Triterpenoid saponins from the stem bark of Catunaregam spinosa

Article abstract of DOI:10.1139/v11-090

Four new triterpenoid saponins, Catunaroside E (1; 3-O-{β-D- glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)] -β-D-glucopyranosyl}-siaresinolic acid), Catunaroside F (2; 3-O-{α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)] -β-D-glucopyranosyl}-28-O-(β-D-glucopyranosyl)-oleanolic acid), Catunaroside G (3; 3-O-[α-L-rhamnopyranosyl-(1→2)-β-D- glucopyranosyl]-28-O-(β-D-glucopyranosyl)-siaresinolic acid), and Catunaroside H (4; 3-O-{β-D-glucopyranosyl-(1→2)-[β-D- glucopyranosyl-(1→3)]-β-D-glucopyranosyl}-28-O-(β-D- glucopyranosyl)-siaresinolic acid), and the known triterpenoid saponin Mussaendoside J (5), were isolated from the stem bark of Catunaregam spinosa. Their structures were elucidated on the basis of their spectral data and chemical evidence.

Full text of DOI:10.1139/v11-090

1
277
Triterpenoid saponins from the stem bark of
Catunaregam spinosa
Guang-Chun Gao, Zhong-Xian Lu, Shu-Hong Tao, Si Zhang, Fa-Zuo Wang, and
Qing-Xin Li
Abstract: Four new triterpenoid saponins, Catunaroside E (1; 3-O-{b-D-glucopyranosyl-(1→2)-[b-D-glucopyranosyl-(1→3)]-
b-D-glucopyranosyl}-siaresinolic acid), Catunaroside F (2; 3-O-{a-L-rhamnopyranosyl-(1→2)-[b-D-glucopyranosyl-(1→3)]-b-
D-glucopyranosyl}-28-O-(b-D-glucopyranosyl)-oleanolic acid), Catunaroside G (3; 3-O-[a-L-rhamnopyranosyl-(1→2)-b-D-
glucopyranosyl]- 28-O-(b-D-glucopyranosyl)-siaresinolic acid), and Catunaroside H (4; 3-O-{b-D-glucopyranosyl-(1→2)-[b-
D-glucopyranosyl-(1→3)]-b-D-glucopyranosyl}-28-O-(b-D-glucopyranosyl)-siaresinolic acid), and the known triterpenoid sap-
onin Mussaendoside J (5), were isolated from the stem bark of Catunaregam spinosa. Their structures were elucidated on
the basis of their spectral data and chemical evidence.
Key words: Catunaregam spinosa, Rubiaceae, triterpenoid saponins, Catunaroside E–H.
Résumé : À partir de l’écorce du tronc de Catunaregam spinosa, on a isolé quatre nouvelles saponines triterpénoïdes, le ca-
tunaroside E (1), l’acide 3-O-{b-D-glucopyranosyl-(1→2)-[b-D-glucopyranosyl-(1→3)]-b-D-glucopyranosyl}siarésinolique, le
catunaroside F (2), l’acide 3-O-{a-L-rhamnopyranosyl-(1→2)-[b-D-glucopyranosyl-(1→3)]-b-D-glucopyranosyl}-28-O-(b-D-
glucopyranosyl)oléanolique, le catunaroside G (3), l’acide 3-O-{a-L-rhamnopyranosyl-(1→2)-[b-D-glucopyranosyl-(1→3)]-
2
8-O-(b-D-glucopyranosyl)siarésinolique et le catunaroside H (4), l’acide 3-O-{b-D-glucopyranosyl-(1→2)-[b-D-glucopyrano-
syl-(1→3)]-28-O-(b-D-glucopyranosyl)siarésinolique et une saponine triterpénoïde connue,le mussaendoside J (5). Leurs
structures ont été élucidées sur la base de leurs propriétés chimiques et de leurs données spectrales.
Mots‐clés : Catunaregam spinosa, rubiacée, saponines triterpénoïdes, caturanoside E-H.
[
Traduit par la Rédaction]
der. Its molecular formula was assigned as C H O based
Introduction
48 78 19
on the high-resolution electrospray ionization mass spectrom-
Catunaregam spinosa T. (Rubiaceae), which is mostly dis-
tributed in tropical and semitropical areas, was used in the
traditional medicinal systems of India and Brazil for its anti-
spasmodic, antidysenteric, anti-inflammatory, immunomodu-
latory, and antifertility properties.
of C. spinosa led to the isolation of triterpene and saponins,
iridoid glucosides, coumarin glucosides, and norneolignans.
In continuation of our studies on the chemical diversity of
mangrove plants on Hainan Island, this plant was investigated
and four new triterpenoid saponins, Catunaroside E–H, to-
gether with Mussaendoside J, were obtained from the n-BuOH
extract of its stem bark (Fig. 1). This paper deals with the iso-
lation and structure elucidation of those triterpenoid saponins.
Mussaendoside J was first obtained from this plant.
+
etry (HR-ESI-MS; m/z: 981.5080 [M + Na] ). The infrared
(
1
IR) spectrum exhibited absorption bands at 3418 (OH),
728 (C=O), and 1644 (C=C) cm . The seven tertiary
–1
methyl groups (d 1.26, 1.08, 0.81, 1.06, 1.66, 1.18, and
.02 ppm) and one olefinic proton (d 5.55, br s) were ob-
1
–4
Chemical investigation
1
4–6
1
served in the H nuclear magnetic resonance (NMR) spec-
1
7
8
13
trum. The C and distortionless enhanced polarization
transfer (DEPT) NMR data of the aglycon part of 1 con-
firmed the presence of seven methyl carbons (d 28.0, 16.7,
1
5.4, 17.5, 24.9, 28.9, and 24.9 ppm), two olefinic carbons
(
d 123.1 and 144.9 ppm), and two oxygenated methine car-
bons (d 89.5 and 81.3 ppm) (Table 2). The comparison of
1H and 13C NMR spectrum data of the aglycon part of 1
with that of Latifolioside H indicated that compound 1 pos-
sessed a 3b,19a-dihydroxyolean-12-ene-28-oic acidic agly-
Results and discussion
9
con. The chemical shifts of C-3 (d 89.5) and C-28 (d 180.9)
Compound 1 was obtained as a colorless amorphous pow-
revealed that 1 was a monodesmosidic glycoside. Eighteen of
Received 16 February 2011. Accepted 10 May 2011. Published at www.nrcresearchpress.com/cjc on 22 September 2011.
G.-C. Gao and Z.-X. Lu. Institute of Plant Protection and Microbiology, Zhejiang Academy of Agriculture Science, 198 Shiqiao Road,
Hangzhou 310021, P. R. China.
S.-H. Tao. Guangdong Pharmaceutical University, University City, Guangzhou 510006, P. R. China.
S. Zhang, F.-Z. Wang, and Q.-X. Li. Key Laboratory of Marine Bio-resources Sustainable Utilization, RNAM Center for Marine
Microbiology, Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of
Sciences, 164 West Xingang Road, Guangzhou 510301, P. R. China.
Corresponding author: Si Zhang (e-mail: zhsimd@scsio.ac.cn).
Can. J. Chem. 89: 1277–1282 (2011)
doi:10.1139/V11-090
Published by NRC Research Press

1278
Can. J. Chem. Vol. 89, 2011
Table 1. H NMR data for the sugar moieties of compounds 1–5 in pyridine-d5 (J in parentheses in Hz).
1
H
1
2
3
4
5
3
-O
Glc
Glc
Glc
Glc
Glc
1
2
3
4
5
6
3
1
2
3
4
5
6
2
1
2
3
4
5
6
2
1
2
3
4
5
6
′
4.83 d (7.5)
4.38
4.25
4.01
3.89
4.83 d (7.5)
4.02
4.20
4.09
4.25
4.94 d (7.0)
4.29
4.28
4.17
4.03
4.83 d (7.5)
4.36
4.23
4.01
3.87
4.94 d (6.0)
4.29
4.28
4.16
4.03
′
′
′
′
′
4.53, 4.24
4.54, 4.39
4.53, 4.37
4.52, 4.24
4.54, 4.36
′-O
′′
5.37 d (7.6)
4.08
4.21
4.01
4.02
5.11 d (7.8)
4.00
4.08
4.18
4.10
5.36 d (8.0)
4.02
4.20
4.12
4.17
′′
′′
′′
′′
′′
4.47
4.24
4.45
′-O
′′′
′′′
′′′
′′′
′′′
′′′
8-O
′′′′
′′′′
′′′′
′′′′
′′′′
′′′′
5.72 d (7.6)
4.03
4.27
4.13
3.84
6.47 br s
4.81
4.60
4.29
4.74
6.56 br s
4.86
4.68
4.34
4.80
5.71 d (7.5)
4.02
4.27
4.14
3.82
6.56 br s
4.86
4.69
4.34
4.80
4.31
1.67 d (6.0)
1.71 d (6.0)
4.31
1.71 d (6.0)
6.31 d (8.1)
4.20
4.28
4.35
3.95
6.38 d (8.0)
4.21
3.98
4.37
4.10
6.36 d (8.0)
4.20
4.01
4.35
4.06
6.34 d (8.0)
4.21
4.03
4.36
3.94
4.47, 4.43
4.48, 4.43
4.45, 4.39
4.47, 4.41
Note: Glc, glucose.
4
8 carbons were assigned to the oligosaccharide moieties.
mula of C H O by its HR-ESI-MS spectrum (m/z:
54 88 22
1
13
+
1
The H and C NMR spectra of 1 showed three sugar
anomeric protons at d 4.83 (d, J = 7.5 Hz), 5.37 (d, J =
1111.5702 [M + Na] ). In the H NMR spectrum, seven
methyl protons (d 1.23, 1.15, 0.83, 1.07, 1.23, 0.89, and
0.87 ppm), one olefinic proton (d 5.41, br s), and four sugar
anomeric protons (d 4.83, d, J = 7.5 Hz; 5.11, d, J = 7.8 Hz;
6.47, br s; and 6.31, d, J = 8.1 Hz) (Table 1) were observed.
The signals of seven methyl carbons at d 28.1, 17.1, 15.7,
17.5, 26.1, 33.1, and 23.7 ppm, two olefinic carbons at d
122.9 and 144.1 ppm, four sugar carbons at d 104.9, 103.9,
101.7, and 95.8 ppm, and two carbons at d 176.4 and
88.7 ppm linked to the glycan part were also shown in the
1
7
1
.6 Hz), and 5.72 (d, J = 7.6 Hz) and carbons at d 105.1,
04.7, and 103.8 (Tables 1 and 2). The monosaccharides
were identified as glucose by comparing the retention factor
Rf) value with that of D-glucose using thin-layer chromatog-
(
raphy (TLC) after acid hydrolysis and a combination of
DEPT, heteronuclear single quantum coherence (HSQC), and
heteronuclear multiple bond correlation (HMBC) experi-
ments, respectively. All glucoses of 1 were in pyranose
forms, as determined by their 1D and 2D NMR experiments.
The b-anomeric configurations of the glucose units were es-
tablished by its J
3C NMR spectrum. The above evidence revealed that 2
was a bisdesmosidic glycoside. The aglycon part of this
3
coupling constants (7.5–7.6 Hz). The
H1,H2
compound was determined to be oleanolic acid by compar-
1
3
signal at d 88.6 in the C NMR indicated a substitution at C-
′ of one of the glucoses. The sequence of the glycan part
1
13
ing the H, C, and DEPT NMR signals of 2 with those of
3
3-O-[b-D-glucopyranosyl-(1→3)-b-D-galactopyranosyl]-olean-12-
connected to C-3 of the aglycon was deduced from the fol-
lowing HMBC correlations: H-1′ (d 4.83) of inner glucose
with C-3 (d 89.5) of sapogenin, H-1′′ (d 5.37) of one terminal
glucose with C-3′ (d 88.6), and H-1′′′ (d 5.72) of another ter-
minal glucose with C-2′ (d 79.3) (Fig. 2). The NMR data of
the glycan part of 1 was in accordance with that of 3-O-
en-3b-ol-28-oic acid.5 Acid hydrolysis of 2 suggested that the
monosaccharides of this compound were L-rhamnose and D-
glucose. In combination with the characteristic proton signal
1
at d 1.67 (3H, d, J = 6.0 Hz) in the H NMR spectrum, 2
could be deduced to contain one rhamnose and three glucose.
By comparing with the NMR spectra of Mussaendoside J (5),
the presence of glucose linked to C-3′ (d 89.6) in 2 was dis-
closed. The sequence of the oligosaccharide chain was con-
[
2′,3′-di-O-(b-D-glucopyranosyl)-b-D-glucopyranosyl] olea-
10
nolic acid. On the basis of the above results, compound 1
was elucidated as 3-O-{b-D-glucopyranosyl-(1→2)-[b-D-
glucopyranosyl-(1→3)]-b-D-glucopyranosyl}-siaresinolic acid,
named as Catunaroside E.
1
1
firmed by HSQC, HMBC, and H- H double quantum filtered
correlation spectroscopy (DQFCOSY) experiments. The key
HMBC correlations were observed: H-1′′′′ (d 6.31) of a glu-
cose and C-28 (d 176.4) of the aglycon part, H-1′ (d 4.83) of
Compound 2 was determined to have the molecular for-
Published by NRC Research Press

Gao et al.
Table 2. 13C NMR data of compounds 1–5 in pyridine-d5.
1279
C
1
2
3
4
5
6
7
8
9
1
2
3
4
5
C
1
Glc
2
Glc
3
Glc
4
Glc
5
Glc
38.5
26.6
89.5
39.6
55.9
18.6
33.3
40.0
48.2
37.1
24.1
123.1
144.9
42.1
29.2
28.4
46.1
44.8
81.3
35.7
29.2
33.7
28.0
16.7
15.4
17.5
24.9
180.9
28.9
24.9
39.1
26.7
88.7
39.5
56.1
18.5
33.1
39.9
48.0
37.0
23.4
122.9
144.1
42.2
28.3
23.8
47.0
41.8
46.3
30.8
34.0
32.6
28.1
17.1
15.7
17.5
26.1
176.4
33.1
23.7
38.9
26.8
88.9
39.5
56.2
18.7
33.0
40.2
48.2
37.1
24.1
123.1
144.3
42.1
29.0
28.0
46.5
44.6
81.1
35.5
29.0
33.2
28.1
17.1
15.5
17.6
24.9
177.2
28.7
24.7
38.6
26.6
89.5
39.6
56.0
18.8
33.1
40.2
48.3
37.1
24.1
39.0
26.8
88.8
39.5
56.1
18.5
33.1
39.9
48.0
37.0
23.4
3-O
1′
105.1
79.3
88.6
70.1
77.7
63.4
Glc
104.7
76.4
78.6
71.6
78.6
62.6
Glc
103.8
75.4
78.6
72.6
77.8
62.4
104.9
79.3
89.6
69.8
77.0
62.7
Glc
103.9
75.2
78.7
71.4
78.4
62.3
Rha
101.7
72.4
72.5
73.9
69.8
18.6
Glc
105.4
78.1
79.8
72.2
79.3
62.9
105.1
79.3
88.7
70.1
77.7
63.5
Glc
104.7
76.4
78.6
71.6
78.6
62.6
Glc
103.9
75.4
78.6
72.7
77.8
62.2
Glc
95.9
74.2
79.0
71.2
77.8
62.4
105.4
78.1
79.9
72.2
79.3
62.9
2′
3′
4′
5′
6′
3′-O
1′′
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
0
2′′
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
3′′
123.2
144.4
42.2
29.1
28.1
46.5
44.6
81.2
35.6
29.0
33.3
28.8
16.7
15.5
17.6
25.0
177.3
28.1
24.7
122.9
144.1
42.2
28.3
23.8
47.0
41.8
46.2
30.8
34.0
32.5
28.1
17.1
15.6
17.5
26.1
176.4
33.1
23.6
4′′
5′′
6′′
2′-O
1′′′
2′′′
3′′′
4′′′
5′′′
6′′′
28-O
1′′′′
2′′′′
3′′′′
4′′′′
5′′′′
6′′′′
Rha
101.7
72.4
72.5
74.2
69.6
18.7
Glc
Rha
101.7
72.4
72.5
74.2
69.6
18.7
Glc
95.8
74.2
78.9
71.1
77.8
62.2
95.8
74.1
78.9
71.1
77.8
62.3
95.9
74.2
78.9
71.1
77.8
62.2
Note: Glc, glucose; Rha, rhamnose.
the inner glucose and C-3 (d 88.7) of the aglycon part, H-1′′
d 5.11) of the terminal glucose and C-3′ (d 89.6) of the inner
glucose, and H-1′′′ (d 6.47) of the terminal rhamnose and C-
′ (d 78.7) of the inner glucose (Fig. 2). The b-anomeric con-
figurations of glucose units were determined by the relatively
the aglycon part at d 81.1 (d, C-19) ppm, which was oxygen-
ated, and 35.5 (s, C-20) ppm.12 The aglycon part of 3 was then
established as siaresinolic acid by comparing the NMR spec-
trum data of 3 with that of 1. From the above evidence, com-
pound 3 was identified as 3-O-[a-L-rhamnopyranosyl-(1→2)-b-
D-glucopyranosyl]-28-O-(b-D-glucopyranosyl)-siaresinolic acid,
named as Catunaroside G.
(
2
3
1
large J
coupling constants (7.5–8.1 Hz). The J
H1,H2
C1,H1
coupling constant (168 Hz) for the rhamnosyl unit confirmed
that the anomeric proton was equatorial (a-pyranoid anomeric
form).11 On the basis of this evidence, compound 2, named as
Catunaroside F, was elucidated as 3-O-{a-L-rhamnopyranosyl-
Compound 4, obtained as an amorphous powder, was de-
duced to have the molecular formula of C H O as indi-
54
88 24
+
cated from the HR-ESI-MS data (m/z: 1143.5586 [M + Na] ).
Acid hydrolysis of 4 showed the existence of D-glucose. The
signals at 177.3 and 89.5 ppm in the 13C NMR spectrum
(
1→2)-[b-D-glucopyranosyl-(1→3)]-b-D-glucopyranosyl}-28-
O-(b-D-glucopyranosyl)-oleanolic acid.
1
Compound 3, also an amorphous powder, has the molecu-
lar formula of C H O as indicated by its HR-ESI-MS
suggested that 4 was a bisdesmosidic glycoside. In the H
and 13C NMR spectrum, four anomeric carbon signals (d
4.83, d, J = 7.5 Hz; 5.36, d, J = 8.0 Hz; 5.71, d, J =
7.5 Hz; and 6.36, d, J = 8.0 Hz and d 105.1, 104.7, 103.9,
95.9 ppm) (Tables 1 and 2) were exhibited. Compound 4
had the same NMR signals as Aralia-saponin V except the
signals of C-19 (d 81.2 ppm) and C-20 (d 35.6 ppm) of the
aglycon part, which is just like the difference between com-
pound 3 and Mussaendoside J.13 Similarly, the aglycon moi-
ety of 4 was determined to be siaresinolic acid. Thus,
compound 4 was elucidated as 3-O-{b-D-glucopyranosyl-
(1→2)-[b-D-glucopyranosyl-(1→3)]-b-D-glucopyranosyl}-28-
O-(b-D-glucopyranosyl)-siaresinolic acid, named as Catunaro-
side H.
4
8
78 17
+
1
spectrum (m/z: 965.5129 [M + Na] ). In the H NMR spec-
trum, seven methyl protons (d 1.26, 1.20, 0.88, 1.13, 1.63,
1
.14, and 0.98 ppm), one olefinic proton (d 5.50, br s), and
three sugar anomeric protons (d 4.94, d, J = 7.0 Hz; 6.56, br
s; and 6.38, d, J = 8.0 Hz) (Table 1) were observed. The sig-
nals of seven methyl carbons at d 28.1, 17.1, 15.5, 17.6, 24.9,
2
1
9
8.,7 and 24.7 ppm, two olefinic carbons at d 123.1 and
44.3 ppm, three sugar carbons at d 105.4, 101.7, and
5.9 ppm, and two carbons at d 177.2 and 88.9 ppm linked
1
3
to the glycan part were also shown in the C NMR spec-
trum. The signals in the NMR spectrum of 3 were in agree-
ment with those of Mussaendoside J, except for the signals of
Published by NRC Research Press

1280
Can. J. Chem. Vol. 89, 2011
Fig. 1. Structures of compounds 1–5.
R3
O
OR2
R O
1
R
1
R
2
R
3
OH
O
1'
HO
HO
2'
O
O
3'
O
1'''
H
OH
1''
HO
O
1
OH
OH
HO
HO
OH
OH
OH
O
1'
OH
HO
O
HO
2'
O
3'
O
1
'''
2
O
H
HO
HO
OH
HO
HO
1''''
O
OH
OH
H
3
C
HO
OH
OH
O
1'
HO
HO 3' 2'
OH
O
1'''
3
O
OH
1
''''
HO
HO
O
OH
OH
H C
3
OH
HO
OH
O
OH
1'
HO
HO
2'
O
O
4
O
3'
O
OH
1''
HO
O
HO
HO
1''''
HO
HO
1'''
OH
OH
OH
OH
OH
OH
O
1'
HO
HO 3' 2'
OH
O
1'''
5
O
H
1
''''
HO
HO
O
OH
OH
H C
3
OH
HO
Compound 5 was elucidated as Mussaendoside J by com-
acquired with a Bruker DRX-500 spectrometer (SiMe as the
4
paring its spectra data with the data reported in the literature
internal standard). Silica gel (Qingdao Haiyang Chemical
Co., Ltd., 100–200 mesh and 200–300 mesh) and LiChro-
prep RP-18 (Merck) were used for the silica gel column
chromatography (CC) and medium-pressure liquid chroma-
tography (MPLC). Sephadex LH-20 (Pharmacia) and macro-
porous resin D101 were also used for column chromatography.
TLC was carried on precoated silica gel 60 F254 plates (Qing-
dao Haiyang Chemical Co., Ltd.), and detection was achieved
by 10% H SO –EtOH for saponins. Semipreparative high-
1
13
12
(
IR, MS, and H and C NMR).
Experimental
General
MS were recorded on a MDS SCIEX API 2000 LC/MS/
MS instrument for ESI and a Bruker BioTOF Q spectrometer
for HR-ESI. IR spectra were measured with a Bruker EQUI-
2
4
1
13
NOX55 infrared spectrometer. H and C NMR spectra were
pressure liquid chromatography (HPLC) was performed using
Published by NRC Research Press

Gao et al.
1281
Fig. 2. Key HMBC correlations for the sugar sequence of 1–4 (from H to C).
HO
O
O
OH
O
OH
OH
1''''
OH
O
O
1'
O
O
HO
O
O
HO
2'
O
HO
OH
OH
HO
3'
O
1
'''
HO
2'
1'
O
O
3'
O
HO
HO
1''
HO
OH
O
HO
HO
OH
OH
OH
OH
1'''
O
H C
3
HO
OH
OH
1
2
HO
HO
O
O
O
O
OH
OH
OH
OH
1''''
1''''
O
O
O
O
O
O
HO
HO
HO
1
'
HO
1'
HO
OH
OH
2'
HO
OH
OH
O
2'
3'
O
O
3'
O
1'''
1''
HO
HO
HO
OH
OH
OH
1'''
O
O
OH
OH
H C
3
OH
HO
3
4
an octadecylsilane (ODS) column (YMC-Pack ODS-5-A,
50 mm × 10 mm inside diameter (i.d.), 5 µm; YMC Co.,
Ltd.) on a Waters-600 HPLC system equipped with a
Waters-996 photodiode array detector.
then fraction 12 was purified by preparative HPLC using
2
MeOH–H O (50:50) to yield compound 4 (70 mg).
2
Catunaroside E (1)
–
1
Colorless amorphous powder. IR (KBr, cm ) ymax: 3418,
1
1
728, 1644. H NMR (500 MHz, pyridine-d ) d: 3.25 (1H,
Plant material
5
dd, J = 4.4, 11.5 Hz, H-3), 5.55 (1H, br s, H-12), 3.62 (1H,
br s, H-18), 3.60 (1H, br s, H-19), 1.26 (3H, s, H-23), 1.08
The stem bark of C. spinosa Tirveng was collected in Feb-
ruary 2006 from Sanya, Hainan Province, P. R. China. The
specimen was identified by professor Si Zhang, South China
Sea Institute of Oceanology, Chinese Academy of Sciences,
Guangzhou, P. R. China. A voucher specimen has been de-
posited in the South China Sea Institute of Oceanology, Chi-
nese Academy of Sciences, Guangzhou, P. R. China
(
3H, s, H-24), 0.81 (3H, s, H-25), 1.06 (3H, s, H-26), 1.66
3H, s, H-27), 1.18 (3H, s, H-29), 1.02 (3H, s, H-30), 4.83
1H, d, J = 7.5 Hz, H-1′), 5.37 (1H, d, J = 7.6 Hz, H-1′′),
(
(
13
5
.72 (1H, d, J = 7.6 Hz, H-1′′′). C NMR (125 MHz, pyri-
+
dine-d ) data, see Table 2. ESI-MS m/z: 981 [M + Na] , 997
5
+
+
[
M + K] . HR-ESI-MS m/z: 981.5080 [M + Na] . Calcd. for
(accession number: GKLMMM020).
C H O Na: 981.5001.
48
78 19
Extraction and isolation
The n-BuOH extract of air-dried and powdered plant mate-
Catunaroside F (2)
–1
Colorless amorphous powder. IR (KBr, cm ) ymax: 3426,
rial (9.0 kg) was extracted by the method as previously re-
1
1
739 1640. H NMR (500 MHz, pyridine-d ) d: 3.38 (1H,
5
8
ported. The n-BuOH soluble fraction (170 g) was passed
dd, J = 3.9, 11.6 Hz, H-3), 5.41 (1H, br s, H-12), 3.18 (1H,
dd, J = 3.4, 13.6 Hz, H-18), 1.23 (3H, s, H-23), 1.15 (3H, s,
H-24), 0.83 (3H, s, H-25), 1.07 (3H, s, H-26), 1.23 (3H, s,
H-27), 0.89 (3H, s, H-29), 0.87 (3H, s, H-30), 4.83 (1H, d,
J = 7.5 Hz, H-1′), 5.11 (1H, d, J = 7.8 Hz, H-1′′), 1.67 (3H,
d, J = 6.0 Hz, H-6′′′), 6.47 (1H, br s, H-1′′′), 6.31 (1H, d,
through a macroporous resin (D101) column eluted with
H O and EtOH–H O (3:7, 6:4, and 95:5), whereas the
2
2
EtOH–H O (95:5, 10 g) eluted portion was subjected to silica
2
gel CC eluted with CHCl –MeOH (98:2–50:50) to give frac-
3
tions A–G. Fraction E was separated by preparative HPLC
using MeOH–H O (68:32) to give compound 1 (10 mg).
13
2
J = 8.1 Hz, H-1′′′′). C NMR (125 MHz, pyridine-d ) data,
5
+
+
Fraction F was recrystallized to afford compound 2 (50 mg).
see Table 2. ESI-MS m/z: 1111 [M + Na] , 1127 [M + K] .
+
The EtOH–H O (6:4, 35 g) eluted portion was subjected to
HR-ESI-MS m/z: 1111.5702 [M + Na] . Calcd. for
2
silica gel CC eluted with CHCl –MeOH (90:10–50:50) to
C H O Na: 1111.5691.
54 88 22
3
give fractions a–f. Fraction b was separated by Sephadex
LH-20 (MeOH), then fractions 25–28 and 53–54 were puri-
Catunaroside G (3)
–
1
fied by preparative HPLC eluting with MeOH–H O (55:45)
Colorless amorphous powder. IR (KBr, cm ) ymax: 3419,
2
1
and MeOH–H O (70:30) to obtain compounds 3 (22 mg)
1730, 1644. H NMR (500 MHz, pyridine-d ) d: 3.34 (1H,
2
5
and 5 (12 mg) separately. Fraction f was rechromatographed
to silica gel CC eluted with CHCl –MeOH–H O (80:20:5),
dd, J = 4.0, 11.5 Hz, H-3), 5.50 (1H, br s, H-12), 3.52 (1H,
br s, H-18), 3.57 (1H, br s, H-19), 1.26 (3H, s, H-23), 1.20
3
2
Published by NRC Research Press

1282
Can. J. Chem. Vol. 89, 2011
(
3H, s, H-24), 0.88 (3H, s, H-25), 1.13 (3H, s, H-26), 1.63
3H, s, H-27), 1.14 (3H, s, H-29), 0.98 (3H, s, H-30), 4.94
1H, d, J = 7.0 Hz, H-1′), 1.71 (3H, d, J = 6.0 Hz, H-6′′′),
EtOAc–MeOH–AcOH–H O (13:3:4:3) solvent system, vi-
2
(
sualized with ethanol and 10% H SO spraying and then
2
4
(
6
heating).
13
.56 (1H, br s, H-1′′′), 6.38 (1H, d, J = 8.0 Hz, H-1′′′′).
C
NMR (125 MHz, pyridine-d ) data, see Table 2. ESI-MS m/z:
Acknowledgments
5
+
+
9
65 [M + Na] , 981 [M + K] . HR-ESI-MS m/z: 965.5129
This work was financially supported by the National Basic
Research Program of China (No. 2010CB833801) and the
Scientific Research Special Funds Project of the Ministry of
Finance (P. R. China, KSCX2-YW-Z-1018).
+
[
M + Na] . Calcd. for C H O Na: 965.5117.
48 78 18
Catunaroside H (4)
–
1
Colorless amorphous powder. IR (KBr, cm ) ymax: 3422,
1
1
738, 1645. H NMR (500 MHz, pyridine-d ) d: 3.26 (1H,
5
References
dd, J = 3.5, 11.5 Hz, H-3), 5.50 (1H, br s, H-12), 3.52 (1H,
(
1) Hamerski, L.; Furlan, M.; Silva, D. H. S.; Cavalheiro, A. J.;
Eberlin, M. N.; Tomazela, D. M.; Bolzani, V. D. S.
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br s, H-18), 3.57 (1H, br s, H-19), 1.26 (3H, s, H-23), 1.09
(
(
(
5
1
3H, s, H-24), 0.84 (3H, s, H-25), 1.12 (3H, s, H-26), 1.63
3H, s, H-27), 1.14 (3H, s, H-29), 0.98 (3H, s, H-30), 4.83
1H, d, J = 7.5 Hz, H-1′), 5.36 (1H, d, J = 8.0 Hz, H-1′′),
(
03)00109-2.
(
(
2) Atal, C. K.; Lamba, S. S. Indian J. Pharm. 1960, 22 (5), 120.
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.71 (1H, d, J = 7.5 Hz, H-1′′′), 6.36 (1H, d, J = 8.0 Hz, H-
13
′′′′). C NMR (125 MHz, pyridine-d ) data, see Table 2.
5
+
+
ESI-MS m/z: 1143 [M + Na] , 1159 [M + K] . HR-ESI-MS
2
009, 1 (3), 036.
(4) Dubois, M.-A.; Benze, S.; Wagner, H. Planta Med. 1990, 56
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+
m/z: 1143.5586 [M + Na] . Calcd. for C H O Na:
5
4
88 24
1
143.5598.
(
(
5) Sati, O. P.; Rana, U.; Chaukiyal, D. C.; Madhusudanan, K. P.;
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2006-962802.
Mussaendoside J (5)
–
1
Colorless amorphous powder. IR (KBr, cm ) ymax: 3422,
1
1
736, 1643. H NMR (500 MHz, pyridine-d ) d: 3.35 (1H,
(6) Sotheeswaran, S.; Bokel, M.; Kraus, W. Phytochemistry 1989,
5
dd, J = 4.0, 11.5 Hz, H-3), 5.43 (1H, br s, H-12), 3.19 (1H,
dd, J = 3.8, 10.5 Hz, H-18), 1.25 (3H, s, H-23), 1.19 (3H, s,
H-24), 0.84 (3H, s, H-25), 1.09 (3H, s, H-26), 1.25 (3H, s,
H-27), 0.91 (3H, s, H-29), 0.88 (3H, s, H-30), 4.94 (1H, d,
J = 6.0 Hz, H-1′), 1.71 (3H, d, J = 6.0 Hz, H-6′′′), 6.56
28 (5), 1544. doi:10.1016/S0031-9422(00)97788-4.
(
7) Sati, S. P.; Chaukiyal, D. C.; Sati, O. P.; Yamada, F.; Ono, M.
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Z. H.; Zhang, S. Helv. Chim. Acta 2010, 93 (2), 339. doi:10.
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(
13
(
1H, br s, H-1′′′), 6.34 (1H, d, J = 8.0 Hz, H-1′′′′).
C
(
9) Ouyang, M. A.; Liu, Y. Q.; Wang, H. Q.; Yang, C. R.
Phytochemistry 1998, 49 (8), 2483. doi:10.1016/S0031-9422
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5
+
+
9
49 [M + Na] , 965 [M + K] . HR-ESI-MS m/z: 949.5178
(
98)00164-2.
+
[
M + Na] . Calcd. for C H O Na: 949.5183.
48 78 17
(
10) Lemmich, E.; Cornett, C.; Furu, P.; Jørstian, C. L.; Knudsen,
A. D.; Olsen, C. E.; Salih, A.; Thiilborg, S. T. Phytochemistry
Acid hydrolysis of 1–5
1
995, 39 (1), 63. doi:10.1016/0031-9422(94)00866-R.
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93. doi:10.1039/p29740000293.
Compounds 1–5 (5 mg each) were refluxed in 5 mL of 4
N HCl for 8 h (kept sealed) in a water bath (100 °C). After
cooling, the reaction mixtures were extracted with EtOAc
(
(
2
12) Zhao, W.-M.; Yang, G.-J.; Xu, R.-S.; Qin, G.-W. Nat. Prod.
Lett. 1996, 8 (2), 119. doi:10.1080/10575639608043250.
(13) Song, S. J.; Nakamura, N.; Ma, C. M.; Hottori, M.; Xu, S. X.
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(00)00379-4.
(
5 mL). The aqueous layers were adjusted to pH 6 with
NaHCO . After being concentrated under reduced pressure,
3
each H O layer (monosaccharide portion) was identified by
comparing the R value with that of the authentic samples on
2
f
the TLC plate (eluted with CHCl –MeOH–H O (8:7:1) and
3
2
Published by NRC Research Press