Four new triterpenoidal saponins acylated with one monoterpenic acid from Gleditsia sinensis

Article abstract of DOI:10.1021/np980441k

Four new oleanane-type triterpenoidal glycosides, named gleditsiosides A-D (1-4), were isolated from the anomalous fruits of Gleditsia sinensis. Using modern NMR techniques, including DQF-COSY, HETCOR, HOHAHA, HMBC, and ROESY experiments and MS analysis as well as chemical methods, their structures were determined as 3-O-β-D-xylopyranosyl-(1→2)-α-L- arabinopyranosyl-(1→6)-β-D-glucopyranosyl oleanolic acid 28-O-β-D- xylopyranosyl-(1→3)-β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)- [(6S,2E)-6-hydroxy-2,6-dimethyl-2,7-octadienoyl-(1→6)]-β-D-glucopyranosyl ester (1); 3-O-β-D-xylopyranosyl-(1→2)-α-L-arabinopyranosyl-(1→6)-β-D- glucopyranosyl oleanolic acid 28-O-β-D-xylopyranosyl-(1→3)-β-D- xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-[(2E)-2-hydroxylmethyl-6- hydroxy-6-methyl-2,7-octadienoyl-(1→6)]-β-D-glucopyranosyl ester (2); 3-O- β-D-xylopyranosyl-(1→2)-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranosyl echinocystic acid 28-O-β-D-xylopyranosyl-(1→3)-β-D-xylopyranosyl-(1→4)- [β-D-galactopyranosyl-(1→2)]-αL-rhamnopyranosyl-(1→2)-[(2E)-2- hydroxylmethyl-6-hydroxy-6-methyl-2,7-octadienoyl-(1→6)]-β-D- glucopyranosyl ester (3); and 3-O-β-D-xylopyranosyl-(1→2)-α-L- arabinopyranosyl-(1→6)-β-D-glucopyranosyl echinocystic acid 28-O-β-D- xylopyranosyl-(1→3)-β-D-xylopyranosyl-(1→4)-[β-D-galactopyranosyl- (1→2)]-α-rhamnopyranosyl-(1→2)-[(6S,2E)-6-hydroxy-2,6-dimethyl-2,7- octadienoyl-(1→6)]-β-D-glucopyranosyl ester (4).

Full text of DOI:10.1021/np980441k

7
40
J . Nat. Prod. 1999, 62, 740-745
F ou r New Tr iter p en oid a l Sa p on in s Acyla ted w ith On e Mon oter p en ic Acid fr om
Gled itsia sin en sis
†
†
†
,†
‡
‡
Zhizhen Zhang, Kazuo Koike, Zhonghua J ia, Tamotsu Nikaido,* Dean Guo, and J unhua Zheng
Department of Pharmacognosy, School of Pharmaceutical Sciences, Toho University, Miyama 2-2-1,
Funabashi, Chiba 274-8510, J apan, and School of Pharmaceutical Sciences, Beijing Medical University,
Beijing 100083, People’s Republic of China
Received October 9, 1998
Four new oleanane-type triterpenoidal glycosides, named gleditsiosides A-D (1-4), were isolated from
the anomalous fruits of Gleditsia sinensis. Using modern NMR techniques, including DQF-COSY,
HETCOR, HOHAHA, HMBC, and ROESY experiments and MS analysis as well as chemical methods,
their structures were determined as 3-O-â-D-xylopyranosyl-(1f2)-R-L-arabinopyranosyl-(1f6)-â-D-glu-
copyranosyl oleanolic acid 28-O-â-D-xylopyranosyl-(1f3)-â-D-xylopyranosyl-(1f4)-R-L-rhamnopyranosyl-
(
1f2)-[(6S,2E)-6-hydroxy-2,6-dimethyl-2,7-octadienoyl-(1f6)]-â-D-glucopyranosyl ester (1); 3-O-â-D-
xylopyranosyl-(1f2)-R-L-arabinopyranosyl-(1f6)-â-D-glucopyranosyl oleanolic acid 28-O-â-D-xylopyranosyl-
1f3)-â-D-xylopyranosyl-(1f4)-R-L-rhamnopyranosyl-(1f2)-[(2E)-2-hydroxylmethyl-6-hydroxy-6-methyl-
,7-octadienoyl-(1f6)]-â-D-glucopyranosyl ester (2); 3-O-â-D-xylopyranosyl-(1f2)-R-L-arabinopyranosyl-
1f6)-â-D-glucopyranosyl echinocystic acid 28-O-â-D-xylopyranosyl-(1f3)-â-D-xylopyranosyl-(1f4)-[â-D-
(
2
(
galactopyranosyl-(1f2)]-R-L-rhamnopyranosyl-(1f2)-[(2E)-2-hydroxylmethyl-6-hydroxy-6-methyl-2,7-
octadienoyl-(1f6)]-â-D-glucopyranosyl ester (3); and 3-O-â-D-xylopyranosyl-(1f2)-R-L-arabinopyranosyl-
(
1f6)-â-D-glucopyranosyl echinocystic acid 28-O-â-D-xylopyranosyl-(1f3)-â-D-xylopyranosyl-(1f4)-[â-D-
galactopyranosyl-(1f2)]-R-L-rhamnopyranosyl-(1f2)-[(6S,2E)-6-hydroxy-2,6-dimethyl-2,7-octadienoyl-
(1f6)]-â-D-glucopyranosyl ester (4).
Gleditsia sinensis Lam. (Leguminosae) is widely distrib-
uted throughout China. Its different partsscalled “Zao J ia”,
“
“
(
Zhu Ya Zao”, “Zao J ia Zi”, “Zao J ia Ye”, “Zao J iao Ci”, and
Zao J ia Gen Pi” from the normal fruits, anomalous fruits
a small fruit), seeds, leaves, thorns, and radix cortexes,
respectivelyspossess different curative effects according to
the theory of traditional Chinese medicine. “Zhu Ya Zao”,
the anomalous fruit produced by old or injured plants, has
long been known in China as a saponin-rich herbal
medicine and used for the treatment of apoplexy, as an
1
expectorant, and for a pesticide. However, until now there
has been no report on the chemical constituents making
up this medicine, though some saponins bearing two
monoterpenic acids were isolated from the fruits of G.
japonica Miquel.2 To shed light on the chemical entity
responsible for its medicinal actions, we conducted a
detailed investigation of the saponins occurring as a very
complex mixture from this source. In this paper, we
describe the isolation and structure elucidation of four new
triterpenoidal saponins, named gleditsiosides A, B, C, and
D (1-4), by using modern NMR techniques, including
DQF-COSY, HETCOR, HOHAHA, HMBC, and ROESY
experiments and MS analysis as well as some chemical
degradation. All four compounds were acylated with one
monoterpenic acid to the C-6 of the glucose directly linked
to C-28 of the aglycon.
-4
Resu lts a n d Discu ssion
Gleditsioside A (1), an amorphous solid, had a molecular
-
formula of C78
H
124
O
35 deduced from the [M - H] ion at
+
m/z 1619.7960 in the negative HRFABMS, [M + Na] ion
+
at m/z 1643, and [M + K] ion at 1659 in the MALDI-TOF
MS (positive ion mode) as well as from its NMR data. The
*
To whom correspondence should be addressed. Tel.: (81) 474-72-1391.
Fax: (81) 474-72-1404. E-mail: nikaido@phar.toho-u.ac.jp.
IR spectrum showed a carbonyl group (1697 cm-1) and a
R,â-unsaturated carbonyl group (1645 cm ) absorption.
†
Toho University.
‡
-1
Beijing Medical University.
1
0.1021/np980441k CCC: $18.00
© 1999 American Chemical Society and American Society of Pharmacognosy
Published on Web 05/06/1999

Gleditsiosides A-D from Gleditsia
J ournal of Natural Products, 1999, Vol. 62, No. 5 741
Ta ble 1. 13C NMR Data for the Aglycon Moieties (125 and 100
MHz in Pyridine-d5)
s) and carbons at δ 94.6, 101.5, 102.4, 105.9, 106.4, 106.7,
and 106.8, respectively (Tables 2 and 3). The C NMR
13
1
2
3
4
1a
1b
2b
1d
2d
spectrum of 1 showed 78 carbon signals, from which 30
were assigned to the aglycon part, 38 to the oligosaccharide
moiety, and the remaining 10 to a monoterpenic acid.
Alkaline hydrolysis of 1 afforded prosapogenins 1a and 1b,
and the monoterpenic acid 1c. Compounds 1a and 1b were
identified as 3-O-â-D-xylopyranosyl-(1f2)-R-L-arabinopy-
ranosyl-(1f6)-â-D-glucopyranosyl oleanolic acid 28-O-â-D-
xylopyranosyl-(1f3)-â-D-xylopyranosyl-(1f4)-R-L-rham-
nopyranosyl-(1f2)-â-D-glucopyranosyl ester and oleano-
2
3
4
5
6
7
8
9
0
1
26.8 26.8 26.8 26.8 26.8 26.8 26.8 28.1 28.2
88.6 88.6 88.8 88.8 88.6 88.5 88.7 78.1 78.1
39.6 39.6 39.6 39.6 39.6 39.6 39.6 39.0 39.1
55.9 55.9 56.1 56.1 55.9 55.9 55.9 55.8 55.9
18.7 18.7 18.8 18.7 18.7 18.5 18.5 18.8 18.9
33.3 33.3 33.5 33.6 33.3 33.2 33.5 33.2 33.6
40.0 40.0 40.2 40.1 40.0 39.8 39.9 39.8 40.0
48.1 48.1 47.2 47.2 48.1 48.1 47.2 48.1 47.3
37.1 37.1 37.2 37.1 37.1 37.1 37.1 37.4 37.5
23.8 23.9 23.9 23.9 23.9 23.9 23.9 23.9 23.9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
lic acid 3-O-â-D-xylopyranosyl-(1f2)-R-L-arabinopyranosyl-
2 122.9 122.9 122.7 122.7 122.8 122.6 122.4 122.6 122.5
3 143.9 143.9 143.8 143.5 144.0 144.8 145.1 144.8 145.1
5
(
1f6)-â-D-glucopyranoside, respectively, based upon their
NMR data (Tables 1, 2, and 3). The monoterpenic acid 1c
was characterized as (6S,2E)-6-hydroxy-2,6-dimethyl-2,7-
octadienoic acid by comparison with the published physi-
cochemical and spectral data.6 Acid hydrolysis of 1 fur-
nished 1d , identified as oleanolic acid, and the monosaccha-
ride components were identified as glucose, xylose, rham-
nose, and arabinose based on GLC analysis. From the
above evidence, it was concluded that 1 was bisdesmosidic
triterpenoid glycoside with glucose, arabinose, and xylose
linked to the C-3 position of the aglycon, and the other four
monosaccharides were connected to C-28 of the aglycon
through an ester bond.
4
5
6
7
8
9
0
1
2
3
4
5
6
7
42.3 42.3 42.1 42.2 42.3 42.1 42.1 42.2 42.1
28.4 28.5 36.2 36.2 28.5 28.3 36.2 28.3 36.2
23.4 23.4 74.1 74.1 23.4 23.7 74.8 23.8 74.8
47.2 47.2 49.4 49.3 47.1 46.7 48.9 46.7 48.9
41.9 41.9 41.5 41.6 42.0 41.9 41.4 42.0 41.5
46.3 46.3 47.3 47.4 46.3 46.4 47.2 46.5 47.3
30.7 30.7 30.8 30.8 30.7 30.7 31.0 31.0 31.1
34.0 34.0 36.0 36.0 34.0 34.3 36.1 34.3 36.1
32.5 32.5 31.9 32.0 32.3 33.3 32.9 33.3 32.9
28.2 28.2 28.3 28.3 28.2 28.2 28.2 28.8 28.8
17.1 17.1 17.1 17.1 17.1 17.1 17.1 16.6 16.6
15.7 15.7 15.8 15.8 15.7 15.5 15.6 15.6 15.7
17.6 17.6 17.6 17.6 17.5 17.4 17.5 17.5 17.6
26.1 26.1 27.1 27.1 26.1 26.3 27.3 26.2 27.3
8 176.5 176.6 175.9 175.9 176.5 180.4 180.0 180.2 179.9
The identity of the monosaccharides and the sequence
of the oligosaccharide chain were determined by a combi-
nation of DQF-COSY, HOHAHA, DEPT, HETCOR, HMBC,
and ROESY experiments. The individual spin systems can
be discerned from the subspectra corresponding to the
anomeric protons or methyl groups (for deoxy sugars) in
the HOHAHA experiment. Starting from the anomeric
proton of each sugar unit, all the hydrogens within each
spin system were assigned using DQF-COSY with the aid
of 2D HOHAHA and ROESY spectra. Information from
COSY, HOHAHA, and ROESY furnished most of the
9
0
33.2 33.2 33.2 33.2 33.2 33.3 33.4 33.3 33.4
23.9 23.9 24.7 24.6 23.7 23.8 24.8 23.8 24.8
After an extensive 2D NMR study, the aglycon was
identified as oleanolic acid (Table 1). The chemical shifts
of C-3 (δ 88.6) and C-28 (δ 176.5) indicated that 1 was a
bisdesmosidic glycoside. The H and C NMR of 1 exhibited
seven sugar anomeric protons at δ 4.83 (d, J ) 7.9 Hz),
.93 (d, J ) 7.0 Hz), 5.03 (d, J ) 7.0 Hz), 5.09 (d, J ) 5.5
Hz), 5.12 (d, J ) 7.4 Hz), 6.09 (d, J ) 8.6 Hz), and 6.34 (br
1
13
4
Ta ble 2. 13C NMR Data for the Sugar Moieties (125 and 100 MHz in Pyridine-d5)
1
2
3
4
1a
1
2
3
4
1a
C3-Glc
1
Rha
1
106.7
157)b
75.7
78.4
72.1
76.2
69.6
106.7
106.7
106.8
106.7
101.5
(175)b
71.6
72.5
84.9
101.5
100.4
100.4
101.3
(
2
3
4
5
6
75.7
78.4
72.3
76.2
69.6
75.7
78.4
72.2
76.1
69.6
75.7
78.4
72.3
76.1
69.6
75.7
78.4
72.3
76.2
69.6
2
3
4
5
6
71.6
72.5
85.0
68.3
18.6
81.2
72.1
83.2
68.2
18.0
81.3
72.1
83.0
68.2
18.6
71.6
72.5
85.1
68.2
18.6
68.3
18.4
Ara
1
Xyl′
1
102.4
163)b
80.7
72.7
67.5
64.4
102.4
102.3
102.3
102.4
106.8
106.8
105.9
106.1
106.8
b
(
(
(
(160)
2
3
4
5
Xyl
1
80.7
72.7
67.5
64.4
80.4
72.6
67.5
64.4
80.6
72.6
67.5
64.3
80.7
72.7
67.5
64.4
2
3
4
75.1c
87.4
68.9
66.9
75.1c
87.4
68.9
66.9
75.0c
87.2
69.0
66.7
75.1c
87.4
69.0
66.8
75.1c
87.4
68.9
66.9
5
Xyl′′
1
106.4
163)b
75.3
77.9
70.9
67.3
106.4
106.2
106.4
106.4
105.9
105.9
105.8
105.8
105.9
b
(161)
c
75.5c
77.9
70.9
75.3c
77.8
70.9
75.5c
77.9
75.3c
77.9
75.2c
78.1
75.2c
78.1
75.2c
78.0
75.2c
78.2
75.2c
78.1
70.8
2
3
4
5
2
3
4
5
Gal
1
2
3
4
5
d
d
d
70.9d
67.3
70.9d
67.3
70.8d
67.4
70.8d
67.4
70.8d
67.2
70.8d
67.3
d
67.3
67.2
67.4
C28-Glc′
1
94.6
168)b
76.8
79.0
71.3
75.8
64.3
94.6
94.5
94.5
94.8
107.6
73.7
74.8
70.2
77.0
62.2
107.6
73.3
74.9
70.2
77.1
62.2
2
3
4
5
6
76.6
79.0
71.5
75.8
64.4
77.2
78.7
71.5
75.8
64.3
77.2
78.8
71.3
75.9
64.3
76.4
79.3
71.3
78.9
62.1
6
a
The assignment was based upon DQF-COSY, HOHAHA, HETCOR, ROESY, and HMBC experiments. b The number in the parentheses
is the 1J C1,H1 coupling constant (Hz).
c,d
The data with the same labels in each column may be interchanged.

7
42 J ournal of Natural Products, 1999, Vol. 62, No. 5
Zhang et al.
Ta ble 3. 1H NMR Data for the Sugar Moieties (500 and 400 MHz in Pyridine-d5)a
1
2
3
4
1a
1
2
3
4
1a
C3-Glc
1
Rha
1
4.83 d
7.9)
4.88 d
(7.6)
4.06
4.20
4.13
4.08
4.25
4.80
4.89 d
(7.9)
4.05
4.16
4.12
4.06
4.22
4.61
4.90 d
(7.8)
4.08
4.15
4.13
4.08
4.25
4.65
4.89 d
(7.6)
4.01
4.21
4.12
4.10
4.25
4.68
6.34
(br. s)
4.80
4.69
4.35
4.40
1.72 d
(6.1)
6.38
(br. s)
4.82
4.66
4.37
4.47
1.77 d
(6.1)
6.45
(br. s)
4.82
4.75
4.36
4.30
1.64 d
(6.1)
6.49
(br. s)
4.85
4.70
4.25
4.38
1.63
(6.0)
6.40
(br. s)
4.82
4.70
4.36
4.52
1.76 d
(6.0)
(
2
3
4
5
6
4.06
4.18
4.12
4.07
4.23
2
3
4
5
6
4
.65
Ara
1
Xyl′
1
5.09 d
5.5)
4.50
4.36
4.34
3.73
5.15 d
(5.2)
4.53
4.37
4.30
5.12 d
(4.9)
4.50
4.35
4.32
5.16 d
(5.6)
4.55
4.38
4.32
5.16 d
(5.2)
4.52
4.37
4.39
5.03 d
(7.0)
4.05
4.01
4.07
5.08 d
(7.3)
4.06
4.03
4.10
5.06 d
(7.3)
4.03
3.96
4.00
5.11 d
5.08 d
(6.8)
4.01
4.03
4.12
(
(
(
(7.1)
4.03
3.96
4.00
3.35
4.10
2
3
4
5
2
3
4
5
3.74
4.30
3.69
4.28
3.75
4.30
3.75
4.31
3.45
4.21
3.52
4.23
3.34
4.07
3.50
4.25
4
.25
Xyl
1
Xyl′′
1
4.93 d
7.0)
4.02
4.05
4.13
3.57
4.92 d
(10.7)
4.04
4.06
4.15
4.97 d
(7.3)
4.03
4.05
4.12
4.98 d
(7.0)
4.03
4.06
4.15
4.98 d
(6.8)
4.01
4.06
4.15
5.12 d
(7.4)
4.05
4.10
4.13
5.19 d
(7.6)
4.06
4.10
4.15
5.09 d
(7.6)
4.03
4.10
4.15
5.13 d
(8.3)
4.03
4.09
4.15
5.18 d
(8.8)
4.01
4.12
4.15
2
3
4
5
2
3
4
5
3.56
4.41
3.56
4.36
3.55
4.43
3.58
4.40
3.65
4.26
3.67
4.31
3.61
4.27
3.65
4.30
3.69
4.28
4
.35
C28-Glc′
1
Gal
1
6.09 d
8.6)
6.13 d
(8.5)
4.37
4.24
4.09
4.06
4.73
4.96
5.93 d
(7.6)
4.25
4.18
4.08
4.10
4.75
4.90
6.09 d
(8.0)
4.27
4.20
4.09
4.10
4.75
4.90
6.20 d
(8.0)
4.40
4.28
4.32
3.97
4.30
4.40
5.15 d
(7.6)
4.45
4.00
4.41
5.18 d
(7.8)
4.51
3.99
4.47
2
3
4
5
6
4.37
4.24
4.10
4.06
4.71
2
3
4
5
6
3.94
4.31
3.95
4.35
4
.88
a
The assignment was based upon DQF-COSY, HOHAHA, HETCOR, ROESY, and HMBC experiments.
aglycon was confirmed by the long-range correlation be-
tween H-1 (δ 4.83) of Glc and C-3 (δ 88.6) of the aglycon.
The sequence of the sugar chain at C-28 was deduced from
the following HMBC correlations: H-1(δ 6.34) of Rha with
C-2 (δ 76.8) of Glc′; H-1(δ 5.03) of Xyl′ with C-4 (δ 84.9) of
Rha; H-1 (δ 5.12) of Xyl′′ with C-3 (δ 87.4) of Xyl′, while
the attachment of the tetrasaccharide chain to C-28 of the
aglycon was based on a correlation of H-1 (δ 6.09) of Glc′
with the C-28 (δ 176.5) of the aglycon. The attachment of
the monoterpenic acid 1c to C-6 of Glc′ was established by
a long-range correlation of H -6 (δ 4.71, 4.88) of Glc′ with
2
C-1 (δ 168.0) of the monoterpenic acid from the HMBC
spectrum of 1. In addition, the downfield shifts of the two
2
H -6 also indicated it was the acylated position. The two
sugar chains of 1 were also deduced from the following
NOE correlations observed in the ROESY spectrum: H-1
(
δ 4.93) of Xyl with H-2 (δ 4.50) of Ara, H-1 (δ 5.09) of Ara
with H-6 (δ 4.65) of Glc, H-1 (δ 5.12) of Xyl′′ with H-3 (δ
.01) of Xyl′, H-1 (δ 5.03) of Xyl′ with H-4 (δ 4.35) of Rha,
4
and H-1 (δ 6.34) of Rha with H-2 (δ 4.37) of Glc′. The major
-
fragmentation peaks at m/z 617 [aglycon + Glc] , 749 [617
-
-
-
+
1
1
Ara] , 881 [749 + Xyl] , 1189 [881 + Glc′ + Rham] ,
-
-
355 [M - Xyl′ - Xyl′′] , 1453 [M - monoterpene] , and
assignments. On the basis of the assigned proton signals,
a HETCOR experiment then gave the corresponding carbon
assignments, and these were further confirmed by an
HMBC experiment. After the assignments of the protons
and protonated carbons were established (Tables 2 and 3),
the seven sugar units were identified as two glucoses, three
xyloses, one rhamnose, and one arabinose and further
confirmed by GLC analysis of the acid hydrolysate. The
linkage of the sugar units at C-3 was established from the
following HMBC correlations: H-1 (δ 5.09) of Ara with C-6
-
487 [M - Xyl′′] in the negative FABMS of 1 were also in
agreement with the structure deduced above.
All the monosaccharides were determined to be in the
pyranose form from their 13C NMR data. The anomeric
configurations for the sugar moieties were fully defined
from their J H1, H2 coupling constants and J C1,H1 coupling
constants as well as from NOE information. Accordingly,
the glucoses and the xyloses were established to be in the
â configuration, while the arabinose and the rhamnose
were in the R configuration. The foregoing evidence led to
the elucidation of the structure of gleditsioside A (1) as 3-O-
3
1
(δ 69.6) of Glc; H-1 (δ 4.93) of Xyl with C-2 (δ 80.7) of Ara.
The attachment of the trisaccharide moiety to C-3 of the

Gleditsiosides A-D from Gleditsia
J ournal of Natural Products, 1999, Vol. 62, No. 5 743
â-D-xylopyranosyl-(1f2)-R-L-arabinopyranosyl-(1f6)-â-D-
glucopyranosyl oleanolic acid 28-O-â-D-xylopyranosyl-(1f3)-
â-D-xylopyranosyl-(1f4)-R-L-rhamnopyranosyl-(1f2)-
monoterpenic acid was deduced from a correlation between
H-6 (δ 4.75, 4.90) of Glc′ and C-1 (δ 167.3) of the mono-
terpenic acid.
[(6S,2E)-6-hydroxy-2,6-dimethyl-2,7-octadienoyl-(1f6)]-â-
The overall structure assignment was accomplished
using the same protocol as in compound 1. The whole sugar
sequence of 3 was deduced from the HMBC information,
from the ROESY experiment, and from the analysis of the
FABMS as depicted for compound 1. All the monosaccha-
rides were determined to be in the pyranose form from their
D-glucopyranosyl ester.
+
Gleditsioside B (2) gave [M + Na] ion at m/z 1659 and
+
[
M + K] ion at 1675 in the MALDI-TOF MS (positive ion
mode), 16 mass units higher than that of 1, implying the
presence of an additional oxygen-bearing function in 2.
Detailed analysis of the 13C NMR spectrum of 2 indicated
the chemical shifts for the aglycon part and sugar moieties
of 2 bore a close resemblance to those of 1, suggesting that
both compounds had a common aglycon and sugar-
substitution pattern. Alkaline hydrolysis of 2 under the
same conditions as that of 1 afforded 1a and 1b as shown
by co-HPLC analysis. Hydrolysis of 2 with 1M HCl gave
1
3
C NMR data. The anomeric configurations for each sugar
3
were evident from their J H1,H2 coupling constants (Tables
and 3) as well as from the ROESY information. On the
2
basis of above evidence, the structure of gleditsioside C (3)
was concluded to be 3-O-â-D-xylopyranosyl-(1f2)-R-L-ara-
binopyranosyl-(1f6)-â-D-glucopyranosyl echinocystic acid
2
8-O-â-D-xylopyranosyl-(1f3)-â-D-xylopyranosyl-(1f4)-[â-
D-galactopyranosyl-(1f2)]-R-L-rhamnopyranosyl-(1f2)-[(2E)-
2-hydroxylmethyl-6-hydroxy-6-methyl-2,7-octadienoyl-(1f6)]-
â-D-glucopyranosyl ester.
1
d , and the sugar units determined by GLC also were
1
D-glucose, D-xylose, L-arabinose, and L-rhamnose. The H
and 1 C NMR data obtained for the monoterpenic acid in
3
+
Gleditsioside D (4) had [M + Na] ion at m/z 1821 and
2
were different from those of 1, in that the methyl group
+
[
M + K] ion at 1837 in the MALDI-TOF MS (positive ion
at C-2 (δ
group (δ
C
12.5, δ
56.2, δ
H
1.88) was replaced by a hydroxylmethyl
4.74). Thus, the monoterpenic acid 2c
mode), 16 mass units lower than that of 3. Detailed
comparison of the NMR data of 4 with those of 3 indicated
both compounds shared a common aglycon and a common
sugar-substitution pattern. The NMR data of the mono-
terpenic acid in 4 were almost superimposable on those of
the monoterpenic moiety in 1. Alkaline hydrolysis of 4,
under the same condition as that of 1, furnished 2b and
C
H
in 2 was shown to be (2E)-2-hydroxylmethyl-6-hydroxy-6-
methyl-2,7-octadienoic acid.3 Unfortunately, attempted
alkaline hydrolysis of 2 did not afford intact 2c, giving only
2
b. Consequently, the absolute configuration of 2c was not
established. The proton and carbon assignments were made
by a combination of DQF-COSY, HOHAHA, DEPT,
HETCOR, and ROESY experiments. From the above
evidence, the structure of 2 was elucidated as 3-O-â-D-
xylopyranosyl-(1f2)-R-L-arabinopyranosyl-(1f6)-â-D-glu-
copyranosyl oleanolic acid 28-O-â-D-xylopyranosyl-(1f3)-
â-D-xylopyranosyl-(1f4)-R-L-rhamnopyranosyl-(1f2)-[(2E)-
1
c as detected by co-HPLC. Acid hydrolysis of 4 afford 2d ,
and the sugar units determined by GLC were D-glucose,
D-xylose, L-arabinose, L-rhamnose, and D-galactose. There-
fore, the structure of 4 was established to be 3-O-â-D-
xylopyranosyl-(1f2)-R-L-arabinopyranosyl-(1f6)-â-D-glu-
copyranosyl echinocystic acid 28-O-â-D-xylopyranosyl-(1f3)-
â-D-xylopyranosyl-(1f4)-[â-D-galactopyranosyl-(1f2)]-R-L-
rhamnopyranosyl-(1f2)-[(6S,2E)-6-hydroxy-2,6-dimethyl-
2
-hydroxymethyl-6-hydroxy-6-methyl-2,7-octadienoyl-(1f6)]-
â-D-glucopyranosyl ester.
Gleditsioside C (3) had a molecular formula of C84
134 42
H O
2
,7-octadienoyl-(1f6)]-â-D-glucopyranosyl ester.
-
determined from the [M - H] ion at m/z 1813.8357 in the
+
negative HRFABMS, an [M + Na] ion at m/z 1837, and
Exp er im en ta l Section
+
an [M + K] ion at 1853 in the MALDI-TOF MS (positive
Gen er a l Exp er im en ta l P r oced u r es. Melting points were
measured with a Yanaco microscope apparatus and are
uncorrected. Optical rotations were performed with a J ASCO
DIP-370 digital polarimeter. IR spectra were carried out on a
J ASCO D-300 FTIR spectrometer. FABMS and MALDI-TOF
MS were conducted using J EOL J MS-700 and Perseptive
Biosystems Voyager DE-STR mass spectrometers, respectively.
ion mode), and from the NMR data. The IR of 3 featured
-
1
absorption of the carbonyl group (1696 cm ) and the R,â-
-
1
unsaturated carbonyl group (1646 cm ). It was apparent
from the chemical shifts of C-3 (δ 88.8) and C-28 (δ 175.9)
of the aglycon in the 13C NMR that 3 was also a bisdes-
1
13
mosidic glycoside. The H and C NMR spectra of 3
displayed eight anomeric proton signals at δ 6.45 (br s,
Rha), 5.93 (d, J ) 7.6 Hz, Glc′), 5.15 (d, J ) 7.6 Hz, Gal),
1
13
The H and C NMR measurements were carried out at 500
MHz (J EOL R-500) or 400 MHz (J EOL EX-400), using an
standard J EOL sequences for 1D and 2D NMR measurement
5
(
.12 (d, J ) 4.9 Hz, Ara), 5.09 (d, J ) 7.6 Hz, Xyl′′), 5.06
d, J ) 7.3 Hz, Xyl′), 4.97 (d, J ) 7.3 Hz, Xyl), and 4.89 (d,
J ) 7.9 Hz, Glc) and carbon signals at δ 100.4 (Rha), 94.5
Glc′), 107.6 (Gal), 102.3 (Ara), 105.8 (Xyl′′), 105.9 (Xyl′),
06.2 (Xyl), and 106.7 (Glc), respectively (Tables 2 and 3).
5
in pyridine-d solution, and chemical shifts were expressed in
δ (ppm) with reference to TMS. ROESY spectra were recorded
with the mixing time set at 250 ms. The spin locking time for
the HOHAHA experiment was 120 ms. The HMBC experiment
was run with the delay time set at 100 ms. Diaion HP-20
(Mitsubishi Chemical), Si gel (Si gel 60, Merck), and ODS
(Chromatorex, 100-200 mesh, Fujisylisia) were used for open
column chromatography. Preparative MPLC and HPLC were
performed using an ODS column (SSC-ODS, 40-60 µm,
detector: UV 210 nm) and an ODS column (PEGASIL ODS-
2, Senshu Pak, 20 mm i.d. × 150 mm, detector: UV 210 nm),
respectively. GLC: Shimadzu GC-7A; column: silicone OV-
(
1
Acid hydrolysis of 2 afforded 2d , identified as echinocystic
3
acid, and the monosaccharide components were identified
as glucose, xylose, arabinose, rhamnose, and galactose
based on the GLC analysis. Alkaline hydrolysis of 3
resulted in the prosapogenin 2b, characterized as echi-
nocystic acid 3-O-â-D-xylopyranosyl-(1f2)-R-L-arabinopy-
ranosyl-(1f6)-â-D-glucopyranoside,5 indicating that the
remaining five sugars were connected to C-28 of the
aglycon. The linkage for sugar units at C-28 was estab-
lished based upon the long-range correlation of H-1 (δ 5.93)
of Glc′ with C-28 (δ 175.9) of the aglycon, appeared in the
HMBC spectrum of 3. The existence of the monoterpenic
acid in compound 3 was indicated from various NMR data
and identified as (2E)-2-hydroxylmethyl-6-hydroxy-6-meth-
yl-2,7-octadienoic acid 2c. The linkage position of the
1
7 on Uniport HP (80-100 mesh), 3 mm i.d. × 2.1 m; column
temperature, 160 °C; carrier gas, N , flow rate 30 mL/min.
2
P la n t Ma ter ia l. The anomalous fruits (Zhu Ya Zao) of G.
sinensis Lam. were purchased from the market of Nanchang,
the capital of J iangxi province, People’s Republic of China, in
J anuary 1998, and were identified by professor Fan Chuishen
(J iangxi College of Traditional Chinese Medicine). A voucher
specimen was deposited in the Division for Pharmacognostical

7
44 J ournal of Natural Products, 1999, Vol. 62, No. 5
Zhang et al.
Biotechnology, School of Pharmaceutical Sciences, Beijing
Medical University, Beijing, China.
Extr a ction a n d Isola tion . The powdered anomalous fruits
3.55 (1H, m, H-3), 1.86, 1.32, 1.11, 1.08, 1.01, 0.93, 0.93 (each
3H, s, H
7.04 (1H, t, J ) 7.6 Hz, H-3), 6.10 (1H, dd, J ) 17.6, 10.8 Hz,
H-7), 5.64 (1H, dd, J ) 17.6, 1.95 Hz, H -8), 5.19 (1H, dd, J )
10.8, 1.95 Hz, H -8), 2.45 (1H, m, H -4), 2.32 (1H, m, H -4),
1.88 (3H, s, H -9), 1.80 (2H, m, H -5), 1.48 (3H, s, H
NMR (pyridine-d , 125 MHz) monoterpenic acid δ 168.0 (C-
3
-27, -23, -26, -30, -24, -25, -29); monoterpenic acid δ
(
Zhu Ya Zao, 4.0 kg) of G. sinensis were refluxed with 95%
EtOH three times for 2 h. The alcoholic extract was concen-
trated (920 g), suspended in H O, and then partitioned
successively with CHCl (45 g) and n-BuOH (480 g). The
n-BuOH-soluble fraction was applied to a column of Diaion
HP-20 (2500 mL) and washed with H O and 30, 50, 70, and
00% MeOH. The 70% MeOH fraction (120 g) was chromato-
2
2
2
2
1
3
2
3
2
3
-10);
C
3
5
1), 146.6 (C-7), 143.5 (C-3), 127.7 (C-2), 111.7 (C-8), 72.2 (C-
6), 41.6 (C-5), 28.6 (C-10), 24.0 (C-4), 12.5 (C-9); other NMR
data are listed in Tables 1, 2, and 3; MALDI-TOF MS (positive
2
1
+
+
graphed over Si gel and ODS columns to give four saponin
fractions of A (5 g), B (22 g), C (2.4 g), and D (60 g). Part of
fraction D (12 g) was chromatographed over ODS columns to
ion mode) m/z [M + Na] 1821, [M + K] 1837.
Alk a lin e Hyd r olysis of Gled itsiosid es A (1), B (2), C
(3), a n d D (4). Gleditsioside A (1, 40 mg) was refluxed with 2
mL 0.8 M NaOH at 80 °C for 4 h. After cooling, the reaction
mixture was neutralized with 1M HCl and then extracted with
n-BuOH (2 mL × 3 times). The organic layers were combined
and then evaporated to dryness in a vacuum. The residue was
subjected to HPLC purification affording prosapogenin 1a (30
mg) and 1c (4 mg). By the same method, 2 (10 mg) afforded
1a (7 mg), 3 afforded 2b, and 4 (10 mg) afforded 2b (3.7 mg)
and 1c (0.5 mg). Compounds 1 and 2 in 2 mL 1 M NaOH were
heated at 100 °C for 4 h, and then isolation and purification
as above depicted furnished 1b.
yield D
repeatedly subjected to MPLC and HPLC purification to afford
(180 mg) and 2 (112 mg). By the same method, fraction C
1 2 3 3
(8.0 g), D (1.5 g), and D (1.1 g). Fraction D was
1
furnished 3 (120 mg) and 4 (65 mg).
Gled itsiosid e A (1): an amorphous solid from MeOH; mp
2
1
2
3
05-206 °C (dec); [R]
D
-11° (c 0.10, MeOH); IR νmax(KBr)
405, 2933, 1697, 1645, 1078 cm-1; 1H NMR (pyridine-d
, 500
5
MHz) aglycon δ 5.44 (1H, br t, H-12), 3.50 (1H, m, H-3), 1.39,
.35, 1.04, 0.98, 0.97, 0.89, 0.87 (each 3H, s, H -23, -27, -26,
24, -30, -25, -29); monoterpenic acid δ 7.02 (1H, t, J ) 7.9 Hz,
H-3), 6.09 (1H, dd, J ) 17.4, 10.7 Hz, H-7), 5.52 (1H, dd, J )
7.4, 1.8 Hz, H -8), 2.45
1H, m, H -4), 2.35 (1H, m, H -9), 1.76 (2H,
-10); C NMR (pyridine-d , 125 MHz)
1
3
-
Acid Hyd r olysis of Gled itsiosid es A (1), B (2), C (3),
a n d D (4). Compound 1 (40 mg) was heated in 1 mL 1M HCl
1
(
2
-8), 5.17 (1H, dd, J ) 10.7, 1.8 Hz, H
-4), 1.88 (3H, s, H
2
2
2
3
(dioxane-H
dioxane was removed, the solution was extracted with EtOAc
O and then
2 2
O, 1:1) at 80 °C for 2 h in a H O bath. After
1
3
m, H
2
-5), 1.46 (3H, s, H
3
5
monoterpenic acid δ 168.0 (C-1), 146.6 (C-7), 143.5 (C-3), 127.7
C-2), 111.7 (C-8), 71.9 (C-6), 41.5 (C-5), 28.6 (C-10), 24.0 (C-
), 12.5 (C-9); other NMR data see Tables 1, 2, and 3; negative
(1 mL × 3). The extraction was washed with H
2
(
concentrated to give an amorphous powder (1d , 10 mg). The
monosaccharide portion was neutralized by passing through
an ion-exchange resin (Amberlite MB-3) column, concentrated
(dried overnight), and then treated with 1-(trimethylsilyl)-
imidazole at room temperature for 2 h. After the excess reagent
4
-
-
FABMS m/z 455 [aglycon - H] , 617 [aglycon + Glc] , 749
-
-
-
[
1
617 + Ara] , 881 [749 + Xyl] , 1189 [881 + Glc′ + Rham] ,
-
-
355 [M - Xyl′ - Xyl′′] , 1453 [M - monoterpene] , 1487 [M
-
-
-
1
Xyl′′] , and 1619 [M - H] ; negative HRFABMS m/z
2
was decomposed with H O, the reaction product was extracted
619.7960 [M - H]- (calcd for 1619.7821); MALDI-TOF MS
with n-hexane (1 mL × 2). The TMSi derivatives of the
monosaccharides were identified as glucose, xylose, arabinose,
and rhamnose by co-GLC analyses with standard monosac-
charides. By the same method, 2 (10 mg) afforded 1d (2.4 mg);
3 (40 mg) afforded 2d (9 mg), and 4 (10 mg) afforded 2d (2.0
mg). GLC analyses showed the monosaccharides of 2 to be
glucose, xylose, arabinose, and rhamnose; those of 3 and 4 were
glucose, xylose, arabinose, rhamnose, and galactose.
+
+
(positive ion mode) m/z [M + Na] 1643, [M + K] 1659.
Gled itsiosid e B (2): an amorphous solid from MeOH; mp
2
1
2
3
03-204 °C (dec); [R]
D
-10.0° (c 0.10, MeOH); IR νmax(KBr)
1
420, 2930, 1709, 1646, 1079 cm-1; H NMR (pyridine-d
, 500
5
MHz) aglycon δ 5.47 (1H, br t, H-12), 3.51, (1H, m, H-3), 1.37,
.34, 1.08, 0.99, 0.97, 0.90, 0.87 (each 3H, s, H -23, -27, -26,
24, -30, -25, -29); monoterpenic acid δ 7.24 (1H, t, J ) 7.9 Hz,
H-3), 6.08 (1H, dd, J ) 17.4, 10.8 Hz, H-7), 5.52 (1H, dd, J )
7.4, 1.8 Hz, H -8), 4.74
2H, br s, H -9), 2.72 (1H, m, H -4), 1.74
2H, m, H -5), 1.45 (3H, s, H , 125
-10); 1 C NMR (pyridine-d
1
3
-
P r osa p ogen in 1a : an amorphous solid from MeOH; mp
1
(
(
2
-8), 5.14 (1H, dd, J ) 10.7, 1.8 Hz, H
-4), 2.60 (1H, m, H
2
255-256 °C (dec); [R]21
-32.0° (c 0.10, MeOH); IR νmax(KBr)
1 1
D
-
2
2
2
3482, 2934, 1744, 1077 cm ; H NMR (pyridine-d
aglycon δ 5.47 (1H, br s, H-12), 3.51 (1H, m, H-3), 1.37, 1.34,
1.08, 0.99, 0.97, 0.90, 0.87 (each 3H, s, H -23, -27, -26, -24,
5
, 400 MHz)
3
2
3
5
MHz) monoterpenic acid δ 167.7 (C-1), 146.6 (C-3), 146.5 (C-
3
7
2
1
1
), 133.0 (C-2), 111.8 (C-8), 72.2 (C-6), 56.2 (C-9), 41.8 (C-5),
8.6 (C-10), 24.0 (C-4); other NMR data are shown in Tables
-30, -25, -29); other NMR data see Tables 1, 2 and 3; MALDI-
+
+
TOF MS (positive ion mode) m/z [M + Na] 1477, [M + K]
+
, 2, and 3; MALDI-TOF MS (positive ion mode) m/z [M + Na]
1493.
+
659, [M + K] 1675.
P r osa p ogen in 1b: an amorphous solid from MeOH; mp
235-236 °C (dec); [R]21
+26° (c 0.10, MeOH); IR νmax(KBr)
3423, 2938, 1695, 1047 cm ; H NMR (pyridine-d
aglycon δ 5.44 (1H, br s, H-12), 3.53 (1H, m, H-3), 1.35, 1.34,
1.01, 1.00, 0.99, 0.95, 0.86 (each 3H, s, H -23, -27, -30, -24,
-26, -29, -25); Glc δ 4.92 (1H, d, J ) 7.8 Hz, H-1), 4.04 (H-2),
4.24 (H-3), 4.12 (H-4), 4.10 (H-5), 4.28 (H -6), 4.67 (H -6); Ara
δ 5.17 (1H, d, J ) 5.1 Hz, H-1), 4.52 (H-2), 4.38 (H-3), 4.40
(H-4), 3.75 (H -5), 4.30 (H -5); Xyl δ 5.00 (1H, d, J ) 6.6 Hz,
H-1), 4.03 (H-2), 4.08 (H-3), 4.14 (H-4), 3.53 (H -5), 4.43 (H
, 100 MHz) Glc δ 106.8 (C-1), 75.7
Gled itsiosid e C (3): an amorphous solid from MeOH; mp
D
2
1
-1 1
2
3
11-212 °C (dec); [R]
D
-15° (c 0.10, MeOH); IR νmax(KBr)
5
, 400 MHz)
412, 2925, 1696, 1646, 1078 cm-1; 1H NMR (pyridine-d
, 500
5
MHz) aglycon δ 5.57 (1H, br t, H-12), 5.05 (1H, br t, H-16),
3
3
3
7
.44 (1H, m, H-3), 1.75, 1.27, 1.06, 1.05, 0.98, 0.95, 0.92 (each
H, s, H -27, -23, -26, -30, -24, -25, -29); monoterpenic acid δ
.15 (1H, t, J ) 7.9 Hz, H-3), 6.05 (1H, dd J ) 17.4, 10.7 Hz,
-8), 5.11 (1H, dd, J )
-9), 2.66 (1H, m, H -4),
-5), 1.42 (3H, s, H
-10); 13
, 125 MHz) monoterpenic acid δ 167.3 (C-
3
2
2
H-7), 5.45 (1H, dd, J ) 17.4, 1.8 Hz, H
1
2
2
2
2
0.7, 1.8 Hz, H
.55 (1H, m, H
2
-8), 4.64 (2H, br s, H
-4), 1.77 (2H, m, H
2
2
2
2
-
13
2
2
3
C
5); C NMR (pyridine-d
5
NMR (pyridine-d
5
(C-2), 78.4 (C-3), 72.3 (C-4), 76.2 (C-5), 69.6 (C-6); Ara δ 102.4
(C-1), 80.6 (C-2), 72.7 (C-3), 67.5 (C-4), 64.4 (C-5); Xyl δ 106.4
(C-1), 75.5 (C-2), 77.8 (C-3), 70.9 (C-4), 67.3 (C-5); MALDI-
1
6
), 146.4 (C-3), 145.9 (C-7), 132.3 (C-2), 111.4 (C-8), 72.1 (C-
), 55.8 (C-9), 41.9 (C-5), 28.3 (C-10), 24.0 (C-4); other NMR
+
+
data are given in Tables 1, 2, and 3; negative FABMS m/z 765
TOF MS (positive ion mode) m/z [M + Na] 905, [M + K]
-
-
[
aglycon + Glc + Ara] , 897 [765 + Xyl] , 1205 [897 + Glc′ +
921.
-
-
-
Rham] , 1337 [1205 + Xyl′] , 1469 [1337 + Xyl′′] ,1499 [1337
P r osa p ogen in 2b: an amorphous solid from MeOH; mp
-
-
-
21
+
Gal] , 1631 [M - monoterpene] , and 1813 [M - H] ;
229-230 °C (dec); [R]
3423, 2938, 1695, 1047 cm ; H NMR (pyridine-d
aglycon δ 5.60 (1H, br s, H-12), 5.26 (1H, br s, H-16), 3.50 (1H,
m, H-3), 1.89, 1.33, 1.19, 1.06, 1.03, 0.99, 0.89 (each 3H, s, H
27, -23, -30, -29, -26, -24, -25); Glc δ 4.91 (1H, d, J ) 7.7 Hz,
H-1), 4.05 (H-2), 4.21 (H-3), 4.15 (H-4), 4.09 (H-5), 4.26 (H
6), 4.68 (H -6); Ara δ 5.16 (1H, d, J ) 5.1 Hz, H-1), 4.52 (H-2),
4.39 (H-3), 4.40 (H-4), 3.75 (H -5), 4.30 (H -5); Xyl δ 5.00 (1H,
D
+12° (c 0.90, MeOH); IR νmax(KBr)
-
-1
1
negative HRFABMS m/z 1813.8357 [M - H] (calcd for
5
, 400 MHz)
1
813.8221); MALDI-TOF MS (positive ion mode) m/z [M +
+
+
Na] 1837, [M + K] 1853.
3
-
Gled itsiosid e D (4): an amorphous solid from MeOH; mp
2
1
2
3
15-216 °C (dec); [R]
D
-19.0° (c 0.10, MeOH); IR νmax(KBr)
2
-
405, 2928, 1708, 1646, 1080 cm-1; H NMR (pyridine-d
1
, 500
5
2
MHz) aglycon δ 5.64 (1H, br t, H-12), 5.01 (1H, br t, H-16),
2
2

Gleditsiosides A-D from Gleditsia
J ournal of Natural Products, 1999, Vol. 62, No. 5 745
d, J ) 7.0 Hz, H-1), 4.03 (H-2), 4.07 (H-3), 4.17 (H-4), 3.60
NMR data are shown in Table 1; MALDI-TOF MS (positive
ion mode) m/z [M + Na] 479.
Ech in ocystic a cid (2d ): an amorphous solid from MeOH;
-5); 1 C NMR (pyridine-d
3
, 100 MHz) Glc δ 106.8
+
(H
2
-5), 4.40 (H
2
5
(C-1), 75.7 (C-2), 78.4 (C-3), 72.2 (C-4), 76.1 (C-5), 69.6 (C-6);
2
1
Ara δ 102.3 (C-1), 80.5 (C-2), 72.6 (C-3), 67.5 (C-4), 64.2 (C-5);
mp 280-281 °C; [R]
D
+52° (c 0.10, MeOH); IR νmax(KBr) 3435,
2942, 1689, 1032; H NMR (pyridine-d , 400 MHz) δ 5.67 (1H,
br s, H-12), 5.26 (1H, br s, H-16), 3.40 (1H, m, H-3), 1.88, 1.24,
1.20, 1.08, 1.07, 1.04, 0.95 (each 3H, s, H -27, -23, -30, -29,
1
Xyl δ 106.3 (C-1), 75.3 (C-2), 77.9 (C-3), 70.9 (C-4), 67.3 (C-5);
5
+
MALDI-TOF MS (positive ion mode) m/z [M + Na] 921.
2
1
Mon oter p en ic a cid 1c: colorless oil; [R]
D
+12.8° (c 1.0,
3
1
13
MeOH); H NMR (pyridine-d
7
5
, 400 MHz) δ 7.20 (1H, t, J )
-26, -24, -25); C NMR data are given in Table 1; MALDI-
TOF MS (positive ion mode) m/z [M + Na] 495.
+
.15 Hz, H-3), 6.10 (1H, dd, J ) 17.2, 10.6 Hz, H-7), 5.53 (1H,
dd, J ) 17.2, 1.8 Hz, H
2
-8), 5.15 (1H, dd, J ) 10.6, 1.8 Hz,
Refer en ces a n d Notes
H
2
-8), 2.52 (1H, m, H
2
-4), 2.44 (1H, m, H
2
-4), 2.04 (3H, s, H -
3
1
3
(1) J iangsu New Medical College, Zhong Yao Da Ci Dian; Shanghai
9
d
2
1
2
2 3
), 1.77 (2H, m, H -5), 1.46 (3H, s, H -10); C NMR (pyridine-
, 100 MHz) δ 170.9 (C-1), 146.6 (C-7), 142.4 (C-3), 129.0 (C-
), 111.7 (C-8), 72.2 (C-6), 41.8 (C-5), 28.5 (C-10), 23.4 (C-4),
People’s Public Health Publishing House: Shanghai, 1977; pp 1144,
5
1
145, 1147, 2198.
(2) Konoshima, T.; Umegaki, Y.; Sawada, T. Chem. Pharm. Bull. 1981,
29, 2695-2699.
(3) Konoshima, T.; Sawada, T. Chem. Pharm. Bull. 1982, 30, 2747-2760.
+
2.8 (C-9); MALDI-TOF MS (positive ion mode) m/z [M + Na]
+
07, [M + K] 223.
(
4) Konoshima, T.; Sawada, T. Chem. Pharm. Bull. 1982, 30, 4082-4087.
Olea n olic a cid (1d ): an amorphous solid from MeOH; mp
(5) Nigam, S. K.; Gopal, M.; Uddin, R.; Yoshikawa, K.; Kawamoto, M.;
Arihara, S. Phytochemistry 1997, 44, 1329-1334.
(6) Yoshikawa, K.; Suzaki, Y.; Tanaka, M.; Arihara, S.; Nigam, S. K. J .
Nat. Prod. 1997, 60, 1269-1274.
2
1
2
2
(
75-276 °C; [R]
D
+82° (c 0.10, MeOH); IR νmax(KBr) 3456,
-1 1
940, 1696, 1036 cm ; H NMR (pyridine-d
1H, br s, H-12), 3.47 (1H, m, H-3), 1.29, 1.25, 1.03, 1.03, 1.02,
.96, 0.91 (each 3H, s, H
5
, 400 MHz) δ 5.51
0
3
-23, -27, -30, -24, -26, -29, -25); 13
C
NP980441K