESIMS m/z 545.3118 [M+HCOO] (C31
3
,
1
and HRESIMS data. Yellow powder. 9c:
00 MHz), d 12.05 (1H, s, 8-OH), 10.24 (1H, s, 7-CHO), 7.86 (1H,
d, J = 9.8 Hz, H-3), 6.31 (1H, d, J = 9.8 Hz, H-2), 4.03 (3H, s, OCH ),
, 100 MHz), d 193.5 (CHO),
3
H NMR (CDCl ,
00 MHz) data, see Table 1; 13C NMR (CDCl
, 100 MHz) data, see
3
4
ꢀ
H
45
O
8
, 545.3120).
3
1
3
4
3 3
.00 (3H, s, OCH ); C NMR (CDCl
4.3.8. Andafocoumarin F (8)
160.0 (C-2), 158.2 (C-7), 137.6 (C-4), 113.8 (C-3), 111.9 (C-6),
106.8 (C-4a), 66.8 (OCH ), 61.6 (OCH ); HRESIMS m/z 251.0552
[M+H] (calcd for C12H11O6, 251.0550).
2
0
Pale yellow oil; [
a]
D
+ 17.5 (c 0.2, CHCl
3
); UV (MeOH) kmax nm
3
3
ꢀ1
+
(
log ): 247 (3.94), 268 (3.99), 312 (3.82); IR (KBr)
e
m
max cm : 3466,
, 400 MHz) data, see
, 100 MHz) data, see Table 2; HRESIMS m/z
] (calcd for C31 , 548.3582).
1
2
925, 2853, 1737, 1593, 1479; H NMR (CDCl
3
1
3
Table 1; C NMR (CDCl
5
3
4.6. Hydrolysis of 12
+
48.3586 [M+NH
4
H50NO
7
Following the reported procedure (Suárez Ortiz et al., 2013),
compound 12 (1.5 mg) was dissolved in MeOH (1 mL), and then
NaOMe (0.2 mg) was added. The reaction mixture was stirred at
4
.3.9. Andafocoumarin G (9)
2
0
D 3
Pale yellow oil; [a] + 39.1 (c 0.1, CHCl ); UV (MeOH) kmax nm
(
log ): 242 (3.88), 248 (3.88), 263 (3.75), 297 (3.62); IR (KBr)
e
m
max
30 °C for 3 h. Saturated aqueous NH Cl solution was added, and
2
the mixture was extracted with CH Cl . The organic layer was col-
lected, filtered, and concentrated under reduced pressure at 40 °C.
4
ꢀ
1
1
cm : 3467, 2923, 2845, 1755, 1445; H NMR (CDCl
data, see Table 1; C NMR (CDCl
HRESIMS m/z 572.3969 [M+NH
3
, 400 MHz)
, 100 MHz) data, see Table 2;
] (calcd for C34 , 572.3946).
2
1
3
3
+
4
H54NO
6
The residue was analyzed by GC–MS.