Five new stilbene glycosides from the roots of Polygonum multiflorum

Article abstract of DOI:10.1080/10286020.2013.837454

Five new stilbene glycosides (1-5), together with six known ones, were isolated from the roots of Polygonum multiflorum. Their structures were elucidated on the basis of spectroscopic analysis and chemical evidence.

Full text of DOI:10.1080/10286020.2013.837454

This article was downloaded by: [Ryerson University]
On: 02 December 2014, At: 10:58
Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered
office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK
Journal of Asian Natural Products
Research
Publication details, including instructions for authors and
subscription information:
http://www.tandfonline.com/loi/ganp20
Five new stilbene glycosides from the
roots of Polygonum multiflorum
abc
bc
bc
bc
Shuo-Guo Li , Li-Li Chen , Xiao-Jun Huang , Bing-Xin Zhao ,
bc
abc
Ying Wang & Wen-Cai Ye
a
Department of Phytochemistry, China Pharmaceutical University,
Nanjing, 210009, China
b
Institute of Traditional Chinese Medicine and Natural Products,
College of Pharmacy, Jinan University, Guangzhou, 510632, China
c
JNU-HKUST Joint Laboratory for Neuroscience and Innovative
Drug Research, Jinan University, Guangzhou, 510632, China
Published online: 11 Nov 2013.
To cite this article: Shuo-Guo Li, Li-Li Chen, Xiao-Jun Huang, Bing-Xin Zhao, Ying Wang & Wen-Cai
Ye (2013) Five new stilbene glycosides from the roots of Polygonum multiflorum, Journal of Asian
Natural Products Research, 15:11, 1145-1151, DOI: 10.1080/10286020.2013.837454
To link to this article: http://dx.doi.org/10.1080/10286020.2013.837454
PLEASE SCROLL DOWN FOR ARTICLE
Taylor & Francis makes every effort to ensure the accuracy of all the information (the
“
Content”) contained in the publications on our platform. However, Taylor & Francis,
our agents, and our licensors make no representations or warranties whatsoever as to
the accuracy, completeness, or suitability for any purpose of the Content. Any opinions
and views expressed in this publication are the opinions and views of the authors,
and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content
should not be relied upon and should be independently verified with primary sources
of information. Taylor and Francis shall not be liable for any losses, actions, claims,
proceedings, demands, costs, expenses, damages, and other liabilities whatsoever
or howsoever caused arising directly or indirectly in connection with, in relation to or
arising out of the use of the Content.
This article may be used for research, teaching, and private study purposes. Any
substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing,
systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &

Conditions of access and use can be found at http://www.tandfonline.com/page/terms-
and-conditions

Journal of Asian Natural Products Research, 2013
Vol. 15, No. 11, 1145–1151, http://dx.doi.org/10.1080/10286020.2013.837454
Five new stilbene glycosides from the roots of Polygonum multiflorum
a,b,c
Shuo-Guo Li , Li-Li Chen , Xiao-Jun Huang , Bing-Xin Zhao , Ying Wang * and
b,c
b,c
b,c
b,c
a,b,c
Wen-Cai Ye
*
a
Department of Phytochemistry, China Pharmaceutical University, Nanjing 210009, China;
Institute of Traditional Chinese Medicine and Natural Products, College of Pharmacy, Jinan
University, Guangzhou 510632, China; JNU-HKUST Joint Laboratory for Neuroscience and
Innovative Drug Research, Jinan University, Guangzhou 510632, China
b
c
(
Received 1 May 2013; final version received 20 August 2013)
Five new stilbene glycosides (1–5), together with six known ones, were isolated from
the roots of Polygonum multiflorum. Their structures were elucidated on the basis of
spectroscopic analysis and chemical evidence.
Keywords: Polygonum multiflorum; Polygonaceae; stilbene glycoside
0
1
.
Introduction
The root of Polygonum multiflorum Thunb
Polygonaceae), He-Shou-Wu in Chinese, is
side (4), and (E)-2,3,5,4 -tetrahydroxystil-
0
0
bene-2-O-(2 -O-b-D-fructofuranosyl)-
b-D-glucopyranoside (5) (Figure 1),
together with six known ones (6–11).
In this paper, we describe the isolation and
structural elucidation of these new stilbene
glycosides.
(
a popular traditional Chinese medicine,
which has been widely used in China and
other Asian countries as a tonic, anti-oxida-
tive, and anti-aging agent for centuries [1].
Stilbene glycosides, one of the main active
constituents of this plant, had been proved to
possess anti-inflammatory, anti-oxidant,
anti-HIV, anti-tumor, and liver protection
activities [2–7]. However, to date, only 13
stilbene glycosides had been isolated from
the plant [8–13]. In our previous study, we
had reported the isolation of a new flavono-
stilbene glycoside and several known
stilbene glycosides from the roots of
P. multiflorum [14]. Our further investiga-
tion of this plant has led to the finding of five
2.
Results and discussion
The molecular formula of 1 was deter-
mined tobeC H O byits HR-ESI-MSat
2
6 32 14
þ
m/z 591.1683 [M þ Na] . The UV spec-
trum showed the absorption maxima at 206
and 314 nm. The IR spectrum implied the
21
presence of hydroxyl group (3407 cm
)
and aromatic ring (1605 and 1508cm ).
21
1
The H NMR spectrum of 1 revealed
proton signals for a para-disubstituted
0
new stilbene glycosides, (E)-2,3,5,4 -tetra-
hydroxystilbene-2-O-(4 -O-a-D-glucopyra-
nosyl)-b-D-glucopyranoside (1), (E)-2,3,
5
glucopyranosyl)-b-D-glucopyranoside (2),
(E)-2,3,5,4 -tetrahydroxystilbene-2-O-b-D-
glucopyranosyl-4 -O-a-D-glucopyranoside
(3), (E)-2,3,5,4 -tetrahydroxystilbene-2-O-
b-D-glucopyranosyl-5-O-a-D-glucopyrano-
benzene ring [d 7.45 (2H, d, J ¼ 8.4 Hz)
H
00
and 6.77 (2H, d, J ¼ 8.4 Hz)], a tetrasubsti-
tuted benzene ring [dH 6.63 (1H, d,
J ¼ 2.4 Hz) and 6.26 (1H, d, J ¼ 2.4 Hz)],
and a trans-disubstitued double bond [dH
7.70 (1H, d, J ¼ 16.4Hz) and 6.92 (1H, d,
0
0
0
,4 -tetrahydroxystilbene-2-O-(6 -O-b-D-
0
0
1
J ¼ 16.4Hz)]. In addition, the H NMR
0
spectrum of 1 displayed the signals of
two anomeric protons [dH 5.22 (1H, d,
*Corresponding authors. Emails: chyewc@gmail.com; wangying_cpu@163.com
q 2013 Taylor & Francis

1
146
S.-G. Li et al.
Figure 1. Chemical structures of compounds 1–5.
J ¼ 4.0 Hz) and 4.53 (1H, d, J ¼ 8.0 Hz)],
suggesting the presence of two sugar
moieties. The above spectral data suggested
HR-ESI-MS
þ
data
(m/z
591.1686
[M þ Na] , calcd for C H O Na,
2
6
32 14
591.1684). Similar to 1, compound 2
showed the characteristic UV (lmax 206
that 1 was a stilbene glycoside with an (E)-
0
2
,3,5,4 -tetrahydroxystilbene aglycone and
and 312 nm) and IR (n
1
3402, 1609,
1508 cm ) absorptions for a stilbene
max
2
two sugar moieties [3]. Acid hydrolysis of 1
afforded D-glucose, which was identified by
high-performance liquid chromatography
1
glycoside. The H NMR spectrum of 2
displayed aromatic and olefinic proton
signals at dH 7.69 (1H, d, J ¼ 16.4Hz),
7.44 (2H, d, J ¼ 8.8 Hz), 6.93 (1H, d,
J ¼ 16.4Hz), 6.77 (2H, d, J ¼ 8.8 Hz), 6.63
(1H, d, J ¼ 2.4 Hz), and 6.25 (1H, d,
J ¼ 2.4 Hz), as well as two anomeric
(HPLC) analysis [15]. The b-configuration
of a D-glucose unit (J ¼ 8.0 Hz) and
a-configuration of another D-glucose unit
(
J ¼ 4.0 Hz) were, respectively, determined
3
based on the J , coupling constants of
H1 H2
1
3
anomeric protons. The C NMR and DEPT
spectra of 1 revealed the presence of 26
carbon signals including 12 aromatic car-
bons, 2 olefinic carbons, and 2 glucose
protons at d 4.54 (1H, d, J ¼ 8.0 Hz) and
H
4.32 (1H, d, J ¼ 7.6 Hz), suggesting that 2
was also a stilbene glycoside with two sugar
1
1
units. The H– H COSY, HSQC, HMBC,
and NOESY spectra of 2 allowed the full
assignments of all proton and carbon signals
(Table 1). Comparison of the NMR data of
the aglycone part of 2 with those of 1
revealed that they were very similar, indica-
1
1
moieties. With the aid of H– H COSY,
HSQC, HMBC, and NOESY experiments,
1
13
all the H and C NMR signals of 1 were
assigned as shown in Table 1. In the HMBC
spectrum of 1, the long-range correlations
0
00
0
between H-1 (d 5.22) of the a-D-glucose
H
ting that 2 possessed the same (E)-2,3,5,4 -
0
0
and C-4 (d 80.1) of the b-D-glucose, as
C
tetrahydroxystilbene aglycone as 1. Acid
hydrolysis of 2 also afforded D-glucose.
The relative anomeric configurations of the
00
well as between H-1 (d 4.53) of the b-D-
H
glucose and C-2 (d 137.8) of the stilbene
C
aglycone were observed. Based on the
above results, the structure of 1 was
two D-glucose moieties were determined to
3
be b on the basis of the large J ,
H1 H2
0
established as (E)-2,3,5,4 -tetrahydroxystil-
0
coupling constants of the anomeric protons.
The sequence and linkage position of the
glucose moieties were deduced by the
HMBCexperiment.IntheHMBCspectrum,
0
bene-2-O-(4 -O-a-D-glucopyranosyl)-b-D-
glucopyranoside.
Compound 2 displayed the same
molecular formula C H O as 1 by its
0
00
the correlations between H-1 (d 4.32) of
H
2
6
32 14

Journal of Asian Natural Products Research
1147
d
d
d
d
d
d
d
d
d
d

1
148
S.-G. Li et al.
00
1
glycoside. The H NMR spectrum of 5
the terminal b-D-glucose and C-6 (d 69.4)
C
of the inner b-D-glucose, as well as between
0
showed proton signals due to a para-
disubstituted benzene ring [d 7.45 (2H, d,
0
H-1 (d 4.54) of the inner b-D-glucose and
H
H
C-2 (d 137.7) of the stilbene aglycone were
C
J ¼ 8.6 Hz) and 6.78 (2H, d, J ¼ 8.6 Hz)], a
observed. Therefore, the structure of 2 was
0
tetrasubstituted benzene ring [d 6.63 (1H,
H
characterized as (E)-2,3,5,4 -tetrahydroxys-
0
d, J ¼ 2.8 Hz)and6.28(1H,d,J ¼ 2.8 Hz)],
0
tilbene-2-O-(6 -O-b-D-glucopyranosyl)-b-
D-glucopyranoside.
a trans-disubstitued double bond [d 7.68
H
(1H, d, J ¼ 16.4Hz) and 6.93 (1H, d,
J ¼ 16.4 Hz)], and an anomeric proton
signal at dH 4.52 (1H, d, J ¼ 7.9Hz).
Besides 12 aromatic carbons and 2 olefinic
The molecular formula of 3 was
established as C H O by its HR-ESI-
2
6 32 14
þ
MS at m/z 591.1686 [M þ Na] , which was
identicaltothatof1and2. Similar to 1and2,
acid hydrolysis of 3 afforded D-glucose.
Comparison of NMR data of 3 with those of
1
3
carbons, the C NMR spectrum of 5
indicated the presence of two sugar units.
All the above information suggested that 5
was also a stilbene glycoside with an (E)-
1
2
suggested they possessed the same (E)-
0
0
,3,5,4 -tetrahydroxystilbene aglycone, as
2,3,5,4 -tetrahydroxystilbene aglycone and
1
1
well as a-D-glucose and b-D-glucose units.
The difference between them was the
linkage position of the a-D-glucose moiety.
In the HMBC spectrum of 3, the correlations
two sugar moieties. Interpretation of H– H
COSY,HSQC,HMBC, andNOESYspectra
of 5 led to the assignment of all proton and
carbon signals as shown in Table 1.
Different from 1 to 4, acid hydrolysis of 5
afforded D-glucose and D-fructose, which
were identified by the gas chromatography
(GC) analysis. The b-configuration of
the D-glucose was determined based on the
0
00
between H-1 (d 5.50) of the a-D-glucose
H
0
and C-4 (d 158.2) of the aglycone, as well
C
00
as between H-1 (dH 4.51) of the b-D-
glucose and C-2 (d 138.1) of the stilbene
C
aglycone were observed. Thus, the structure
0
3
of 3 was confirmed as (E)-2,3,5,4 -tetrahy-
0
JH1,H2 coupling constant (J ¼ 8.0 Hz) of
droxystilbene-2-O-b-D-glucopyranosyl-4 -
O-a-D-glucopyranoside.
the anomeric proton. The b-configuration of
the D-furanose was deduced by comparison
of the NMR data with those reported in the
literature [16]. In addition, the HMBC
Compound 4 showed the same molecu-
lar formula as 1–3 by its HR-ESI-MS data.
Acid hydrolysis of 4 also afforded
0
00
correlation between H-5 (dH 3.74) of
000
1
13
D-glucose. The H and C NMR data of 4
were similar to those of 3, suggesting that
they possessed the same stilbene aglycone
and glucose units. Different from 3, in the
HMBC spectrum of 4, the correlation
fructose and C-2 (dC 105.2) of fructose
was observed, which confirmed the exist-
ence of b-D-fructofuranose. Furthermore, in
the HMBC spectrum of 5, the correlations
0
0
between H-2 (d 3.56) of b-D-glucose and
H
0
00
000
between H-1 (dH 5.45) of a-D-glucose
and C-5 (dC 156.2) of aglycone was
observed, indicating that the a-D-glucose
unit in 4 was attached to the C-5 position of
aglycone. Hence, the structure of 4 was
C-2 (d 105.2) of b-D-fructofuranose, as
C
0
0
well as between H-1 (d 4.52) of b-D-
H
glucoseandC-2(d 137.9)ofaglyconewere
C
observed. Hence, the structure of 5 was
0
established to be (E)-2,3,5,4 -tetrahydrox-
00
0
determined as (E)-2,3,5,4 -tetrahydroxystil-
bene-2-O-b-D-glucopyranosyl-5-O-a-D-
glucopyranoside.
ystilbene-2-O-(2 -O-b-D-fructofuranosyl)-
b-D-glucopyranoside.
The six known stilbene glycosides were
0
The molecular formula of 5 was also
deduced as C H O by its HR-ESI-MS
identified as (E)-2,3,5,4 -tetrahydroxystil-
0
bene-2-O-b-D-glucopyranosyl-4 -O-b-D-
0
2
6 32 14
þ
at m/z 591.1686 [M þ Na] . The UV and
IR spectra of 5 showed the characteristic
absorptions corresponding to a stilbene
glucopyranoside (6) [17], (E)-2,3,5,4 -tetra-
0
0
hydroxystilbene-2-O-(6 -O-a-D-glucopyr-
anosyl)-b-D-glucopyranoside (7) [12],

Journal of Asian Natural Products Research
1149
0
polygonimitin (8) [13], (E)-2,3,5,4 -tetrahy-
droxystilbene-2-O-b-D-glucopyranoside
silica gel GF254 plates (Yantai Chemical
Industry Research Institute, Yantai, China).
0
(9) [3], (E)-2,3,5,4 -tetrahydroxystilbene-2-
O-(3 -O-galloyl)-b-D-glucopyranoside (10)
11], and (Z)-2,3,5,4 -tetrahydroxystilbene-
-O-b-D-glucopyranoside (11) [18],
respectively, by comparison of their physi-
cal and spectroscopic data with literature
values.
0
0
0
3.2 Plant material
[
2
The roots of P. multiflorum were collected
in Deqing County, Guangdong Province of
China, in October of 2010 and authenti-
cated by Prof. Guang-Xiong Zhou (College
of Pharmacy, Jinan University). A voucher
specimen (No. 20101003) had been depo-
sited in the Institute of Traditional Chinese
Medicine & Natural Products, Jinan
University, Guangzhou, China.
3
.
Experimental
3
.1 General experimental procedures
Optical rotations were measured on a
JASCO P-1020 digital polarimeter at room
temperature (JASCO, Tokyo, Japan). UV
spectra were obtained on a JASCO-V-550
UV/VIS spectrophotometer (JASCO).
IR spectra were recorded on a JASCO
FT/IR-480 plus Fourier transform infrared
spectrometer with KBr pellets (JASCO).
HR-ESI-MS data were obtained on an
Agilent 6210 LC/MSD TOF mass spec-
trometer (Agilent, Pala Alto, CA, USA). 1D
and 2D NMR experiments were performed
on a Bruker AV-400 spectrometer (Bruker,
Billerica, MA, USA). GC–MS analyses
were performed on a Shimadzu GCMS-
QP2010 plus gas chromatograph–mass
spectrometer (Shimadzu, Kyoto, Japan).
Analytical HPLC was performed on an
Agilent 1260 chromatography equipped
withaG1311Cpump,aG1315Dphotodiode
array detector (Agilent) and a Cosmosil
3
.3 Extraction and isolation
The air-dried and powdered roots of
P. multiflorum (14.5 kg) were extracted
with 95% (v/v) EtOH under reflux for two
times (2 £ 25 liters, 2 h each). The com-
bined EtOH solution was concentrated
under vacuum to yield a residue (1200 g).
The crude extract was suspended in water
and then partitioned with petroleum
ether (b.p. 60–908C), ethyl acetate, and
n-butanol, respectively. The n-butanol sol-
uble fraction (144g) was redissolved in H O
2
and subjected to macroporous resin HP-20
column eluted with EtOH–H O mixtures.
2
The 30% EtOH eluate (14g) was subjected
to Sephadex LH-20 column and eluted with
MeOH–H Omixtures(70:30 ! 100:0,v/v)
2
to obtain nine fractions (1–9). Fraction 5
(2.0g) was separated by silica gel column
with gradient mixtures of CHCl –MeOH
5
C -MS-II Waters column (4.6 £ 250 mm,
18
5
mm; Nacalai Tesque, Inc., Kyoto, Japan).
3
Preparative HPLC was carried on an
Agilent 1260 chromatography equipped
with a G1310B pump, a G1365D detector
(95:5, 90:10, 85:15, 80:20, 75:25, v/v) as
eluents to yield five subfractions (5a–5e).
Subfraction 5d (0.5 g) was further separated
by preparative HPLC on a reversed-phase
C18 column (20 mm £ 250 mm, 5 mm)
(Agilent), and a Cosmosil 5C -MS-II
18
Waters column (20 £ 250 mm, 5 mm;
Nacalai Tesque, Inc.). Column chromatog-
raphies were performed on macroporous
resin Diaion HP-20 (Mitsubishi Chemical
Corporation, Tokyo, Japan), silica gel(300–
using MeOH–H O (25:75, v/v, 6 ml/min,
2
320 nm) as mobile phase to afford 3 (8.0mg,
t ¼ 37.7min), 4 (12.0 mg, t ¼ 25.3min),
R
R
6 (24.4 mg, t ¼ 33.8min), and 8 (25.0 mg,
R
4
00 mesh; Qingdao Marine Chemical Co.
Ltd, Qingdao, China), and Sephadex LH-20
Pharmacia Biotech AB, Uppsala, Sweden).
TLC analyses were carried on precoated
t ¼ 51.4 min), respectively. Fraction 7
R
(1.0 g) was separated by silica gel
column (CHCl –MeOH, 90:10, v/v)
and then further purified by preparative
(
3

1
150
S.-G. Li et al.
HPLC on a reversed-phase C18 column
(log 1): 208 (4.39), 314 (4.26) nm; IR
(KBr) n : 3407, 1605, 1508, 1459, 1018,
(20 mm £ 250 mm, 5 mm) using MeOH–
max
2
1
1
13
839 cm ; H and C NMR spectral data
H O (35: 65, v/v, 6 ml/min, 320 nm) as
2
eluent to yield 2 (15.4 mg, t ¼ 20.7min), 5
(see Table 1); ESI-MS m/z: 591
þ
R
2
[M þ Na] , 567 [M 2 H] ; HR-ESI-MS
(
12.0 mg, t ¼ 29.9min), and 7 (26.0 mg,
R
þ
t ¼ 18.1 min), respectively. Fraction 8
m/z: 591.1686 [M þ Na] (calcd for
R
(0.8g) was separated by silica gel column
with gradient mixtures of CHCl –MeOH–
C H O Na, 591.1684).
26 32 14
3
H O (85:15:1.5, 80:20:2, 75:25:3, 70:30:5,
2
3
.3.4 Compound 4
v/v) as eluents to yield four subfractions
(8a–8d). Subfraction 8a (0.25g) was puri-
2
D
0
Amorphous powder;
(
½aꢀ þ 47.3
c ¼ 0.60, MeOH); UV (MeOH) l
max
fied by Sephadex LH-20 column (MeOH)
to yield 10 (220mg). Subfraction 8c (0.2g)
was reseparated by preparative HPLC
(
log 1): 208 (4.09), 314 (3.94) nm; IR
(KBr) n : 3387, 1609, 1516, 1456, 1018,
max
2
28 cm ; H and C NMR spectral data
1
1
13
8
on
a
reversed-phase C18 column
(
[
see Table 1); ESI-MS m/z: 591
þ
(
20 mm £ 250 mm, 5 mm) using MeOH–
2
M þ Na] , 567 [M 2 H] ; HR-ESI-MS
H O (38:62, v/v, 6 ml/min, 320 nm) as
2
þ
m/z: 591.1683 [M þ Na] (calcd for
eluent to affford 1 (11.0 mg, t ¼ 23.5min),
R
C H O Na, 591.1684).
26 32 14
9
(34.0 mg, t ¼ 39.5 min), and 11
R
(22.0 mg, t ¼ 21.9min), respectively.
R
3.3.5 Compound 5
2
D
0
Amorphous powder;
½aꢀ 2 16.3
3
.3.1 Compound 1
(c ¼ 0.45, MeOH); UV (MeOH) l
(log 1): 216 (4.09), 322 (4.10) nm; IR
max
20
Amorphous powder; ½aꢀ þ 79.5 (c ¼
D
0
.28, MeOH); UV (MeOH) lmax (log1):
(
KBr) n : 3431, 1609, 1512, 1036,
max
21
28 cm ; H and C NMR spectral data
2
06 (4.54), 314 (4.26) nm; IR (KBr) n
max
407, 1605, 1508, 1463, 1038, 828 cm
:
;
1
13
8
21
3
(
[
see Table 1); ESI-MS m/z: 591
2
1
13
H and C NMR spectraldata (see Table 1);
þ
þ
M þ Na] , 567 [M 2 H] ; HR-ESI-MS
ESI-MS m/z: 591 [M þ Na] , 567
þ
m/z: 591.1686 [M þ Na] (calcd for
2
M 2 H] ; HR-ESI-MS m/z: 591.1683
[
C H O Na, 591.1684).
26 32 14
þ
[
M þ Na] (calcd for C H O Na,
2
6
32 14
5
91.1684).
3.4 Acid hydrolysis and HPLC analysis
of 1–4
3
.3.2 Compound 2
Each solution of compounds 1–4 (each
2 mg) in 2 mol/l HCl (5ml) was heated in
water bath (808C) for 4 h. The solution was
evaporated under reduced pressure. Each
residue was dissolved in pyridine (1.0ml)
and stirred with L-cysteine methyl ester
hydrochloride (2mg) for 1 h at 608C, and
then O-tolyl isothiocyanate (20 ml) was
added to the mixture and heated at 608C
for another 1 h. The reaction mixtures
were analyzed by HPLC and detected
at 250 nm. Analytical HPLC was per-
formed on a Cosmosil 5C -MS-II column
2
D
0
Amorphous powder; ½aꢀ 2 19.4 (c ¼
0
.35, MeOH); UV (MeOH) l
(log1):
max
2
06 (4.30), 312 (4.11) nm; IR (KBr) n
max
402, 1609, 1508, 1455, 1058, 828 cm
:
;
21
3
1
13
H and C NMR spectraldata (see Table 1);
þ
ESI-MS m/z: 591 [M þ Na] , 567
2
M 2 H] ; HR-ESI-MS m/z: 591.1686
[
þ
[
M þ Na] (calcd for C H O Na,
2
6
32 14
5
91.1684).
3
.3.3 Compound 3
1
8
2
0
Amorphous powder;
(
c ¼ 0.43, MeOH); UV (MeOH) lmax
[a ] þ 68.2
(4.6 mm £ 250 mm, 5 mm) at 208C
D
using CH CN–0.05% CH COOH (25:75,
3
3

Journal of Asian Natural Products Research
1151
1.0 ml/min) as the mobile phase. Peaks were
detected with a G1315D photodiode array
Excellent Talents in University (No. NCET-11-
857), and the Science and Technology
Planning Project of Guangzhou (No.
011J2200046).
0
detector. D-glucose (t ¼ 16.36 min) was
R
2
identified as sugar moiety of 1–4 based
on comparisons with authentic samples of
D-glucose (t ¼ 16.36 min) and L-glucose
R
References
(
t ¼ 14.99 min).
R
[
1] P.G. Xiao, S.T. Xing, and L.W. Wang,
J. Ethnopharmacol. 38, 167 (1993).
[
2] Y.Z. Zhang, J.F. Shen, J.Y. Xu, J.H. Xiao,
and J.L. Wang, J. Asian. Nat. Prod. Res. 9,
355 (2007).
3] Y. Chen, M.F. Wang, R.T. Rosen, and C.T.
Ho, J. Agric. Food Chem. 47, 2226 (1999).
4] H.W. Lin, M.X. Sun, Y.H. Wang, L.
Meng, Y.R. Yang, N. Ning, L.J. Xuan,
Y.M. Xu, D.L. Bai, Y.T. Zhang, and K.
Xiao, Planta Med. 76, 889 (2010).
3
.5 Acid hydrolysis and GC analysis of 5
Compound 5 (3.7 mg) was heated in an
ampoule with 4.0 ml of 2 mol/l HCl at
[
[
8
with a steam of N to yield a residue,
08C for 6 h. The solution was evaporated
2
which was dissolved in H O and extracted
2
with CHCl . The aqueous layer was
3
[
[
5] M.J. Wang, R.H. Zhao, W.G. Wang, X.J.
Jian, and J. Yu, J. Ethnopharmacol. 139,
evaporated by N2 and treated with
anhydrous pyridine (3 ml) and L-cysteine
methyl ester hydrochloride (4 mg), fol-
lowed by heating at 608C for 2 h and then
2
87 (2012).
6] W.H. Park, S.J. Lee, and H.I. Moon,
Antimicrob. Agents Chemother. 52, 3451
(2008).
concentrated to dryness with N . The
2
[
[
7] K. Likhitwitayawuid and B. Sritularak,
J. Nat. Prod. 64, 1457 (2001).
8] W.S. Chen, W.D. Zhang, G.J. Yang, H.S.
Chen, and C.Z. Qiao, Chin. Chem. Lett.
12, 503 (2001).
residue was added to N-(trimethylsilyl)
imidazole (0.3 ml) and kept at 608C for 1 h.
Subsequently, the solution was diluted
with H O (1 ml) and extracted with hexane
2
(2 ml). The organic layer was analyzed
using GC under the following conditions:
[9] J.N. Grech, Q. Li, B.D. Roufogalis, and
C.C. Duke, J. Nat. Prod. 57, 1682 (1994).
10] K. Xiao, L. Xuan, Y. Xu, and D. Bai, Acta
Bot. Sin. 44, 1491 (2002).
11] H.K. Kim, Y.H. Choi, J.S. Choi, S.U. Choi,
Y.S. Kim, K.R. Lee, Y.K. Kim, and S.Y.
Ryu, Arch. Pharm. Res. 31, 1225 (2008).
12] W.S. Chen, G.J. Yang, W.D. Zhang, C.Z.
Qiao, and H.S. Chen, Acta. Pharmacol.
Sin. 35, 273 (2000).
13] L.X. Zhou, M. Lin, and J.B. Li, Acta.
Pharmacol. Sin. 29, 107 (1994).
[14] L.L. Chen, X.J. Huang, M.M. Li, G.M.
Ou, B.X. Zhao, M.F. Chen, Q.W. Zhang,
Y. Wang, and W.C. Ye, Phytochem. Lett.
[
[
column: HT-SE-30 (0.32 mm £ 30 mm,
0
.5 mm), detector: FID, column tempera-
ture: 200–2508C (58C/min), detector tem-
perature: 2808C, injector temperature:
[
[
2
508C, and carrier gas: N . The standard
2
D-glucose and L-glucose were subjected to
the same reaction. As a result, D-glucose
(
t ¼ 18.86 min) and D-fructose (t ¼
R
R
16.06 min) were identified as sugar moiety
of 5 based on comparisons with authentic
samples of D-glucose (t ¼ 18.86 min) and
R
5, 756 (2012).
D-fructose (t ¼ 16.06 min).
R
[15] T. Tanaka, T. Nakashima, T. Ueda, K.
Tomii, and I. Kouno, Chem. Pharm. Bull.
5
5, 899 (2007).
16] C.Z. Zhang, X.Z. Xu, and C. Li,
Acknowledgments
[
Financial support of this work was provided by
the Program for Changjiang Scholars and
Innovative Research Team in the University
Phytochemistry 41, 975 (1995).
[17] S.R. Oh, S.Y. Ryu, S.H. Park, K.Y. Jung,
I.S. Lee, K.S. An, J.J. Lee, and H.K. Lee,
Arch. Pharm. Res. 21, 703 (1998).
(
No. IRT0965), the Joint Fund of NSFC-
Guangdong Province (No. U0932004), the
National Natural Science Foundation of China
[18] M.L. Xu, M.S. Zheng, Y.K. Lee, C.S. Lee,
M.H. Woo, B.S. Jeong, E.S. Lee, and J.K.
Son, Arch. Pharm. Res. 29, 946 (2006).
(
No. 81172946), the Program for New Century