Glycosidic constituents from the roots and rhizomes of Melicope pteleifolia

Article abstract of DOI:10.1016/j.carres.2012.08.011

Seven new diglycosidic constituents, named pteleifosides A-G (1-7), along with ten known glycosides, were isolated from the roots and rhizomes of Melicope pteleifolia (Champ. ex Benth.) T. Hartley. The structures of the isolated compounds were established on the basis of chemical and spectroscopic methods, mainly 1D and 2D NMR data and mass spectrometry.

Full text of DOI:10.1016/j.carres.2012.08.011

Carbohydrate Research 361 (2012) 114–119
Contents lists available at SciVerse ScienceDirect
Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres
Glycosidic constituents from the roots and rhizomes of Melicope pteleifolia
Yang Zhang a,b,c, , Li-Jun Yang a, , Kun Jiang a, Chang-Heng Tan a, , Jun-Jie Tan a, Pei-Ming Yang b,
,
⇑
⇑
Da-Yuan Zhu a
a Department of Natural Medicinal Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, PR China
b Department of Chinese Traditional Medicine, Shanghai Institute of Pharmaceutical Industry, 1320 Beijing West Road, Shanghai 200040, PR China
c Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiaotong University, Shanghai 200240, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 8 July 2012
Received in revised form 10 August 2012
Accepted 21 August 2012
Available online 30 August 2012
Seven new diglycosidic constituents, named pteleifosides A–G (1–7), along with ten known glycosides,
were isolated from the roots and rhizomes of Melicope pteleifolia (Champ. ex Benth.) T. Hartley. The struc-
tures of the isolated compounds were established on the basis of chemical and spectroscopic methods,
mainly 1D and 2D NMR data and mass spectrometry.
Ó 2012 Elsevier Ltd. All rights reserved.
Keywords:
Melicope pteleifolia
Rutaceae
Diglycosidic constituents
Pteleifosides A–G
1. Introduction
3,9-diol 9-O-
citroside A 60-O-b-
apiofuranosyl(1?6)-b-
4-O-b- -apiofuranosyl(1?6)-b-
dihydrokaempferol 3-O-b- -apiofuranosyl(1?6)-b-
side (6), and (+)-lyoniresinol -glucopyranosyl
90-O-b-
(1?3)-b- -glucopyranoside (7), together with ten known glyco-
sides, tachioside (8),6 isotachioside (9),6 (+)-lyoniresinol 90-O-b-
glucopyranoside (10),7 cuneataside D (11),8 canthoside D (12),9
1-O-(b- -apiosyl(1?6)-b- -glucopyranosyl)-3-O-methylphloro-
glucinol (13),10 vanillyl alcohol 4-O-b- -glucopyranoside (14),11
melia-ionoside B (15),12 syrigoylglycerol 9-O-b-
-glucopyranoside
(16),13 and dihydrophaseic acid 3-O-b- -glucopyranoside (17).14
a
-
L
-rhamnopyranosyl(1?2)-b-
-apiofuranoside (3), 2-pinen-5,10-diol 5-O-b-
-glucopyranoside (4), homovanillyl alcohol
-glucopyranoside (5), (2R,3R)-
-glucopyrano-
D-glucopyranoside (2),
D
D-
Melicope pteleifolia (Champ. ex Benth.) T. Hartley (= Melicope
ptelefolia, Rutaceae) is a deciduous shrub or arbor distributed in
southern China and Southeast Asia. Its fresh leaves are a traditional
vegetable among the Malaysian community,1 and are also a main
material of Guangdong Herbal Tea—a popular healthy drink in China,
whereas its roots and barks, known as ‘San-ya-ku’ in traditional Chi-
nese medicines, are acted as an antipyretic, anti-inflammatory, and
analgesic agent to treat trauma, abscess, eczema, dermatitis, and
hemorrhoids.2 M. pteleifolia had been taxonomically assigned as
Euodia genus with Latin name of Euodia lepta (Spreng.) Merr. (= Evo-
dia lepta = E. roxburghiana Pierre) before 2009.3 Previous studies on
the title plant disclosed numbers of ester-soluble constituents, such
as benzopyrans, alkaloids, acetophenones, sesquiterpenes, and
flavonoids.4,5 However, little is known about the water-soluble
chemical profiles despite its water decoction is a main ingredient
of ‘999 weitai’—a famous traditional Chinese medicine formula
for the treatment of various gastritis. Our investigation focused
on the water-soluble constituents of the roots and rhizomes,
which led to the isolation of seven new diglycosidic
compounds, named pteleifosides A–G (1–7), structurally as
D
D
D
D
D
D
D
D-
D
D
D
D
D
The above compounds were reported from the title plant for the first
time.
2. Results and discussion
Pteleifoside A (1) was isolated as an amorphous powder and its
molecular formula was determined to be C25H46O12 by HR-ESI-MS.
Acid hydrolysis and GC analysis of 1 gave an
D
L-rhamnose and a
-glucose as sugar moiety. The 1H and 13C NMR spectra of 1 (Tables
(3S,5R,6R,9S)-megastigman-3,6,9-triol
9-O-
a-L-rhamnopyrano-
1 and 2) showed signals for a b-glucopyranosyl at dH 4.41 (1H, d,
J = 7.7 Hz) and dC 100.6, 79.5, 78.4, 77.8, 71.8, and 62.8, an
syl(1?2)-b-
D-glucopyranoside (1), (3R,9S)-megastigman-5-ene-
a-rhamnopyranosyl at dH 5.24 (1H, d, J = 1.4 Hz) and 1.22 (3H, d,
J = 6.2 Hz) as well as dC 101.9, 74.1, 72.1, 72.0, 69.5, and 17.9, and
a C13-aglycone unit which comprised four methyls at dH 1.22 (d,
J = 6.2 Hz), 1.02 (s), 0.99 (s), and 0.96 (d, J = 6.6 Hz), four methyl-
enes at dC 47.6, 40.7, 31.9, 32.1, three methines at dC 76.0, 67.3,
⇑
Corresponding authors. Tel.: +86 21 50806728; fax: +86 21 50807088.
E-mail addresses: chtan@mail.shcnc.ac.cn (C.-H. Tan), yangpeiming@yahoo.cn
(P.-M. Yang).
These authors contributed equally to this work.
0008-6215/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.carres.2012.08.011

Y. Zhang et al. / Carbohydrate Research 361 (2012) 114–119
115
Table 1
1H NMR spectroscopic data for compounds 1–4a
No.
1 (500 MHz)
2 (500 MHz)
3 (400 MHz)
4 (400 MHz)
1
2
—
—
—
2.05 dd (6.6 2.1)
—
1.63 m, 1.29 m
1.67 br d (12.1)
1.39 t (12.0)
3.85 m
2.19 dd (16.3, 5.2)
1.93 dd (16.3, 11.1)
—
2.06 td (13.3, 5.0)
1.95 td (13.3, 5.0)
1.69 m, 1.58 m
3.94 m
1.91 ddd (12.5, 3.7, 1.7)
1.37 dd (12.5, 11.0)
4.29 tt (11.2, 3.8)
2.46 ddd (13.4, 3.7, 1.7)
1.40 dd (13.4, 11.4)
—
3
4
3.76 tt (11.4, 4.6)
1.60 m, 1.39 m
5.62 br s
2.39 dd (17.0, 2.0)
2.52 dd (17.0, 2.0)
—
2.71 dd (8.4, 6.6)
1.53 d (8.4)
1.32 s
5
7
1.92 dqd (10.4, 6.6, 4.4)
1.64 m, 1.61 m
—
8
9
1.74 m, 1.51 m
3.88 m
5.95 s
—
0.93 s
10
11
12
13
10
1.22 d (6.2)
0.99 s
1.02 s
0.96 d (6.6)
1.25 d (5.6)
2.20 s
1.16 s
1.37 s
1.47 s
3.91 br d (1.2)
1.05 s
1.073/1.068 sb
1.65 s
4.41 d (7.7)
4.440/4.438 d (7.7)b
3.35 dd (9.3, 7.8)
3.45 t (9.0)
4.50 d (7.7)
3.13 dd (9.3, 7.8)
3.34 m
3.22 dd (9.5, 9.2)
3.32 m
3.94 dd (10.9, 1.2)3.50 dd (10.9, 6.0)
4.31 d (7.8)
3.14 dd (8.6, 7.8)
3.35 m
3.23 dd (9.4, 9.0)
3.32 m
20
3.37 dd (8.9, 7.6)
3.46 dd (8.9, 8.5)
3.26 dd (9.7, 8.5)
3.21 ddd (9.7, 5.8, 1.6)
3.85 dd (11.9, 1.6)
3.65 dd (11.9, 5.8)
5.24 d (1.4)
3.89 dd (3.4, 1.4)
3.71 dd (9.6, 3.4)
3.36 t (9.7)
30
40
3.27 dd (9.8, 8.8)
3.22 ddd (9.8, 5.6, 2.1)
3.85 dd (11.8, 2.1)3.65 dd (11.8, 5.6)
50
60
3.95 dd (11.0, 1.5)3.55 dd (11.0, 6.5)
100
200
300
400
500
600
5.18 br s
3.92 br s
3.64 br d (9.1)
3.37 dd (9.7, 9.3)
4.04 dq (9.4, 6.2)
1.20 d (6.2)
4.93 d (2.4)
3.89 d (2.4)
—
3.95 d (9.6), 3.76 d (9.6)
3.56 s
4.99 d (2.2)
3.88 d (2.2)
—
3.96 d (9.6), 3.76 d (9.6)
3.58 s
4.14 dq (9.7, 6.2)
1.22 d (6.2)
a
Measured in methanol-d4, residue solvent peak at dH 3.31 ppm, J in Hz.
Splitted signals caused by atropisomerism at 25 °C.
b
Table 2
chemical conversion.15 Besides an additional rhamnopyranosyl
13C NMR spectroscopic data for compounds 1–4
unit presented in 1, the main differences between the 13C NMR
No.
1a
2a
3a
3b
4a
data of
d1–1a = +3.3, ꢀ2.1 and +1.2, respectively), suggesting 1 to be
20- -rhamnopyranoside of C-9 diastereomer of 1a.15
Previous studies disclosed an empirical rule that b-
1
and 1a were observed at C-20, C-8, and C-10
1
2
3
4
5
6
7
8
41.5 (s)
47.6 (t)
67.3 (d)
40.7 (t)
35.1 (d)
76.8 (s)
31.9 (t)
32.1 (t)
76.0 (d)
21.1 (q)
26.5 (q)
25.6 (q)
16.9 (q)
100.6 (d)
78.4 (d)
79.5 (d)
71.8 (d)
77.8 (d)
62.8 (t)
101.9 (d)
72.0 (d)
72.1 (d)
74.1 (d)
69.5 (d)
17.9 (q)
38.8 (s)
49.3 (t)
65.7 (d)
42.9 (t)
36.8 (s)
49.7 (t)
63.6 (d)
48.3 (t)
78.3 (s)
118.5 (s)
212.9 (s)
101.2 (d)
200.7 (s)
26.5 (q)
32.8 (q)
30.0 (q)
26.2 (q)
98.5 (d)
75.0 (d)
78.2 (d)
71.3 (d)
76.4 (d)
68.7 (t)
110.8 (d)
77.9 (d)
79.8 (s)
74.8 (t)
65.5 (t)
36.1
50.1
62.3
47.7
77.8
118.0
211.6
100.6
197.6
26.9
32.1
29.7
26.3
98.4
75.0
78.9
71.4
76.6
68.8
110.9
77.7
80.2
74.9
65.4
40.4 (d)
148.2 (s)
121.2 (d)
37.3 (t)
82.0 (s)
44.7 (s)
39.2 (t)
22.5 (q)
20.1 (q)
65.3 (t)
(D
a
-L
D
-glucosyla-
tion on chiral secondary alcohols would cause non-equivalent shift
changes on b-carbons, which was also appropriable to some digly-
cosides such as ginsenoside-Rd.16 Namely, 10-methyl would
125.6 (s)
138.2/138.1 (s)c
24.8/24.7 (t)c
36.6/36.4 (t)c
75.6/75.4 (d)c
20.92/20.88 (q)c
29.0 (q)
subject to greater shielding effect than C-8 when b-
occurred on 9R-hydroxymegastigmanes, and vice versa.15–18 The
-glucosylation shielding effects on C-8 and C-10 of
D-glucosylation
9
10
11
12
13
10
20
30
40
50
60
100
200
300
400
500
600
D
1
30.4 (q)
20.2 (q)
(
D
d1–1b = ꢀ3.7 and ꢀ2.5 ppm, respectively) were opposite trends
to those of 1a (
D
d1a–1b = ꢀ1.6 and ꢀ3.7 ppm for C-8 and C-10,
100.6/100.5 (d)c
79.25/79.19 (d)c
79.3 (d)
99.8 (d)
75.0 (d)
78.2 (d)
71.9 (d)
76.6 (d)
68.9 (t)
111.0 (d)
78.1 (d)
80.5 (s)
75.0 (t)
65.8 (t)
respectively), indicating 9S-configuration of 1. The absolute struc-
ture of pteleifoside (1) was therefore determined to be
(3S,5R,6R,9S)-megastigman-3,6,9-triol 9-O- -rhamnopyrano-
syl(1?2)-b- -glucopyranoside.
A
71.9 (d)
77.9 (d)
62.8 (t)
102.3 (d)
72.1 (d)
72.4 (d)
73.8 (d)
69.8 (d)
17.9 (q)
a-L
D
Pteleifoside B (2) was also obtained as an amorphous powder
whose elemental composition C25H44O11 was deduced from the
HR-ESI-MS. The acid hydrolysis, 1H and 13C NMR spectra (Tables
1 and 2) disclosed 2 to have the same sugar moiety of
a-L-rhamno-
pyranosyl(1?2)-b-
D
-glucopyranose with that of 1. The 1H and 13C
NMR spectra (Tables 1 and 2) of the aglycone of 2 showed 13 car-
bon atoms and 22 carbon-bearing proton signals for a doublet
methyl at dH 1.25 (d, J = 5.6 Hz), three singlet methyls at dH 1.65,
1.07, and 1.05, four methylenes at dC 49.3, 42.9, 36.6, and 24.8,
two oxygen-bearing methines at dC 75.6 and 65.7, and three qua-
ternary carbons at dC 138.2, 125.6, and 38.8, suggesting a dihydr-
oxymegastigman-5-ene diglycoside.17 The 1H and 13C NMR data
of the aglycone portion of 2 were nearly superposed with those
of linarionoside B (2a) except for the side chain carbons (C-8 to
C-10), suggesting a 9-isomer aglycone with that of 2a.17 The ob-
a
Measured in methanol-d4, residue solvent peak at dC 49.0 ppm.
Measured in pyridine-d5, residue solvent peaks at dC 149.64, 135.31, and
b
123.29 ppm.
c
Splitted signals caused by atropisomerism at 25 °C.
and 35.1, and two quaternary carbons at dC 76.8 and 41.5, indicat-
ing trihydroxylated megastigmane diglycoside. The three
hydroxyls of the aglycone were assigned on C-3, C-6, and C-9 based
on the NMR data comparison with those of dihydroalangionoside A
(1a) and dihydrodendranthemoside A (1b), which both have
3S,5R,6R,9R-configuration established by X-ray diffraction and
a
served b-
D
-glucosylation trend on C-8 (ꢀ4.2) and C-10 (ꢀ2.4) of
2 in contrast with those of linarionoside A (2b) was opposite to

116
Y. Zhang et al. / Carbohydrate Research 361 (2012) 114–119
H
H
OR1
OR1
C
C
9
8
O9
O
6
(S)
( S)
1
3
OH
OH
3
5
HO
HO
HO
HO
RO
OR
OR3
2
3
R= R2
3b
1
3a R= R3
OR'
OR'
HOH21C0
(R)
( R)
7
2
5
OR2
HO
6
4' OH
RO
1
8
9
1'
O
7
4
1a R = H, R' = R3
1b R = R3, R' = H
2a R = H, R' = R3
2b R = R3, R' = H
3
OR2
5
OH
OH
HO
HO
OH
O
OH
R2 =
O
HO
6
O
O
HO
O
OH
OH
O
HO
OH
OH
OH
OH
R1 =
O
R3=
O
O
OH OH
9
OH
OMe
O
HO
HO
MeO
3
1
6
CH2OH
7
OH
OH
O
O
HO
HO
1'''
OH
OH
9'
O
7'
1'
OH
HO
1''
O
OH
OMe
O
HO
O
MeO
OMe
4'
OH
HMBC (H to C)
OH
OH
5
7
Figure 1. Structures of compounds 1–7, 1–3a and 1–3b.
that of 2a and 2b (
D
d2a–2b = ꢀ1.8 and ꢀ3.5 ppm for C-8 and C-10,
ion peaks at m/z 563 [M+HCOO]ꢀ and 1035 ([2MꢀH]ꢀ, indicating
the molecular formula of C24H38O12. The IR absorptions displayed
the existences of hydroxyls (3417 cmꢀ1), an allenic moiety
(1940 cmꢀ1) and a conjugated carbonyl group (1664 cmꢀ1).20 After
respectively). Thus, pteleifoside B (2) was assigned to be (3R,9S)-
megastigman-5-ene-3,9-diol 9-O- -rhamnopyranosyl(1?2)-b-
-glucopyranoside.
It is interesting that some 1H and 13C signals of 2 exist in pairs
a-L
D
acid hydrolysis, 3 gave a D-apiose and a D-glucose as sugar moiety.
(Table 2), whereas it is not found in the NMR spectra of 1. A reason-
able explanation was that double bond between C-5 and C-6 of 2
obstructed the rapid rotation of the sugar unit around the gluco-
sidic linkage with the aglycone to result into equilibrium of two
conformational isomers,19 which was confirmed by the observa-
tion that the shift differences between those paired carbons tended
to zero (Fig. 2) when 13C NMR spectrum of 2 was measured at rising
temperatures.
The 1H and 13C NMR spectra (Tables 1 and 2) of 3 exhibited signals
for a b-apiofuranosyl(1?6)-b-glucopyranose at dH 4.93 (1H, d,
J = 2.4 Hz) and 4.50 (1H, d, J = 7.7 Hz), and dC 98.5, 78.2, 76.4,
75.0, 71.3, and 68.7, as well as 110.8, 79.8, 77.9, 74.8, and 65.5,
an allenic unit at dH 5.95 (s) and dC 212.9 (s), 118.5 (s) and 101.2
(d), an a,b-unsaturated carbonyl at dC 200.7, four tertiary methyls
at dH 2.20, 1.47, 1.37, and 1.16, two sp3 quaternary carbons at dC
78.3 and 36.8, one oxy-bearing methine at dC 63.6, and two meth-
ylenes at dC 49.7 and 48.3, diagnostic of grasshopper ketone.20 The
diglycosylation position of 3 was reasonably assigned on C-5 rather
than C-3 on the basis of downfield shift of a quaternary carbon at
78.3 ppm. The 13C NMR data (in CD3OD) of 3 were very similar to
those of citrosides A (3a) and B (3b)20 of which represented two
kinds of diastereomeric grasshopper ketone glucosides on C-8.
The 13C NMR data of 3a and 3b were nearly superposed except
for slight differences on C-9 and C-13 (d 197.6/199.0 and 26.5/
27.6 for 3a/3b, respectively, in pyridine-d5). Furthermore, the origi-
nal assignments for C-7 and C-9, C-2 and C-4, and four methyls of
3a and 3b in the literature20 were reversed as indicated by Hao and
co-workers.21 The 13C NMR data of the aglycone portion of 3 mea-
sured in pyridine-d5 (Table 2) were identical with those of 3a,
especially C-9 and C-13 at d 197.6 and 26.3, respectively, demon-
Pteleifoside C (3) was isolated as an amorphous powder, whose
ESI-MS (negative-ion mode) spectrum exhibited quasi-molecular
0.25
25 °C
40 °C
0.2
60 °C
0.15
0.1
0.05
0
strating 3 to be citroside A 60-O-b-
D-apiofuranoside.
Pteleifoside C (4) was obtained as an amorphous powder and its
molecular formula was determined to be C21H34O11 by the HR-ESI-
MS. The acid hydrolysis, 1H and 13C NMR spectra (Tables 1 and 2)
Δδ(C)
6
7
8
9
10
1'
2'
Figure 2. The shift differences of the paired carbons of 2 at different temperatures.

Y. Zhang et al. / Carbohydrate Research 361 (2012) 114–119
117
Table 3
1H and 13C NMR spectroscopic data (400 and 100 MHz) for 5–7a
No.
5
6
7
dC
dH (J in Hz)
dC
dH (J in Hz)
dC
dH (J in Hz)
1
2
3
4
5
6
7
135.5
114.5
150.7
146.4
118.4
122.6
39.8
—
130.1
107.7
148.6
138.9
147.6
126.3
33.8
—
6.58 s
—
—
—
—
6.88 d (1.8)
—
—
7.07 d (8.2)
6.78 dd (8.2, 1.8)
2.76 t (7.0)
83.5
77.2
195.7
165.5
96.5
5.29 d (10.0)
4.91 d (10.0)
—
—
5.90 d (2.3)
—
169.6
2.72 dd (15.0, 5.0)
2.62 dd (15.0, 11.4)
1.69 m
3.64 dd (11.9, 4.9)
3.54 dd (11.9, 6.2)
8
9
64.2
3.73 t (7.0)
96.5
164.1
5.87 d (2.3)
—
40.6
66.2
10
102.5
128.5
130.5
116.3
159.3
—
—
10
134.4
106.8
148.9
139.3
42.7
—
6.43 s
—
—
4.42 d (6.2)
2.08 m
20 & 60
30 & 50
40
70
80
90
7.36 d (8.5)
6.81 d (8.5)
—
46.7
71.4
3.88 dd (11.8, 5.3)
3.46 dd (11.8, 3.9)
3.86 s
3.35 s
3.74 s
4.35 d (7.8)
3.44 dd (9.0, 7.8)
3.56 dd (8.9, 8.7)
3.41 dd (9.8, 9.0)
3.28 m
3.85 dd (11.9, 2.0)
3.67 dd (11.9, 5.4)
4.57 d (7.8)
3.26 dd (9.2, 7.8)
3.39 dd (9.0, 8.6)
3.28 dd (8.5, 8.7)
3-OMe
56.7
3.85 s
56.5
60.1
56.8
104.4
74.5
88.1
70.0
77.6
62.6
5-OMe
30&50-OMe
100
200
300
400
500
600
103.1
74.9
78.0
71.6
77.0
68.7
4.79 d (7.2)
102.3
74.6
77.6
71.4
77.1
68.4
3.84 d (7.7)
3.22 dd (9.0, 7.8)
3.10 dd (9.1, 8.9)
3.23 dd (9.5, 9.1)
3.10 m
3.89 dd (11.0, 2.0)
3.54 dd (11.0, 5.0)
4.98 d (2.5)
3.89 d (2.5)
—
3.46 dd (7.7, 7.2)
3.44 dd (9.2, 7.7)
3.34 dd (9.8, 9.2)
3.53 ddd (9.8, 6.7, 1.5)
3.99 dd (11.1, 1.5)
3.61 dd (11.1, 6.5)
4.97 d (2.4)
3.89 d (2.4)
—
3.94 d (9.7)
3.74 d (9.7)
1000
2000
3000
4000
111.0
77.8
80.5
74.9
110.8
77.9
80.5
74.9
105.3
75.5
77.8
71.5
3.94 d (9.7)
3.74 d (9.7)
3.55 s
5000
6000
65.5
3.57 s
65.5
78.2
62.6
3.30 m
3.87 dd (11.5, 2.6)
3.63 dd (11.5, 6.6)
a
Measured in methanol-d4, residue solvent peak at dH 3.31 ppm and dC 49.0 ppm.
disclosed 4 to have the same sugar linkage of b-
D
-apiofurano-
also observed, which corroborated further 5 to be homovanillyl
alcohol 4-O-b- -apiofuranosyl-(1?6)-b- -glucopyranoside.
syl(1?6)-b-
D
-glucopyranose with that of 3. The 1H and 13C NMR
D
D
spectra (Table 1) of the aglycone of 4 showed signals for a trisub-
stituted double bond at dH 5.62 (br s) and dC 148.2 and 121.2,
two quaternary carbons at dC 82.0 and 44.7, one methine at dC
40.4, three methylenes at dC 65.3, 39.2, and 37.3, and two methyls
at dH 1.32 (s) and 0.93 (s), which were similar to those of 5-hydrox-
ynopol (4a)22 except for a hydroxymethyl signals at dH 3.91 (br d,
J = 1.2 Hz, 2H) and dC 65.3 in 4 instead of the hydroxyethyl of 4a,
Pteleifoside F (6) was isolated as an amorphous powder with
the molecular formula of C26H30O15 based on the HR-ESI-MS. The
1H and 13C NMR spectra of 6 (Table 2) exhibited signals for a b-
D-
apiofuranosyl(1?6)-b-D-glucopyranosyl group and a 2,3-trans-
dihydrokaempferol characterized with a 1,2,3,5-tetrasubstituted
aromatic ring at dH 5.90 and 5.87 (each 1H, d, J = 2.3 Hz), a 1,4-
disubstituted phenyl at dH 7.36 and 6.81 (each 2H, d, J = 8.5 Hz),
an isolated spin system of trans-1,2-diol at dH 5.29 and 4.91 (each
1H, d, J = 10.0 Hz). Compared the 1H and 13C NMR data of 6 with
and downfield shift at C-5 (Dd4–4a = 7.0 ppm). The above evidences
suggested 4 to be a 5-O-diglycoside of 2-pinen-5,10-diol. In the
HMBC spectrum of 4, obvious cross-peaks of H-10/C-5 and H2-60/
those of (2R,3R)-dihydrokaempferol 3-O-b-
main differences were that 6 had a downfield shift at C-60 and an
additional apiofuranosyl, suggesting 6 to be a 60-O-b-
-apiofur-
D
-glucoside (6a),23 the
C-100 were observed, substantiating a disaccharide of b-
D
-apiofur-
-glucopyranosyl to be connected with C-5 of the
aglycone. Compound 4 was therefore elucidated to be 2-pinen-
5,10-diol 5-O-b- -apiofuranosyl(1?6)-b- -glucopyranoside.
anosyl(1?6)-b-
D
D
anosylated derivative of 6a. The CD curve of 6 showed positive
Cotton effect at 329 nm and negative absorption at 290 nm, in
agreement with those of engeletin,24 indicating the same
2R,3R-configuration of 6. The structure of 6 was thus determined
D
D
Pteleifoside E (5) had the molecular formula of C20H30O12 as
suggested by the HR-ESI-MS. The acid hydrolysis experiment and
1H and 13C NMR data (Table 3) demonstrated a disaccharide se-
to be (2R,3R)-dihydrokaempferol 3-O-b-D-apiofuranosyl(1?6)-b-
quence of b-
D
-apiofuranosyl(1?6)-b-
D
-glucopyranosyl in the mol-
D-glucopyranoside.
ecule of 5. The aglycone of 5 was determined to be homovanillyl
alcohol based on the diagnostic 1H signals for a 1,2,4-trisubstitued
aromatic ring at dH 7.07 (d, J = 8.2 Hz), 6.88 (d, J = 1.8 Hz), and 6.78
(dd, J = 8.2, 1.8 Hz), a hydroxyethyl at dH 3.73 and 2.76 (each 2H, t,
J = 7.0 Hz), and a methoxyl at dH 3.85 (s, 3H), as well as the HMBC 3J
correlations (Fig. 1) of H2-8/C-1, MeO/C-3 and H-6/C-4. Further-
more, obvious HMBC cross-peaks of H-10/C-4 and H-100/C-60 were
The molecular formula of pteleifoside G (7) was determined to
be C34H48O18 by the HR-ESI-MS. The 1H NMR spectrum of 7 (Table
3) showed signals for two b-glucopyranosyls at dH 4.57 (d,
J = 7.8 Hz) and 4.35 (d, J = 7.8 Hz), two aromatic rings at d 6.43
H
(s, 2H) and 6.58 (s, 1H), and four methoxyls at d
3.86, 3.74,
H
3.74, and 3.35 (each s). In the 13C NMR spectrum (Table 3), the
carbon signals of 7 were in accord with those of ( )-lyoniresinol

118
Y. Zhang et al. / Carbohydrate Research 361 (2012) 114–119
90-O-b-
D
-glucopyranoside25 except for a downfield shift at C-30, to-
3.4. Identification
gether with the occurrence of an additional glucosyl unit, which
suggested 7 to be lyoniresinol 90-O-b-
D
-glucopyranosyl(1?3)-b-
D
-
3.4.1. Pteleifoside A (1)
glucopyranoside. The HMBC cross-peaks (Fig. 1) of H-70 with C-5,
C-20, and C-90, H-100 with C-90, and H-1000 with C-300 confirmed such
a planar structure. The absolute configuration of 7 was established
as 70S,8R,80R based on the same CD orientations at 242, 274, and
White amorphous powder (MeOH); ½a D22
ꢀ61.5 (c 0.13, MeOH);
ꢂ
IR (KBr) mmax 3411, 2972, 2933, 1637, 1452, 1383, 1257, 1130,
1070, 1043, 987, 642 cmꢀ1 1H NMR (500 MHz, CD3OD), and 13C
;
NMR (125 MHz, CD3OD) data, see Tables 1 and 2; ESI-MS m/z (po-
sitive) 561 [M+Na]+, (negative) 1075 [2MꢀH]ꢀ; HR-ESI-MS m/z
561.2891 [M+Na]+ (calcd for C25H46O12Na, 561.2887).
286 nm with that of (+)-lyoniresinol 90-O-b-
D
-glucopyranoside
(10).25 Therefore, 7 was elucidated to be (+)-lyoniresinol 90-O-b-
-glucopyranosyl(1?3)-b- -glucopyranoside.
D
D
3.4.2. Pteleifoside B (2)
White amorphous powder (MeOH); ½a D22
ꢂ
ꢀ66.5 (c 0.2, MeOH);
3. Experimental
IR (KBr) mmax 3415, 2967, 2927, 1639, 1454, 1381, 1259, 1130,
1043, 814, 636 cmꢀ1 1H NMR (500 MHz, CD3OD), and 13C NMR
;
3.1. General experimental procedures
(125 MHz, CD3OD) data, see Tables 1 and 2; ESI-MS m/z (positive)
543 [M+Na]+, (negative) 1039 [2MꢀH]ꢀ; HR-ESI-MS m/z 543.2783
[M+Na]+ (calcd for C25H44O11Na, 543.2781).
Optical rotations were determined on a Perkin–Elmer 341
polarimeter. CD curves were obtained on a Jasco J-810 spectropo-
larimeter. IR spectra were recorded on a Nicolet Magna-750-FTIR
spectrometer with KBr tablet. NMR spectra were acquired on a
Bruker AM-400 or a Bruker AV-500 spectrometer. ESI-MS and
HR-ESI-MS were obtained on an Esquire 3000plus and a Q-TOF-Ul-
tima mass spectrometer, respectively. Semipreparative HPLC was
carried out on a Waters 515 pump with a Waters 2487 (UV) detec-
3.4.3. Pteleifoside C (3)
White amorphous powder (MeOH);
MeOH); IR (KBr) mmax 3417, 2927, 1940, 1664, 1457, 1365, 1245,
1070, 819 cmꢀ1 1H NMR (500 MHz, pyridine-d5) d 6.15 (1H, s, H-
½
a 2D5
ꢂ
ꢀ110.3 (c 0.31,
;
8), 5.69 (1H, d, J = 2.2 Hz, H-100), 5.09 (1H, d, J = 7.7 Hz, H-10), 4.74
(1H, d, J = 2.2 Hz, H-200), 4.67 (1H, br d, J = 10.5 Hz, H-60a), 4.60
(1H, d, J = 9.3 Hz, H-400a), 4.37 (1H, d, J = 9.3 Hz, H-400b), 4.19 (2H,
s, H2-500), 4.18 (1H, dd, J = 9.3, 8.9 Hz, H-30), 4.04 (1H, dd, J = 10.6,
6.2 Hz, H-60b), 4.01–3.93 (3H, m, H-20, H-40 and H-50), 3.00 (1H,
br d, J = 13.3 Hz, H-4a), 2.27 (1H, br d, J = 12.6 Hz, H-2a), 2.21
(3H, s, Me-10), 1.77 (1H, dd, J = 13.5, 11.4 Hz, H-4b), 1.74 (3H, s,
Me-12), 1.71 (3H, s, Me-13), 1.22 (3H, s, Me-11); 1H NMR
(400 MHz, CD3OD), and 13C NMR (100 MHz, CD3OD or 125 MHz,
pyridine-d5) data, see Tables 1 and 2; ESI-MS m/z (positive) 541
[M+Na]+, (negative) 563 [M+HCOO]ꢀ, 1035 [2MꢀH]ꢀ; HR-ESI-MS
m/z 541.2270 [M+Na]+ (calcd for C24H38O12Na, 541.2261).
tor and a Kromasil 100-5-C18 column (250 ꢁ 10 mm, 5
lm). D-
1400 macroporous resin (Yangzhou Pharmaceutical Factory,
Yangzhou, China), Rp-18 reversed phase silica gel (150–200 mesh,
Fuji Silysia Chemical LTD, Aichi, Japan), MCI gel (CHP20P, 75–
150 lm, Mitsubishi Chemical Industries Ltd, Tokyo, Japan), and
Sephadex LH-20 gel (Pharmacia Biotech AB, Uppsala, Sweden)
were used for column chromatography (CC), and precoated silica
gel GF254 plates (Qingdao Haiyang Chemical Plant, Qingdao, Chi-
na) were used for TLC. All solvents used were of analytical grade
(Shanghai Chemical Plant, Shanghai, China).
3.2. Plant material
3.4.4. Pteleifoside D (4)
White amorphous powder (MeOH); ½a D25
ꢀ20.4 (c 0.43, MeOH);
ꢂ
The roots and rhizomes of M. pteleifolia were purchased from
Bozhou Chinese Materia Medica Market, Bozhou, Anhui Province,
P.R. China, in July 2009, which was identified by Professor Da-Yuan
Zhu of Shanghai Institute of Materia Medica. A voucher specimen
(No. 09-1008) was deposited with the Herbarium of Shanghai
Institute of Materia Medica.
CD (MeOH) kmax
(D
e
) 188 (ꢀ17.8), 205 (+54.1) nm; IR (KBr) mmax
3405, 2933, 1646, 1384,1319, 1216, 1049, 823 cmꢀ1 1H NMR
;
(400 MHz, CD3OD), and 13C NMR (100 MHz, CD3OD) data, see Ta-
bles 1 and 2; ESI-MS m/z (positive) 485 [M+Na]+, (negative) 461
[MꢀH]ꢀ, 923 [2MꢀH]ꢀ; HR-ESI-MS m/z 485.2001 [M+Na]+ (calcd
for C21H34O11Na, 485.1999).
3.3. Extraction and isolation
3.4.5. Pteleifoside E (5)
White amorphous powder; ½a D25
ꢂ
ꢀ74.3 (c 0.075, MeOH); IR (KBr)
1H NMR
Dried roots and rhizomes of M. pteleifolia (5 kg) were powdered
and extracted with 95% ethanol at room temperature three times.
After removal of the solvents by evaporation, the extract was sus-
pended in H2O and then partitioned successively with petroleum
ether, EtOAc, and n-BuOH. The n-BuOH-soluble part (180 g) was
subjected to macroporous resin column chromatography (CC, i.d.
10 ꢁ 80 cm) with gradient EtOH/H2O (0%, 20%, 50%, 75%, and
95%, v/v) as eluant to give fractions A–E. Fraction C (30% EtOH frac-
tion, 65 g) was separated by CC of MCI gel (MeOH/H2O 0%, 20%,
40%, 60%, and 95%, v/v) to afford sub-fractions B1-B5. Fr. B4 was
purified by CC of Rp-18 (MeOH/H2O, 50%) and Sephadex LH-20
(MeOH) to obtain 8 (20 mg), 9 (18 mg), 10 (34 mg), 16 (18 mg),
and 17 (52 mg); Fr. B3 was isolated by CC of Rp-18 (MeOH/H2O,
20–60%) and Sephadex LH-20 (MeOH/H2O, 75%) to furnish 3
(118 mg), 4 (36 mg), and 14 (30 mg); Fr. B2 was separated by CC
of Rp-18 (MeOH/H2O, 10–50%) and Sephadex LH-20 (MeOH/H2O,
50%) to offer 7 (77 mg), 6 (19 mg), and 13 (18 mg). Semiprepared
HPLC was applied to isolate 15 (9 mg, CH3OH/H2O, 50%) from Fr.
B4, 2 (9 mg, CH3OH/H2O, 40%) from Fr. B3, and 1 (8 mg, CH3OH/
H2O, 25%), 5 (9 mg, CH3OH/H2O, 20%), 11 (3 mg, CH3OH/H2O,
20%), and 12 (6 mg, CH3OH/H2O, 20%) from Fr. B2.
mmax 3404, 2932, 1603, 1516, 1272, 1051, 822 cmꢀ1
.
(400 MHz, CD3OD) and 13C NMR (100 MHz, CD3OD) data, see Table
3; ESI-MS (positive) m/z (positive) 485 [M+Na]+, (negative) 461
[MꢀH]ꢀ, 923 [2MꢀH]ꢀ; HR-ESI-MS m/z 485.1639 [M+Na]+ (calcd
for C20H30O12Na, 485.1635).
3.4.6. Pteleifoside F (6)
White amorphous powder; ½a D25
ꢀ22.1 (c 0.26, MeOH); IR (KBr)
ꢂ
mmax 3415, 2927, 1641, 1519, 1467, 1261, 1166, 1083, 833 cmꢀ1
;
CD (MeOH) kmax
(
D
e
) 216 (+115.1), 231 (ꢀ4.8), 252 (+24.0), 290
(ꢀ113.9), 329 (+42.5) nm; 1H and 13C NMR (400 and 100 MHz,
CD3OD) data, see Table 3; ESI-MS (positive) m/z (positive) 605
[M+Na]+, (negative) 581 [MꢀH]ꢀ; HR-ESI-MS m/z 605.1468
[M+Na]+ (calcd for C26H30O15Na, 605.1482).
3.4.7. Pteleifoside G (7)
White amorphous powder; ½a D25
ꢂ
+50.5 (c 0.05, MeOH); CD
(MeOH) kmax
(
D
e
) 206 (+248.6), 215 (ꢀ129.6), 243 (+110.7), 274
(+31.0), 286 (ꢀ10.0) nm; 1H and 13C NMR (400 and 100 MHz,
CD3OD) data, see Table 3; ESI-MS m/z (positive) 767 [M+Na]+,

Y. Zhang et al. / Carbohydrate Research 361 (2012) 114–119
119
(negative) 743 [MꢀH]ꢀ; HR-ESI-MS m/z 767.2732 [M+Na]+ (calcd
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Supplementary data
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