J.-H. Yi et al. / Phytochemistry 51 (1999) 825±828
827
3.3. Extraction and separation
ml) and heated at 908C for 2.5 h. After cooling, the
reaction mixture was extracted with EtOAc. The or-
ganic layer was concentrated to dryness. 2-(4-Hydroxy-
3-methoxyphenyl)ethanol and caeic acid were ident-
i®ed in the EtOAc layer by TLC on Kieselgel 60 F254
(CHCl3:EtOH, 9:1). Glucose, rhamnose and apiose
were detected in the aqueous layer by TLC on silica
gel G (lower phase of CHCl3:MeOH:H2O 15:6:2±
HOAc (9:1)) and PC (n-BuOH±HOAc±H2O (4:1:5),
upper phase).
The dried and powdered roots (2.5 kg) were
extracted with 80% EtOH under re¯ux (10 l  3, each
2 h). After the removal of solvent, the residue was suc-
cessively fractionated with petroleum ether (b.p. 60±
908C), EtOAc and n-BuOH. The n-BuOH extract was
chromatographed
on
silica
gel
with
CHCl3:MeOH:H2O (3:6:2 lower layer) to give frs A±F.
Fr. D was subjected to HPLC gradient-eluted with
H2O/MeOH (40±60%) at a ¯ow rate of 5 ml/min to
yield compounds 1 (30 mg), 2 (65 mg) and 3 (28 mg).
3.7. Permethylation of compound 2 and lamiophlomisde
A (4)
3.4. Compound 1
Compound 2 and lamiophlomiside A (ca. 10 mg),
Me2SO4 (0.2 ml) and K2CO3 (30 mg) in Me2CO (6 ml)
were stirred for 24 h. After ®ltration, the ®ltrate was
concentrated and analysed by TLC. They both gave
the same permethyl (tetramethyl) compound (5), which
was obtained as an o-white amorphous powder by
preparative TLC (CHCl3:MeOH:H2O, 13:5:2 lower
layer). 1H NMR (CD3OD) was identical to those
reported (Yi et al., 1995).
O-white amorphous powder. UV nmax (nm): 332,
290, 220; IR nmax cm 1: 3450 (OH), 1700 (C.O), 1630
(C.C), 1602 and 1510 (aromatic rings), 1445, 1265,
1155, 1035, 810; H, 13C NMR (CD3OD) and FABMS
1
were identical to those reported for cistanoside C
(Kobayashi et al., 1984).
3.5. Compound 2
O-white amorphous powder. UV lmax (nm): 333,
219; IR nmax cm 1: 3420 (OH), 1700 (C.O), 1630
(C.C), 1600 and 1510 (aromatic rings), 1445, 1265,
1155, 1035, 810; 1H NMR(CD3OD) d ppm: 6.85 (d,
J=2.0 Hz, 1H, H-2), 6.70 (d, J=8.0 Hz, 1H, H-5),
6.67 (dd, J=8.0/2.0 Hz, 1H, H-6), 4.04 (m, 1H, H-a),
3.72 (m, 1H, H-a), 2.85 (m, 2H, H-b), 3.84 (s, 3H,
OMe), 7.05 (d, 1H, J=2.0 Hz, H-20), 6.77 (d, 1H,
J=8.3 Hz, H-50), 6.95 (dd, 1H, J=8.3/2.0 Hz, H-60),
6.27 (d, 1H, J=16.0 Hz, H-a'), 7.59 (d, 1H, J=16.0
Hz, H-b'), 4.37 (d, 1H, J=8.0 Hz, H-1'), 3.38 (dd, 1H,
J=9.3/8.0 Hz, H-2'), 3.80 (t, 1H, J=9.3 Hz, H-3'),
4.94 (t, 1H, J=9.3 Hz, H-4'), 5.18 (d, 1H, J=1.0 Hz,
H-1 of Rha), 3.29 (t, 1H, J=9.5 Hz, H-4 of Rha), 1.09
(d, 3H, J=6.0 Hz, H-6 of Rha), 4.90 (d, 1H, J=2.4
Hz, H-1 of Api), 3.86 (d, 1H, J=2.4 Hz, H-2 of Api),
3.53 (s, 3H, H-3' of Api). 13C NMR (CD3OD) d ppm:
132.4 (C-1), 114.5 (C-2), 149.6 (C-3), 146.7 (C-4), 116.9
(C-5), 123.2 (C-6), 73.1 (C-a), 37.6 (C-b), 57.2 (OMe),
128.4 (C-10), 115.5 (C-20), 150.6 (C-30), 148.8 (C-40),
117.3 (C-50), 124.0 (C-60), 116.0 (C-a'), 147.6 (C-b'),
169.0 (C.O), 105.0 (C-1'), 76.9 (C-2'), 82.4 (C-3'), 71.7
(C-4'), 75.4 (C-5'), 69.3 (C-6'), 103.8 (C-1 of Rha),
73.1 (C-2 of Rha), 72.8 (C-3 of Rha), 74.6 (C-4 of
Rha), 71.2 (C-5 of Rha), 19.2 (C-6 of Rha), 111.9 (C-1
of Api), 78.9 (C-2 of Api), 81.4 (C-3 of Api), 75.9 (C-4
of Api), 66.4 (C-3' of Api). FABMS (m/z): 809
[M+K]+, 793 [M+Na]+.
3.8. Compound 3
O-white amorphous powder. UV lmax (nm): 331,
289, 220; IR nmax cm 1: 3440 (OH), 1670 (C.O), 1630
(C.C), 1595 and 1510 (aromatic rings), 1445, 1425,
1
1265, 1155, 1030, 810. H NMR (CD3OD) dppm: 6.85
(d, 1H, J=2.0 Hz, H-2), 6.70 (d, 1H, J=8.0 Hz, H-5),
6.67 (dd, 1H, J=8.0/2.0 Hz, H-6), 4.04 (m, 1H, H-a0),
3.72 (m, 1H, H-a) 2.85 (m, 2H, H-b), 3.83 (s, 3H,
OMe), 7.87 (d, 1H, J=2.0 Hz, H-20), 6.77 (d, 1H,
J=8.3 Hz, H-50), 7.15 (dd, 1H, J=8.3/2.0 Hz, H-60),
5.79 (d, 1H, J=13.0 Hz, H-a'), 6.93 (d, 1H, J=13.0
Hz, H-b'), 3.89 (s, 3H, OMe), 4.35 (d, 1H, J=8.0 Hz,
H-1'), 3.38 (dd, 1H, J=9.3/8.0 Hz, H-2'), 3.80 (t, 1H,
J=9.3 Hz, H-3'), 4.94 (t, 1H, J=9.3 Hz, H-4'), 5.18
(d, 1H, J=1.0 Hz, H-1 of Rha), 3.29 (t, 1H, J=9.5
Hz, H-4 of Rha), 1.08 (d, 3H, J=6.0 Hz, H-6 of Rha),
4.90 (d, 1H, J=2.4 Hz, H-1 of Api), 3.87 (d, 1H,
J=2.4 Hz, H-2 of Api), 3.53 (s, 3H, H-3' of Api). 13C
NMR (CD3OD) d ppm: 132.3 (C-1), 114.5 (C-2), 149.6
(C-3), 146.7 (C-4), 116.9 (C-5), 123.2 (C-6), 73.1 (C-a),
37.6 (C-b), 57.2 (OMe), 128.7 (C-10), 116.5 (C-20),
150.6 (C-30), 149.1 (C-40), 116.4 (C-50), 128.2 (C-60),
116.2 (C-a'), 148.6 (C-b'), 167.6 (C.O), 57.3 (OMe),
105.0 (C-1'), 76.9 (C-2'), 82.7 (C-3'), 71.6 (C-4'), 75.4
(C-5'), 69.2 (C-6'), 104.0 (C-1 of Rha), 73.1 (C-2 of
Rha), 72.9 (C-3 of Rha), 74.6 (C-4 of Rha), 71.2 (C-5
of Rha), 19.2 (C-6 of Rha), 111.7 (C-1 of Api), 78.9
(C-2 of Api), 81.4 (C-3 of Api), 75.8 (C-4 of Api), 66.4
(C-3' of Api). FABMS (m/z ) 823 [M+K]+, 807
[M+Na]+.
3.6. Hydrolysis of compound 2
Compound 2 (10 mg) was dissolved in 5 M HCl (2