Aromatic glycosides from the flower buds of Lonicera japonica

Article abstract of DOI:10.1080/10286020.2013.785531

Six new glycosides (1-6) have been isolated from the flower buds of Lonicera japonica. Their structures including the absolute configurations were determined by spectroscopic and chemicalmethods as (2)-2-hydroxy-5- methoxybenzoic acid 2-O-β-D-(6-O-benzoyl

Full text of DOI:10.1080/10286020.2013.785531

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Aromatic glycosides from the flower
buds of Lonicera japonica
a
a
a
Fang Wang , Yue-Ping Jiang , Xiao-Liang Wang , Su-Juan Wang
a
a
a
a
a
,
Peng-Bin Bu , Sheng Lin , Cheng-Gen Zhu & Jian-Gong Shi
a
State Key Laboratory of Bioactive Substance and Function of
Natural Medicines, Institute of Materia Medica, Chinese Academy
of Medical Sciences and Peking Union Medical College , Beijing ,
100050 , China
Published online: 24 Apr 2013.
To cite this article: Fang Wang , Yue-Ping Jiang , Xiao-Liang Wang , Su-Juan Wang , Peng-Bin
Bu , Sheng Lin , Cheng-Gen Zhu & Jian-Gong Shi (2013) Aromatic glycosides from the flower
buds of Lonicera japonica , Journal of Asian Natural Products Research, 15:5, 492-501, DOI:
10.1080/10286020.2013.785531
To link to this article: http://dx.doi.org/10.1080/10286020.2013.785531
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Journal of Asian Natural Products Research, 2013
Vol. 15, No. 5, 492–501, http://dx.doi.org/10.1080/10286020.2013.785531
Aromatic glycosides from the flower buds of Lonicera japonica
Fang Wang, Yue-Ping Jiang, Xiao-Liang Wang, Su-Juan Wang, Peng-Bin Bu, Sheng Lin*,
Cheng-Gen Zhu and Jian-Gong Shi*
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of
Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing
1
00050, China
(
Received 22 January 2013; final version received 10 March 2013)
Six new glycosides (1–6) have been isolated from the flower buds of Lonicera
japonica. Their structures including the absolute configurations were determined
byspectroscopicandchemicalmethodsas(2)-2-hydroxy-5-methoxybenzoicacid2-O-b-
D-(6-O-benzoyl)-glucopyranoside (1), (2)-4-hydroxy-3,5-dimethoxybenzoic acid 4-O-
b-D-(6-O-benzoyl)-glucopyranoside (2), (2)-(E)-3,5-dimethoxyphenylpropenoic acid
4
9
-O-b-D-(6-O-benzoyl)-glucopyranoside (3), (2)-(7S,8R)-(4-hydroxyphenylglycerol
-O-b-D-[6-O-(E)-4-hydroxy-3,5-dimethoxyphenylpropenoyl]-glucopyranoside (4),
(2)-(7S,8R)-(4-hydroxy-3-methoxyphenylglycerol 9-O-b-D-[6-O-(E)-4-hydroxy-3,5-
dimethoxyphenylpropenoyl]-glucopyranoside (5), and (2)-4-hydroxy-3-methoxyphenol
b-D-{6-O-[4-O-(7S,8R)-(4-hydroxy-3-methoxyphenylglycerol-8-yl)-3-methoxyben-
zoyl]}-glucopyranoside (6), respectively.
Keywords: flower buds; Lonicera japonica; Caprifoliaceae; aromatic glycosides
1
.
Introduction
phenylpyruvic acid-derived moieties-
coupled (loniphenyruviridosides A–D)
homosecoiridoids, together with 7 known
iridoid derivatives, were reported from the
water extract of the flower buds of
L. japonica [14,15]. Focusing our investi-
gation on the components of this plant
material, after the flower buds were
extracted by water, the residue was further
extracted with EtOH (95%). From the
EtOH extract, six new glycosides (1–6)
have been characterized (Figure 1). The
isolation and structure determination of the
new isolates have been reported in this
study.
Lonicera japonica Thunb. (Caprifolia-
ceae) is widely cultivated in the Shandong
and Henan provinces of China. Its flower
buds and stems are used in traditional
Chinese medicine for the treatment of
various diseases, and are among the most
common ingredients of formulations used
for treating influenza, cold, fever, and
infections [1–5]. Chemical and pharma-
cological studies have resulted in the
characterization of constituents with
different structural features and biological
activities from the flower buds of
L. japonica, including caffeoyl quinic
acids, secoiridoids, flavonoids, saponins,
cerebrosides, polyphenols, and nitrogen-
containing iridoids [6–13]. As part of a
program to assess the chemical and
biological diversity of traditional Chinese
medicines, 14 pyridinium alkaloid-
coupled (lonijaposides A–N) and 4
2
.
Results and discussion
Compound 1 was obtained as a white
2
0
D
amorphous solid with ½aꢀ 251.5 (c 0.18,
MeOH). The IR spectrum of 1 showed
absorption
bands
(3307 cm ), carbonyl (1708 cm ), and
for
hydroxyl
21
2
1
*Corresponding authors. Email: lsznn@126.com; shijg@imm.ac.cn
q 2013 Taylor & Francis

Journal of Asian Natural Products Research
493
3
"
O
O
HO
HO
O
7
1
7
"
O
"
O
7 OH
1
MeO 3
1
OH
OH
5
"
5'
O
HO
O
1
' O
O
O
5
3'
HO
OH
5
3
OH
OMe
OMe
1
2
MeO
OH HO
MeO
O
O
O
9
O
O
HO
HO
7
HO
1
5
MeO 3
1"
7
1
9
O
HO
1' O
OH
O
3
R
O
HO
5
OMe
OH
OH
4=H
3
5=OMe
HO
OH
OMe
3
8
O
'"
O
O
HO
HO
1
"
OH
1'"
MeO
HO
O
5
MeO
1'
O 1
6
OH
Figure 1. The structures of compounds 1–6.
2
1
aromatic (1603 and 1516 cm ) functional
groups. The positive mode electrospray
ionization mass spectrometry (ESIMS) of
an authentic sugar sample. The glucose
isolated from the hydrolysate gave a
2
0
positive optical rotation, ½aꢀ þ 40.7 (c
13
glucose [15]. The C NMR and DEPT
D
1
gave a quasi-molecular ion peak at m/z
þ
0.10, H O), indicating that it was the b-D-
2
4
C H O was indicated by high resol-
57 [M þ Na] . The molecular formula
spectra of 1 showed carbon signals (Table
1) corresponding to the above units,
including two oxygen-bearing aromatic
2
1 22 10
ution electrospray ionization mass spec-
trometry (HRESIMS) at m/z 457.1106
þ
1
[
M þ Na] . The H NMR spectrum of 1
carbons [d 150.2 (C-5) and 151.0 (C-2)]
C
0
0
in CD OD showed signals attributed to a
3
and a carbonyl carbon [d 167.8 (C-7 )].
C
meta-ortho-trisubstituted phenyl group at
d 7.07 (1H, d, J ¼ 8.4 Hz, H-3), 7.40 (1H,
These spectroscopic data suggested that 1
was a meta-ortho-substituted benzolic acid
b-glucopyranoside with the benzoyl unit
H
d, J ¼ 8.4 Hz, H-4), and 7.55 (1H, s, H-6); a
0
benzoyl unit at d 7.94 (2H, d, J ¼ 7.2 Hz,
esterified at C-6 . The suggestion was
H
0
0
00
00 00
H-2 /6 ), 7.43 (2H, t, J ¼ 7.2 Hz, H-3 /5 ),
and 7.55 (1H, t, J ¼ 7.2 Hz, H-4”); and an
aromatic methoxy group at d 3.82 (s). In
addition, it showed signals assignable to a
b-glucopyranosyl unit, of which the
supported by comparing of the NMR data
of 1 with those of the co-occurring vanillic
acid 4-O-b-D-(6-O-benzoyl)-glucopyrano-
side [16], and further confirmed by 2D
NMR data analysis. The HMQC and
1
1
anomeric proton was resonated at d 4.98
H
H– H correlation spectroscopy (COSY)
0
(
d, J ¼ 7.2 Hz, H-1 ), whereas the oxy-
spectra of 1 provided unambiguous assign-
ments of proton and carbon signals in the
NMR spectra. In the HMBC spectrum,
methylene protons were at d 4.66 (dd,
H
0
0
J ¼ 12.0 and 1.8 Hz, H-6 a) and 4.32 (dd,
0 00 00
correlations from H -6 and H-2 /H-6 to
2
J ¼ 12.0 and 7.8 Hz, H-6 b). The presence
0
0
of a b-glucopyranosyl unit was confirmed
by acid hydrolysis of 1 that produced
glucose identified by a thin-layer liquid
chromatography (TLC) comparison with
C-7 confirmed that the benzoyl unit was
0
located at C-6 of the b-glucopyranosyl
moiety. In addition, correlations of C-2
with H-4 and H-6 and C-5 with H-3 and

4
94
F. Wang et al.
d
d
d
d
d
d
d
d
d
d
d
d
d

Journal of Asian Natural Products Research
495
d
d
d

4
96
F. Wang et al.
OCH , together with their shifts and the
3
C-3/C-5, C-4, and C-7; from H-7 to C-1,
C-2/C-6, and C-9; and from H-8 to C-1 and
C-9, in the HMBC spectrum of 3, in
combination with the shifts of the proton
and carbon resonances. In addition, the
molecular composition, demonstrated that
the meta-ortho-substituted benzolic acid
moiety was 2-oxy-5-methoxybenzolic acid
though the signals of C-1 and C-7 were not
1
3
0
displayed in the C NMR spectrum of 1.
Meanwhile, a HMBC correlation of C-2
HMBC correlations from H-1 to C-4 and
00
0
from H -6 to C-7 proved the linkage of
2
0
with H-1 revealed that the b-glucopyr-
the three moieties in 3. The D-configur-
ation of the b-glucopyranosyl moiety was
also confirmed by the acid hydrolysis of 3
using the same protocol as described
earlier. Thus, the structure of compound
3 was designated as (E)-3,5-dimethoxy-
phenylpropenoic acid (2)-4-O-b-D-(6-O-
benzoyl)-glucopyranoside.
anosyl moiety was located at C-2 in 1.
Therefore, the structure of compound 1
was determined as (2)-2-hydroxy-5-meth-
oxybenzoic acid 2-O-b-D-(6-O-benzoyl)-
glucopyranoside.
Compound 2 was obtained as a white
amorphous powder with ½aꢀ 231.8 (c
20
D
0
.09, MeOH). The molecular formula
C H O was indicated from HRESIMS
Compound 4 was obtained as a yellow-
2
0
D
ish amorphous powder with ½aꢀ 210.5 (c
2
2
24 11
þ
at m/z 487.1212 [M þ Na] , combined
with the NMR spectral data. The UV, IR,
and NMR spectroscopic features of 2 (see
Section 3 and Table 1) were similar to
those of 1. Comparison of the NMR
spectral data of 1 and 2 indicated that the
0.20, CH CN), and has the molecular
3
formula C H O as indicated by HRE-
2
6 32 13
þ
SIMS at m/z 575.1745 [M þ Na] and the
NMR spectral data (Table 1). The NMR
spectra of 4 showed the presence of (E)-4 -
0
0
0
0
00
hydroxy-3 ,5 -dimethoxyphenylprope-
noyl, 4-hydroxyphenyl, and b-glucopyra-
nosyl moieties in the molecule. In addition,
it demonstrated the presence of two
oxygen-bearing methines (CH-7 and CH-
2
-oxy-5-methoxybenzolic acid moiety in 1
was replaced by a 4-oxy-3,5-dimethox-
ybenzoic acid unit in 2. This demonstrated
that 2 was 4-hydroxy-3,5-dimethoxyben-
zoic acid 4-O-b-D-(6-O-benzoyl)-gluco-
pyranoside, which was confirmed by 2D
NMR data analysis and acid hydrolysis.
Particularly, the HMBC spectrum showed
8) and an oxygen-bearing methylene (CH -
2
1
9) in 4. The H– H gCOSY spectrum of 4
1
displayed vicinal coupling correlations of
H-7/H-8/H -9, whereas the HMBC spec-
2
0
correlations from H-1 and H-2/H-6 to C-4,
trum showed correlations from H-7 to C-1
and C-2/C-6 of the 4-hydroxyphenyl
moiety. These 2D NMR data revealed
that there was a 4-hydroxyphenylglycerol
indicating the b-D-glucopyranosyl moiety
at C-4 in 2. Therefore, the structure of
compound 2 was assigned as (2)-4-
hydroxy-3,5-dimethoxybenzoic acid 4-O-
b-D-(6-O-benzoyl)-glucopyranoside.
moiety. In addition, the HMBC spectrum
0
exhibited correlations from H -9 to C-1
2
0
Compound 3, a white amorphous
powder with ½aꢀ
and from H-1 to C-9, indicating that the
b-glucopyranosyl was located at C-9.
20
210.1 (c 0.37,
MeOH), has the molecular formula
C H O as indicated by HRESIMS at
D
00
Moreover, HMBC correlations of C-9
0
00
00
00
with H -6 , H-7 , and H-8 and of C-3 /C-
2
4
26 11
2
þ
00
00 00
with OCH -3 /OCH -5 , together
3 3
m/z 513.1374 [M þ Na] and the NMR
spectral data (Table 1). Comparison of
the NMR spectral data of 3 and 2 indicated
the substitution of the 4-oxy-3,5-
dimethoxybenzoic acid unit in 2 by a
trans-4-oxy-3,5-dimethoxyphenylprope-
noic acid moiety in 3. This was confirmed
by correlations from H-2/H-6 to C-1,
5
with their shifts, demonstrated that the
0
0
00 00
(E)-4 -hydroxy-3 ,5 -dimethoxyphenyl-
propenoyloxy moiety was substituted at C-
0
6 in 4. The Dd
value in DMSO was
C8–C7
smaller than 0.5 ppm, suggesting a 7,8-
erythro configuration for 4 [17–19]. In
the CD spectrum of 4, a negative Cotton

Journal of Asian Natural Products Research
497
effect at 227 nm indicated the 8R configur-
ation [19]. The D-configuration of the b-
glucopyranosyl moiety in 4 was proved
by acid hydrolysis using the aforemen-
tioned method. Therefore, the structure
of compound 4 was determined as
moiety, as well as two oxygen-bearing
0
00
000
methines (CH-7 and CH-8 ), an oxygen-
0
00
bearing methylene (CH -9 ), and an ester
2
0
0
carbonyl carbon (d 167.6, C-7 ). In the
C
HMBC spectrum of 6, correlations of H-
2/C-1, C-3, C-4, and C-6; H-5 and/or H-
(
2)-(7S,8R)-(4-hydroxyphenylglycerol 9-
6/C-1, C-3, and C-4; and OCH -3/C-3;
3
O-b-D-[6-O-(E)-4-hydroxy-3,5-dimethox-
yphenylpropenoyl]-glucopyranoside.
Compound 5, a yellowish amorphous
together with their shifts, suggested the
presence of a 4-hydroxy-3-methoxyphe-
noxy unit, which was supported by a
2
0
powder with ½aꢀ 225.1 (c 0.12, MeOH),
correlation between H-2 and OCH -3 in the
3
D
has the molecular formula C H O as
2
NOESY spectrum. The HMBC spectrum
00 00
7 34 14
indicated by HRESIMS at m/z 605.1854
also showed correlations of H-2 /C-1 , C-
00 00 00 00 00 00 00
3 , C-4 , C-6 , and C-7 ; H-5 /C-1 , C-3 ,
þ
[
(
M þ Na] and the NMR spectral data
00 00 00 00 00 00
Table 1). Comparison of the NMR spectral
C-4 , and C-6 ; H-6 /C-1 , C-2 , C-4 , C-
0
0
00
00
00
data of 4 and 5 indicated the replacement
of the 4-hydroxyphenylglycerol moiety
in 4 by a 4-hydroxy-3-methoxyphenylgly-
cerol moiety in 5. This was confirmed
by 2D NMR data analysis of 5. Particularly,
5 , and C-7 ; and OCH -3 /C-3 . The
3
correlations in combination with shifts of
these proton and carbon resonances indi-
0
0
00
cated that there was a 4 -hydroxy-3 -
methoxybenzoyloxy moiety. In addition,
1
1
000 000 000 000
H– H gCOSY correlations of H-7/
H-8/H -9 and HMBC correlations from
correlations of H-7 /C-1 , C-2 , C-6 , C-
000 000 000 000 000 000
8 and C-9 ; H-2 /C-1 , C-3 , C-4 , C-
2
000 000 000 000 000 000
H-7 to C-1, C-2, and C-6 and from OCH -3
3
6 , and C-7 ; H-5 /C-1 , C-3 , C-4 , and
000 000 000 000 000 000
C-6 ; H-6 /C-1 , C-2 , C-4 , C-5 , and
to C-3, together with their shifts, proved
the presence of 4-hydroxy-3-methoxyphe-
nylglycerol moiety. HMBC correlations
0
00
000
000
C-7 ; and OCH -3 /C-3 , together with
3
their shifts, demonstrated the presence of
000 000
0
0
from H -9 to C-1 , from H-1 to C-9, and
2
a 4 -hydroxy-3 -methoxyphenylglycerol
0
00
1
moiety. This was supported by H– H
1
from H -6 to C-9 confirmed the linkage
2
0
00
000
000
of the moieties in 5. The DdC8–C7 value in
DMSO was similar to that of 4, indicating
the 7,8-erythro configuration for 5 [17–
gCOSY correlations of H-7 /H-8 /H -9 .
2
Furthermore, HMBC correlations of
0
0
00
H-1 /C-1 and H -6 /C-7 indicated that
2
00
1
9]. The 8R configuration was supported
the 4-hydroxy-3-methoxyphenoxy and 4 -
00
hydroxy-3 -methoxybenzoyloxy units
by the negative Cotton effect at 230 nm
in the CD spectrum [19], and the D-
configuration of the b-glucopyranosyl
moiety in 5 was confirmed by acid
hydrolysis using the method as described
earlier. Thus, the structure of compound 5
was assigned as (2)-(7S,8R)-(4-hydroxy-
0
0
were connected at C-1 and C-6 of the b-
glucopyranosyl moiety, respectively,
0
00
00
whereas a linkage between C-8 and C-4
was established by an HMBC correlation
of H-8 /C-4 . The Dd 000
0
00
00
000 value
C8 – C7
00 000
0
(11.8 ppm) suggested an 7 ,8 -erythro
configuration for 6 [17–19]. In the CD
spectrum of 6, a negative Cotton effect at
228 nm indicated the 8R configuration [19].
Acid hydrolysis of 6 produced glucose
identified by TLC comparison with an
authentic sugar sample. The glucose iso-
lated from the hydrolysate gave a positive
3
-methoxyphenylglycerol 9-O-b-D-[6-O-
E)-4-hydroxy-3,5-dimethoxyphenylpro-
(
penoyl]-glucopyranoside.
Compound 6 was obtained as a white
amorphous powder. The molecular formula
C H O was indicated by HRESIMS at
3
1 36 15
m/z 671.1950 and the NMR spectral data.
The NMR spectra of 6 (Table 1) showed the
presence of three meta-ortho-trisubstituted
phenyl groups and a b-glucopyranosyl
2
0
optical rotation, ½aꢀ þ 42.3 (c 0.08,
D
H O), indicating that it was the b-D-
2
glucose [7]. Therefore, compound 6 was

4
98
F. Wang et al.
determined as (2)-4-hydroxy-3-methoxy-
phenol b-D-{6-O-[4-O-(7S,8R)-(4-
hydroxy-3-methoxyphenylglycerol-8-yl)-
3.2 Plant material
The flower buds of Lonicera japonica
were collected in May 2005 from Shang-
qiu, Henan Province, China. Plant identity
was verified by Mr Lin Ma (Institute of
Materia Medica, Beijing, China). A
voucher specimen (No. ZH02273) was
deposited at the herbarium of the Depart-
ment of Chemistry of Natural Products,
Institute of Materia Medica.
3
-methoxybenzoyl]}-glucopyranoside.
3
.
Experimental
3
.1 General experimental procedures
Optical rotations were measured on P-
000 polarimeter (JASCO, Tokyo, Japan).
UV spectra were measured on a JASCOP-
50 spectrometer (JASCO). CD spectra
2
3.3 Extraction and isolation
6
were recorded on a JASCO J-815 CD
spectrometer (JASCO). IR spectra were
recorded on a Nicolet 5700 FT-IR
microscope instrument (FT-IR microscope
transmission; Thermo Electron Corpor-
ation, Madison, WI, USA). NMR spectra
The air-dried flower buds of L. japonica
(40 kg) were extracted with H O
2
(120 liters, 3 £ 24 h) at room temperature,
then the flower buds’ residue was extracted
by sonication with 95% EtOH (40 liters,
3 £ 1 h). The ethanol extract was evapor-
ated under reduced pressure to yield a dark
brown residue (1536 g), which was sus-
1
were obtained at 500 MHz for H and
1
1
3
25 MHz for C, respectively, on an
Inova 500 spectrometer (Varian Associ-
ates Inc., Palo Alto, CA, USA) in MeOH-
d4 with solvent peaks used as references.
ESIMS and HRESIMS data were
measured using an AccuToFCS JMS-
T100CS spectrometer (Agilent Technol-
ogies Ltd. Santa Clara, CA, USA). Column
chromatography (CC) was performed with
silica gel (200–300 mesh; Qingdao
Marine Chemical Inc. Qingdao, China)
and Sephadex LH-20 (Pharmacia Biotech
AB, Uppsala, Sweden). HPLC separation
was performed on an instrument consisting
of a Waters 600 controller, a Waters 600
pump, and a Waters 2487 dual l
absorbance detector (Waters Corporation,
Milford, PA, USA), with an Prevail
pended in H O (5 liters) and partitioned
2
with EtOAc (6 £ 5 liters). After removing
the solvent, the EtOAc fraction (815 g) was
chromatographed over silica gel, eluting
with increasing amounts of acetone (0–
100%) in petroleum ether, to afford 10
fractions (A–J) based on TLC analysis.
Fraction I (90 kg) was separated by CC over
MCI gel CHP 20P, successively eluted by
H O (10 liters), 50% EtOH (20 liters), 90%
2
EtOH (10 liters), and Me CO (8 liters), to
2
give corresponding fractions I1 2 I4. Frac-
tion I1 (19 g) was separated via reverse
phase medium pressure liquid chromatog-
raphy (RP-MPLC) eluting with a gradient
of MeOH (5–100%) in H O to give
2
fractions I1-1 to I1-6. Fraction I1-4
(7.52 g) was chromatographed over Sepha-
dex LH-20 eluting with chloroform/metha-
nol (1:1) to yield fractions I1-4-1 to I1-4-4.
Separation of fraction I1-4-3 (3.6 g) by a
normal-phase silica gel column (CHCl3/-
MeOH, 40:1) yielded fractions I1-4-3-1 to
I1-4-3-3. Fraction I1-4-3-2 (286 mg) was
separated by RP flash CC (20–80% MeOH
(
250 £ 10 mm i.d.) column packed with
C
(5 mm) (Alltech Associates Inc.,
1
8
Deerfield, IL, USA). TLC was carried
out in glass precoated silica gel GF254
plates (Qingdao Marine Chemical Inc.).
Spots were visualized under UV light or by
spraying with 7% H SO in 95% EtOH
2
4
followed by heating. Unless otherwise
noted, all chemicals were obtained from
commercially available sources, and were
used without further purification.
in H O) to afford fractions I1-4-3-2-1 to I1-
2
4-3-2-3, and purification of fraction I1-4-3-
2-1 (63 mg) by RP-HPLC (17% MeCN in

Journal of Asian Natural Products Research
499
H O) gave 2 (4.2 mg). Fraction I1-4-3-2-2 tral data (see Table 1); ESIMS: m/z 487
2
þ
2
[M þ Na] , 463 [M 2 H] . (þ)-HRE-
(
(
103 mg) was isolated by preparative TLC
mobile phase: CHCl /MeOH, 6:1), fol-
þ
SIMS: m/z 487.1212 [M þ Na] (calcd for
3
lowed by RP HPLC separation (24%
MeCN in H O) to yield 3 (5.7 mg) and 1
C H O Na, 487.1211).
2
2
24 11
2
(
3.6 mg). Fraction I1-5 (3.76 g) was chro-
3.3.3. (2)-(E)-3,5-
Dimethoxyphenylpropenoic acid 4-O-b-D-
matographed over Sephadex LH-20 eluting
with methanol to yield fraction I1-5-1 to I1-
(
6-O-benzoyl)-glucopyranoside (3)
20
5
by RP flash CC (20–90% MeOH in H O) to
-4, of which I1-5-2 (1.63 g) was isolated
White amorphous powder; ½aꢀ 210.1 (c
D
2
0
2
.37, MeOH); UV (MeOH) l (log 1)
max
02 (3.95), 227 (4.17), 288 (3.87) nm; IR
afford fractions I1-5-2-1 to I1-5-2-5.
Fraction I1-5-2-3 (414 mg) was separated
by silica gel CC (CHCl /MeOH, 20:1) to
nmax 3394, 3188, 3010, 2921, 2850, 1719,
3
1
1
6
646, 1587, 1503, 1468, 1420, 1393, 1333,
279, 1246, 1124, 1070, 1026, 820, 719,
give fractions I1-5-2-3-1 to I1-5-2-3-7, of
which fraction I1-5-2-3-6 (158 mg) was
isolated by preparative TLC (mobile phase:
21 1
34 cm ; H NMR (DMSO-d , 500 MHz)
6
13
and
spectral data (see Table 1); ESIMS: m/z
C NMR (DMSO-d , 125 MHz)
6
CHCl / MeOH, 4.5:1), followed by RP-
3
HPLC separation (15% MeCN in H O) to
2
þ
2
5
13 [M þ Na] , 489 [M 2 H] . (þ)-
yield 4 (3.7 mg), 5 (6.7 mg), and 6 (8.2 mg).
þ
HRESIMS: m/z 513.1374 [M þ Na]
(calcd for C H O Na, 513.1367).
24 26 11
3
.3.1. (2)-2-Hydroxy-5-methoxybenzoic
acid 2-O-b-D-(6-O-benzoyl)-
glucopyranoside (1)
3
.3.4. (2)-(7S,8R)-(4-
Hydroxyphenylglycerol 9-O-b-D-[6-O-
(E)-4-hydroxy-3,5-dimethoxyphenyl
propenoyl]-glucopyranoside (4)
2
D
0
White amorphous solid; ½aꢀ 251.5 (c
0.18, MeOH); UV (MeOH) l (log 1)
206 (4.21), 232 (3.93), 284 (3.29) nm; IR
max
20
D
Yellowish amorphous powder; ½aꢀ
nmax 3307, 2919, 1708, 1603, 1516, 1452,
210.5 (c 0.20, MeCN); UV (MeOH)
lmax (log 1) 204 (4.43), 225 (4.39), 329
1
1
7
415, 1346, 1315, 1281, 1224, 1185, 1116,
070, 1027, 972, 914, 883, 822, 762,
(4.27) nm; CD (MeOH) D1227 nm 23.97,
2
10 cm ; H NMR (CD OD, 600 MHz)
1 1
3
D1252 nm 21.16, D1275 nm 21.64, D1306 nm
20.38; IR nmax 3318, 2934, 1703, 1632,
1
3
and C NMR (CD OD, 150 MHz) spec-
3
tral data (see Table 1); ESIMS: m/z 457
þ
1
1
8
612, 1599, 1515, 1458, 1427, 1386, 1340,
258, 1230, 1173, 1115, 1049, 1026, 1005,
2
[
M þ Na] , 433 [M 2 H] ; (þ)-HRE-
þ
2
1
1
SIMS: m/z 457.1106 [M þ Na] (calcd for
35, 765 cm
;
H NMR (CD OD,
3
1
3
C H O Na, 457.1105).
2
1
22 10
600 MHz) and
150 MHz) spectral data (see Table 1);
C NMR (CD OD,
3
þ
ESIMS: m/z 575 [M þ Na] , 551
3
.3.2. (2)-4-Hydroxy-3,5-
2
2
[
M 2 H] , 587 [M þ Cl] ; HRESIMS:
dimethoxybenzoic acid 4-O-b-D-(6-O-
benzoyl)-glucopyranoside (2)
þ
m/z 575.1745 [M þ Na] (calcd for
C H O Na, 575.1735).
26 32 13
20
White amorphous powder; ½aꢀ 231.8 (c
D
0
2
.09, MeOH); UV (MeOH) l
(log 1)
07 (4.37), 230 (sh, 3.94), 255 (3.67) nm;
max
3
.3.5. (2)-(7S,8R)-(4-Hydroxy-3-
methoxyphenylglycerol 9-O-b-D-[6-O-
E)-4-hydroxy-3,5-dimethoxy
IR nmax 3394, 2938, 1717, 1566, 1502,
(
1
1
7
454, 1394, 1319, 1280, 1224, 1189, 1125,
071, 1026, 968, 920, 877, 790, 740,
phenylpropenoyl]-glucopyranoside (5)
2
1
17 cm ; H NMR (CD OD, 500 MHz)
1
20
D
Yellowish amorphous powder; ½aꢀ
3
1
3
and C NMR (CD OD, 125 MHz) spec-
3
225.1 (c 0.12, MeOH); UV (MeOH)

5
00
F. Wang et al.
lmax (log 1) 205 (4.80), 225 (4.87), 329
4.79) nm; CD (MeOH) D1218 nm 21.35,
D1230 nm 20.48, D1277 nm 20.61, D1329 nm
0.10, D1324 nm 20.26; IR nmax 3405,
TLC identification of glucose (retention
factor, 0.31).
(
2
2
1
1
5
1
940, 1693, 1631, 1611, 1515, 1458, 1427,
340, 1286, 1257, 1227, 1176, 1157, 1114,
Acknowledgments
The authors acknowledge the financial support
from the National Natural Sciences Foundation
of China (NNSFC; grant nos. 20772156,
30825044, and 20932007), the Program for
Changjiang Scholars and Innovative Research
Team in University (PCSIRT, grant no.
IRT1007), and the National Science and
Technology Project of China (no.
2012ZX09301002-002).
2
044, 835, 623 cm ; H NMR (CD OD,
1 1
3
1
3
00 MHz) and C NMR (CD OD,
3
25 MHz) spectral data (see Table 1);
þ
ESIMS: m/z 605 [M þ Na] , 581
2
2
[
M 2 H] , 617 [M þ Cl] ; (þ)-HRE-
þ
SIMS: m/z 605.1854 [M þ Na] (calcd
for C H O Na, 605.1841).
7 34 14
2
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.3.6. (2)-4-Hydroxy-3-methoxyphenol
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3
20
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D
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2
CHCl –MeOH–H O (8:5:1) was used for
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2

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