Study on chemical constituents of Cyclocarya paliurus

Article abstract of DOI:10.1080/10286020.2013.825254

A new dammarane triterpenoid glycoside named cyclocarioside J (1) and other three known triterpenoid glycosides were isolated from the leaves of Cyclocarya paliurus. Based on ESI-MS, HR-ESI-MS, ¹H NMR, ¹³C NMR, and 2D NMR techniques

Full text of DOI:10.1080/10286020.2013.825254

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Study on chemical constituents of
Cyclocarya paliurus
Yue Liua, Man-Qi Zhangb, Xia-Lei Lia, Tun-Hai Xuc, Sheng-Xu Xiea,
Ya-Juan Xua & Dong-Ming Xua
a Department of Phytochemistry, Jilin Academy of Chinese
Medicine Sciences, Changchun, 130021, China
b School of Pharmacy, China Pharmaceutical University, Nanjing,
211198, China
c Department of Traditional Chinese Medicine Chemistry, School
of Traditional Chinese Medicine, Beijing University of Chinese
Medicine, Beijing, 100102, China
Published online: 29 Nov 2013.
To cite this article: Yue Liu, Man-Qi Zhang, Xia-Lei Li, Tun-Hai Xu, Sheng-Xu Xie, Ya-Juan Xu &
Dong-Ming Xu (2014) Study on chemical constituents of Cyclocarya paliurus, Journal of Asian
Natural Products Research, 16:2, 206-209, DOI: 10.1080/10286020.2013.825254
To link to this article: http://dx.doi.org/10.1080/10286020.2013.825254
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Journal of Asian Natural Products Research, 2014
Vol. 16, No. 2, 206–209, http://dx.doi.org/10.1080/10286020.2013.825254
Study on chemical constituents of Cyclocarya paliurus
Yue Liua, Man-Qi Zhangb, Xia-Lei Lia, Tun-Hai Xuc*, Sheng-Xu Xiea,
Ya-Juan Xua* and Dong-Ming Xua
aDepartment of Phytochemistry, Jilin Academy of Chinese Medicine Sciences, Changchun 130021,
China; bSchool of Pharmacy, China Pharmaceutical University, Nanjing 211198, China;
cDepartment of Traditional Chinese Medicine Chemistry, School of Traditional Chinese Medicine,
Beijing University of Chinese Medicine, Beijing 100102, China
(Received 14 December 2012; final version received 11 July 2013)
A new dammarane triterpenoid glycoside named cyclocarioside J (1) and other three
known triterpenoid glycosides were isolated from the leaves of Cyclocarya paliurus.
Based on ESI-MS, HR-ESI-MS, 1H NMR, 13C NMR, and 2D NMR techniques including
1H–1H COSY, HMBC, HMQC, and NOESY correlations, the structure of cyclocarioside J
was elucidated as (20S,24R)-epoxydammarane 3b,12b,25-trihydroxy-12-O-b-D-quinovo-
pyranosyl-3-O-a-L-arabinopyranoside.
Keywords: Cyclocarya paliurus; dammarane; triterpenoid glycosides; cyclocarioside J
1. Introduction
777.4753 [M þ Na]þ. ESI-MS showed
the major ion peak at m/z 777 which was
assigned to [M þ Na]þ. MS/MS of
1 showed fragment ion peaks at m/z 613
[M þ Na-164]þ, corresponding to the loss
of a deoxyhexose unit, and at m/z 481
[M þ Na-164-132]þ, corresponding to the
loss of a pentose unit. The 13C NMR
spectrum of 1 exhibited 41 carbon signals,
of which two anomeric C-atoms at dC 102.0
(C-10) and dC 102.0 (C-100) supported the
presence of two sugar moieties in 1. By
comparing the 13C NMR data of 1 with
those of cyclocarioside C [7] and cyclocar-
ioside I [8], it was suggested that 1 has a
similar aglycone as those of cyclocarioside
C and cyclocarioside I. The almost identical
13C NMR data ascribable to C-3 (dC 81.5),
C-12 (dC 77.4), C-20 (dC 86.6), and C-24
(dC 84.3) in 1 with those of cyclocarioside
C and cyclocarioside I proposed that the
stereochemistry of C-12, C-20, and C-24 in
compound 1 should be R-, S-, and
R-configurations, respectively. In addition,
Cyclocarya paliurus (Batal.) Iljinsk
(Juglandaceae) is an endemic species
growing in southern China [1]. Its leaves
possess an intensive sweet taste and are
used as a tea which is reported to have
beneficial effects of lowering blood sugar
[2,3]. Previously, several sweet dammarane
triterpenoid glycosides had been reported
from C. paliurus [4–6]. In order to obtain a
large amount of triterpenoid glycosides for
pharmacological study and quality control,
the leaves of C. paliurus were investigated
to afford one new dammarane triterpenoid
glycoside and three known triterpenoid
glycosides. This study deals with the
isolation and characterization of cyclocar-
ioside J (1).
2. Results and discussion
Compound 1 was obtained as a white
amorphous solid with a molecular formula
of C41H70O12, as determined by the data of
the positive-ion HR-ESI-MS at m/z
*Corresponding authors. Email: thxu@163.com; xyj6492@sohu.com
q 2013 Taylor & Francis

Journal of Asian Natural Products Research
207
Table 1. 1H and 13C NMR spectral data of 1 (400/100 MHz in pyridine-d5; d in ppm, J in Hz).
Aglycone of 1
dH (J, Hz)
Sugar moieties of 1
No.
dC
No.
dC
d
H (J, Hz)
1
2
3
4
5
6
35.7
26.8
81.5
38.2
51.1
18.4
3.09 (br d, 13.2), 2.00 (m)
3-O-Ara ( p)
2.00 (m), 1.72 (m)
3.60 (m)
–
10
20
30
40
50
102.0
72.5
74.8
69.2
66.3
4.70 (d, 6.4)
4.37 (m)
4.17 (m)
1.56 (m)
1.46 (m)
4.34 (m)
4.28 (dd, 12.0, 3.6),
3.73 (br d, 12.0)
7
8
9
36.5
41.6
54.1
40.1
34.6
77.4
41.3
50.2
31.7
22.0
49.3
17.1
16.9
86.6
24.6
34.2
26.4
84.3
71.3
26.2
27.8
23.3
30.1
16.9
1.51 (m), 1.17 (m)
–
12-O-Qui
1.86 (br d, J ¼ 10.0 Hz)
100
200
300
400
500
600
102.0
75.6
78.5
77.0
72.9
18.7
4.99 (d, 7.6)
3.93 (m)
4.14 (t-like, 8.8)
3.65 (m)
3.79 (m)
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
–
2.85 (br d, 12.0), 1.46 (m)
4.38 (m)
1.78 (m)
–
1.37 (m), 0.96 (m)
1.86 (m), 0.98 (m)
1.84 (m)
1.59 (d, 6.0)
1.06 (s)
1.39 (s)
–
1.14 (s)
1.73 (m), 1.54 (m)
1.93 (m), overlapped
3.92 (dd, 13.2, 7.6)
–
1.42 (s)
1.37 (s)
0.98 (s)
1.23 (s)
0.62 (s)
Note: Assignments based on 1H–1H COSY, HMQC, and HMBC experiments.
carbon signals for D-quinovopyranose and
L-arabinopyranose were observed in the
13C NMR spectrum of 1 (Table 1). Acidic
hydrolysis of 1 with mineral acid afforded
quinovose and arabinose, which were
identified by comparison with authentic
samples.
NOESY spectrum, correlations between
H-3 (dH 3.60) and H-5 (dH 1.56), H-28 (dH
0.98), correlations between H-12 (dH 4.38)
and H-11a (dH 2.85, a-orientation), H-9 (dH
1.86), and correlations between H-21 (dH
1.14) and H-24 (dH 3.92), H-17 (dH 1.84)
indicated the a-orientations of H-3, H-12,
and H-24. All 1H and 13C NMR data
(Table 1) were assigned by 1H–1H COSY,
HMQC, and HMBC experiments. In the
HMBC spectrum of 1 (Figure 1), the
correlations between H-10 (dH 4.70) and
C-3 (dC 81.5) suggested that the arabino-
pyranose was linked at C-3, while the
correlations between H-100 (dH 4.99)
1
In the H NMR spectrum of 1, eight
methyl signals (d 1.42, 1.39, 1.37, 1.23,
1.14, 1.06, 0.98, and 0.62, each s)
assignable to the aglycone moiety were
observed. There were also two anomeric
proton signals at dH 4.99 (d, J ¼ 7.6 Hz)
for the quinovose and at dH 4.70
(d, J ¼ 6.4 Hz) for arabinose units. In the

208
Y. Liu et al.
liquid chromatography (HPLC) (Shi-
madzu Company, Kyoto, Japan) was
carried out using an octadecylsilane-
bonded silica column (HC-ODS,
250 mm £ 4.6 mm, 5 mm). Prep-HPLC
(Waters 600E, Waters Company, Billerica,
MA, USA) was carried out using an
octadecylsilane-bonded silica column
(Agilent-ZORBAX PrepHT Eclipse
XDB-C18, 250 mm £ 21.2 mm, 7 mm).
Column chromatography (CC) was carried
out on silica gel (200–300 mesh, Qingdao
Oceanic Chemical Industry, Qingdao,
China), reversed silica gel (50 mm, YMC
Co., Ltd., Kyoto, Japan), and macroporous
resin D21 made in Shandong Lu Neng Gel
Co. (Shandong, China). TLC was carried
out on aluminium sheets (20 £ 20 cm,
RP-18F254 S, MERCK KGaA, Darmstadt,
Germany). Spots were detected after
spraying 10% H2SO4.
Figure 1. Structure and key HMBC
correlations (H ! C) of compound 1.
and C-12 (dC 77.4) indicated that the
quinovopyranose was present at C-12.
Consequently, the structure of 1 was
determined as (20S,24R)-epoxydammarane
3b,12b,25-trihydroxy-12-O-b-D-quinovo-
pyranosyl-3-O-a-L-arabinopyranoside.
The other compounds were identified
as cyclocarioside I (2) [8], arjunglucoside
II (3) [9], and quadranoside IV (4) [10].
3.2 Plant material
The leaves used in this experiment were
collected from Xiushui County, Jiangxi
Province, China, in September 2009 and
identified by Professor Minglu Deng,
Changchun College of Traditional Chinese
Medicine. A voucher specimen has been
deposited in the phytochemistry laboratory
of Jilin Academy of Chinese Medicine
Sciences.
3. Experimental
3.1 General experimental procedures
Melting points were determined on an X-6
microscope apparatus and are uncorrected
(Beijing Fukai Instrument Co., Ltd.,
Beijing, China). Optical rotations were
determined on a WZZ-2SS auto-polari-
meter (Shanghai Jingke Scientific Instru-
ment Co., Ltd., Shanghai, China). IR
spectrum was measured as film on KBr
pellet using a Bruker VERTEX70 spec-
trometer (Bruker Corporation, Billerica,
MA, USA). NMR spectra were obtained
on a Bruker AV400 instrument (Bruker
Corporation, Rheinstetten, Switzerland),
using tetramethylsilane as internal stan-
dard. The HR-MS was recorded on an
autoflex III MALDI/TOF/TOF-MS instru-
ment (Bruker Corporation, Ettlingen,
Germany). ESI-MS were recorded on a
Finnigan Mat LCQ mass spectrometer
(Thermo Fisher Scientific Inc., West Palm
Beach, FL, USA). High-performance
3.3 Extraction and isolation
The dried leaves (10 kg) were extracted
three times with 60% EtOH for 2 h under
reflux. The extract was concentrated under
vacuum to give a residue, which was
suspended in H2O and extracted with
CHCl3 and n-BuOH, respectively. The
n-BuOH fraction (205 g) was chromato-
graphed on 1.5 kg D21 porous resin, eluting
with water until the eluate was colorless
and then with 70% EtOH (12 liters). The
70% EtOH eluate was further subjected to
neutral resin to remove most of the color
material and then evaporated to dryness to

Journal of Asian Natural Products Research
209
give crude glycosides (74 g). Part of crude
glycosides were chromatographed on
silica gel (200–300 mesh) with CHCl3:
MeOH (100:1 to 1:1) to give Fr. 1–Fr. 14.
Fr. 11 (6.7 g) was subjected to reversed-
phase silica gel CC [CHCl3:MeOH:H2O:
n-BuOH (25:4:2:0.5)] to give 91 fractions.
Fr. 64–Fr. 67 were separated by HPLC
[MeOH:H2O (85:15, v/v)] to give com-
pound 1 (82 mg). Fr. 45– Fr. 55 (41 mg)
were recrystallized to give compound
2 (32 mg). Fr. 9–Fr. 11 were separated
by HPLC [MeOH:H2O (82:18, v/v)] to
give compounds 3 (66 mg) and 4 (34 mg).
corresponding to those of L-arabinose and
with Rf [CHCl3:MeOH:H2O (6:4:1)
and (EtOAc:CHCl3:MeOH:H2O, 3:2:2:1)]
20
and specific rotation (½aꢀD þ 69.2
(c ¼ 0.13, H2O) corresponding to those
of D-quinovose, respectively.
Acknowledgments
The authors were financially supported by the
International Science – Technology Cooperation
Program of China (2010DFB33260) and the
Cooperation Program of Beijing Municipal
Education Commission and the Innovative
team of Beijing TCM University (No. 2011-
CXTD-19).
3.3.1 Compound 1
References
Amorphous powder; m.p. 148.0–148.78C;
20
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Academiae Sinicae Edita, Flora Repub-
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Beijing, 1979), Vol. 21, p. 18.
[2] M.Y. Xie, L. Li, S.P. Nie, X.R. Wang, and
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202 (2006).
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Y. Toyoda, H. Shibata, Y. Matsui, S.
Asami, and T. Tanaka, Biosci. Biotechnol.
Biochem. 67, 877 (2003).
½aꢀD : 21.29 (c ¼ 0.98, MeOH); IR (KBr)
nmax (cm21): 3428, 2969, 1055. H NMR
1
(400 MHz, pyridine-d5) and 13C NMR
(100 MHz, pyridine-d5) spectral data are
given in Table 1. ESI-MS: m/z 777
[M þ Na]þ, 613 [M þ Na-164]þ, 481
[M þ Na-164-132]þ. HR-ESI-MS: m/z
777.4753 [M þ Na]þ (calcd for
C41H70O12Na, 777.4759).
[4] R.G. Shu, C.R. Xu, L.N. Li, and Z.L. Yu,
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3.4 Acid hydrolysis
Compound 1 (15 mg) in a mixture of
MeOH (5 ml) and 5% H2SO4 (5.0 ml) was
refluxed for 2 h. After cooling, the reaction
mixture was diluted twofold with H2O,
and extracted with CHCl3 (3 £ 30 ml). The
aqueous layer was neutralized with aq.
Ba(OH)2 and concentrated in vacuo to give
a residue, then chromatographed on silica
gel with CHCl3:MeOH:H2O (70:30:5) to
give two monosaccharides with Rf
[CHCl3:MeOH:H2O (6:4:1) and (EtOAc:
CHCl3:MeOH:H2O, 3:2:2:1)] and specific
20
rotation (½aꢀD þ 110.9 (c ¼ 0.11, H2O)