732
X. Wu et al.
using silica gel (200–300 mesh, Qingdao
Haiyang Chemical Group Corporation,
Qingdao, China) and Sephadex LH-20
(Pharmacia Biotech AB, Uppsala, Swe-
den). TLC analysis was performed on
precoated silica gel GF254 plates (Yantai
Chemical Industrial Institute, Yantai,
China). Analytical high-performance
liquid chromatography (HPLC) was car-
ried out on a Dionex chromatography
equipped with a P680 pump, a PDA-100
photodiode array detector, and a Cosmosil
5C18-MS-II reversed-phase column
(4.6 mm £ 250 mm, 5.0 mm, Nacalai
Tesque, Kyoto, Japan). Preparative HPLC
was carried out on a Varian instrument
equipped with a Prostar 215 pump, a
Prostar 325 UV/VIS detector, and a
Cosmosil 5C18-MS-II reversed-phase col-
umn (20 mm £ 250 mm, 5.0 mm, Nacalai
Tesque, Kyoto, Japan).
(100:0 ! 0:100) as eluent to give four
fractions (Fr. 1–4). Fr. 2 (30 g) was
separated by silica gel column using
CHCl3–acetone (100:0 ! 0:100) as eluent
to yield three subfractions (Fr. 2a–2c). Fr.
2a (14 g) was subjected to preparative
HPLC [CH3OH–H2O (40: 60)] to give
compounds 4 (188mg), 5 (131mg), and 6
(336mg). Fr. 2b (11 g) was re-separated by
Sephadex LH-20 column (CH3OH) and
preparative HPLC [CH3OH-H2O (25: 75)]
to yield compounds 1 (84 mg), 2 (23 mg), 3
(34 mg), 7 (1170 mg), 8 (129 mg), 9
(12 mg), 10 (14 mg), 11 (20 mg), 12
(21 mg), 13 (150 mg), and 14 (30 mg),
respectively.
3.3.1 Compound 1
Colorless oil: ½aꢀ2D1 234.8 (c 0.1, CH3OH);
UV (CH3OH) lmax (log1) 208 (3.88), 251
(2.58)nm; IR (KBr): nmax 3325, 2932,
2886, 1724, 1637, 1455, 1383, 1318, 1210,
1160, 1079, 1046, 1019, 905, 751,
3.2 Plant material
1
The flowers of H. undatus were collected in
Zhaoqing city, Guangdong Province of
China, in February 2010, and authenticated
by Prof. Guang-Xiong Zhou (Institute of
Traditional Chinese Medicine & Natural
Products, Jinan University). A voucher
specimen (No. 20100215) has been depos-
ited in the Institute of Traditional Chinese
Medicine & Natural Products, Jinan Uni-
versity, Guangzhou, China.
700 cm21; H and 13C NMR spectral data,
see Table 1; HR-ESI-MS m/z 437.1423
[M þ Na]þ (calcd for C19H26O10Na,
437.1418).
3.3.2 Compound 2
Colorless oil: ½aꢀ2D1 229.8 (c 0.2, CH3OH);
UV (CH3OH) lmax (log1) 211 (3.74), 258
(2.52)nm; IR (KBr): nmax 3377, 2944, 2874,
1729, 1687, 1452, 1354, 1323, 1209, 1158,
1078, 1049, 1019, 912, 742, 700 cm21; 1H
and 13C NMR spectral data, see Table 1;
HR-ESI-MS m/z 451.1584 [M þ Na]þ
(calcd for C20H28O10Na, 451.1575).
3.3 Extraction and isolation
The air-dried flowers of H. undatus (15 kg)
were powdered and extracted with 95%
(V/V) EtOH at room temperature, and the
solution was evaporated under vacuum to
yield a residue (5700 g). The crude EtOH
extract was subsequently suspended in
distilled water and partitioned successively
with petroleum ether, ethyl acetate, and
n-butanol, respectively. After removing
the solvent, the ethyl acetate extract (90 g)
was subjected to silica gel CC using
gradient mixtures of CHCl3 –CH3OH
3.3.3 Compound 3
21
Colorless oil: ½aꢀD 212.9 (c 0.3,
CH3OH); UV (CH3OH) lmax (log 1) 212
(3.59), 258 (2.38) nm; IR (KBr): nmax
3331, 2938, 2881, 1723, 1637, 1455, 1381,
1318, 1202, 1158, 1085, 1017, 908, 748,
700 cm21; 1H and 13C NMR spectral data,
see Table 1; HR-ESI-MS m/z 451.1584