New triterpenoid saponins from Stenocereus eruca

Article abstract of DOI:10.3987/COM-12-12458

Eight new triterpenoid saponins, stellatosides C (1), D (2), and E (3), stellatoside B methyl ester (4), stellatoside C methyl ester (5), thurberoside A (6), phillyriside A (7), and treleaseside A (8) were isolated from Stenocereus eruca A. C. Gibson & K. E. Horak (Machaerocereus eruca Br. & R.). The structures of these compounds were elucidated on the basis of spectroscopic evidence and their physicochemical properties.

Full text of DOI:10.3987/COM-12-12458

HETEROCYCLES, Vol. 85, No. 6, 2012, pp. © 2012 The Japan Institute of Heterocyclic Chemistry
Received, 2nd March, 2012, Accepted, 25th April, 2012, Published online, 11th May, 2012
DOI: 10.3987/COM-12-12458
NEW TRITERPENOID SAPONINS FROM STENOCEREUS ERUCA
Kazutaka Kakuta, Takeshi Koike, Kaoru Kinoshita, Satoru Ito, Kiyotaka
Koyama, and Kunio Takahashi
Department of Pharmacognosy and Phytochemistry, Meiji Pharmaceutical
University, Noshio 2-522-1, Kiyose-shi, Tokyo 204-8588, Japan. E-mail address:
diamonds@my-pharm.ac.jp
Abstract – Eight new triterpenoid saponins, stellatosides C (1), D (2), and E (3),
stellatoside B methyl ester (4), stellatoside C methyl ester (5), thurberoside A (6),
phillyriside A (7), and treleaseside A (8) were isolated from Stenocereus eruca A.
C. Gibson & K. E. Horak (Machaerocereus eruca Br. & R.). The structures of
these compounds were elucidated on the basis of spectroscopic evidence and their
physicochemical properties.
INTRODUCTION
The cactus plant is a succulent plant in North and South America which grows wild during the dry season
and during breaks in the rainy season. Cacti are classified by their forms into tree-form cactus
(Pereskioideae), bushy cactus and columnar cactus (Cactioideae), and globular cactus (Opuntioideae).
After screening numerous cacti, we discovered that some bushy or columnar cacti (Cactioideae)
contained many saponins. Further research led us to isolate seventeen new and twenty known triterpene
sapogenins,1-16 in addition to fifteen new saponins and two known ones from several cacti.18-21 Some
triterpene sapogenins from cacti showed antinociceptive22 and antitumor-promotion activities.23 In the
previous study, we have reported the structures of stellatoside B and erucasaponin A from Stenocereus
eruca A. C. Gibson & K. E. Horak (Machaerocereus eruca Br. & R.). 20 In this study, newly we
isolated eight new triterpenoid saponins, stellatosides C (1), D (2), and E (3), stellatoside B methyl ester
(4), stellatoside C methyl ester (5), thurberoside A (6), phillyriside A (7), and treleaseside A (8) from
Stenocereus eruca. Thurberoside A (6), phillyriside A (7) and treleaseside A (8) were the first saponins
with thurberogenin, 27-desoxyphillyrigenin, and treleasegenic acid as aglycons. This paper discusses
the isolation and structure elucidation of these new saponins (1-8).

RESULTS AND DISCUSSION
Dried Stenocereus eruca was extracted repeatedly with chloroform and then with methanol. The
methanol extract was passed through a Diaion HP-20 column to adsorb the saponins. The methanolic
eluate was separated by silica gel and octadecyl silyl silica gel (ODS) column chromatography, yielding
stellatosides C, D, and E (1-3), stellatosides B and C methyl esters (4 and 5), thurberoside A (6),
phillyriside A (7), and treleaseside A (8). The structures of compounds 1-8 were determined
spectroscopically using DEPT, HMQC, HMBC, DQF-COSY, HSQC-TOCSY, phase sensitive TOCSY
and phase sensitive NOESY experiments (Figure 1).
CH2OAc
HO
O
OH
H
O
O
H
O
O
H
H
H
H
H
COO rha
O
H
H
H
H
RO
RO
RO
RO
H
H
H
H
2
2
2
2
8
glcA rha
6
glcA
3
7
glcA rha
3
1
rha
glcA rha
3
rha
rha
rha
2
3
2
3
4
5
glcA rha glc
2
glcA rha
2
2
glcA(Me)
glc xyl
2
glcA(Me)
rha
3
rha
Figure 1. Structures of 1-8
Stellatoside C (1), a colorless powder, has a lupane-type triterpene skeleton, stellatogenin as the aglycon
and three sugars, glucuronic acid and two rhamnoses. Its molecular formula C48H76O18 was determined
with its positive ion HRFABMS (m/z 941.5127, [M+H]+). The IR spectrum of 1 showed absorptions at
1
3410 cm-1 (hydroxy) and 1740 cm-1 (lactone). In the aglycon moiety, H-NMR spectrum indicated the
presence of seven methyl groups characterized by singlets at H 0.70-1.16. Since 1 did not have an
olefinic proton, had two methine carbon signals, the aglycon moiety of compound 1 was predicted to be a
lupane-type triterpene. From the dara of HMQC, HMBC, DQF-COSY, and phase sensitive NOESY
spectral correlations, revealed that –OH was in the C-20 and lactone ring was existed between C-21 and
C-28, the aglycon moiety of 1 was determined to be stellatogenin (Figure 1, 2). Addition, the aglycon
1
was obtained by 3.5%HCl hydrolysis of 1 and was identified by H- and 13C-NMR spectral data.13,26
About the sugar unit, in the 1H- and 13C-NMR spectra, three anomeric proton signals [H 4.35 (1H, d, J =

7.3 Hz), 4.73, 4.88 (each 1H, br s)] and carbon signals [C 100.6 (CH), 100.9 (CH), 103.3 (CH)] were
recognized. Detailed analysis of 1 was conducted with HMQC, HMBC, DQF-COSY, phase sensitive
NOESY, TOCSY, and phase sensitive HSQC-TOCSY experiments, each sugar units were determined as
one -glucuronic acid and two -rhamnoses. About the linkage of the each sugar units, between H-1’
and C-3 correlated in the HMBC experiment indicated that the glucuronic acid unit binds at C-3 of the
aglycon moiety. From the anomeric proton of rhamnose-1 to C-2’ of glcuronic acid HMBC correlation
indicates that rhamnose-1 unit binds at C-2’ of the glucuronic acid unit. Similarly, the HMBC
correction from anomeric proton of rhamnose-2 to C-3’ of glcuronic acid indicated that rhamnose-2 unit
binds at C-3’ of glucuronic acid unit (Figure 2). The absolute configuration of each sugar units as
D-glucuronic acid and L-rhamnose were deternined by measuring ODS HPLC retention times of each
sugar derivatives.24 Thus, the structure of 1 was determined to be stellatogenin 3-O--L-
rhamnopyranosyl-(1→2)-[-L-rhamnopyranosyl-(1→3)]--D-glucuronopyranoside.
HO
stellatogenin
O
glcA
O
COOH
3
O
HO
O
rha-2
HO
O
O
O
HO
OH
DQF-COSY
NOESY
1''
O
HO
rha-1
HMBC
HO
OH
Figure 2. 2D-NMR Correlations of Stellatoside C (1)
Stellatoside D (2) was isolated as a colorless powder. Its molecular formula C48H76O19 was determined
from its negative ion HRFABMS (m/z 955.4916, [M-H]-). The IR spectrum of 2 showed absorptions at
1
3400 cm-1 (hydroxy) and 1760 cm-1 (lactone). About the aglycon moiety, the H- and 13C-NMR
spectrum data was confortable with 1, the aglycon moiety of 2 was determined to be stellatogenin.
Regarding the sugar moieties, three anomeric proton signals [H 4.28 (1H, d, J = 7.6 Hz), H 4.29 (1H, d,
J = 6.1 Hz), H 5.26 (1H, br s)] and carbon signals [C 99.7 (CH), 103.3 (CH), 104.8 (CH)] suggested the
presence of three sugar units. In addition, carbonyl carbon signal (C 174.2), methylene signal (C 60.8)
and methyl signal (C 17.8) indicated the presence of glucuronic acid, glucose and rhamnose units,
respectively. From the analysis of the HMQC, HMBC, DQF-COSY, Phase sensitive NOESY, TOCSY

and Phase sensitive TOCSY experiments all the protons and carbons of the sugar moiety were assigned as
shown as Table 1 and Table 2-1. The linkage of the sugar units at C-3 of stellatogenin and the sequence
of the sugar chains were established by the following HMBC and NOESY spectra. Analysis of the
HMBC spectrum of 2 established that these were cross-peaks between anomeric proton of glucuronic acid
at  4.29 and C-3 of stellatogenin at  88.0. This result indicated that the glucuronic acid was connected
to C-3 of the aglycon. The linkage of the rhamnose and glucuronic acid was indicated by HMBC
cross-peak between anomeric proton of rhamnose at  5.26 and C-2' of glucuronic acid at  75.8.
Similarly, the linkage of the glucose and rhamnose was indicated by HMBC cross-peak between
anomeric proton of glucose at  4.28 and C-3"" of rhamnose at  81.7. In the NOESY spectrum, NOE
correlations between the methine proton at H-3 of the aglycon ( 3.00) and the anomelic proton of
glucuronic acid ( 4.29), between H-2' of glucuronic acid ( 3.20) and the anomelic proton ( 5.26) of
rhamnose, and between H-3"" ( 3.65) of rhamnose and the anomelic proton of glucose (4.28). On the
basis of the above, the structure of 2 was determined as stellatogenin 3-O--D-glucopyranosyl-
(1→3)--L-rhamnopyranosyl-(1→2)--D-glucuronopyranoside.
3
H
HOOC
HO
HO
O
O
O
H
H
O
H
H
H
H
O
H
HO
HOH2C
HO
HO
H
H
OH
DQF-COSY
NOESY
O
OH
HMBC
H
H
Figure 3. 2D Correlations of Stellatoside D (2)
Stellatoside E (3) was obtained as a colorless powder. Its molecular formula C42H66O14 was determined
from its negative ion HRFABMS (m/z 793.4370 [M-H]-). The IR spectrum of 3 showed absorptions at
1
13
3410 cm-1 (hydroxy) and 1760 cm-1 (lactone). The H- and C-NMR spectrum data were similar with
compound 2 (stellatoside D). Although compound 2 had a glucuronic acid, rhamnose and glucose,
compound 3 had glucuronic acid and rhamnose as sugar moiety. The linkage of the oligosaccharide
moiety was determined by HMBC correlations between anomeric proton of glucuronic acid ( 4.25) and
C-3 of aglycon ( 87.9), and anomeric proton of rhamnose ( 5.24) and C-2' of glucuronic acid ( 76.3).
In conclusion, the structure of 3 was determined as stellatogenin 3-O--L-rhamnopyranosyl-(1→2)-
-D-glucuronopyranoside.

Stellatoside B methyl ester (4) was obtained as a colorless powder. Its molecular formula C48H76O19
was determined from its negative ion HRFABMS (m/z 955.4915, [M-H]-). The IR spectrum of 5
showed absorptions at 3410 cm-1 (hydroxy) and 1750 cm-1 (lactone). From the 1H- and 13C-NMR signal
1
13
patterns, the aglycon moiety was determined to be stellatogenin. The H- and C-NMR spectrum data
showed that compound 4 has three sugars, glucuronic acid methyl ester, glucose, and xylose. Since
resonance C-6’ of glucuronic acid was upfield shifted and a new methyl proton signal atH3.64 (3H, s)
had HMBC correlation with carbonyl carbon of glucuronic acid, the structure of 4 was determined as
methyl ester of stellatoside B.20 Thus, 4 was elucidated as stellatogenin 3-O--D-xylopyranosyl-(1→2)-
-D-glucopyranosyl-(1→2)-6-O-methyl--D-glucuronopyranoside.
Stellatoside C methyl ester (5) was obtained as a colorless powder. Its molecular formula C49H78O18 was
determined from its positive ion HRFABMS (m/z 955.5283, [M+H]+). The IR spectrum of 5 showed
absorptions at 3410 cm-1 (hydroxy) and 1760 cm-1 (lactone). Compared to the 1H- and 13C-NMR data of
1, a proton signal at H 3.66 (3H, s) and two carbon signals at C 51.9 (CH3) and C 169.4 (C) were
different. Since these signals indicated that 5 has a methyl ester in its structure, the oligosaccharide
moiety of 5 was methyl ester of 1. Thus, 5 was determined to be stellatogenin 3-O--L-rhamnopyranosyl-
(1→2)-[-L-rhamnopyranosyl-(1→3)]-6-O-methyl--D-glucuronopyranoside. Although 5 was methyl
ester of 1, we considered compound 5 is not artifact from compound 1. Because if compound 5 was
artifact, probably, we had isolated methyl esters of saponins from cacti. In our previous study, we have
isolated same saponins from cacti17-21 as same methods, and this is first isolation as methyl ester of
glucuronic acid on saponin from cacti. Then, we considered this methyl ester of stellatoside C was new
compound from S. eruca.
Thurberoside A (6) was isolated as a colorless powder. Its molecular formula C48H74O17 was
determined from its positive ion HRFABMS (m/z 923.4998, [M+H]+). The IR spectrum of 6 showed
absorptions at 3400 cm-1 (hydroxy) and 1760 cm-1 (lactone). The compound displayed 48 signals in its
13C-NMR spectrum. Almost all the signals in the 13C-NMR spectrum were similar with 1, however,
exomethylene signals [H 4.87, 4.88 (each 1H, br s), C 112.3 (CH2), 150.3 (C)] were newly appeared in 6.
Then, aglycon moiety of 6 was considered to be thurberogenin.25 The sugar moiety was same with its of
1. From the data of 1D and 2D NMR data, the structure of 6 was determined to be thurberogenin
3-O--L-rhamnopyranosyl-(1→2)-[-L-rhamnopyranosyl-(1→3)]--D-glucuronopyranoside.
Phirylliside A (7) was obtained as a colorless powder. Its molecular formula C48H76O17 was determined
from its positive ion HRFABMS (m/z 925.5161, [M+H]+). The IR spectrum showed absorptions at 3410
1
cm-1 (hydroxy) and 1740 cm-1 (lactone). In the H-NMR spectrum, six methyl signals as singlet (H
0.71–1.21) and one methyl signal as doublet [H 0.91 (3H, d, J = 6.7 Hz)] were observed. The aglycon

of 7 was identified as 27-desoxyphillyrigenin by comparison with our NMR data.16 The sugar moieties
were determined same as 1. Finally, the structure of 7 was determined to be 27-desoxyphillyrigenin
3-O--L-rhamnopyranosyl-(1→2)-[-L-rhamnopyranosyl-(1→3)]--D-glucuronopyranoside.
Treleaseside A (8) was isolated as a colorless powder. Its molecular formula C50H78O20 was determined
from its negative ion HRFABMS (m/z 997.5016, [M-H]-). The IR spectrum showed absorptions at 3400
cm-1 (hydroxy) and 1720 cm-1 (carbonyl). The compound displayed 50 signals in its 13C-NMR spectrum.
1
Its H-NMR spectrum indicated the presence of six methyl groups characterized by singlets at H 0.65,
0.73, 0.84, 1.09 (each 3H, s), and 0.94, (6H, s). The signal patterns of 1H- and 13C-NMR spectrum data
of aglycon moiety were similar with treleasegenic acid which had already reported as triterpene
sapogenin from S. eruca. 16 The different part of the aglycon of 8 and treleasegenic acid was 8 had
newly acetyl group [c 20.7 (CH3) and c 170.6 (C=O)]. The position of the acetyl group was
determined by HMBC correlations between H2-30 of aglycon (H 3.91 and 4.26, each 1H, d, J = 11.3 Hz)
and C 170.6 (carbonyl carbon of acethyl group). From this information, the acetyl group was
considered to be bound to position 30 of treleasegenic acid.
Figure 4. NOESY Correlations of Treleaseside A (8)
The structure of treleasegenic acid 30 acetate16 as aglycon was identified by NOESY correlations. In
NOESY spectrum, the methine proton of H-18 ( 2.82) had cross-peaks with the methine proton of H-12
( 5.18) and the methylene protons of H2-30 ( 3.91 and 4.26). The three anomeric carbons of the sugar
moieties [C 93.7 (CH), 99.8 (CH), 103.7 (CH)] had HMQC correlations with anomeric protons at H
5.72 (1H, br s),H 5.26 (1H, br s) and 4.25 (1H, d, J = 7.6 Hz), respectively. The sugar moiety at C-3
was same with 3, and the rhamnose unit was bound at C-28 of treleasegenic acid 30-acetate. In
conclusion, the structure of
8
was determined to be treleasegenic acid 30-acetate
3-O--L-rhamnopyranosyl-(1→2)--D-glucuronopyranosyl 28-O--L-rhamnopyranoside. The absolute
configuration of each sugar units of compounds 2-8 as D-glucuronic acid, D-xylose and L-rhamnose were
deternined by measuring ODS HPLC retention times of each sugar derivatives.24

Table 1. 13C-NMR Spectral Data of 1-8 (125 MHz, in DMSO-d6)
Position
C-1
1
2
38.6
25.5
88.0
38.7
55.2
17.7
34.0
40.5
49.7
36.3
20.7
27.8
40.2
42.5
26.0
23.4
51.4
41.6
54.8
68.6
81.0
43.9
27.4
16.0*
16.1*
15.7
13.8
178.7
29.9
30.2
3
38.7
25.5
87.9
38.6
55.2
17.6
34.0
40.5
49.7
36.3
20.7
27.8
40.2
42.5
26.0
23.3
51.4
41.6
54.7
68.9
81.0
43.9
27.3
16.0
16.0
15.7
13.8
178.7
29.9
30.2
4
5
38.5
25.6
88.1
38.6
55.0
17.6
34.0
40.5
49.6
36.4
20.7
27.8
40.1
42.5
26.0
23.3
51.4
41.6
54.7
68.5
81.0
43.9
27.2
15.9*
16.0*
15.7
13.8
178.7
29.9
30.1
6
7
8
38.7
25.5
87.8
38.6
55.2
17.6
34.0
40.5
49.7
36.7
20.7
27.8
40.2
42.5
26.0
23.3
51.4
41.6
54.7
68.6
81.0
43.9
27.3
15.9*
16.1*
15.7
13.8
178.7
29.9
30.2
38.2
25.6
88.4
38.7
54.9
17.6
34.0
40.5
49.6
36.3
20.7
27.8
40.1
42.5
26.0
23.3
51.4
41.6
54.7
68.6
81.0
43.9
27.3
15.9
15.9
15.7
13.9
178.7
29.9
30.1
38.5
25.7
88.1
38.7
55.2
17.6
33.7
40.5
49.7
36.4
20.2
25.7
40.3
41.9
26.0
22.7
50.9
44.0
52.5
150.3
81.0
43.8
27.2
15.9
16.0
15.5
13.6
177.7
112.3
23.0
38.8
25.5
87.8
38.6
55.3
17.6
33.5
40.0
49.9
36.4
20.5
24.5
42.5
40.6
26.6
27.1
41.4
47.3
41.5
83.8
26.7
31.2
27.2
15.9
16.2
15.3
13.9
175.9
18.3
23.6
38.4
25.4
87.8
38.5
55.2
17.7
32.4
38.8
46.9
36.2
22.9
122.6
142.1
41.2
27.2
23.8
47.9
40.1
40.0
40.0
70.6
39.8
16.2
27.4
15.2
16.9
25.3
173.6
23.9
63.4
170.6
20.7
C-2
C-3
C-4
C-5
C-6
C-7
C-8
C-9
C-10
C-11
C-12
C-13
C-14
C-15
C-16
C-17
C-18
C-19
C-20
C-21
C-22
C-23
C-24
C-25
C-26
C-27
C-28
C-29
C-30
30-CH3CO
30-CH3CO
glcA
1’
103.3
77.2
82.0
70.9
74.1
172.6
103.3
75.8
77.6
72.5
74.0
174.2
103.7
76.3
72.3
77.8
74.1
172.7
103.5
79.2
76.2
71.1
75.1
102.8
76.2
81.0
70.1
74.9
103.3
76.2
82.0
70.5
75.2
170.2
103.4
77.1
82.1
71.0
74.1
173.3
103.7
76.4
73.7
78.0
73.7
173.6
2’
3’
4’
5’
COOH
COOCH3
COOCH3
glc
169.4
51.8
169.4
51.9
1’’
2’’
3’’
4’’
5’’
6’’
104.8
73.9
76.3
76.8
69.8
60.8
100.5
82.9
76.5
70.0
76.0
61.1
xyl
1’’’
2’’’
3’’’
4’’’
104.5
74.1
75.7
69.3
65.8
5’’’
C-3 rha 1
1’’’’
2’’’’
3’’’’
4’’’’
5’’’’
6’’’’
C-3 rha 2
1’’’’’
2’’’’’
3’’’’’
4’’’’’
5’’’’’
6’’’’’
C-28 rha
1’’’’’’
2’’’’’’
3’’’’’’
4’’’’’’
5’’’’’’
6’’’’’’
100.6
70.0
70.4
71.7
68.8
17.7
99.7
69.3
81.7
70.8
67.8
17.8
99.8
70.3
70.3
72.0
68.9
17.8
100.1
70.5
70.4
71.6
68.9
17.7
100.3
70.4
70.4
71.8
68.8
17.7
100.6
70.0
70.4
71.7
68.8
17.7
99.8
70.3
70.4
72.5
68.0
17.8*
100.9
70.4
70.6
71.9
68.5
17.7
100.6
70.4
70.0
71.8
68.8
17.7
101.0
70.2
70.1
71.6
68.5
17.7
101.0
70.3
70.3
71.9
68.5
17.7
93.7
70.5
71.1
71.4
69.5
17.9*
*: may be interchanged

Table 2-1. 1H-NMR Spectroscopic Data [H, mult. (J in Hz)] of 1-4 (500 MHz, in DMSO-d6)
Position
1
2
3
4
1
0.87 (1H, m), 1.53 (1H, m)
 1.92 (1H, br d, 11.5)
1.46 (1H, m)
3.03 (1H, dd, 11.5, 3.6)
0.65 (1H, br d, 11.3)
 1.47 (1H, m)
 1.32 (1H, m)
1.35 (2H, m)
1.21 (1H, m)
0.87, 1.54 (each 1H, m)
 1.99 (1H, br d, 11.3)
1.47 (1H, m)
3.00 (1H, dd, 11.3, 3.7)
0.65 (1H, br d, 10.7)
 1.44 (1H, m)
 1.33 (1H, m)
1.33 (2H, m)
1.21 (1H, m)
0.82, 1.58 (each 1H, m)
 1.92 (1H, br d, 10.4)
1.49 (1H, m)
3.00 (1H, dd, 10.4, 3.7)
0.66 (1H, br d, 11.3)
 1.44 (1H, m)
1.32 (1H, m)
1.33 (2H, m)
1.22 (1H, m)
0.82, 1.51 (each 1H, m)
 1.62 (1H, m)
1.49 (1H, m)
3.01 (1H, m)
0.64 (1H, br d, 11.3)
 1.46 (1H, m)
1.30 (1H, m)
1.32 (2H, m)
2
3
5
6
7
9
1.17(1H, m)
11
 1.15 (1H, m)
 1.14 (1H, m)
 1.40 (1H, m)
1.17, 1.47 (each 1H, m)
1.22 (1H, m)
 1.15 (1H, m)
1.41 (1H, m)
 1.14 (1H, m)
1.40 (1H, m)
1.14, 1.49 (each 1H, m)
1.22 (1H, m)
 1.42 (1H, m)
1.15, 1.47 (each 1H, m)
1.21 (1H, m)
12
13
15
1.15, 1.47 (each 1H, m)
1.22 (1H, br d, 11.5)
 0.99 (1H, m)
 2.29 (1H, dt, 13.1, 4.7)
 1.60 (1H, br dd, 13.1, 4.7)
 1.81 (1H, m)
1.86 (1H, dd, 11.5, 5.8)
1.57 (1H, br d, 5.8)
4.67 (1H, br s)
 0.99 (1H, m)
 2.28 (1H, ddd, 13.0, 12.8, 4.7)
 1.61 (1H, m)
 1.81 (1H, m)
1.84 (1H, dd, 11.2, 5.6)
1.56 (1H, br d, 5.6)
4.66 (1H, br s)
 0.97 (1H, m)
0.99 (1H, m)
 2.29 (1H, dt, 13.1, 4.6)
1.60 (1H, m)
1.82 (1H, m)
1.86 (1H, dd, 11.9, 5.7)
1.57 (1H, br d, 5.7)
4.68 (1H, br s)
 2.06 (1H, d, 10.2)
1.79 (1H, d, 10.2)
0.92 (3H, s)
 2.29 (1H, dt, 11.9, 3.7)
 1.60 (1H, br dd, 12.5, 3.7)
 1.83 (1H, m)
1.86 (1H, dd, 11.2, 5.6)
1.57 (1H, br d, 5.6)
4.67 (1H, br s)
16
18
19
21
22
 2.04 (1H, d, 10.4)
1.77 (1H, d, 10.4)
0.91 (3H, s)
2.05 (1H, d, 10.1)
1.78 (1H, br d, 10.1)
0.93 (3H, s)
 2.05 (1H, d, 10.2)
1.78 (1H, d, 10.2)
0.91 (3H, s)
23
24
0.70 (3H, s)
0.72 (3H, s)
0.71 (3H, s)
0.71 (3H, s)
25
0.74 (3H, s)
0.75 (3H, s)
0.75 (3H, s)
0.74 (3H, s)
26
0.85 (3H, s)
0.86 (3H, s)
0.86 (3H, s)
0.86 (3H, s)
27
0.93 (3H, s)
0.94 (3H, s)
0.94 (3H, s)
0.94 (3H, s)
29
1.16 (3H, s)
1.17 (3H, s)
1.17 (3H, s)
1.17 (3H, s)
30
1.07 (3H, s)
1.08 (3H, s)
1.08 (3H, s)
1.07 (3H, s)
glcA
1’
2’
3’
4’
4.35 (1H, d, 7.3)
3.24 (1H, dd, 7.9, 7.3)
3.44 (1H, m)
3.30 (1H, dd, 9.7, 8.9)
3.37 (1H, d, 9.7)
4.29 (1H, d, 6.1)
3.20 (1H, m)
3.34 (1H, m)
3.14 (1H, br t, 9.8)
3.28 (1H, d, 9.8)
4.25 (1H, d, 7.3)
3.22 (1H, br t, 7.3)
3.16 (1H, m)
3.29 (1H, dd, 9.5, 8.9)
3.32 (1H, d, 9.5)
4.38 (1H, d, 5.2)
3.43 (1H, m)
3.36 (1H, m)
3.35 (1H, m)
3.75 (1H, d, 9.5)
3.64 (3H, s)
5’
COOCH3
glc
1’’
2’’
3’’
4’’
5’’
6’’
4.28 (1H, d, 7.6)
3.04 (1H, m)
3.13 (1H, m)
3.06 (1H, m)
3.07 (1H, m)
4.74 (1H, d, 6.0)
3.16 (1H, m)
3.06 (1H, m)
3.06 (1H, m)
3.42 (1H, m)
3.45 (1H, br d, 10.7)
3.62 (1H, dd, 10.7, 5.2)
3.41, 3.63 (each 1H, m)
xyl
1’’’
2’’’
3’’’
4’’’
5’’’
4.45 (1H, d, 6.4)
3.01 (1H, m)
3.11 (1H, m)
3.27 (1H, m)
3.05 (1H, m)
3.73 (1H, dd, 11.3, 5.2)
C-3 rha 1
1’’’’
4.88 (1H, br s)
3.62 (1H, br s)
3.40 (1H, m)
3.15 (1H, m)
3.71 (1H, dq, 9.5, 6.1)
1.04 (1H, d, 6.1)
5.26 (1H, br s)
3.93 (1H, br s)
3.65 (1H, dt, 9.2, 2.1)
3.40 (1H, dt, 9.2, 2.1)
3.83 (1H, dq, 9.2, 6.1)
1.07 (1H, d, 6.1)
5.24 (1H, br s)
3.67 (1H, br s)
3.46 (1H, br d, 9.2)
3.16 (1H, br t, 9.2)
3.88 (1H, dq, 9.2, 6.1)
1.04 (1H, d, 6.1)
2’’’’
3’’’’
4’’’’
5’’’’
6’’’’
C-3 rha 2
1’’’’’
2’’’’’
3’’’’’
4’’’’’
5’’’’’
6’’’’’
4.73 (1H, br s)
3.66 (1H, br s)
3.40 (1H, m)
3.15 (1H, m)
3.91 (1H, dq, 9.2, 6.1)
1.04 (1H, d, 6.1)

Table 2-2. 1H-NMR Spectroscopic Data [dH, mult. (J in Hz)] of 5-8 (500 MHz, in DMSO-d6)
Position
5
6
7
8
1
0.81, 1.55 (each 1H, m)
0.88 (1H, m)
0.88, 1.46 (each 1H, m)
0.86 (1H, m)
1.54 (1H, d, 12.5)
1.53 (1H, m)
2
1.04, 1.73 (each 1H, m)
1.70 (1H, m)
1.93 (1H, br d, 11.3)
1.49 (1H, m)
3.03 (1H, dd, 11.3, 3.4)
0.66 (1H, br d, 8.2)
1.28 (1H, m)
1.47 (1H, m)
 1.51 (1H, m)
3.00 (1H, dd, 11.0, 4.0)
0.70 (1H, m)
 (1H, m)
3.04 (1H, dd, 11.6, 4.0)
0.66 (1H, br d, 11.3)
1.44 (1H, m)
3
5
6
3.03 (1H, dd, 11.3, 4.0)
0.67 (1H, br d, 10.7)
1.43 (1H, d, 8.5)
1.34 (1H, m)
1.30, 1.46 (each 1H, m)
1.33 (1H, m)
1.43 (1H, m)
7
1.33 (2H, m)
1.29 (2H, m)
1.41, 1.43 (each 1H, m)
1.34 (1H, m)
 (1H, m)
9
1.22 (1H, m)
1.25 (1H, m)
1.27 (1H, m)
1.45 (2H, m)
1.50 (1H, m)
1.76 (2H, m)
11
1.15, 1.39 (each 1H, m)
1.13 (1H, m)
1.40 (1H, m)
12
13
15
1.15, 1.46 (each 1H, m)
1.22 (1H, br d, 11.6)
0.99 (1H, m)
2.29 (1H, dt, 13.1, 5.1)
1.62 (1H, br dd, 13.1, 4.6)
1.83 (1H, m)
1.86 (1H, dd, 11.6, 6.0)
1.57 (1H, br d, 6.0)
4.68 (1H, br s)
0.99, 1.22 (each 1H, m)
1.26 (1H, dd, 12.0, 4.5)
1.00 (1H, m)
1.60 (2H, m)
5.18 (1H, br t, 3.6)
1.00 (1H, br d, 11.3)
0.99 (1H, m)
1.83 (1H, dd, 11.3, 3.4)
1.20 (1H, m)
1.67 (1H, m)
1.08 (1H, m)
1.01, 1.51 (each 1H, m)
2.19 (1H, dt, 13.0, 4.2)
1.69 (1H, m)
 1.83 (1H, br d, 11.3)
1.90 (1H, dd, 12.0, 6.9)
2.09 (1H, br d, 6.9)
4.56 (1H, br s)
16
1.59 (1H, m)
1.64 (1H, m)
2.82 (1H, dd, 13.7, 4.0)
1.59 (1H, m)
18
19
21
1.47 (1H, m)
3.46 (1H, m)
1.47 (1H, m)
1.89 (1H, d, 10.4)
1.60 (1H, m)
.47 (1H , m)
0.91 (3H, s)
0.71 (3H, s)
0.75 (3H, s)
0.81 (3H, s)
0.87 (3H, s)
22
1.72 (1H, d, 10.1)
1.95 (1H, d, 10.1)
0.91 (3H, s)
0.71 (3H, s)
0.74 (3H, s)
0.85 (3H, s)
0.94 (3H, s)
4.87, 4.88 (each 1H, br s)
2.05 (1H, d, 10.1)
 1.78 (1H, d, 10.1)
0.90 (3H, s)
0.70 (3H, s)
0.75 (3H, s)
0.86 (3H, s)
0.94 (3H, s)
1.18 (3H, s)
1.38 (1H, m)
1.61 (1H, m)
0.73 (3H, s)
0.94 (3H, s)
0.84 (3H, s)
0.65 (3H, s)
1.09 (3H, s)
0.94 (3H, s)
3.91, 4.26 (each 1H, d,
11.3)
23
24
25
26
27
29
0.91 (3H, d, 6.7)
30
1.05 (3H, s)
1.78 (3H, s)
1.21 (3H, s)
30-CH3CO
glcA
2.03 (3H, s)
1’
2’
3’
4’
4.57 (1H, d, 6.7)
3.38 (1H, t, 6.7)
3.54 (1H, t, 6.7)
3.61 (1H, m)
3.90 (1H, d, 9.2)
3.66 (3H,s)
4.35 (1H, d, 7.2)
3.35 (1H, t, 7.2)
3.52 (1H, m)
3.30 (1H, m)
3.69 (1H, m)
4.35 (1H, d, 7.5)
3.25 (1H, t, 7.5)
3.47 (1H, m)
3.31 (1H, t, 9.6)
3.38 (1H, d, 9.6)
4.25 (1H, d, 7.6)
3.17 (1H, m)
3.25 (1H, m)
3.30 (1H, m)
3.21 (1H, m)
5’
6-COOCH3
C-3 rha 1
1’’’’
4.85 (1H, br s)
3.64 (1H, br s)
3.40 (1H, m)
3.17 (1H, t, 9.3)
3.67 (1H, m)
4.88 (1H, br s)
3.64 (1H, br s)
3.40 (1H, m)
3.18 (1H, m)
3.69 (1H, m)
1.07 (1H, d, 6.1)
4.89 (1H, br s)
3.62 (1H, br s)
3.40 (1H, m)
3.16 (1H, m)
3.73 (1H, dq, 9.4, 6.1)
1.05 (1H, d, 6.1)
5.26 (1H, br s)
3.68 (1H, m)
3.45 (1H, m)
3.14 (1H, m)
3.79 (1H, m)
1.05 (1H, d, 6.1)
2’’’’
3’’’’
4’’’’
5’’’’
6’’’’
1.07 (1H, d, 6.1)
C-3 rha 2
1’’’’’
2’’’’’
3’’’’’
4’’’’’
5’’’’’
6’’’’’
C-28 rha
1’’’’’’
2’’’’’’
3’’’’’’
4’’’’’’
5’’’’’’
6’’’’’’
4.71 (1H, br s)
3.64 (1H, br s)
3.40 (1H, m)
3.19 (1H, t, 9.1)
3.74 (1H, dq, 9.1, 6.1)
1.07 (1H, d, 6.1)
4.72 (1H, br s)
3.67 (1H, br s)
3.39 (1H, m)
3.19 (1H, m)
3.76 (1H, dq, 9.2, 6.1)
1.04 (1H, d, 6.1)
4.74 (1H, br s)
3.67 (1H, br s)
3.40 (1H, m)
3.16 (1H, m)
3.92 (1H, dq, 9.5, 6.1)
1.04 (1H, d, 6.1)
5.72 (1H, br s)
3.44 (1H, m)
3.47 (1H, m)
3.23 (1H, m)
3.55 (1H, m)
1.10 (1H, d, 6.4)

EXPERIMENTAL
General experimental procedures
The []D values were determined with a JASCO DIP-140 digital polarimeter. IR spectra were measured
with a JASCO A-102 IR spectrophotometer. 1H- and 13C-NMR spectra were recorded using a JNM
LA-500 spectrometer in DMSO-d6 with tetramethylsilane as an internal standard. Kieselgel-60F254
(MERCK) or RP-18F254S (MERCK)-precoated plates were employed for thin-layer chromatography
(TLC). Column chromatography was carried out on 70-230 mesh silica gel (MERCK) and prep
ODS-7515-12A. HREIMS and HRFABMS were obtained using a JEOL JMS-700 spectrometer.
Plant materal
Stenocereus eruca A.C. Gibson & K. E. Horak was cultivated originally at the Hokoen (Iga City, Mie,
Japan).
Extraction and isolation
The dried and powdered whole plants of S. eruca (432.7 g) were extracted three times with CHCl3 and
then extracted three times with MeOH. The MeOH extract (83.0 g, 19.2%) was applied to a column of
Diaion HP-20 and eluted with 30% MeOH in H2O (1.4 g), 70% MeOH in H2O (20.4 g), and 100% MeOH
(20.8 g). Since the same spot was observed in the 70% MeOH and 100% MeOH eluted fractions, these
fractions were mixed (41.2 g, 9.5%). The mixed fraction was separated by silica gel column
chromatography using a stepwise gradient [Silica gel: 800 g, CHCl3–MeOH–H2O 90:12:1→70:10:1→
60:10:1→50:10:1→40:10:1→35:10:1→30:10:1→25:10:1→20:10:1→15:6:1→13:6:1→10:5:1→8:5:1→
6:4:1→MeOH] to give seven fractions [Fr. A (1.3 g), Fr. B (8.6 g), Fr. C (6.0 g), Fr. D (7.8 g), Fr. E (6.8
g), Fr. F (6.1 g), and Fr. G (3.6 g)].
Fraction C (6.0 g) was further subjected to silica gel column chromatography using a stepwise gradient
[Silica gel: 220 g, CHCl3–MeOH–H2O 70:10:1→60:10:1→50:10:1→40:10:1→30:10:1→20:10:1→
15:10:1→MeOH] to give seven fractions [Fr. C1 (39.1 mg), Fr. C2 (775.8 mg), Fr. C3 (2031.6 mg), Fr.
C4 (1170.0 mg), Fr. C5 (1024.6 mg), Fr. C6 (752.2 mg), and Fr. C7 (42.6 mg)]. Fraction C2 (775.8 mg)
was chromatographed on silica gel column chromatography using a stepwise gradient [Silica gel: 70 g,
CHCl3–MeOH–H2O 80:10:1→70:10:1→60:10:1→50:10:1→MeOH] to give four fractions [Fr. C2-1
(140.8 mg), Fr. C2-2 (132.8 mg), Fr. C2-3 (138.7 mg) and Fr. C2-4 (358.0 mg)]. Fraction C2-4 (358.0
mg) was separated by octadecyl silylated silica gel (ODS) column chromatography using a stepwise
gradient [50% MeOH→55% MeOH→65% MeOH→80% MeOH→MeOH] to afford stellatoside C (1,
109.9 mg, 0.03%).
Fraction C6 (752.2 mg) was separated by ODS column chromatography using a stepwise gradient [30%
MeOH→40% MeOH→50% MeOH→60% MeOH→MeOH] to afford stellatoside D (2, 64.1 mg 0.01%).
Fraction B (8.6 g) was separated by silica gel column chromatography using a stepwise gradient [Silica

gel: 400 g, CHCl3–MeOH–H2O 60:10:1→55:10:1→45:10:1→40:10:1→30:10:1→20:10:1→MeOH] to
afford four fractions [Fr. B1 (155.9 mg), Fr. B2 (696.4 mg), Fr. B3 (2217.6 mg), and Fr. B4 (5487.6 mg)].
Fraction B2 (696.4 mg) was separated by ODS column chromatography using a stepwise gradient [60%
MeOH→70% MeOH→80% MeOH→MeOH] to give twelve fractions [Fr. B2-1 (323.1 mg), Fr. B2-2
(26.9 mg), Fr. B2-3 (21.1 mg), Fr. B2-4 (17.7 mg), Fr. B2-5 (4.4 mg), Fr. B2-6 (3.4 mg), Fr. B2-7 (13.4
mg), Fr. B2-8 (19.2 mg), Fr. B2-9 (155.2 mg), Fr. B2-10 (50.1 mg), Fr. B2-11 (3.3 mg), and Fr. B2-12
(65.9 mg)]. Fr. B2-9 (155.2 mg) was subjected to silica gel column chromatography using a stepwise
gradient [Silica gel: 50 g, CHCl3–MeOH–H2O 60:10:1→MeOH] to afford stellatoside C methyl ester (5,
73.6 mg, 0.02%). Furthermore, fraction B4 (5487.6 mg) was separated on silica gel column
chromatography using a stepwise gradient [Silica gel: 250 g, CHCl3–MeOH–H2O 55:10:1→50:10:1→
45:10:1→40:10:1→30:10:1→20:10:1→15:10:1→MeOH] to give four fractions [B4-1 (48.3 mg), B4-2
(4228.0 mg), B4-3 (282.8 mg), and B4-4 (977.2 mg)]. Fr. B4-2 (4228.0 mg) was chromatographed on
silica gel column chromatography using a stepwise gradient [Silica gel: 150 g, CHCl3–MeOH–H2O
70:10:1→60:10:1→50:10:1→40:10:1→30:10:1→MeOH] to give five fractions [B4-2-1 (49.4 mg),
B4-2-2 (1389.0 mg), B4-2-3 (1860.2 mg), B4-2-4 (479.0 mg), and B4-2-5 (276.3 mg)]. Fraction B4-2-4
(479.0 mg) was separated by ODS column chromatography to afford five fractions [B4-2-4-1 (20.6 mg),
B4-2-4-2 (79.2 mg), B4-2-4-3 (161.9 mg), B4-2-4-4 (129.9 mg), and B4-2-4-5 (42.0 mg)]. Fraction
B4-2-4-3 (161.9 mg) was identified as stellatoside C (1, 161.9 mg, 0.04%).
Fraction B3 (2217.6 mg) was separated by silica gel column chromatography using a stepwise gradient
[Silica gel: 80 g, CHCl3–MeOH–H2O 70:10:1→60:10:1→50:10:1→40:10:1→30:10:1→20:10:1→
MeOH] to afford three fractions [Fr. B3-1 (184.7 mg), Fr. B3-2 (1848.4 mg) and Fr. B3-3 (21.3 mg)].
Fraction B3-2 (1848.4 mg) was separated by ODS column chromatography using a stepwise gradient
[60% MeOH→80% MeOH→MeOH] to give three fractions [Fr. B3-2-1 (348.7 mg), Fr. B3-2-2 (1523.0
mg) and Fr. B3-2-3 (45.8 mg)]. Fraction B3-2-2 (1523.0 mg) underwent ODS column chromatography
using a stepwise gradient [60% MeOH→80% MeOH→MeOH] to give seven fractions [Fr. B3-2-2-1
(86.0 mg), Fr. B3-2-2-2 (124.3 mg), Fr. B3-2-2-3 (74.8 mg), Fr. B3-2-2-4 (184.1 mg), Fr. B3-2-2-5
(203.8 mg) Fr. B3-2-2-6 (327.9 mg), and Fr. B3-2-2-7 (385.9 mg)]. Fraction B3-2-2-3 (74.8 mg) was
purified by ODS column chromatography using a stepwise gradient [60% MeOH→MeOH] to give
stellatoside E (3, 21.9 mg, 0.005%) and stellatoside B methyl ester (4, 49.8 mg, 0.01%).
Fraction B3-2-2-7 (385.9 mg) was separated by ODS column chromatography using a stepwise gradient
[70% MeOH→MeOH] to give four fractions [Fr. B3-2-2-7-1 (49.0 mg), Fr. B3-2-2-7-2 (8.0 mg), Fr.
B3-2-2-7-3 (151.9 mg), and Fr. B3-2-2-7-4 (126.8 mg)] and thurberoside A (6, 71.1 mg, 0.02%).
Fraction B3-2-2-7-3 (151.9 mg) was repeatedly chromatographed on silica gel using a stepwise gradient
[Silica gel: 50 g, CHCl3–MeOH–H2O 60:10:1→50:10:1→40:10:1→30:10:1→MeOH] to afford four

fractions [Fr. 3-2-2-7-3-1 (4.6 mg), Fr. B3-2-2-7-3-2 (8.7 mg), Fr. B3-2-2-7-3-3 (120.5 mg), and Fr.
B3-2-2-7-3-4 (9.6 mg)]. Fraction B2-2-2-7-3-3 (120.5 mg) was subjected to ODS column
chromatography using a stepwise gradient [50% MeOH→60% MeOH→65% MeOH→75% MeOH→
MeOH] to obtain phillyriside A (7, 20.3 mg, 0.005%).
Fraction E (6.85 g) was separated by silica gel column chromatography using a stepwise gradient [Silica
gel: 240 g, CHCl3–MeOH–H2O 40:10:1→30:10:1→25:10:1→20:10:1→15:10:1→10:6:1→6:4:1→
MeOH] to give eight fractions [Fr. E1 (81.3 mg), Fr. E2 (272.4 mg), Fr. E3 (714.8 mg), Fr. E4 (1158.2
mg), Fr. E5 (882.7 mg), Fr. E6 (689.4 mg), Fr. E7 (1906.7 mg), and Fr. E8 (3.6 g)]. Fraction E3 (714.8
mg) was repeatedly chromatographed on silica gel using a stepwise gradient [Silica gel: 77 g,
CHCl3–MeOH–H2O 30:10:1→25:10:1→20:10:1→MeOH] to give four fractions [Fr. E3-1 (5.6 mg), Fr.
E3-2 (88.0 mg), Fr. E3-3 (429.9 mg), and Fr. B3-4 (22.0 mg)]. Fraction E3-3 (429.9 mg) was separated
by ODS column chromatography using a stepwise gradient [50% MeOH→60% MeOH→MeOH] to give
four fractions [Fr. E3-3-1 (92.8 mg), Fr. E3-3-2 (144.2 mg), Fr. E3-3-3 (22.8 mg), and Fr. B3-3-4 (161.4
mg)]. Fraction E3-3-1 (92.8 mg) was separated by silica gel column chromatography using a stepwise
gradient [Silica gel: 25 g, CHCl3–MeOH–H2O 30:10:1→20:10:1→MeOH] to give two fractions [Fr.
E3-3-1-1 (26.5 mg) and Fr. E3-3-1-2 (39.0 mg)]. Fraction E3-3-1-2 (39.0 mg) was separated by ODS
column chromatography using a stepwise gradient [40% MeOH→50% MeOH→MeOH] to give
treleaseside A (8, 13.8 mg, 0.003%).
Stellatoside C (1)
27
Colorless powder (279.9 mg); []D -110.4 (c 1.00, MeOH); IR max (KBr): 3410, 2950, 1740, 1600,
1450, 1390, 1040 cm-1; positive FABMS m/z: 941 [M+H]+, 795 [M-rha+H]+; negative FABMS m/z: 939
[M-H]-, 793 [M-rha-H]-; positive HRFABMS m/z: 941.5127 (calcd. for C48H77O18 [M+H]+, 941.5110);
1H- and 13C-NMR, see Tables 1 and 2.
Stellatoside D (2)
Colorless powder (64.1 mg); []D27 -114.1 (c 1.00, MeOH); IR max (KBr): 3400, 2950, 1760, 1630, 1040
cm-1; negative FABMS m/z: 955 [M-H]-, 793 [M-glc-H]-, 647 [M-glc-rha-H]-; negative HRFABMS m/z:
955.4916 (calcd. for C48H75O19 [M-H]-, 955.4903); 1H- and 13C-NMR, see Tables 1 and 2.
Stellatoside E (3)
Colorless powder (21.9 mg); []D22 -32.2 (c 1.00, MeOH); IR max (KBr): 3410, 2950, 1760, 1600, 1450,
1390, 1040 cm-1; positive FABMS m/z: 795 [M+H]+; negative FABMS m/z: 793 [M-H]-, 647 [M-rha-H]-;
1
negative HRFABMS m/z: 793.4370 (calcd. for C42H65O14 [M-H]-, 793.4374); H- and 13C-NMR, see
Tables 1 and 2.
Stellatoside B methyl ester (4)

Colorless powder (49.8 mg); []D23 -16.1 (c 1.00, MeOH); IR max (KBr): 3410, 2950, 1750, 1640, 1450,
1360, 1040 cm-1; positive FABMS m/z: 957 [M+H]+, 825 [M-xyl+H]+; negative FABMS m/z: 955 [M-H]-,
1
823 [M-xyl-H]-; negative HRFABMS m/z: 955.4915 (calcd. for C48H75O19 [M-H]-, 955.4903); H- and
13C-NMR, see Tables 1 and 2.
Stellatoside C methyl ester (5)
Colorless powder (73.6 mg); []D27 -144.5 (c 1.00, MeOH); IR max (KBr): 3410, 2950, 1760, 1640, 1450,
1390, 1040 cm-1; positive FABMS m/z: 955 [M+H]+, 794 [M-glc+H]+, 647 [M-glc-rha+H]+; positive
HRFABMS m/z: 955,5283 (calcd. for C49H79O18 [M+H]+, 955.5266); 1H- and 13C-NMR, see Tables 1 and
2.
Thurberoside A (6)
Colorless powder (71.1 mg); []D27 -31.7 (c 1.00, MeOH); IR max (KBr): 3400, 2950, 1760, 1640, 1450,
1390, 1050 cm-1; positive FABMS m/z: 923 [M+H]+; negative FABMS m/z: 921 [M-H]-, 775 [M-rha-H]-;
1
positive HRFABMS m/z: 923.4998 (calcd. for C49H79O18 [M+H]+, 923.5004); H- and 13C-NMR, see
Tables 1 and 2.
Phillyriside A (7)
Colorless powder (20.3 mg); []D23 -18.5 (c 1.00, MeOH); IR max (KBr): 3410, 2950, 1740, 1600, 1450,
1400, 1040 cm-1; positive FABMS m/z: 925 [M+H]+, 779 [M-rha+H]+; negative FABMS m/z: 923 [M-H]-,
1
778 [(M-rha-H)-; positive HRFABMS m/z: 925.5161 (calcd. for C48H77O17 [M+H]+, 925.5161); H- and
13C-NMR, see Tables 1 and 2.
Treleaseside A (8)
23
Colorless powder (13.8 mg); []D -8.7 (c 0.30, MeOH); IR max (KBr): 3400, 2930, 1720, 1610, 1460,
1390, 1250, 1140, 1040 cm-1; negative FABMS m/z: 997 [M-H]-, 851 [M-rha-H]-; negative HRFABMS
m/z: 997.5016 (calcd. for C50H77O20 [M-H]-, 997.5008); 1H- and 13C-NMR, see Tables 1 and 2.
Acidic hydrolysis of 1
o
1 (20 mg) was hydrolyzed with 5 mL of 3.5 % HCl and heated at 110 C for 3 h. The reaction mixture
was neutralized with 1M NaOH, extracted with CHCl3. CHCl3 extract was chromatographed on silica gel
column chromatography (CHCl3:MeOH = 100:1), to afford aglycon of 1 (7.6 mg). The aglycon was
identified as stellatogenin by TLC with authentic sample and comparison with 1H- and 13C-NMR.13,26
Acidic hydrolysis of 6, 7
6, 7 (8 mg) were hydrolyzed and separated with similar way as in the case of 1, to afford thurberogenin25
(2.3 mg) and 27-desoxyphillyrigenin16 (2.8 mg). These aglycons were identified by TLC with authentic
samples and comparison with 1H- and 13C-NMR, respectively.16,25
Determination of sugar configuration

Saponins (1-8) (each 2 mg) were hydrolyzed by 3.5% HCl (0.4 mL) at 110 °C for 2.5 h. The reaction
mixture was neutralized with 0.5 M NaOH. After drying under vacuum, the residue was dissolved in
0.4 mL pyridine containing 2 mg of L-cysteine methyl ester hydrochloride and heated at 60 °C for 1 h.
O-Tolylisothiocyanate (2 L) was then added and the mixture was heated at 60 °C for 1 h. The reaction
mixture was directly analyzed by reversed-phase HPLC using a SSC-3461 HPLC Pump with a SSC-5410
UV/VIS detector (Senshu Scientific Co., Ltd.). A Senshu Pak PEGASIL 4.6   250 mm HPLC column
was used (temp, 35 °C; flow, 0.8 mL/min; eluate, MeCN-H2O 25:75 containing 50 mM H3PO4). The
HPLC column was washed with MeOH after each injection. The reaction conditions for D, L-glucose,
D-glucuronic acid, D, L-xylose and L-rhamnose were the same as described above. In the case of the
L-glucuronic acid and D-rhamnose, D-glucuronic acid or L-rhamnose dissolved in 0.4 mL pyridine
containing 2 mg D-cysteine methyl ester hydrochloride was used for the reaction, respectively. The
derivative was used for determining the retention time instead of L-glucuronic acid, D-rhamnose. The
retention times for the authentic sugar derivatives, D-glucose (19.49), L-glucose (17.98), D-xylose (22.66),
L-xylose (21.10), D-glucuronic acid (20.34), L-glucuronic acid (derived from D-glucuronic acid and
D-cysteine methyl ester, 19.51), L-rhamnose (33.10) and D-rhamnose (derived from L-rhamnose and
D-cysteine methyl ester, 17.69) were used for comparison with the retention times from the reaction
mixtures for each saponin.24 The peaks at 20.28 and 33.09 of the sugar derivatives from 8 coincided
with derivatives of D-glucuronic acid and L-rhamnose, respectively.21 For the other saponins (1-7), the
same results as for compound 8 were obtained.
ACKNOWLEDGEMENTS
This work was partly supported by a Sasakawa Scientific Research Grant from The Japan Science Society.
The authors would like to thank Mrs. S. Kubota and Mrs. T. Koseki for the spectroscopic (NMR or MS)
measurements. This work was partially supported by a grant from the High-Tech Research Center
Project, the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan (No.
S0801043).
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25. 1H-NMR data of thurberogenin in C5D5N  ppm; 0.83 (3H, s, H3-25), 0.98 (3H, s, H3-27), 1.03 (6H,
s, H3-24, H3-26), 1.23 (3H, s, H3-23), 1.80 (3H, s, H3-30), 3.45 (1H, br t, J = 8.1 Hz, H-3), 4.59 (1H.
d, J = 1.9 Hz, H-21), 4.91, 4.97 (each 1H, d, J = 1.1 Hz, H2-29).
13C-NMR data of thurberogenin in C5D5N  ppm; 39.1 (C-1), 28.2 (C-2), 78.0 (C-3), 39.4 (C-4),
55.8 (C-5), 18.6 (C-6), 34.5 (C-7), 40.9 (C-8), 50.6 (C-9), 37.4 (C-10), 20.8 (C-11), 26.9* (C-12),

40.4 (C-13), 42.5 (C-14), 26.5* (C-15), 23.7 (C-16), 51.5 (C-17), 44.9 (C-18), 53.1 (C-19), 144.2
(C-20), 81.7 (C-21), 44.4 (C-22), 28.6 (C-23), 16.0 (C-24), 16.3 (C-25), 16.3(C-26), 14.0 (C-27),
178.2 (C-28), 112.5 (C-29), 23.3 (C-30), (*: may be interchanged).
26. 1H-NMR data of stellatogenin in C5D5N  ppm; 3.43 (1H, brt, J = 8.0 Hz, H-3), 5.01 (1H. brs, H-21),
1.21 (3H, s, H3-23), 1.01 (3H, s, H3-24), 0.83 (3H, s, H3-25), 1.08 (3H, s, H3-26), 0.95 (3H, s, H3-27),
1.48 (3H, s, H3-29), 1.39 (3H, s, H3-30).