3
30
X.-T. Zhang et al.
Norwalk, CT, USA). IR spectra were
measured on a Nicolet Impact 410 FT-IR
instrument (Nicolet Instrument Ltd, Madi-
son, WI, USA). UV spectra were recorded
(3 £ 20 liters) under reflux. The EtOH
extract was suspended in water and then
successively extracted with petroleum
ether, EtOAc, and n-BuOH. The n-BuOH
solution was concentrated and given a
residue (207 g), which was separated by a
on a Shimadzu UV-2501 spectrophotometer
(
1
Shimadzu Ltd, Kyoto, Japan). The H and
1
3
C NMR spectra were obtained on a Bruker
1
AV500 Avance spectrometer ( H,
silica gel column using CHCl –MeOH
3
(1:0 ! 1:1) as an eluent, affording five
fractions (frs A–F). Fr. B (3.5 g) was
purified by CC (Sephadex LH-20, MeOH):
Fr. B1–B4. Fr. B4 (385 mg) was subjected
1
00 MHz; C, 125 MHz; Bruker Ltd,
3
5
Karlsruhe, Germany) and chemical shifts
were given in d (ppm) with TMS as a
reference. HR-ESI-MS were obtained on an
Applied Biosystems Mariner 5140 spec-
trometer(LifeTechnologiesLtd, NewYork,
NY, USA). Column chromatography (CC)
was performed on silica gel (Qingdao
Marine Chemical Ltd, Qingdao, China),
Sephadex LH-20 (Pharmacia Ltd, New
York, NY, USA), and ODS (Merck Ltd,
Darmstadt, Germany). Thin-layer chroma-
tography was performed on precoated silica
gel GF254 plates (Qingdao Marine Chemical
Ltd). Preparative HPLC was carried out
using a Zobax XDB-18 column (10 mm
i.d. £ 15 cm, Agilent Technologies Ltd,
Wilmington, DE, USA). GC experiments
werecarriedoutonanHP-1TCDinstrument
to reversed-phase CC (ODS, MeOH/H O
2
60:40–80:20 (v/v)), followed by HPLC
(MeCN/H O 34:66 (v/v)) to give com-
2
pounds 1 (45 mg, t 17 min) and 2 (67 mg,
R
tR 26 min), respectively.
3
.3.1 3-O-b-D-Glucopyranosyl (1 ! 4)-
b-D-fucopyranosyl-(22S,24Z)-cycloart-
4-en-3b, 22,26,30-tetraol 26-O-b-D-
2
glucopyranoside (1)
A white powder, ½aꢀ 8.80 (c ¼ 0.68,
D
MeOH); mp 258–2598C; IR (KBr) v
3416, 2937, 1614, 1384, 1363, 1105, 1078,
max
2
1
1
13
773, 625, 476 cm ; H and C NMR
spectral data see Table 1; ESI-MS
2
(negative ion mode) m/z 943 [M–H] ,
(Hewlett-Packard Ltd, Palo Alto, CA,
USA) using an HP-Chiral column
2
2
781 [M-163] , 619 [M-325] ; HR-ESI-
2
MS: m/z 943.5280 [M–H] (calcd for
(
30 m £ 0.25 mm £ 1.0 mm, 20% per-
methylated b-cyclodextrin; Agilent Tech-
nologies Ltd). All chemical reagents were
purchased from Nanjing Reagent Co., Ltd
C H O , 943.5266).
48 79 18
3
.3.2 3-O-b-D-Glucopyranosyl (1 ! 4)-
b-D-fucopyranosyl-(22S,24Z)-cycloart-
4-en-3b,22,26,29-tetraol 26-O-b-D-
(Nanjing, China).
2
3
.2 Plant material
glucopyranoside (2)
The aerial parts of T. fortunei were collected
in Wuhu City, Anhui Province, China, in
April 2004, and authenticated by Dr Ming-
Jian Qin of China Pharmaceutical Univer-
sity. A voucher specimen (No. 040192) has
been deposited in the herbarium of China
Pharmaceutical University, Nanjing.
A white powder, ½aꢀ 21.3 (c ¼ 0.25,
D
MeOH); mp 235–2378C; IR (KBr) v
21 1
3434, 2929, 1637, 1383, 1072 cm ; H
max
1
3
and C NMR spectral data see Table 1;
2
HR-ESI-MS: m/z 943.5280 [M–H]
(calcd for C H O , 943.5266).
48 79 18
3.4 Acid hydrolysis
3
.3 Extraction and isolation
A solution of the compound (10 mg) in
15 ml of 1 M HCl (MeOH–H O, 1:1) was
heated under reflux for 3 h. After the
The dried aerial parts (4.8 kg) of T.
fortunei were extracted with 95% EtOH
2