Steroidal saponins from fresh stem of Dracaena cochinchinensis

Article abstract of DOI:10.1016/j.steroids.2003.11.004

A further phytochemical investigation on the fresh stem of Dracaena cochinchinensis yielded 18 steroidal saponins. Fourteen of which are new compounds, designated as 25(R,S)-dracaenosides E-H, M, O-Q, dracaenosides I-L, R, and 25(S)-dracaenoside N. Their structures were determined by detailed spectroscopic analysis, including extensive 1D and 2D NMR data, and the result of hydrolytic reaction.

Full text of DOI:10.1016/j.steroids.2003.11.004

Steroids 69 (2004) 111–119
Steroidal saponins from fresh stem of Dracaena cochinchinensis
∗
Qing-An Zheng, Ying-Jun Zhang , Hai-Zhou Li, Chong-Ren Yang
State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany,
Chinese Academy of Sciences, Kunming 650204, PR China
Received 27 September 2003; accepted 21 November 2003
Abstract
A further phytochemical investigation on the fresh stem of Dracaena cochinchinensis yielded 18 steroidal saponins. Fourteen of which
are new compounds, designated as 25(R,S)-dracaenosides E–H, M, O–Q, dracaenosides I–L, R, and 25(S)-dracaenoside N. Their structures
were determined by detailed spectroscopic analysis, including extensive 1D and 2D NMR data, and the result of hydrolytic reaction.
©
2004 Elsevier Inc. All rights reserved.
Keywords: Dracaena cochinchinensis; Agavaceae; Steroidal saponins; Dracaenosides E–R
1
. Introduction
2. Experimental
2.1. General methods
The genus Dracaena (Agavaceae) containing about 60
species is distributed from the Old World tropic region to
Canary Island. The resins from stems of several species
of this genus as a source of dragon’s blood were used as
traditional medicine from ancient time. In China, the red
resin of Dracaena cochinchinensis (Lour.) S.C. Chen, called
Optical rotations were measured on a SEPA-3000 auto-
matic digital polarimeter. NMR spectra were run on Bruker
1
13
AM-400 (for H NMR and C NMR) and DRX-500 (for
D NMR) instruments with TMS as internal standard; IR
2
spectra were measured on a Bio-Rad FTS-135 spectrom-
eter with KBr pellets. FAB-MS spectra were recorded
on a VG Auto Spec-300 spectrometer. UV spectra were
obtained on a Shimadzu double-beam 210A spectropho-
tometer. GC–MS was run on Fisons MD-800 GC/MS in-
struments using 30QC2/AC-5 fused silica capillary column
“
longxuejie”, was used as Chinese dragon’s blood for pro-
moting blood circulation and the treatment of traumatic and
visceral hemorrhages [1]. In previous study, a lot of phe-
nolic compounds were isolated from the resins [2–6], and
steroids, such as protogracillin, gracillin, protodioscin and
dioscin, were obtained from the fruits of this plant [7]. Re-
cently, we reported four new pregnane glycosides, dracaeno-
sides A–D (1–4) from the fresh stem of D. cochinchinen-
sis [8,9]. In a continuation of a study on Chinese dragon’s
blood and its origin plants, we have further phytochemical
investigated the fresh stem, resulting in the isolation of 14
new steroidal saponins, designated as 25(R,S)-dracaenosides
E–H (5–8), M (13), and O–Q (15–17), dracaenosides I–L
(
30 m × 0.25 mm) in following conditions—filament cur-
◦
rent: 4.2 A; column temperature: 180/260 C, programmed
◦
increase, 5 C/min; carrier gas: He; head pressure: 12 psi;
EI-MS: 70 eV; ion source temperature: 250 C. Silica gel
◦
(
200–300 mesh and 10–40 ␮m), RP-18 (40–63 ␮m) and
Sephadex LH-20 were used for column chromatography.
2
.2. Plant material
(9–12) and R (18), and 25(S)-dracaenoside N (14), together
with four known 25(R,S)-steroid saponins epimers (19–22).
The present paper deals with the experimental details of sep-
aration and structure elucidation of these new compounds on
the basis of the spectroscopic analysis, including 2D NMR
techniques, and the result of hydrolytic reaction.
The fresh stems of D. cochinchinensis were collected from
Xishuangbanna, Yunnan, China, and a voucher specimen is
deposited at the Herbarium of Kunming Institute of Botany,
Chinese Academy of Sciences.
2
.3. Extraction and isolation
∗
Corresponding authors. Tel.: +86-871-5223424;
fax: +86-871-5150124.
The chipped fresh stem of D. cochinchinensis (25.0 kg)
E-mail addresses: cryang@public.km.yn.cn,
zhangyj@mail.kib.ac.cn (Y.-J. Zhang).
was extracted with hot MeOH. The MeOH extract was
0
039-128X/$ – see front matter © 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.steroids.2003.11.004

1
12
Q.-A. Zheng et al. / Steroids 69 (2004) 111–119
condensed under reduced pressure. The viscous concentrate
was partitioned between H2O and n-BuOH successively.
The n-butanol fraction was submitted through a column
chromatography of high porous absorption resin (Diaion
HP-20), eluting with H2O and methanol. The methanol
fraction (150 g) was repeatedly column chromatographed
(CC) over normal and reverse phase silica gel to afford three
fractions (Frs. I–III). Fr. I was subjected to ODS CC eluting
with MeOH–H2O and silica gel with CHCl3–MeOH–H2O
to give compounds 1 (40 mg), 2 (26 mg), 3 (36 mg), 4
(25 mg), 5 (30 mg) and 6 (17 mg). Fr. II was subjected to
ODS CC eluting with MeOH–H2O and silica gel CC with
CHCl3–MeOH–H2O to give 19 (50 mg), 20 (56 mg), 21
(
1
(
200 mg), 22 (67 mg), 7 (33 mg), 8 (150 mg), 9 (677 mg),
0 (56 mg), 11 (100 mg), 12 (200 mg), 13 (123 mg), 14
70 mg), 15 (15 g) and 16 (2.5 mg). Fr. III was subjected
to ODS CC eluting with MeOH–H2O and silica gel CC
with CHCl3–MeOH–H2O to give 15 (4.0 g), 16 (1.5 g), 17
(35 mg) and 18 (18 mg).
13
d, J = 5.50 Hz, 27-CH3, 25R). C NMR (pyridine-d ): see
5
Tables 1 and 2.
2
.3.2. 25(R,S)-Dracaenoside F (6)
White amorphous powder, C39H O13, [α] − 45.17
2
8
◦
62
d
(
[
[
MeOH, c = 0.2). Negative ion HRFAB-MS, m/z: 737.4126
−
M−H] (calcd. 737.4112). Negative ion FAB-MS, m/z: 737
−
+
KBr
max
1
−1
M−H] , 592 [M−146] . IR,ν
(cm ): 3420 (br, OH),
2
938, 2866, 1642, 915, 894, 870; H NMR (pyridine-d ), δ
5
ꢀꢀ
ppm): 6.32 (1H, brs, 1 -H), 5.39 (1H, brs, 6-H), 4.96 (1H,
(
ꢀ
ꢀꢀ
2
.3.1. 25(R,S)-Dracaenoside E (5)
d, J = 6.4 Hz, 1 -H), 1.76 (3H, d, J = 6.16 Hz, 6 -CH3),
1.19 (d, J = 6.25 Hz, 21-CH3, 25R), 1.18 (3H, d, J =
6.0 Hz, 21-CH3, 25S), 1.07 (3H, s, 18-CH3), 0.99 (3H, s,
White amorphous powder, C39H O13, [α] − 71.55◦
28
62
d
(
[
7
MeOH, c = 0.2). Negative ion HRFAB-MS, m/z: 737.4066
−
M − H] (calcd. 737.4112). Negative ion FAB-MS, m/z:
19-CH3), 1.08(3H, d, J = 2.10 Hz, 27-CH3, 25S), 0.68 (d,
+
+
−
13
38 [M] , 592 [M − 146] , 429 [M − 146 − 162 − H] .
J = 5.50 Hz, 27-CH3, 25R). C NMR (pyridine-d ): see
5
KBr
−1
IR,ν
(cm ): 3428 (br, OH), 2940 (CH), 2868 (CH),
Tables 1 and 2.
max
1
1
645, 917, 898, 878; H NMR (pyridine-d ), δ (ppm): 5.89
5
ꢀ
ꢀ
(
7
1H, brs, 1 -H), 5.40 (1H, brs, 6-H), 4.94 (1H, d, J =
2.3.3. 25(R,S)-Dracaenoside G (7)
ꢀ
ꢀꢀ
20
◦
.70 Hz, 1 -H), 1.72 (3H, d, J = 5.85 Hz, 6 -CH3), 1.18
White amorphous powder, C H72O17, [α] − 102.63
45
d
(
6
3H, d, J = 6.25 Hz, 21-CH3, 25R), 1.19 (3H, d, J =
(MeOH, c = 0.2). Negative ion HRFAB-MS, m/z: 883.4687
+
.0 Hz, 21-CH3, 25S), 1.07 (3H, s, 18-CH3), 0.99 (3H, s,
9-CH3), 1.07 (3H, d, J = 2.10 Hz, 27-CH3, 25S), 0.67 (3H,
[M − H] (calcd. 883.4691). Negative ion FAB-MS, m/z:
+
+
KBr
−1
(cm ): 3428
1
883 [M − H] , 737 [M − 146] . IR,ν
max

Table 1
1
3C NMR signals of the aglycon moieties of the dracaenosides 5–22
Position
5 (R/S)
6 (R/S)
7 (R/S)
8 (R/S)
9
10
37.6
11
37.5
12
37.637.8
13
37.5
14
37.9
15
37.7
16
37.6
17
37.5
18
37.2
19 (R/S)
20 (R/S)
21
37.3
22
1
2
3
4
5
6
7
8
9
37.5
30.3
78.2
39.4
140.5
122.5
26.8
35.7
43.7
37.8
20.5
32.1
45.2
86.5
39.9
82.0
37.6
30.1
78.3
39.1
140.4
122.5
26.8
35.7
43.7
37.8
20.5
32.0
45.3
86.6
40.0
82.1
82.2
60.0
59.9
20.1
19.5
42.6
42.4
15.5
15.4
109.7
110.2
32.1
27.7
29.5
26.7
30.1
26.4
66.9
65.2
17.5
16.5
37.6
30.1
78.3
39.1
140.5
122.5
26.9
35.8
43.7
37.9
20.5
32.1
45.2
86.6
40.0
82.1
82.2
60.0
59.9
20.2
19.5
42.6
42.4
15.4
15.4
109.8
110.3
32.1
27.7
29.5
26.7
30.1
26.4
67.0
65.2
17.5
16.4
37.6
30.2
77.9
37.5
30.1
78.5
38.8
37.3
30.3
78.5
39.0
140.9
121.9
32.3
31.7
50.4
37.6
21.1
39.9
40.5
56.7
32.2
81.2
81.5
62.9
62.8
16.4
19.5
42.0
42.5
15.1
15.0
109.3
109.8
31.9
26.5
29.3
26.3
30.7
27.6
66.9
65.1
17.4
16.4
37.2
30.2
78.7
38.8
140.9
122.0
32.5
31.8
50.5
37.6
21.2
40.0
40.9
56.7
32.6
81.3
81.5
62.9
62.8
16.6
19.6
42.0
42.4
15.1
15.0
109.4
109.7
30.2
26.6
28.4
26.4
30.6
27.5
66.8
65.0
17.5
16.3
30.2
78.5
39.4
140.3
122.5
26.8
35.7
43.7
37.8
20.5
32.0
45.2
86.5
39.9
82.0
30.3
78.2
39.1
140.4
122.4
26.7
35.7
43.7
37.8
20.4
29.9
45.1
86.5
39.9
82.4
30.1
78.5
39.9
140.3
122.5
26.8
35.7
43.6
37.8
20.5
31.9
45.1
86.5
38.8
82.4
30.2
78.6
40.0
140.4
122.4
26.7
35.7
43.7
37.8
20.4
31.9
45.5
86.4
39.0
82.2
30.2
78.3
40.0
140.3
122.4
26.7
35.5
43.7
37.9
20.4
31.8
45.2
86.2
39.0
82.2
30.3
78.2
39.1
140.4
122.6
26.9
35.8
43.8
37.9
20.5
30.7
45.6
86.5
40.2
81.9
30.3
78.6
39.9
140.5
122.7
26.9
35.8
43.8
37.9
20.6
32.0
45.7
86.6
38.9
82.0
30.4
78.6
39.2
140.4
122.6
26.9
35.2
43.8
37.8
20.7
32.1
48.1
85.6
42.6
85.0
30.2
78.1
39.0
140.4
122.4
26.7
35.6
43.7
37.8
20.4
31.9
45.2
86.3
39.9
82.0
30.2
78.6
39.0
140.9
122.0
32.4
31.7
50.4
37.5
21.1
39.8
40.8
56.6
32.4
81.4
30.3
78.7
38.8
140.9
122.0
32.5
31.8
50.5
37.6
21.2
40.0
40.9
56.7
32.6
81.3
38.8
140.4
122.6
26.8
35.8
43.7
37.8
20.5
32.1
45.2
86.6
40.0
82.0
82.1
59.9
59.8
20.1
19.5
140.8
121.9
32.2
31.8
50.3
37.2
21.2
39.9
40.6
56.7
32.4
81.5
1
1
1
1
1
1
1
0
1
2
3
4
5
6
8
2.1
60.0
9.9
1
7
62.9
59.9
59.5
59.5
60.1
60.1
60.7
60.8
62.2
59.7
64.2
63.9
5
1
1
2
8
9
0
20.2
19.4
42.6
16.4
19.5
41.9
20.1
19.4
39.2
19.9
19.4
42.7
20.1
19.4
42.7
20.3
19.5
40.1
18.7
19.4
40.1
20.3
19.5
40.9
20.4
19.6
40.9
18.1
19.6
105.3
19.9
19.4
38.7
16.3
19.5
40.8
16.6
19.6
40.9
42.2
42.7
15.5
15.3
4
2.0
15.5
5.3
109.9
21
22
23
24
25
26
27
15.1
109.5
33.3
15.4
110.1
26.8
24.2
42.2
64.5
64.1
15.2
111.8
36.3
66.6
35.9
64.6
9.8
15.3
111.9
36.2
66.6
35.9
64.6
9.8
16.7
110.3
31.2
27.7
34.5
75.3
17.7
16.7
110.3
31.0
27.7
34.3
75.3
17.6
16.9
111.2
32.1
28.5
34.6
75.3
17.7
16.9
111.3
32.2
28.6
34.5
75.1
17.7
12.1
154.4
31.9
26.9
31.2
75.3
17.9
15.6
120.9
33.9
28.8
88.4
66.9
65.4
16.4
110.9
30.2
28.2
34.5
75.3
17.6
16.6
110.9
30.2
28.4
34.5
75.4
17.6
1
109.7
110.2
32.05
26.69
29.46
26.38
30.72
27.67
66.96
65.21
17.46
16.45
110.2
32.1
2
7.7
29.4
6.7
30.7
6.4
66.9
5.2
17.45
6.43
29.0
2
144.5
65.0
2
6
108.8
1

1
14
Q.-A. Zheng et al. / Steroids 69 (2004) 111–119
Table 2
1
3C NMR signals of the sugar moieties of the saponins 5–22 (in C5D5N)
Position
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
Glc-1
102.4 100.4 100.3
99.9 100.0
99.9 100.3
99.9 102.1 100.3 100.3 100.0 100.3 100.2 100.3
99.8 100.4 100.0
2
3
4
5
6
75.6
76.8
78.4
77.1
61.6
78.3
79.7
71.9
78.6
62.8
78.1
76.9
77.9
78.0
61.3
77.2
89.5
69.7
77.8
62.5
78.7
89.5
69.7
77.9
62.5
78.5
89.5
77.1
78.8
62.5
78.9
77.9
78.0
76.9
61.4
78.5
89.5
77.1
78.7
62.5
75.3
77.0
78.3
77.9
61.2
78.3
79.7
71.8
78.7
61.3
78.7
78.0
78.2
77.0
61.4
78.8
89.4
77.3
77.9
62.6
78.8
89.4
77.0
78.1
61.5
78.6
77.9
78.0
77.0
61.3
78.2
77.9
78.7
76.9
61.3
78.5
89.4
77.0
78.7
62.4
78.6
77.8
78.1
76.9
61.3
78.5
77.8
78.6
77.2
62.5
Rha-1
102.2 102.1 102.3 102.3 102.3 102.0 102.3
102.2 102.1 102.3 102.2 102.1 102.1 102.2 102.2 102.3
2
3
4
5
6
72.7
72.9
74.2
69.6
18.8
72.6
72.8
74.2
69.6
18.7
72.5
72.9
74.2
69.7
18.8
72.5
72.8
74.1
69.7
18.8
72.5
72.8
74.2
69.6
18.8
72.5
72.8
74.2
69.5
18.7
72.5
72.8
74.1
69.7
18.8
72.8
72.9
74.2
69.6
18.7
72.6
72.8
74.2
69.6
18.6
72.6
72.9
74.2
69.8
18.9
72.7
72.9
74.2
69.7
18.7
72.6
72.8
74.2
70.0
18.6
72.6
72.8
73.9
69.6
18.7
72.5
72.8
74.1
69.6
18.7
72.6
72.8
74.2
69.6
18.7
72.5
72.9
74.2
69.7
18.8
ꢀ
Glc -1
104.6 104.6 104.6
104.6
75.0
77.6
71.5
77.9
62.5
104.6 104.9
104.6
75.0
77.7
71.5
77.9
62.4
104.9
75.0
77.2
71.6
77.8
62.5
2
3
4
5
6
75.0
78.5
71.6
78.7
62.5
75.0
77.1
71.5
77.7
62.4
75.0
77.8
71.6
77.9
62.5
75.1
78.6
71.9
78.6
62.6
75.3
78.6
71.8
78.7
62.9
ꢀ
Rha -1
102.8
72.7
72.9
74.1
70.4
18.6
102.9
72.6
72.9
73.9
70.5
18.6
102.9
72.5
72.9
73.9
70.5
18.5
102.9
72.6
72.8
74.0
70.5
18.5
102.9
72.6
72.9
74.0
70.5
18.8
102.9 102.9
103.0
72.6
72.9
73.9
70.5
18.6
2
3
4
5
6
72.6
72.9
74.0
70.5
18.7
72.6
72.9
74.1
70.5
18.6
ꢀ
ꢀ
Glc -1
105.1 105.1 105.0 105.0 105.1
105.1 105.2
2
3
4
5
6
75.3
78.7
71.8
78.6
62.7
75.3
78.7
71.8
78.6
62.9
75.3
78.7
71.8
78.6
62.9
75.3
78.7
71.8
78.5
62.9
75.1
78.7
71.8
78.6
62.9
75.0
78.7
71.8
78.6
62.9
75.3
78.6
71.7
78.5
62.9
1
(
br, OH), 2940 (CH), 2868 (CH), 1645, 917, 898, 878; H
2.3.5. Dracaenoside I (9)
ꢀꢀ
20
◦
NMR (pyridine-d ), δ (ppm): 6.34 (1H, brs, 1 -H), 5.81 (1H,
Amorphous solid, C H70O17, [α] − 62.50 (MeOH,
5
45
d
ꢀ
ꢀꢀ
brs, 1 -H), 5.38 (1H, brs, 6-H), 4.90 (1H, d, J = 6.0 Hz,
c = 0.2). Negative ion HRFAB-MS, m/z: 881.4500 [M −
ꢀ
ꢀꢀ
−
1
5
(
(
2
-H), 1.74 (3H, d, J = 5.55 Hz, 6 -CH3), 1.59 (3H, d, J =
H] (calcd. 881.4534). Negative ion FAB-MS, m/z: 881
ꢀ
ꢀꢀ
−
−
KBr
−1
.40 Hz, 6 -CH3), 1.17 (1H, d, J = 5.56 Hz, 21-CH3), 1.16
[M − H] , 719 [M − 162 − H] . IR,ν
(cm ): 3428
max
1
1H, d, J = 5.56 Hz, 21-CH3), 1.11 (3H, s, 18-CH3), 1.06
(br, OH), 2937 (CH), 2904, 1652, 1048, 920; H NMR
ꢀ
ꢀ
3H, s, 19-CH3), 1.06 (3H, brs, 27-CH3, 25S), 0.68 (3H, brs,
(pyridine-d ), δ (ppm): 6.35 (1H, brs, 1 -H), 5.48 (1H, brs,
5
7-CH3, 25R). 13C NMR (pyridine-d ): see Tables 1 and 2.
27-H), 5.34 (1H, brs, 6-H), 5.09 (1H, brs, 27-H), 5.08 (1H,
5
ꢀ
ꢀꢀ
ꢀ
d, J = 7.76 Hz, 1 -H), 4.90 (1H, d, J = 6.0 Hz, 1 -H),
ꢀ
ꢀ
2
.3.4. 25(R,S)-Dracaenoside H (8)
1.76 (3H, d, J = 6.0 Hz, 6 -CH3), 1.11 (3H, d, J = 6.8 Hz,
2
0
◦
13
Amorphous solid, C H72O18, [α] − 69.77 (MeOH,
21-CH3), 1.09 (3H, s, 18-CH3), 1.06 (3H, s, 19-CH3).
C
45
d
c = 0.2). Negative ion HRFAB-MS, m/z: 899.4681
NMR (pyridine-d ): see Tables 1 and 2.
5
−
[
9
(
8
M − H] (calcd. 899.4640). Negative ion FAB-MS, m/z:
+
−
+
KBr
00 [M] , 737 [M − 162 − H] , 754 [M − 146] . IR,ν
2.3.6. Dracaenoside J (10)
max
−
1
20
d
◦
cm ): 3424 (br, OH), 2943 (CH), 2337, 1652, 1050, 920,
97. H NMR (pyridine-d , 400 MHz), δ (ppm): 6.39 (1H,
Amorphous solid, C H72O19, [α] − 85.00 (MeOH,
45
1
c = 0.2). Negative ion HRFAB-MS, m/z: 915.4572
5
ꢀ
ꢀ
+
brs, 1 -H), 5.40 (1H, brs, 6-H), 5.07 (1H, d, J = 7.76,
[M − H] (calcd. 915.4589). Negative ion FAB-MS, m/z:
ꢀꢀꢀ
ꢀ
−
−
−
1
-H), 4.90 (1H, d, J = 6.0 Hz, 1 -H), 1.74 (3H, d,
915 [M − H] , 769 [M − 146 − H] , 753 [M − 162 − H] .
ꢀ
ꢀ
KBr 1
−1
(cm ): 3428 (br, OH), 2935 (CH), 1652. H
max
J = 5.55 Hz, 6 -CH3), 1.17 (1H, d, J = 5.56 Hz, 21-CH3,
IR,ν
ꢀꢀ
NMR (pyridine-d ), δ (ppm): 6.39 (1H, brs, 1 -H), 5.39
5
2
5S), 1.19 (1H, d, J = 5.56 Hz, 21-CH3, 25R), 1.12 (3H,
s, 18-CH3), 1.06 (3H, s, 19-CH3), 0.66 (brs, 27-CH3,
5R), 1.07 (brs, 27-CH3, 25S). 13C NMR (pyridine-d ): see
ꢀ
ꢀꢀ
(1H, brs, 6-H), 5.08 (1H, d, J = 7.76 Hz, 1 -H), 4.90 (1H,
ꢀ
ꢀꢀ
2
d, J = 6.0 Hz, 1 -H), 1.76 (3H, d, J = 6.0 Hz, 6 -CH3),
1.18 (3H, d, J = 6.80 Hz, 21-CH3), 1.12 (3H, s, 18-CH3),
5
Tables 1 and 2.

Q.-A. Zheng et al. / Steroids 69 (2004) 111–119
115
.99 (3H, s, 19-CH3). 13C NMR (pyridine-d ): see Tables 1
(3H, d, J = 7.44 Hz, 27-CH3). C NMR (pyridine-d ): see
13
0
5
5
and 2.
Tables 1 and 2.
Enzymatic hydrolysis of mixture of ophipojaponin A and
2
.3.7. Dracaenoside K (11)
2
5(S)-dracaenoside N. Mixture of ophipojaponin A and
25(S)-dracaenoside N (5 mg) was treated with ␤-glucosidase
10 mg) in HOAc–NaOAC buffer (pH 5, 5 ml) at room
White amorphous powder, C H72O18, [α] − 100.00◦
20
45
d
(
[
9
MeOH, c = 0.2). Negative ion HRFAB-MS, m/z: 899.4678
(
−
M − H] (calcd. 899.4640). Negative ion FAB-MS, m/z:
temperature for 12 h to obtain 6.
+
−
KBr
−1
(cm ): 3434 (br,
max
00 [M] , 753 [M − 146 − H] . IR,ν
1
OH), 2931 (CH), 1635. H NMR (pyridine-d ), δ (ppm):
2.3.11. 25(R,S)-Dracaenoside O (15)
5
ꢀ
ꢀ
ꢀꢀꢀ
Amorphous solid, C H84O23, [α] − 0.08◦ (MeOH,
20
6
6
1
2
1
.35 (1H, brs, 1 -H), 5.08 (1H, brs, 1 -H), 5.39 (1H, brs,
-H), 4.91 (1H, d, J = 6.76 Hz, 1 -H), 3.90 (1H, m, 3-H),
51
d
ꢀ
c = 0.2). Negative ion HRFAB-MS, m/z: 1063.5337
ꢀꢀ
+
.75 (3H, d, J = 6.0 Hz, 6 -CH3), 1.30 (3H, d, J = 6.96 Hz,
7-CH3), 1.21 (3H, d, J = 6.84 Hz, 21-CH3), 1.05 (3H, s,
[M − H] (calcd. 1063.5325). Negative ion FAB-MS, m/z:
+
+
+
1064 [M] , 918 [M − 146] , 772 [M − 146 − 146] , 610
8-CH3), 1.11 (3H, s, 19-CH3). 13C NMR (pyridine-d ): see
[M − 146 − 146 − 162] . IR,ν
+
KBr
(cm ): 3431 (br, OH),
−1
5
max
1
Tables 1 and 2.
2935 (CH), 1651. H NMR (pyridine-d ), δ (ppm): 6.38 (1H,
5
ꢀ
ꢀ
ꢀꢀꢀ
brs, 1 -H), 5.84 (1H, brs, 1 -H), 5.41 (1H, brs, 6-H), 4.91
ꢀ
ꢀꢀꢀꢀ
2
.3.8. Dracaenoside L (12)
(1H, d, J = 6.0 Hz, 1 -H), 4.83 (1H, d, J = 7.76, 1 -H),
White amorphous powder, C H72O19, [α] − 66.67◦
2
d
8
1.77 (3H, d, J = 6.0 Hz, 6 -CH3), 1.62 (3H, d, J = 6.0 Hz,
ꢀꢀ
45
ꢀ
(
[
MeOH, c = 0.2). Negative ion FAB-MS, m/z: 915
6 -CH3), 1.36 (3H, d, J = 7.0 Hz, 21-CH3), 1.14 (3H, s,
−
−
−
M − H] , 753 [M − 162 − H] , 769 [M − 146 − H] .
18-CH3), 1.13 (3H, s, 19-CH3), 1.01 (3H, d, J = 7.44 Hz,
KBr
−1
1
13
IR,ν
(cm ): 3434 (br, OH), 2931 (CH), 1635. H NMR
27-CH3). C NMR (pyridine-d ): see Tables 1 and 2.
max
5
ꢀꢀ
(pyridine-d ), δ (ppm): 6.35 (1H, brs, 1 -H), 5.08 (1H, d,
Enzymatic hydrolysis of 25-(R,S)-dracaenoside O. 25(R,
S)-Dracaenoside O (2 mg) was treated with ␤-glucosidase
(4 mg) in HOAc–NaOAC buffer (pH 5, 3 ml) at room tem-
perature for 12 h to afford 7 and glucose.
5
ꢀ
ꢀꢀ
J = 7.56 Hz, 1 -H), 5.39 (1H, brs, 6-H), 4.91 (1H, d, J =
ꢀ
6
6
.76 Hz, 1 -H), 3.90 (1H, m, 3-H), 1.75 (3H, d, J = 6.0 Hz,
ꢀ
ꢀ
-CH3), 1.30 (3H, d, J = 6.84 Hz, 27-CH3), 1.21 (3H, d,
J = 6.84 Hz, 21-CH3), 1.05 (3H, s, 18-CH3), 1.12 (3H, s,
1
9-CH3). 13C NMR (pyridine-d ): see Tables 1 and 2.
2.3.12. 25(R,S)-Dracaenoside P (16)
5
2
0
◦
Amorphous solid, C H84O24, [α] − 45.00 (MeOH,
51
d
2
.3.9. 25(R,S)-Dracaenoside M (13)
c = 0.2). Negative ion HRFAB-MS, m/z: 1079.5211
2
0
◦
+
Amorphous solid, C H74O19, [α] − 74.90 (MeOH,
[M − H] (calcd. 1079.5274). Negative ion FAB-MS, m/z:
45
d
+
+
+
c = 0.2). Negative ion HRFAB-MS, m/z: 917.4753 [M −
1080 [M] , 918 [M−162] , 756 [M−162−162−H] , 610
+
+
KBr −1
(cm ): 3424 (br, OH),
max
H] (calcd. 917.4746). Negative ion FAB-MS, m/z: 932
[M − 162 − 146 − 162] . IR,ν
+
−
KBr
−1
1
[
3
M + 14] , 769 [M + 14 − 146 − H] . IR,ν
(cm ):
2930 (CH), 1647. H NMR (pyridine-d ), δ (ppm): 6.35
max
5
1
ꢀꢀ
ꢀꢀꢀ
428 (br, OH), 2935 (CH), 1652. H NMR (pyridine-d ), δ
(1H, brs, 1 -H), 5.85 (1H, brs, 1 -H), 5.34 (1H, brs, 6-H),
5
ꢀ
ꢀ
ꢀꢀꢀꢀ
(ppm): 6.39 (1H, brs, 1 -H), 5.39 (1H, brs, 6-H), 4.90 (1H,
5.08 (1H, brs, 27-H), 5.08 (1H, d, J = 7.76 Hz, 1 -H),
ꢀ
ꢀꢀꢀ
ꢀ
d, J = 6.0 Hz, 1 -H), 4.81 (1H, d, J = 7.76, 1 -H), 1.76
4.90 (1H, d, J = 6.0 Hz, 1 -H), 1.76 (3H, d, J = 6.0 Hz,
ꢀꢀ
ꢀꢀ
(
2
3H, d, J = 6.0 Hz, 6 -CH3), 1.18 (3H, d, J = 6.80 Hz,
6 -CH3), 1.35 (3H, d, J = 7.85 Hz, 21-CH3), 1.13 (3H, s,
1-CH3), 1.12 (3H, s, 18-CH3), 0.99 (3H, s, 19-CH3), 1.01
18-CH3), 1.11 (3H, s, 19-CH3), 1.02 (3H, d, J = 6.50 Hz,
13
13
(
3H, d, J = 7.44 Hz, 27-CH3). C NMR (pyridine-d ): see
27-CH3). C NMR (pyridine-d ): see Tables 1 and 2.
5
5
Tables 1 and 2.
Enzymatic hydrolysis of 25-(R,S)-dracaenoside P. 25(R,
S)-Dracaenoside P (2 mg) was treated with ␤-glucosidase
(4 mg) in HOAc–NaOAC buffer (pH 5, 3 ml) at room tem-
perature for 12 h to afford 8 and glucose.
Enzymatic hydrolysis of 25-(R,S)-dracaenoside M. 25(R,
S)-Dracaenoside M (5 mg) was treated with ␤-glucosidase
(10 mg) in HOAc–NaOAC buffer (pH 5, 5 ml) at room tem-
perature for 12 h to obtain 5.
2
.3.13. 25(R,S)-Dracaenoside Q (17)
2
0
◦
2
2
.3.10. Mixture of ophipojaponin A and
Amorphous solid, C H82O23, [α] − 65.53 (MeOH,
51
d
5(S)-dracaenoside N (14)
c = 0.2). Negative ion HRFAB-MS, m/z: 1061.5221 [M −
2
0
◦
+
Amorphous solid, C H74O19, [α] − 74.90 (MeOH,
H] (calcd. 1061.5168). Negative ion FAB-MS, m/z: 1062
45
d
+
+
+
c = 0.2). Negative ion HRFAB-MS, m/z: 917.4754 [M −
[M] , 915 [M − 146 − H] , 769 [M − 146 − 162 − H] .
+
KBr 1
−1
H] (calcd. 917.4746). Negative ion FAB-MS, m/z: 932
IR,ν
(cm ): 3419 (br, OH), 2932 (CH), 1653. H NMR
max
+
−
KBr
−1
ꢀꢀ
[
3
M + 14] , 769 [M + 14 − 146 − H] . IR,ν
(cm ):
(pyridine-d ), δ (ppm): 6.35 (1H, brs, 1 -H), 5.82 (1H, brs,
max
5
1
ꢀꢀꢀ
ꢀ
428 (br, OH), 2935 (CH), 1652. H NMR (pyridine-d ), δ
1 -H), 5.34 (1H, brs, 6-H), 4.90 (1H, d, J = 6.0 Hz, 1 -H),
5
ꢀ
ꢀ
ꢀꢀ
ꢀꢀꢀ
(ppm): 6.39 (1H, brs, 1 -H), 5.39 (1H, brs, 6-H), 4.90 (1H,
1.78 (3H, d, J = 6.0 Hz, 6 -CH3), 1.74 (3H, s, 21-CH3),
ꢀ
ꢀꢀꢀ
d, J = 6.0 Hz, 1 -H), 4.81 (1H, d, J = 7.76, 1 -H), 1.76
1.60 (3H, d, J = 6.0 Hz, 6 -CH3), 1.08 (3H, s, 18-CH3),
ꢀꢀ
(
2
3H, d, J = 6.0 Hz, 6 -CH3), 1.18 (3H, d, J = 6.80 Hz,
1.05 (3H, s, 19-CH3), 1.02 (3H, d, J = 6.50 Hz, 27-CH3).
13
1-CH3), 1.12 (3H, s, 18-CH3), 0.99 (3H, s, 19-CH3), 1.01
C NMR (pyridine-d ): see Tables 1 and 2.
5

1
16
Q.-A. Zheng et al. / Steroids 69 (2004) 111–119
1
3
2
.3.14. Dracaenoside R (18)
d, J = 6.20 Hz, 27-CH3). C NMR (pyridine-d ): see
5
2
0
◦
Amorphous solid, C H72O19, [α] − 75.13 (MeOH,
Tables 1 and 2.
45
d
c = 0.2). Negative ion HRFAB-MS, m/z: 915.4513
+
[
9
3
M − H] (calcd. 915.4589). Negative ion FAB-MS, m/z:
2.3.18. 26-O-␤-d-Glucopyranosyl
+
+
KBr
−1
15 [M − H] , 769 [M − 146 − H] . IR,ν
(cm ):
25(R,S)-spirost-5-en-3,22␰,26-triol 3-O-␣-l-
rhamnopyranosyl-(1,2)-[␣-l-rhamnopyranosyl-(1,4)]-␤-d-
glucopyranoside (mixture of protogracillin and
protoneogracillin) (22)
max
1
434 (br, OH), 2937 (CH), 1652. H NMR (pyridine-d ),
5
ꢀꢀ
ꢀꢀꢀ
ꢀ
δ (ppm): 6.38 (1H, brs, 1 -H), 5.84 (1H, brs, 1 -H), 5.36
(
(
6
1
1H, brs, 6-H), 4.90 (1H, d, J = 6.0 Hz, 1 -H), 1.76
ꢀ
ꢀ
3H, d, J = 6.0 Hz, 6 -CH3), 1.62 (3H, d, J = 6.0 Hz,
White amorphous powder, C H84O23. Negative ion
51
ꢀꢀꢀ
+
−
-CH3), 1.16 (3H, d, J = 6.84 Hz, 21-CH3), 1.11 (3H, s,
FAB-MS, m/z: 1064 [M] , 901 [M − 162 − H] , 755
8-CH3), 1.05 (3H, s, 19-CH3). 1 C NMR (pyridine-d ): see
3
[M − 162 − 146 − H] , 593 [M − 162 − 146 − 162 − H] ,
−
−
5
+
1
Tables 1 and 2.
432 [M −162 −146−162 −162] . H NMR (pyridine-d ,
5
ꢀꢀ
4
00 MHz), δ (ppm): 6.33 (1H, brs, H-1 ), 5.32 (1H, brs,
ꢀ
ꢀꢀ
2
3
.3.15. 25(R,S)-Spirost-5-en-3-ol
-O-␣-l-rhamnopyranosyl-(1,2)-[␣-l-rhamnopyranosyl-
H-6), 5.08 (1H, d, J = 6.50 Hz, H-1 ), 4.92 (1H, d, J =
ꢀ
ꢀ
6.20 Hz, H-1 ), 4.87 (1H, d, J = 6.60 Hz, H-1 ), 1.73 (3H, d,
ꢀ
ꢀ
(1,4)]-␤-d-glucopyranoside (mixture of dioscin and
J = 5.85 Hz, H-6 ), 1.31 (3H, d, J = 6.40 Hz, H-21), 1.05
collettinside III) (19)
(3H, s, CH3-19), 0.88 (3H, s, CH3-18), 0.99 (3H, d,
13
White amorphous powder, C H72O . Negative ion FAB-
J
=
6.20 Hz, CH3-27).
C NMR (pyridine-d ): see
5
4
5
16
−
−
1
MS, m/z: 867 [M − H] , 721 [M − H − 146] . H NMR
Tables 1 and 2.
(
(
(
(
pyridine-d , 500 MHz), δ (ppm): 3.90 (1H, m, H-3), 4.57
5
1H, m, H-16), 5.10 (1H, d, J = 7.7 Hz, H-Glc-1), 5.53
2.3.19. Sugar analysis of compounds 5–22
Each compound (2 mg) in 1 M HCl (dioxane–H2O, 1:1,
1 ml) was heated at 100 C for 2 h. After removing the solu-
ꢀ
1H, d, J = 6.9 Hz, H-Glc -1), 6.01 (1H, s, H-Rha-1), 1.79
◦
3H, d, J = 6.5 Hz, H-Rha-6), 0.81 (3H, s, H-18), 1.05 (3H,
s, H-19), 1.13 (3H, d, J = 6.7 Hz, H-21, 25R), 1.12 (3H,
tion under reduced pressure, the residue was extracted with
chloroform three times. The monosaccharide fraction was
condensed. The residue was dried and solved in pyridine
(5 ml). The 0.5 ml trimethylchlorosilane was added into
the pyridine solution and reacted 30 min in room temper-
ature. The solution was condensed under reduce pressure;
the residue was washed with 0.5 ml ether. The product
was analyzed by means of GC–MS. The retention time
of l-rhamnose and d-glucose were 7.17 and 11.43 min,
respectively.
d, J = 6.7 Hz, H-21, 25S), 1.00 (3H, d, J = 6.20 Hz, 27-
1
3
CH3, 25R), 0.69 (3H, d, J = 9.1 Hz, H-27, 25S). C NMR
(pyridine-d ): see Tables 1 and 2.
5
2
3
.3.16. 25(R,S)-Spirost-5-en-3-ol
-O-␣-l-rhamnopyranosyl-(1,2)-[␤-d-glucopyranosyl-
(
1,3)]-␤-d-glucopyranoside (mixture of gracillin and
collettinside IV) (20)
White amorphous powder, C H72O17. Negative ion FAB-
45
−
−
1
MS, m/z: 883 [M − H] , 721 [M − H − 162] . H NMR
ꢀꢀ
pyridine-d , 500 MHz), δ (ppm): 6.39(1H, brs, 1 -H), 5.34
5
(
(
ꢀ
ꢀꢀ
1H, brs, 6-H), 5.04 (1H, d, J = 7.76 Hz, 1 -H), 4.90 (1H,
3. Result and discussion
ꢀ
ꢀꢀ
d, J = 6.0 Hz, 1 -H), 1.73 (3H, d, J = 6.0 Hz, 6 -CH3),
1
5
1
.18 (1H, d, J = 5.56 Hz, 21-CH3, 25R), 1.17 (1H, d, J =
.56 Hz, 21-CH3, 25S), 1.12 (3H, s, 18-CH3), 1.06 (3H, s,
9-CH3), 1.00 (3H, d, J = 6.20 Hz, 27-CH3, 25R), 0.66
The fresh stem of D. cochinchinensis was extracted
with hot methanol. After removal of the solvent under re-
duced pressure, the residue was partitioned between H2O
and n-butanol. The n-butanol fraction was supplied to a
high porous absorption resin (Diaion HP-20) column chro-
matography, eluting with H2O and methanol. The methanol
fraction was repeatedly chromatographed on silica gel and
octadecylsilanized (ODS) silica gel to afford 18 steroidal
saponins (5–22).
Compounds 19–22 were known 25(R,S)-steroidal
saponin epimers, identified as 25(R,S)-spirost-5-en-3-ol
3-O-␣-l-rhamnopyranosyl-(1,2)-[␣-l-rhamnopyranosyl-(1,
)]-␤-d-glucopyranoside (dioscin and collettinside III) (19)
[7,10], 25(R,S)-spirost 5-en-3-ol 3-O-␣-l-rhamnopyranosyl-
1,2)-[␤-d-glucopyranosyl-(1,3)]-␤-d-glucopyranoside (gr-
1
3
(
3H, d, J = 6.20 Hz, 27-CH3, 25S). C NMR (pyridine-
d ): see Tables 1 and 2.
5
2
2
.3.17. 26-O-␤-d-Glucopyranosyl
5(R,S)-furost-5-en-3,22␰,26-triol 3-O-␣-l-
rhamnopyranosyl-(1,2)-[␤-d-glucopyranosyl(1,3)]-␤-d-
glucopyranoside (mixture of protodioscin and
protoneodioscin) (21)
White amorphous powder, C H84O22. Negative ion FAB-
51
+
+
1
+
MS, m/z: 1048 [M] , 902 [M−146] , 756 [M−146−146] ,
4
+
5
5
1
94 [M − 146 − 146 − 162] . H NMR (pyridine-d ,
5
00 MHz), δ (ppm): 3.90 (1H, m, H-3), 4.57 (1H, m, H-
(
ꢀ
ꢀꢀ
6), 5.32 (1H, brs, H-6), 5.80 (1H, brs, H-1 ), 6.32 (1H,
acillin and collettinside IV) (20) [7,10], 26-O-␤-d-
glucopyranosyl 25(R,S)-furost-5-en-3,22␰,26-triol 3-O-␣-l-
rhamnopyranosyl-(1,2)-[␤-d-glucopyranosyl(1,3)]-␤-d-glu-
copyranoside (protodioscin and protoneodioscin) (21)
ꢀ
ꢀ
ꢀ
brs, H-1 ), 4.90 (1H, d, J = 6.60 Hz, H-1 ), 4.82 (1H, d,
ꢀꢀꢀꢀ
J = 6.60 Hz, H-1 ), 0.87 (3H, s, 18-CH3), 1.04 (3H, s,
9-CH3), 1.31 (3H, d, J = 6.35 Hz, 21-CH3), 1.00 (3H,
1

Q.-A. Zheng et al. / Steroids 69 (2004) 111–119
117
[
2
7,11], 26-O-␤-d-glucopyranosyl 25(R,S)-spirost-5-en-3,
2␰,26-triol 3-O-␣-l-rhamnopyranosyl-(1,2)-[␣-l-rhamno-
[␣-l-rhamnopyranosyl-(1,4)]-␤-d-glucopyranoside, named
25(R,S)-dracaenoside G.
pyranosyl-(1,4)]-␤-d-glucopyranoside (protogracillin and
protoneogracillin) (22) [7,11], respectively, by detailed
spectral analysis and compared with reference data.
Saponins 5–8 were obtained as white amorphous pow-
der. The negative ion HRFABMS determined their molec-
The NMR signals due to the sugar moiety of saponin
8 were in good agreement with those of gracillin [7].
Since then, the structure of 8 was formulated as 25-(R,S)-
spirost-5-en-3␤,14␣-diol 3-O-␣-l-rhamnopyranosyl-(1,2)-
[␤-d-glucopyranosyl-(1,3)]-␤-d-glucopyranoside, named
25(R,S)-dracaenoside H.
ular formula as C39H O13, C39H O13, C H72O17, and
6
2
62
45
1
13
C H72O18, respectively. NMR data due to the H and
C
Dracaenoside I (9) was obtained as a white amorphous
4
5
2
0
◦
signals around C-25 of their aglycone moieties revealed that
–8 were spirostanol monodesmosides, existed as mixtures
powder, [α] − 62.50 in MeOH. Negative ion HRFABMS
d
5
gave a quasi-molecular ion peak at m/z = 881.4500
−
of C-25 R and S epimers. As far as we know, the isolation of
a steroidal mixture of C-25 R and S epimers is very difficult
and could not be done by the present chromatographic tech-
nique [12]. Thus, their structures were elucidated as C-25
epimeric mixtures.
([M − H] ; calcd. 881.4534) corresponding to an empir-
ical molecular formula C H70O17. Acid hydrolysis of 9
45
gave d-glucose and l-rhamnose by GC analysis. Compared
the NMR feature with that of 20 suggested that saponin
9 contained a same sugar chain as 20, the difference be-
tween these two saponins only appeared in F-ring of their
aglycons. Compound 9 showed an exomethylene group at
δC = 108.8 (CH2) and 144.5 (C), which was ascribed to
C-25 and C-27 by compared with the reference data [15].
On the basis of the above evidence, dracaenoside I (9)
was formulated as spirost-5, 25(27)-dien-3␤-ol 3-O-␣-l-
rhamnopyranosyl-(1,2)-[␤-d - glucopyranosyl - (1,3)] - ␤ - d-
glucopyranoside.
The NMR spectral features of 5–8 were nearly identical to
each other, except for the sugar moieties. Comparing the 13
C
NMR data of their aglycone moiety with those of diosgenin
and yamogenin [13], indicated that the aglycone moiety of
5
–8 had one more hydroxyl group attached at C-14, which
let the methine carbon signal of C-14 (δ = 56.7) in diosgenin
and yamogenin changed to a quaternary carbon signal at
δ = 86.6 in 5–8. Thus, the sapogenin moiety of 5–8 was
identified as 25(R,S)-spirost-5-en-3␤,14␣-diol (prazerigenin
A and neoprazerigenin A) [14].
Saponin 10 gave a molecular formula as C H72O19 by
4
5
+
negative ion HRFABMS (m/z = 915.4572, [M − H] ;
The negative ion FABMS of saponin 5 showed frag-
calcd. 915.4589) and it had one more oxygen atom than
−
13
ment ions at m/z = 591 [M − 146 − H] and 429 [M −
8. The C NMR feature is quite similar to that of 8 with
−
1
46 − 162 − H] , suggested that 5 contained a deoxy-
exceptions of the signals due to the F-ring carbons. In
the case of saponin 10, the C-27 methyl of 8 was substi-
tuted to a hydroxymethyl group, which was deduced as
eq orientation based on the IR characteristic absorptions
hexosyl as terminal unit and a hexosyl as inner unit at
the sugar moiety. Acid hydrolysis of 5 with 1 mol/l HCl
in dioxane (v/v, 1:1) afforded d-glucose and l-rhamnose
as sugar residue. HMBC spectrum showed the correlations
of anomeric proton (δ = 4.94 (d, J = 7.70 Hz)) of d-
glucopyranosyl unit with C-3 (δ = 78.2) of the aglycone,
and anomeric proton (δ = 5.89, brs) of l-rhamnopyranosyl
unit with C-4 (δ = 78.4) of glucopyranosyl unit, clearly indi-
cated the location of sugar moieties. Therefore, the structure
of 5 was determined as 25(R,S)-spirost-5-en-3␤,14␣-diol 3-
O-␣-l-rhamnopyranosyl-(1,4)-␤-d-glucopyranoside, named
−
1
at 992 and 1018 cm [16]. So, 10 was deduced to be a
prazerigenin d-glycoside as (25S)-spirost-5-en-3␤,14␣,27-
triol (prazerigenin D) 3-O-␣-l-rhamnopyranosyl-(1,2)-[␤-
d-glucopyranosyl-(1,3)]-␤-d-glucopyranoside, named dra-
caenoside J.
Dracaenoside K (11) was obtained as a white amorphous
2
0
◦
powder, [α] −100.00 in MeOH. Negative ion HRFABMS
d
gave a quasi-molecular ion peak at m/z = 899.4678
−
2
5(R,S)-dracaenoside E.
([M − H] ; calcd. 899.4640) corresponding to an empiri-
The 13C NMR spectral data together with the result of
cal molecular formula C H72O18. The C NMR chemical
13
45
acidic hydrolysis, showed that saponin 6 had similar struc-
ture to that of 5, except for the linkage position of the l-
rhamnopyranosyl unit to the d-glucopyranosyl unit, which
was determined on the basis of the glycosylation shift effects
of the C-2 (δ = 79.7) of d-glucopyranosyl unit comparing
with the reference data (δ = 75.6) [13]. Thus, the structure
of 6 was established as 25(R,S)-spirost-5-en-3␤,14␣-diol 3-
O-␣-l-rhamnopyranosyl-(1,2)-␤-d-glucopyranoside, named
shifts of 11 are closely related to those of 7 with exceptions
of the signals due to the F-ring. A hydroxyl group located
on C-24 (δ = 66.6) position caused significant down-field
shift of the C-27 methyl group (δ = 9.76) by comparing
with the reference data [17]. In addition, the 26-H2 ap-
peared at δ = 3.53 (Heq, brd, J = 10.4 Hz) and δ = 4.05
(Hax, 1H, dd, J = 3.15, 10.4 Hz). The J values implicated
that the 25-H was in equatorial orientation. In the NOESY
spectrum of 11, correlation peak of 24-H (δ = 4.60)
with 26-Hax indicated the equatorial orientation of C-24
OH. Thus, the structure of dracaenoside K (11) was de-
duced as (24S,25R)-spirost-5-en-3␤,14␣,24␤-triol 3-O-␣-l-
rhamnopyranosyl-(1,2)-[␣-l-rhamnopyranosyl-(1,4)]-␤-d-
glucopyranoside.
2
5(R,S)-dracaenoside F.
Acid hydrolysis of 7 also afforded d-glucose and
l-rhamnose. The 1H and C NMR spectra (pyridine-
d5) indicated that it had a same sugar chain as that in
dioscin [7]. Therefore, 7 was characterized as 25(R,S)-
spirost-5-en-3␤,14␣-diol 3-O-␣-l-rhamnopyranosyl-(1,2)-
13

1
18
Q.-A. Zheng et al. / Steroids 69 (2004) 111–119
The 13C NMR spectrum of saponin 12 was quite similar
structure rings A–E (C-1–C-21) of 18 was similar to that
of 7. Significant differences were the carbon signals of F-
ring. In the spectrum of 18, the downfield of C-26 and C-
27 to δ = 66.9 and 65.4, indicated that these positions
were oxygenated. The furospirostane skeleton was deduced
from an easily distinguished tertiary carbon signal in δ =
120.9, which was assigned to C-22 [13]. Thus, the struc-
ture of dracaenoside R (18) was determined as furospirost-5-
en-3␤,14␣,26,27-tetrol 3-O-␣-l-rhamnopyranosyl-(1,2)-[␣-
l-rhamnopyranosyl-(1,4)]-␤-d-glucopyranoside.
to 11 except the sugar moiety, which was identical to that
of 8 by comparing their spectral data. Thus, the structure
of 12 was deduced as (24S,25R)-spirost-5-en-3␤,14␣,24␤-
triol 3-O-␣-l-rhamnopyranosyl-(1,2)-[␤-d-glucopyranosyl-
(
1,3)]-␤-d-glucopyranoside, named dracaenoside L.
5(R,S)-Dracaenosides M–P (13–16) were obtained as
white amorphous powders. The negative ion HRFABMS
determined their molecular formula as C H74O19 for
2
45
1
3 and 14, C H84O23 for 15, and C H84O24 for 16,
51 51
respectively. The positive color reaction in Ehrlich test
and the characteristic 13C NMR signals suggested that
all of these four compounds are furostanol glycoside
with same aglycon [18]. Enzymatic hydrolysis of 13 and
It is well known that the C-27 steroidal glycosides were
widely distributed in plant kingdom, especially in the fami-
lies of Liliaceae and Agavaceae [21]. The molecular diver-
sity of steroidal saponins was exhibited not only on the sugar
moiety, but also the structure of aglycons. The biological
activities and physiological functions of steroidal saponins
were dependent on both of their glycosylated level of the
sugar moiety and the oxidized level of sapogenins [22].
Generally, the sapogenins occurring in the genus Dracaena
are characterized with 1,3-dihydroxyl substituents [15,17].
However, most of the saponins isolated from D. cochinchi-
nensis were composed of 1,14-dihydroxyl substitutions. It
is suggested that as a northernmost species of Dracaena
in East Asia, D. cochinchinensis involved in a specifically
secondary metabolism pathway of steroidal saponins. More-
over, it is noticed that the chemical constituents of the fresh
stems are quite other with those of the resins. Most com-
pounds isolated from the latter are polyphenols, including
chalcones and their oligomers, flavans, flavonoids, stilbenes,
as well as lignans. It is proposed that the formation of the
red resin may due to a complex physiological, ecological
and biochemical process.
1
4 gave 5 and 6, respectively, together with d-glucose.
Therefore, the structures of 13 and 14 were character-
ized as 26-O-␤-d-glucopyranosyl 25(R,S)-furost-5-en-
3
␤,14␣,22␰,26-tetrol 3-O-␣-l-rhamnopyranosyl-(1,4)-␤-
d-glucopyranoside [25(R,S)-dracaenoside M (13)], and
2
2
6-O-␤-d-glucopyranosyl 25(R,S)-furost-5-en-3␤,14␣,22,
6-tetrol 3-O-␣-l-rhamnopyranosyl-(1,2)-␤-d-glucopyra-
noside (14), respectively. The 25-(R) epimer of 14 was a
known saponin once isolated from Ophipogon japonicus
[19], named ophipojaponin A, and its 25-(S)-epimer was
given the trivial name of 25(S)-dracaenoside N.
Enzymatic hydrolysis of 15 and 16 gave 7 and 8, respec-
tively, together with d-glucose. Therefore, the structures
of 15 and 16 were elucidated as 26-O-␤-d-glucopyranosyl
2
5(R,S)-furost-5-ene-3␤,14␣,22␰,26-tetrol 3-O-␣-l-rhamn-
opyranosyl-(1,2)-[␣-l-rhamnopyranosyl-(1,4)]-␤-d-glucop-
yranoside [25(R,S)-dracaenoside O (15)], and 26-O-␤-d-glu-
copyranosyl 25(R,S)-furost-5-en-3␤,14␣,22␰,26-tetrol 3-O-
␣
-l-rhamnopyranosyl-(1,2)-[␤-d-glucopyranosyl-(1,3)]-␤-
d-glucopyranoside [25(R,S)-dracaenoside P (16)], respec-
tively.
Acknowledgements
The molecular formula C H82O23 was deduced for
51
saponin 17 from its quasi-molecular ion peak at m/z =
We are grateful to the staffs of the analysis group of our
institute for the measurement of spectroscopic data.
1
061.5222 (calcd. for C H82O23 1061.5169) in the
51
1
13
HRFABMS. Comparison of the H and C NMR spec-
tra of 17 with those of compound 16 showed that both
structures are very similar except an additional double
bond signals at the aglycon of 17. The downfield chemical
shifts of C-22 (δ = 154.4) and C-20 (δ = 105.3) indi-
cated the olefinic substitution located between C-22 and
C-20 [20]. Acid hydrolysis of 17 gave l-rhamnose and
d-glucose as sugar residue by GC analysis. Therefore, 17
was identified as 26-O-␤-d-glucopyranosyl 25(R,S)-furost-
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