Polyphenolic glycosides and oligosaccharide multiesters from the roots of Polygala dalmaisiana

Article abstract of DOI:10.1021/np010434n

Four new polyphenolic glycosides, dalmaisiones A-D (1-4), 16 new oligosaccharide multiesters, dalmaisioses A-P (5, 7-21), and one known tetrasaccharide multiester, reiniose G (6), were isolated from the roots of Polygala dalmaisiana. The structures of the new compounds were elucidated on the basis of chemical and spectroscopic evidence.

Full text of DOI:10.1021/np010434n

J . Nat. Prod. 2002, 65, 319-328
319
P olyp h en olic Glycosid es a n d Oligosa cch a r id e Mu ltiester s fr om th e Roots of
P olyga la d a lm a isia n a
Satoko Kobayashi, Toshio Miyase,* and Hiroshi Noguchi
School of Pharmaceutical Sciences, University of Shizuoka, 52-1, Yada, Shizuoka 422-8526, J apan
Received September 5, 2001
Four new polyphenolic glycosides, dalmaisiones A-D (1-4), 16 new oligosaccharide multiesters,
dalmaisioses A-P (5, 7-21), and one known tetrasaccharide multiester, reiniose G (6), were isolated
from the roots of Polygala dalmaisiana. The structures of the new compounds were elucidated on the
basis of chemical and spectroscopic evidence.
In the course of conducting a research program on the
oligosaccharide esters from Polygala species,1 we have
investigated P. dalmaisiana (P. oppositifolia × P. myrti-
folia) (Polygalaceae). No previous investigation has been
reported on the oligosaccharide esters of P. dalmaisiana.
We now report on an investigation of the roots of P.
dalmaisiana, which led to isolation of four new compounds
(1-4), 16 new oligosaccharide esters (5, 7-21), and a
known oligosaccharide ester, reiniose G (6) (Chart 1).
Known compounds were identified by comparison of their
spectral data with reported data.2
Glc and between the proton signal due to H-1 of Glc and
the carbon signal due to C-2′ (δ 143.8), respectively. Thus,
the structure of dalmaisione A was determined to be 1.
Dalmaisione B (2) gave a pseudo molecular ion peak at
m/z 979 [M + Na]+ in a FABMS compatible with molecular
formula C45H48O23. The 1H NMR spectrum showed the
presence of an aglycone (1a ), three monosaccharides [δ 5.56
(1H, d, J ) 2 Hz), 5.18 (1H, d, J ) 7.5 Hz), 4.43 (1H, d, J
) 7.5 Hz)], and a sinapoyl residue [δ 7.20 (1H, d, J ) 16
Hz), 6.56 (2H, s), 6.02 (1H, d, J ) 16 Hz), 3.76 (6H, s)].
Acid hydrolysis afforded 1a , D-glucose, L-rhamnose, and
sinapinic acid. The 1H-1H COSY and HOHAHA difference
spectra on irradiation at each anomeric proton signal and
ROE experiments involving irradiation at each anomeric
proton signal enabled us to assign all proton signals. In
the ROE difference spectra of 2, when the proton signals
at δ 5.18 due to H-1 of Glc (inner), at δ 4.43 due to H-1 of
Glc (terminal), and at δ 5.56 due to H-1 of Rha were
irradiated, ROEs were observed at δ 7.79 due to H-3′, at δ
4.08 due to H-6 of Glc (inner), and at δ 3.85 due to H-2 of
Glc (inner), respectively. In the HMBC spectrum of 2, long-
range correlations were observed between the proton signal
at δ 5.18 due to H-1 of Glc (inner) and the carbon signal at
δ 145.9 due to C-2′; between the proton signal δ 5.56 due
to H-1 of Rha and the carbon signal δ 77.3 due to C-2 of
Glc (inner); between the proton signal δ 4.43 due to H-1 of
Glc (terminal) and the carbon signal δ 71.6 due to C-6 of
Glc (inner); between the proton signals δ 4.15 and 4.78 due
to H2-6 of Glc (terminal) and the sinapoyl carbonyl carbon
signal δ 168.5, respectively. Thus, the structure of dalmai-
sione B was determined to be 2.
Resu lts a n d Discu ssion
The roots of P. dalmaisiana were extracted with MeOH.
The MeOH extract was partitioned between H2O and
diethyl ether. The H2O layer was passed through a porous
polymer gel (Mitsubishi Diaion HP-20) column and eluted
with a mixture of H2O and MeOH. The 70% MeOH elute
was separated further to afford compounds 1-21.
The FABMS of dalmaisione A (1) gave pseudo molecular
ion peaks at m/z 589 [M + H]+ and 611 [M + Na]+,
compatible with a molecular formula C28H28O14. The 1H
1
NMR and H-1H COSY spectra showed the presence of a
1,2-disubstituted benzene ring [δ 7.86 (1H, dd, J ) 7.5, 1.5
Hz), 7.90 (1H, td, J ) 7.5, 1.5 Hz), 7.58 (1H, td, J ) 7.5,
1.5 Hz), 8.12 (1H, dd, J ) 7.5, 1.5 Hz) in this order] and a
1,2,3-trisubstituted benzene ring [δ 7.66 (1H, dd, J ) 7.5,
1.5 Hz), 7.46 (1H, t, J ) 7.5 Hz), 7.92 (1H, dd, J ) 7.5, 1.5
Hz) in this order] and two anomeric protons [δ 5.45 (1H,
1
d, J ) 7.5 Hz), 5.24 (1H, d, J ) 1.5 Hz)]. The H-1H COSY
and HOHAHA difference spectra on irradiation at each
anomeric proton signal and ROE experiments involving
irradiation at each anomeric proton signal enabled us to
assign all proton signals. Acid hydrolysis afforded 1a as
an aglycone and D-glucose and L-rhamnose as sugar
moieties.3 Compound 1a was methylated by diazomethane
to afford the methyl ether 1b. Compound 1b was identified
as frutinone B by comparison of spectral data with reported
data.4 The sugar sequence and the glycosidic site were
decided by ROE and HMBC. In the ROE difference spectra
of 1, when the proton signals at δ 5.45 due to H-1 of Glc
and δ 5.24 due to H-1 of Rha were irradiated, ROEs were
observed at δ 7.66 due to H-3′ and δ 3.68 due to H-2 of
Glc, respectively. In the HMBC spectrum of 1, long-range
correlations were observed between the proton signal due
to H-1 of Rha and the carbon signal due to C-2 (δ 76.2) of
Dalmaisione C (3) gave NMR data similar to those of 1.
In the 1H-1H COSY spectrum, a 1,2-disubstituted benzene
ring [δ 7.43 (1H, dd, J ) 8, 1.5 Hz), 7.84 (1H, td, J ) 8, 1.5
Hz), 7.55 (1H, td, J ) 8, 1.5 Hz), 8.45 (1H, dd, J ) 8, 1.5
Hz) in this order] and a 1,2,3-trisubstituted benzene ring
[δ 7.70 (1H, dd, J ) 8, 1.5 Hz), 7.49 (1H, t, J ) 8 Hz), 7.91
(1H, dd, J ) 8, 1.5 Hz) in this order] were observed other
than two anomeric protons [δ 5.47 (1H, d, J ) 7.5 Hz), 5.38
(1H, d, J ) 2 Hz)]. Compound 3 afforded D-glucose and
L-rhamnose on acid hydrolysis. After assignment of all
1
proton signals by H-1H COSY and HOHAHA difference
spectra, the sugar sequence was determined to be R-L-
rhamnopyranosyl-(1f2)-â-D-glucopyranosyl using ROE and
HMBC spectra. On irradiation of the glucosyl anomeric
proton signal at δ 5.47, ROE was observed at the aromatic
proton signal at δ 7.70, which had an HMBC correlation
to the carbon signal at δ 120.3. The carbon signal at δ 120.3
was correlated to the proton signal δ 7.91 in the HMQC
* To whom correspondence should be addressed. Tel/Fax: +81-54-264-
5661. E-mail: miyase@ys7.u-shizuoka-ken.ac.jp.
10.1021/np010434n CCC: $22.00
© 2002 American Chemical Society and American Society of Pharmacognosy
Published on Web 02/19/2002

320 J ournal of Natural Products, 2002, Vol. 65, No. 3
Kobayashi et al.
Ch a r t 1
spectrum. The proton signal at δ 7.91 had a correlation to
the carbon signal at δ 175.3 due to C-4 of the aglycone.
Thus, the structure of dalmaisione C was determined to
be 3.
correlations were observed between a proton signal at δ
7.10 and carbon signals at δ 177.1, 123.1 (W-type long-
range coupling5), 160.4, 120.6, 155.3, and an anomeric
proton signal at δ 5.10 and carbon signal at δ 116.0, and
aromatic proton signals at δ 7.73 and 8.06 (W-type long-
range coupling) and a carbonyl carbon signal at δ 177.1
(Figure 2). Sugar sequence was decided by the ROE
spectrum to be glucopyranosyl-(1f6)-glucopyranosyl. These
data led us to assign the structure of dalmaisione D as 4.
Dalmaisiose A (5) has a molecular formula of C38H46O20
on the basis of FABMS [M + Na]+ m/z 845. The UV and
1H NMR spectra showed the presence of an acetyl, a
cinnamoyl, and a p-coumaroyl residue. Hydrolysis with
alkali and acid afforded acetic acid, p-coumaric acid, and
cinnamic acid as ester moieties and D-glucose and D-
fructose as sugar moieties.3,6 All proton signals were
assigned by the aid of 1H-1H COSY and HOHAHA
difference spectra. In the HMBC spectrum, correlations
were observed at the signals H2-6 (δ 4.14, 4.09) of Glc1/
acetyl carbonyl carbon (δ 172.7), H-4 (δ 4.85) of Glc1/
carbonyl carbon at δ 167.5 which was assigned to the
Dalmaisione D (4) was shown to have molecular formula
C
27H30O13 by FABMS ([M + H]+ m/z 563), which was
consistent with the 13C NMR spectrum. Acid hydrolysis
afforded 4a and D-glucose. Two 1,2-disubstituted benzene
rings [δ 8.06 (1H, dd, J ) 7.5, 1.5 Hz), 7.50 (1H, td, J )
7.5, 1.5 Hz), 7.82 (1H, td, J ) 7.5, 1.5 Hz), 7.73 (1H, dd, J
) 7.5, 1.5 Hz) in this order; 7.92 (1H, dd, J ) 7.5, 1.5 Hz),
7.22 (1H, td, J ) 7.5, 1.5 Hz), 7.57 (1H, td, J ) 7.5, 1.5
Hz), 7.49 (1H, dd, J ) 7.5, 1.5 Hz) in this order] and an
olefinic proton signal at δ 7.10 (1H, s) were observed other
than two monosaccharides in the 1H NMR spectrum. All
1
proton signals were assigned by the aid of H-1H COSY
and HOHAHA difference spectra. In the ROE spectra,
irradiation at the olefinic proton signal at δ 7.10 enhanced
the aromatic proton signal at δ 7.92, and irradiation at the
anomeric proton signal at δ 5.10 enhanced the aromatic
proton signal at δ 7.49. In the HMBC spectrum, long-range

Polyphenolic Glycosides from the Roots of Polygala
J ournal of Natural Products, 2002, Vol. 65, No. 3 321
Ta ble 1. 1H NMR and 13C NMR Data of Compound 1-3 at 35 °Cd
1a
2b
3b
1H
13C
1H
13C
1H
13C
aglycon moiety
2
3
164.8G
104.7
166.6G
105.4
166.9G
105.7
4
172.5A
125.5
174.9A
127.1
175.3A
120.3
5
8.12 (dd, 7.5, 1.5)A
7.58 (td, 7.5, 1.5)B
7.90 (td, 7.5, 1.5)C
7.86 (dd, 7.5, 1.5)D
8.15 (dd, 8, 2)A
7.52 (td, 8, 2)B
7.83 (td, 8, 2)C
7.65 (dd, 8, 2)D
7.91 (dd, 8, 1.5)A
7.49 (t, 8)B
6
126.5
127.7
127.6
123.8
7
135.0
136.4A
119.3
7.70 (dd, 8, 1.5)C,#
8
118.4
147.3B,C,H
147.2C
126.5B
158.6
9
154.1A,C
124.0B,D
154.9
155.9A
125.2B,D
158.4
10
11
1′
2′
143.6E,G
143.8F,H
145.4E,G
145.9F,H
124.4
155.6D,F,G
118.1
7.43 (dd, 8, 1.5)D
7.84 (td, 8, 1.5)E
7.55 (td, 8, 1.5)F
8.45 (dd, 8, 1.5)G
3′ 7.66 (dd, 7.5, 1.5)E,† 120.1
7.79 (dd, 8, 2)E,#
7.31 (t, 8)F
137.1
4′ 7.46 (t 7.5)F
124.5
116.5
114.2F
126.3
126.7
5′ 7.92 (dd, 7.5, 1.5)G
6′
7.70 (dd, 8, 2)G
118.3
126.4
115.2F
114.8D,F,G
sugar moiety
Glc-1 5.45 (d, 7.5)H,#
97.3
76.2I
77.6
69.6
77.0
60.4
5.18 (d, 7.5)H,#
3.85 (dd, 9, 7.5)*
3.71 (dd, 9, 9)
3.35 (dd, 9, 9)
3.95 (m)
101.2
77.3L
79.7
72.4
77.1
71.6
5.47 (d, 7.5)H,#
3.86 (dd, 9.5, 7.5)*
3.69 (dd, 9.5, 9.5)
3.51 (dd, 9.5, 9.5)
(3.46 (m)
101.4
80.8I
78.6
71.4
78.3
62.4
2
3
4
5
6
3.68 (dd, 8.5, 7.5)*
3.54 (dd, 8.5, 8.5)
3.27 (dd, 8.5, 8.5)
3.42 (m)
3.47c
3.90 (br d, 12)I
4.08 (br d, 12)I,§
4.43 (d, 7.5)§
3.28c
3.69 (dd, 12.5, 5)
3.83 (dd, 12.5, 2.5)
3.66c
Glc (terminal)-1
105.9I
75.5
77.9
71.2
75.5
63.0
2
3
4
5
6
3.40 (dd, 9, 9)
3.52 (dd, 9, 9)
3.45 (m)
4.15 (dd, 12.5, 3.5)J
4.78c,K
Rha-1 5.24 (d, 1)I,*
100.1
70.4
70.4
72.0
68.5
18.0
5.56 (d, 2)L,*
3.99 (dd, 3.5, 2)
3.85 (dd, 10, 3.5)
3.42 (dd, 10, 10)
4.20 (m)
101.2
72.3
72.4
74.4
70.3
18.5
5.38 (d, 2)I,*
4.04 (dd, 3, 2)
3.64 (dd, 10, 3)
3.26 (dd, 10, 10)
3.78 (m)
103.0
72.2
72.2
73.9
70.2
17.9
2
3
4
5
6
3.73 (dd, 3, 1)
3.34 (dd, 9, 3)
3.15 (dd, 9, 9)
3.73 (m)
1.14 (d, 6)
1.34 (d, 6)
0.92 (d, 6)
acid
(at C-6 of Glc2)
R
â
168.5J ,K,N
115.6
6.02 (d, 16)M,†
7.20 (d, 16)N,†
γ
146.5O
126.3M
106.5
1
2,6
3,5
4
6.56 (s)O,†,‡
149.0P
139.4O
56.9
OMe
3.76 (s)P,‡
a
b
d
In DMSO-d6. In CD3OD. c Overlapped with other signals. Assigned with the aid of HOHAHA difference, 1H-1H COSY, HMQC,
and HMBC spectra. ROEs were observed between protons that have the same (#,*,§,†,‡,¶ ) in each column. Long-range correlations were
observed between protons and carbons that have the same letter (A,B,...,P) in the same compounds.
F igu r e 2. 1H-13C long-range correlations and ROEs of 4.
carbonyl carbon at δ 167.8 which was assigned to the
R-carbon of a cinnamoyl residue, and H-1 (δ 5.42) of Glc1/
anomeric carbon (δ 107.2) of Fru and H-1 (δ 4.43) of Glc2/
C-4 (δ 84.4) of Fru. Accordingly, 5 was confirmed to be â-D-
glucopyranosyl-(1f4)-(3-O-cinnamoyl)-â-D-fructofuranosyl-
(2f1)-(6-O-acetyl-4-O-p-coumaroyl)-R-D-glucopyranoside.
The NMR spectra of dalmaisiose B (7) were similar to
those of reiniose G (6)2 except for the presence of a
rhamnosyl residue. After assignment of all proton signals
F igu r e 1. 1H-13C long-range correlations and ROEs of 7.
R-carbon of a p-coumaroyl residue by HMBC correlation
with sequence of the aromatic proton, H-3 (δ 5.77) of Fru/

322 J ournal of Natural Products, 2002, Vol. 65, No. 3
Kobayashi et al.
Ta ble 2. 1H NMR Data of Compounds 7-11 in CD3OD at 35 °Cb
7
8
9
10
11
sugar moiety
Glc1-1 5.85 (d, 3)A
-2 3.81 (dd, 8.5, 3)#
5.86 (d, 3.5)A
5.86 (d, 3.5)A
5.86 (d, 3.5)A
3.81 (dd, 9, 3.5)
4.03 (dd, 9, 9)
5.00 (dd, 9, 9)B
4.39 (m)
5.86 (d, 5)A
3.82 (dd, 10, 3.5)#
3.81 (dd, 10, 3.5)#
4.03 (dd, 10, 10)*
5.00 (dd, 10, 8.5)B
4.39 (m)
3.81 (dd, 10, 5)#
4.03 (dd, 10, 10)*
5.00 (dd, 10, 10)B
4.39 (m)
-3 3.95 (dd, 8.5, 8.5)* 3.97 (dd, 10, 10)*
-4 5.01 (dd, 10, 8.5)B
-5 4.39 (m)
5.02 (dd, 10, 9)B
4.39 (m)
-6 4.14 (dd, 12, 5.5)C
4.20a
4.14 (dd, 12.5, 5.5)C 4.14 (dd, 12.5, 5)C
4.14 (dd, 12.5, 5.5)C 4.14 (dd, 12.5, 5)C
4.20 (dd, 12.5, 3.5)
4.60 (d, 7.5)D,#
3.32a
4.20 (dd, 12.5, 3.5)C 4.21 (dd, 12.5, 3) 4.20a
Glc2-1 4.60 (d, 7.5)D,#
-2 3.32a
4.59 (d, 7.5)D,#
3.31a
4.59 (d, 7.5)D
3.32a
4.59 (d, 7.5)D,#
3.32a
-3 3.32a
3.32a
3.31a
3.32a
3.32a
-4 3.32a
3.32a
3.31a
3.32a
3.32a
-5 3.32a
3.32a
3.31a
3.31a
3.32a
-6 3.71 (m)
3.72 (dd, 12.5, 5.5)
3.71 (d, 12)
3.93 (d, 12)
4.46 (d, 7.5)E,*
3.01 (dd, 8.5, 7.5)
3.20 (dd, 8.5, 8.5)
3.17 (dd, 8.5, 8.5)
3.02a
3.71 (d, 12)
3.72a
3.94a
3.95a
3.93a
3.93 (d, 11)
4.46 (d, 7.5)E,*
3.01 (dd, 8.5, 7.5)
3.20 (dd, 8.5, 8.5)
3.16 (dd, 8.5, 8.5)
3.02a
Glc3-1 4.49 (d, 8)E,*
-2 2.99 (dd, 9, 8)
-3 3.17 (dd, 9, 9)
-4 3.18a
4.50 (d, 7.5)E,*
3.00 (dd, 8.5, 7.5)
3.18 (dd, 8.5, 8.5)
3.21 (dd, 8.5, 8.5)
3.10 (m)
4.46 (d, 7.5)E
3.01 (dd, 8.5, 7.5)
3.20 (dd, 8.5, 8.5)
3.15 (dd, 8.5, 8.5)
3.03 (m)
-5 3.08 (m)
-6 3.95a,F
3.98 (dd, 12.5, 5.5)F
4.07 (dd, 12.5, 3.5)
4.21 (d, 12.5)G
4.72 (d, 12.5)G
5.73 (d, 8)H
4.43 (dd, 8, 8)
4.07a
3.43 (dd, 12, 5)
3.58 (dd, 12, 4)
4.20 (d, 12.5)F
4.71 (d, 12.5)F
5.74 (d, 8)G
4.43 (dd, 8, 8)
4.07 (m)
3.44 (dd, 12, 5)
3.61 (dd, 12, 3)
4.19 (d, 12.5)F
4.70 (d, 12.5)F
5.74 (d, 8)G
4.43 (dd, 8, 8)
4.07 (m)
3.43 (dd, 12.5, 5)
3.58 (dd, 12.5, 2.5)
4.20 (d, 12)F
4.70a,F
4.03a
Fru-1 4.20 (d, 12)G
4.70 (d, 12)G
-3 5.73 (d, 8)H
-4 4.43 (dd, 8, 8)
-5 4.07 (m)
5.74 (d, 8)G
4.44 (dd, 8, 8)
4.08 (m)
-6 3.86a
3.86a
3.86a
3.86a
3.87a
3.86a
3.86a
3.86a
3.86a
3.87a
Rha-1 5.53 (d, 2)I,§
-2 4.02 (dd, 3, 2)
-3 3.85 (dd, 9, 3)
-4 3.48 (dd, 9, 9)
-5 3.62 (m)
-6 1.23 (d, 6)
5.48 (d, 2)I,§
4.09 (dd, 3, 2)
3.91a
5.52 (d, 2)H,§
4.02 (dd, 3, 2)
3.85 (dd, 9, 3)
3.48 (dd, 9, 9)
3.63 (m)
5.46 (d, 2)H,§
4.08 (dd, 3, 2)
3.90 (dd, 9, 3)
3.47 (dd, 9, 9)
3.75 (m)
5.52 (d, 2)H,§
4.02 (dd, 3, 2)
3.85 (dd, 9, 3)
3.48 (dd, 9, 9)
3.63 (m)
3.47 (dd, 9, 9)
3.75 (m)
1.24 (d, 6)
1.24 (d, 6)
1.24 (d, 6)
1.24 (d, 6)
ester moiety
(at C-6 of Glc1)
(at C-6 of Glc3)
(at C-4 of Glc1)
Ac 2.06 (s)J
Ac 1.59 (s)K
2.06 (s)J
2.07 (s)I
2.07 (s)I
2.07 (s)I
1.58 (s)K
â
6.31 (d, 16)L
7.59 (d, 16)M
7.56 (d, 8)N
7.15 (d, 8)O
7.15 (d, 8)O,§
7.56 (d, 8)N
6.34 (d, 16)L
7.59 (d, 16)M
7.27 (d, 2)N,†
6.41 (d, 16)J
7.62 (d, 16)K
7.60 (d, 8)L
7.15 (d, 8)M,§
7.15 (d, 8)M,§
7.60 (d, 8)L
6.44 (d, 16)J
7.61 (d, 16)K
7.31 (d, 1.5)L,†
6.41 (d, 16)J
7.62 (d, 16)K
7.60 (d, 8)L
7.15 (d, 8)M,§
7.15 (d, 8)M,§
7.60 (d, 8)L
γ
2
3
5
6
7.19 (d, 8)O,§
7.13 (dd, 8, 2)P
3.93 (s)Q,†
7.19a,M
7.19a,N,§
3.93 (s)O,†
OMe
(at C-6 of Glc3)
â
γ
2
3
5
6
OMe
(at C-1 of Fru)
â
6.37 (d, 16)P
7.68 (d, 16)Q
7.43 (d, 8)R
6.81 (d, 8)S
6.81 (d, 8)S
7.43 (d, 8)R
6.40 (d, 16)R
7.48 (d, 16)S
7.20 (d, 2)T,‡
6.36 (d, 16)N
7.68 (d, 16)O
7.43 (d, 8)P
6.81 (d, 8)Q
6.81 (d, 8)
6.36 (d, 16)P
7.68 (d, 16)Q
7.43 (d, 8)R
6.81 (d, 8)S
6.81 (d, 8)
6.41 (d, 16)N
7.68 (d, 16)O
7.20 (d, 2)P,†
γ
2
3
5
6
6.81 (d, 8)U
6.82 (d, 8)Q
7.02 (dd, 8, 2)V
3.91 (s)W,‡
7.43 (d, 8)
7.43 (d, 8)
7.04 (dd, 8, 2)R
3.91 (s)S,†
OMe
(at C-3 of Fru)
2, 6 8.18 (dd, 7.5, 1)T
8.18 (dd, 7.5, 1)X
7.59 (t, 7.5)Y
7.61 (t, 7.5)
8.16 (dd, 8, 2)R
7.58 (t, 8)S
7.65 (tt, 8, 2)
8.16 (dd, 7.5, 1)T
7.58 (t, 7.5)U
7.61 (tt, 7.5, 1)
8.16 (dd, 7.5, 1)T
7.58 (t, 7.5)U
7.63 (tt, 7.5, 1)
3, 5 7.59 (t, 7.5)U
4
7.71 (tt, 7.5, 1)
a
Overlapped with other signals ROEs were observed between protons that have the same letter (#,*,§,†,‡,¶ ) in each column. Long-range
correlations were observed between protons and carbons that have the same letter (A,B,...,γ,δ) in the same compounds in Table 5. Assigned
b
with the aid of HOHAHA difference, 1H-1H COSY, HMQC, and HMBC spectra.
by 1H-1H COSY and HOHAHA difference spectra, irradia-
tion of a rhamnosyl anomeric proton signal at δ 5.53 (1H,
d, J ) 2 Hz) enhanced the signal area of an aromatic proton
signal at δ 7.15 (2H, d, J ) 8 Hz), which was assigned to
H-3 and H-5 of the p-coumaroyl residue attached to C-4 of
R-D-glucose (Figure 1). Hydrolysis with alkali and acid gave
acetic acid, p-coumaric acid, and benzoic acid as ester
moieties and D-glucose, D-fructose, and L-rhamnose as
sugar moieties.
The NMR spectra of dalmaisioses C-G (8-12) were
similar to those of 7, showing acetyl(s), a benzoyl, and two
oxycinnamoyl residues as ester moieties and a rhamnosyl,
a fructosyl, and three glucosyl residues as sugar moieties.
The binding sites of these residues were decided with the
aid of ROE and HMBC after assignment of all proton
signals by 1H-1H COSY and HOHAHA difference spectra.
The structures of dalmaisioses C-G were determined to
be 8-12, respectively.

Polyphenolic Glycosides from the Roots of Polygala
J ournal of Natural Products, 2002, Vol. 65, No. 3 323
Ta ble 3. 1H NMR Data of Compounds 12-16 in CD3OD at 35 °Cb
12
13
14
15
16
sugar moiety
Glc1-1
5.86 (d, 3.5)A
3.82 (dd, 9, 3.5)#
4.03 (dd, 9, 9)*
5.00 (dd, 10.5, 9)B
4.39 (m)
5.84 (d, 3.5)A
3.82 (dd, 10, 3.5)#
3.99 (dd, 10, 10)*
5.02 (dd, 10, 9)B
4.38 (m)
5.84 (d, 3.5)A
3.82 (dd, 10, 3.5)#
3.97 (dd, 10, 10)*
5.03 (dd, 10, 10)B
4.38 (m)
5.82 (d, 3.5)A
3.79 (dd, 9, 3.5)#
3.99 (dd, 9, 9)*
5.03 (dd, 9, 9)B
4.20a
5.84 (d, 3)A
3.83 (dd, 9, 3)
3.97 (dd, 9, 9)
5.03 (dd, 9, 9)B
4.38 (m)
-2
-3
-4
-5
-6
4.14 (dd, 12.5, 5.5)C
4.21 (dd, 12.5, 3)C
4.60 (d, 7.5)D,#
3.32a
4.13a,C
4.14a,C
3.54 (dd, 12.5, 5)
3.66 (dd, 12.5, 3.5)
4.62 (d, 7.5)C,#
3.33a
4.14a,C
4.13a
4.14a
4.14a
Glc2-1
4.62 (d, 7.5)D,#
3.35a
4.63 (d, 8)D,#
3.34a
4.63 (d, 7.5)D
3.34 (dd, 10, 7.5)
3.31a
-2
-3
-4
-5
-6
3.32a
3.35a
3.34a
3.33a
3.32a
3.35a
3.34a
3.33a
3.31a
3.32a
3.35a
3.34a
3.33a
3.31a
3.72 (dd, 12, 3.5)
3.72a
3.72a
3.73 (dd, 12.5, 5.5)
3.93 (dd, 12.5, 5)
3.72 (dd, 12, 3)
3.94 (br d, 12)
4.53 (d, 8)E
3.06 (dd, 9, 8)
3.21 (dd, 9, 9)
3.33 (dd, 9, 9)
3.13 (m)
3.93a
3.94 (dd, 12, 2)
4.52 (d, 8)E,*
3.06 (dd, 9, 8)
3.21 (dd, 9, 9)
3.30 (dd, 9, 9)
3.15 (m)
3.94 (br d, 12.5)
4.52 (d, 8)E,*
3.06 (dd, 9, 8)
3.21 (dd, 9, 9)
3.34 (dd, 9, 9)
3.12 (m)
Glc3-1
-2
4.47 (d, 8)E,*
3.01 (dd, 8.5, 8)
3.20 (dd, 8.5, 8.5)
3.15 (dd, 8.5, 8.5)
3.05 (m)
4.50 (d, 8)D,
*
3.06 (dd, 9, 8)
3.20 (dd, 9, 9)
3.33a
-3
-4
-5
--6
3.10 (m)
3.44 (dd, 12, 5)
3.61 (dd, 12, 3)
4.19 (d, 12)F
4.72 (d, 12)F
5.u75 (d, 8)G
4.44 (dd, 8, 8)
4.08a
4.09 (dd, 12.5, 3)
4.18 (dd, 12.5, 4)F
4.22 (d, 12)G
4.71 (d, 12)G
5.73 (d, 8)H
4.43 (dd, 8, 8)
4.06 (m)
4.07a,F
4.08 (dd, 12.5, 2.5)
4.19 (dd, 12.5, 3.5)E
4.22 (d, 12)F
4.72a
3.04 (dd, 12, 2.5)
4.23 (dd, 12, 3.5)F
4.22 (d, 2.5)G
4.72a,G
4.23a,F
Fru-1
4.22 (d, 12)G
4.70 (d, 12)G
5.73 (d, 8)H
4.44 (dd, 8, 8)
4.06a
-3
-4
-5
-6
5.72 (d, 8)G
4.50 (dd, 8, 8)
4.03a
5.74 (d, 8)H
4.43 (dd, 8, 8)
4.07a
3.86a
3.86a
3.86a
3.86a
3.85a
3.86a
3.86a
3.86a
3.86a
3.87a
Rha-1
-2
5.46 (d, 2)H,§
4.09 (dd, 3, 2)
3.91a
5.42 (d, 2)I,§
4.07 (dd, 3, 2)
3.90 (dd, 9, 3)
3.48 (dd, 9, 9)
3.79 (m)
5.42 (d, 2)I,§
4.07 (dd, 3.5, 2)
3.90 (dd, 9, 3.5)
3.48 (dd, 9, 9)
3.78 (m)
-3
-4
-5
3.48 (dd, 9, 9)
3.76 (m)
-6
1.24 (d, 6.5)
1.27 (d, 6)
1.26 (d, 6)
ester moiety
(at C-6 of Glc1)
(at C-6 or Glc3)
(at C-4 of Glc1)
Ac
Ac
2.07 (s)I
2.04 (s)I
2.04 (s)J
2.04 (s)J
â
6.44 (d, 16)J
7.16 (d, 16)K
7.30 (d, 1)L,†
6.18 (d, 15.5)J
7.50 (d, 15.5)K
7.31 (d, 8)L
6.71 (d, 8)M
6.71 (d, 8)
6.28 (d, 16)K
7.50 (d, 16)L
7.04 (d, 2)M,†
6.21 (d, 16)H
7.49 (d, 16)I
7.00 (d, 2)J
6.28 (d, 16)K
7.50 (d, 16)L
7.04 (d, 2)M,†
γ
2
3
5
6
7.19a,M,§
7.19a,N
7.03 (d, 8)N,§
6.97 (dd, 8, 2)O
3.76 (s)P,†
6.69 (d, 8)K,§
6.90 (dd, 8, 2)L
3.81 (s)M,§
7.03 (d, 8)N,§
6.97 (dd, 8, 2)O
3.76 (s)P,†
7.31 (d, 8)
OMe
3.92 (s)O,†
(at C-6 of Glc3)
â
γ
2
3
5
6
5.98 (d, 15.5)N
7.35 (d, 15.5)O
7.24 (d, 8)P
6.77 (d, 8)Q
6.77 (d, 8)
5.89 (d, 16)Q
7.26 (d, 16)R
7.20 (d, 8)S
6.76 (d, 8)T
6.76 (d, 8)
5.92 (d, 16)N
7.27 (d, 16)O
7.20 (d, 8)P
6.76 (d, 8)Q
6.76 (d, 8)
5.89 (d, 16)Q
7.26 (d, 16)R
7.21 (d, 8)S
6.76 (d, 8)
6.76 (d, 8)
7.21 (d, 8)
7.24 (d, 8)
7.20 (d, 8)
7.20 (d, 8)
OMe
(at C-1 of Fru)
â
6.41 (d, 16)P
7.68 (d, 16)Q
7.20 (d, 2)R,‡
6.36 (d, 15.5)R
7.68 (d, 15.5)S
7.42 (d, 8)T
6.81 (d, 8)U
6.81 (d, 8)
6.36 (d, 16)U
7.78 (d, 16)V
7.42 (d, 8)W
6.81 (d, 8)X
6.81 (d, 8)
6.36 (d, 16)R
7.68 (d, 16)S
7.42 (d, 8)T
6.81 (d, 8)U
6.81 (d, 8)
6.41 (d, 16)U
7.68 (d, 16)V
7.20 (d, 2)W,‡
γ
2
3
5
6
6.81 (d, 8)S
7.04 (dd, 8, 2)T
3.90 (s)U,‡
8.16 (dd, 8, 2)V
7.58a,W
6.82 (d, 8)X
7.42 (d, 8)
7.42 (d, 8)
7.42 (d, 8)
7.02 (dd, 8, 2)Y
3.91 (s)Z,‡
OMe
2, 6
3, 5
4
(at C-3 of Fru)
8.17 (dd, 7.5, 1)V
7.57 (t, 7.5)W
7.68 (tt, 7.5, 1)
8.18 (dd, 7.5, 1)Y
7.60 (t, 7.5)Z
7.71 (tt, 7.5, 1)
8.20 (dd, 8, 1)V
7.60 (t, 8)W
7.70 (tt, 8, 1)
8.18 (dd, 7.5, 1)R
7.60 (t, 7.5)â
7.71 (tt, 7.5, 1)γ
7.60a
a
Overlapped with other signals ROEs were observed between protons that have the same letter (#,*,§,†,‡,¶ ) in each column. Long-range
correlations were observed between protons and carbons that have the same letter (A,B,...,γ,δ) in the same compounds in Table 5. Assigned
b
with the aid of HOHAHA difference, 1H-1H COSY, HMQC, and HMBC spectra.
Dalmaisioses H-K (13-16) are tetrasaccharides having
the same sugar sequence as that of reiniose G (6) and with
a p-coumaroyl residue at C-6 of Glc3. Dalmaisioses L-P
(17-21) are also tetrasaccharides, but these have a feruloyl
residue at C-6 of Glc3. In these compounds, the positions
of each residue were confirmed by the same method as for
previously mentioned compounds.
D-glucopyranosyl-(1f3)]-R-D-glucopyranoside are most widely
distributed in Polygala species. The sugar sequence of â-D-
glucopyranosyl-(1f4)-â-D-fructofuranosyl-(2f1)-R-D-glu-
copyranoside in dalmaisiose A (5) is the first example in
Polygala species.
Exp er im en ta l Section
Tetrasaccharide multiesters having a sugar sequence of
â-D-fructofuranosyl-(2f1)-[ â-D-glucopyranosyl-(1f2)]-[â-
Gen er a l Exp er im en ta l P r oced u r es. Optical rotations
were measured on a J ASCO DIP-1000 digital polarimeter at

324 J ournal of Natural Products, 2002, Vol. 65, No. 3
Kobayashi et al.
Ta ble 4. 1H NMR Data of Compounds 17-21 in CD3OD at 35 °Cb
17
18
19
20
21
sugar moiety
Glc1-1
5.82 (d, 3)A
3.79a,#
3.98 (dd, 9, 9)*
5.02 (dd, 10.5, 9)B
4.23a
5.84 (d, 3.5)A
3.82 (dd, 10, 3.5)#
3.98 (dd, 10, 10)*
5.02 (dd, 10, 8.5)B
4.38 (m)
5.85 (d, 3)A
3.82 (dd, 9, 3)#
3.97 (dd, 9, 9)*
5.03 (dd, 10.5, 9)B
4.38 (m)
5.85 (d, 3.5)A
3.82 (dd, 10, 3.5)#
3.97 (dd, 10, 10)*
5.03 (dd, 10, 9)B
4.38 (m)
5.83 (d, 3.5)A
-2
-3
-4
-5
-6
3.79 (dd, 9, 3.5)#
3.98 (dd, 9, 9)*
5.04 (dd, 9, 9)B
4.22 (m)
3.54 (dd, 12.5, 5)
3.66 (dd, 12.5, 2.5)
4.62 (d, 7.5)C,#
3.33a
4.13a,C
4.14a,C
4.14a,C
3.54 (dd, 12.5, 5.5)
3.67 (dd, 12.5, 3.5)
4.63 (d, 7.5)C,#
3.33a
4.13a
4.14a
4.14a
Glc2-1
4.62 (d, 8)D,#
3.32a
4.63 (d, 7.5)D,#
3.35 (dd, 7.5, 7.5)
3.33a
4.63 (d, 7.5)D,#
3.37a
-2
-3
-4
-5
-6
3.33a
3.32a
3.34a
3.33a
3.33a
3.32a
3.33a
3.34a
3.33a
3.33a
3.32a
3.33a
3.34a
3.33a
3.73 (dd, 12, 6)
3.93 (dd, 12, 2)
3.72 (br d, 12.5)
3.93 (br d, 12.5)
4.53 (d, 8)E,*
3.06 (dd, 9, 8)
3.21 (dd, 9, 9)
3.35 (dd, 9, 9)
3.12 (m)
3.72a
3.73a
3.73a
3.94 (dd, 11, 2.5)
4.53 (d, 7.5)E,*
3.06 (dd, 9, 7.5)
3.22 (dd, 9, 9)
3.36 (dd, 9, 9)
3.13 (m)
3.94 (dd, 11, 2.5)
4.53 (d, 7.5)E,*
3.07 (dd, 9, 7.5)
3.22 (dd, 9, 9)
3.35 (dd, 9, 9)
3.14 (m)
3.94a
Glc3-1
4.51 (d, 7.5)D,
*
4.51 (d, 7.5)D,
*
-2
-3
-4
-5
-6
3.06 (dd, 9, 7.5)
3.20 (dd, 9, 9)
3.35 (dd, 9, 9)
3.10 (m)
3.06 (dd, 9, 7.5)
3.20 (dd, 9, 9)
3.35 (dd, 9, 9)
3.11 (m)
4.07 (dd, 12, 3)
4.20 (dd, 12, 3)
4.23 (d, 12)F
4.79 (d, 12)F
5.72 (d, 8)G
4.50 (dd, 8, 8)
4.03 (m)
4.08a
4.03 (dd, 12, 2)F
4.25 (dd, 12, 3.5)
4.22 (d, 12)G
4.71 (d, 12)G
5.73 (d, 8)H
4.43 (dd, 8, 8)
4.08a
4.03 (dd, 12, 2)
4.25 (dd, 12, 3)F
4.22 (d, 12)G
4.72 (d, 12)G
5.74 (d, 8)H
4.44 (dd, 8, 8)
4.09 (m)
3.84a
4.21 (dd, 12, 3.5)F
4.21 (d, 12)G
4.72 (d, 12)G
5.73 (d, 8)H
4.42 (dd, 8, 8)
4.07a
4.03a,E
Fru-1
4.32 (d, 12)F
4.73 (d, 12)F
5.72 (d, 8)G
4.51 (dd, 8, 8)
4.04a
-3
-4
-5
-6
3.85a
3.85a
3.86a
3.85a
3.86a
3.85a
3.85a
3.86a
3.85a
3.86a
Rha-1
-2
5.42 (d, 2)I,§
4.07 (dd, 3, 2)
3.89 (dd, 9, 3)
3.48 (dd, 9, 9)
3.77 (m)
5.42 (d, 2)I,§
4.06 (dd, 3.5, 2)
3.89 (dd, 9, 3.5)
3.48 (dd, 9, 9)
3.77 (m)
5.42 (d, 2)H,§
4.06 (dd, 3, 2)
3.91a
-3
-4
-5
3.47 (dd, 9, 9)
3.76a
-6
1.25 (d, 6)
1.26 (d, 6)
1.25 (d, 6)
ester moiety
(at C-6 of Glc1)
(at C-6 of Glc3)
(at C-4 of Glc1)
Ac
Ac
2.04 (s)I
2.04 (s)J
2.05 (s)J
â
6.20 (d, 16)H
7.48 (d, 16)I
6.97 (d, 2)J ,§
6.19 (d, 16)J
7.48 (d, 16)K
6.98 (d, 2)L,§
6.28 (d, 16)K
7.50 (d, 16)L
7.01 (d, 2)M,†
6.28 (d, 16)K
7.50 (d, 16)L
7.01 (d, 2)M,†
6.28 (d, 16)I
7.50 (d, 16)J
7.00 (d, 2)K,†
γ
2
3
5
6
6.67 (d, 8)K
6.84 (dd, 8, 2)L
3.88 (s)M,§
6.68 (d, 8)M
6.89 (dd, 8, 2)N
3.80 (s)O,§
7.03 (d, 8)N,§
6.97 (dd, 8, 2)O
3.72 (s)P,†
7.03 (d, 8)N,§
6.97 (dd, 8, 2)O
3.73 (s)P,†
7.03 (d, 8)L,§
6.97 (dd, 8, 2)M
3.72 (s)
OMe
(at C-6 of Glc)
â
5.95 (d, 16)N
7.23 (d, 16)O
6.90 (d, 2)P,†
5.96 (d, 16)P
7.24 (d, 16)Q
6.90 (d, 2)R,†
5.93 (d, 16)Q
7.23 (d, 16)R
6.92 (d, 2)S,‡
5.94 (d, 16)Q
7.23 (d, 16)R
6.92 (d, 2)S,‡
5.93 (d, 16)O
7.23 (d, 16)P
6.92 (d, 2)Q,‡
γ
2
3
5
6
6.77 (d, 8)Q
6.88 (dd, 8, 2)R
3.80 (s)S,†
6.76 (d, 8)S
6.84 (dd, 8, 2)T
3.87 (s)U,†
6.78 (d, 8)T
6.85 (dd, 8, 2)U
3.87 (s)V,‡
6.78 (d, 8)T
6.78 (d, 8)R
6.86 (dd, 8, 2)S
3.88 (s)T,‡
6.85 (dd, 8, 2)U
3.87 (s)V,‡
OMe
(at C-1 of Fru)
â
γ
2
3
5
6.36 (d, 16)T
7.68 (d, 16)U
7.42 (d, 8)V
6.81 (d, 8)W
6.81 (d, 8)
6.35 (d, 16)V
7.68 (d, 16)W
7.42 (d, 8)X
6.81 (d, 8)Y
6.81 (d, 8)
6.36 (d, 16)W
7.68 (d, 16)X
7.42 (d, 8)Y
6.81 (d, 8)Z
6.81 (d, 8)
6.41 (d, 16)W
7.69 (d, 16)X
7.20 (d, 2)Y,¶
6.41 (d, 16)U
7.69 (d, 16)V
7.21 (d, 2)W,¶
6.82 (d, 8)Z
6.82 (d, 8)X
7.03 (dd, 8, 2)Y
3.91 (s)Z,¶
6
7.42 (d, 8)
7.42 (d, 8)
7.42 (d, 8)
7.01 (dd, 8, 2)R
3.91 (s)â,¶
OMe
2, 6
3, 5
4
(at C-3 of Fru)
8.20 (dd, 8, 1)X
7.61 (t, 8)Y
7.71 (tt, 8, 1)
8.18 (dd, 7.5, 1)Z
7.60 (t, 7.5)R
7.71 (tt, 7.5, 1)
8.18 (dd, 7.5, 1)R
7.60 (t, 7.5)â
7.71 (tt, 7.5, 1)
8.18 (dd, 7.5, 1)γ
7.60 (t, 7.5)δ
7.71 (tt, 7.5, 1)
8.20 (dd, 8, 1)R
7.60 (t, 8)â
7.71 (tt, 8, 1)
a
Overlapped with other signals ROEs were observed between protons that have the same letter (#,*,§,†,‡,¶ ) in each column. Long-range
correlations were observed between protons and carbons that have the same letter (A,B,...,γ,δ) in the same compounds in Table 5. Assigned
b
with the aid of HOHAHA difference, 1H-1H COSY, HMQC, and HMBC spectra.
23 °C. UV spectra were recorded on a Hitachi U-3410 spec-
trophotometer. 1H (400 MHz) and 13C (100 MHz) NMR spectra
were recorded on a J EOL R-400 FT-NMR spectrometer with
TMS as an internal standard. Inverse-detected heteronuclear
HEWLETT PACKARD 5890 gas chromatograph. HPLC was
performed using a J ASCO System 800.
P la n t Ma ter ia l. The seedling of Polygala dalmaisiana (P.
oppositifolia × P. myrtifolia) was purchased from Sakata Seed
(Yokohama, J apan) in April 1998 and grown in the botanical
garden of the University of Shizuoka. The roots of P. dalmai-
siana were harvested in November 2000.
1
correlations were measured using HMQC (optimized for J C-H
n
) 145 Hz) and HMBC (optimized for J C-H ) 8 Hz) pulse
sequences with a pulse-field gradient. Positive-mode FABMS
were recorded on a J EOL J MS-SX102 spectrometer, using a
m-nitrobenzyl alcohol as matrix. GC was carried out with a
Extr a ction a n d Isola tion . The plant material (fresh roots,
1.7 kg) was extracted twice with hot MeOH. After evaporation

Polyphenolic Glycosides from the Roots of Polygala
J ournal of Natural Products, 2002, Vol. 65, No. 3 325

326 J ournal of Natural Products, 2002, Vol. 65, No. 3
Kobayashi et al.
of MeOH under reduced pressure, the extract was suspended
in H2O and extracted with diethyl ether. The H2O layer was
passed through a Mitsubishi Diaion HP-20 column (9 cm ×
36 cm). The absorbed material was eluted with 50% aqueous
MeOH, 70% aqueous MeOH, and MeOH, successively. The
70% MeOH eluate (16 g) was submitted to column chroma-
tography on silica gel (500 g) and eluted with CHCl3-MeOH-
H2O (72:25:3) to afford 13 fractions (A-M). Fractions E + F
(1215 mg) were chromatographed over ODS [5 cm × 50 cm,
CH3CN-H2O (15:85 to 23:77) linear gradient] to afford 1 (412
mg), 3 (6.4 mg), 4 (106 mg), 5 (32 mg), 6 (14 mg), and fraction
N (128 mg). Fraction N (128 mg) was chromatographed over
Ph-A [2 cm × 25 cm, MeOH-H2O (60:40)] to give fraction N-a
(19 mg) and N-b (13 mg). Fraction N-a was chromatographed
over Ph-A [2 cm × 25 cm, MeOH-H2O (52:48)] to afford 13
(26 mg), and fraction N-b was chromatographed over Ph-A [2
cm × 25 cm, MeOH-H2O (60:40)] to afford 18 (6.9 mg).
Fractions G + H (1074 mg) were chromatographed over ODS
[5 cm × 50 cm, CH3CN-H2O (15:85 to 31:69) linear gradient]
to afford 19 fractions (a-s). Fractions j (187 mg), n (316 mg),
and p (22 mg) were chromatographed over Ph-A [2 cm × 25
cm, MeOH-H2O (50:50)] to afford 8 (19 mg) from fraction j,
16 (79 mg) and 20 (184 mg) from fraction n, and 15 (3.3 mg)
and 17 (4.7 mg) from fraction p. Fraction f (17 mg) was
chromatographed over Ph-A [2 cm × 25 cm, CH3CN-H2O (45:
55)] to afford 2 (5.4 mg). Fraction l (63 mg) was chromato-
graphed over ODS [2 cm × 25 cm, CH3CN-H2O (22.5:77.5)]
to afford 19 (5.1 mg). Fraction I (2047 mg) was chromato-
graphed over ODS [5 cm × 100 cm, CH3CN-H2O (21:79)] to
afford 10 fractions (O-X). Fraction P (248 mg) was chromato-
graphed over Ph-A [2 cm × 25 cm, CH3CN-H2O (22.5:77.5)]
to afford 9 (36 mg), 10 (24 mg), and 21 (15 mg). Fractions Q
(271 mg), S (229 mg), and U (244 mg) were chromatographed
over Ph-A [2 cm × 25 cm, MeOH-H2O (50:50)] to afford 11
(51 mg) and 12 (103 mg) from fractions Q, 7 (50 mg) from
fraction S, and 14 (94 mg) from fraction U. Fraction R (208
mg) was separated by HPLC [Ph-A, 2 cm × 25 cm, CH3CN-
H2O (20:80)] to afford 12 (18 mg). Fraction V (115 mg) was
separated successively by Ph-A [2 cm × 25 cm, CH3CN-H2O
(22.5:77.5)] and Ph-A [2 cm × 25 cm, MeOH-H2O (50:50)] to
afford 14 (6.6 mg).
Da lm a ision e A (1): amorphous powder, [R]23 -62.0° (c
D
1.0, pyridine); UV (MeOH) λmax (log ꢀ) 245 (4.26), 266 (4.23),
305 (4.20); 1H and 13C NMR, see Table 1; FABMS m/z 589 [M
+ H]+, 611 [M + Na]+.
Da lm a ision e B (2): amorphous powder, [R]23 -42.4° (c
D
0.59, pyridine); UV (MeOH) λmax (log ꢀ) 240 (4.33), 308 (4.25);
1H and 13C NMR, see Table 1; FABMS m/z 979 [M + Na]+.
Da lm a ision e C (3): amorphous powder, [R]23 -110.2° (c
D
0.64, pyridine); UV (MeOH) λmax (log ꢀ) 272 (4.09), 314 (3.85);
1H and 13C NMR, see Table 1; FABMS m/z 589 [M + H]+, 611
[M + Na]+.
Da lm a ision e D (4): amorphous powder, [R]23D -19.7° (c 1.0,
pyridine); UV (MeOH) λmax (log ꢀ) 235 (3.97), 248 (4.01), 306
(4.04); 1H NMR (DMSO-d6) δ 8.06 (1H, dd, J ) 7.5, 1.5 Hz,
H-7), 7.92 (1H, dd, J ) 7.5, 1.5 Hz, H-6′), 7.82 (1H, td, J )
7.5, 1.5 Hz, H-5), 7.73 (1H, dd, J ) 7.5, 1.5 Hz, H-4), 7.57 (1H,
td, J ) 7.5, 1.5 Hz, H-4′), 7.50 (1H, td, J ) 7.5, 1.5 Hz, H-6),
7.49 (1H, dd, J ) 7.5, 1.5 Hz, H-3′), 7.22 (1H, td, J ) 7.5, 1.5
Hz, H-5′), 7.10 (1H, s, H-10), 5.10 (1H, d, J ) 7 Hz, H-1 of
Glc1), 4.24 (1H, d, J ) 7.5 Hz, H-1 of Glc2), 4.02 (1H, br d, J
) 11 Hz, H-6 of Glc1), 3.68 (1H, dd, J ) 9, 7.5 Hz, H-4 of Glc1),
3.65 (1H, dd, J ) 12.5, 4 Hz, H-6 of Glc2), 3.63 (1H, br d, J )
11 Hz, H-6 of Glc1), 3.42 (1H, dd, J ) 12.5, 6 Hz, H-6 of Glc2),
3.42 (overlapped, H-5 of Glc1), 3.34 (1H, dd, J ) 9, 7 Hz, H-2
of Glc1), 3.20 (1H, dd, J ) 9, 9 Hz, H-3 of Glc1), 3.12 (1H, dd,
J ) 8, 8 Hz, H-3 of Glc2), 3.06 (1H, dd, J ) 8, 8 Hz, H-4 of
Glc2), 3.03 (overlapped, H-5 of Glc2), 2.99 (1H, dd, J ) 8, 7.5
Hz, H-2 of Glc2); 13C NMR (DMSO-d6) δ 177.1 (C-3), 160.4 (C-
2), 155.9 (C-9), 155.3 (C-2′), 134.0 (C-5), 132.9 (C-4′), 128.9 (C-
6′), 125.2 (C-6), 124.6 (C-7), 123.1 (C-8), 121.9 (C-5′), 120.6 (C-
1′), 118.4 (C-4), 116.0 (C-3′), 112.0 (C-10), 103.3 (C-1 of Glc2),
100.2 (C-1 of Glc1), 76.8 (C-3 of Glc2), 76.8 (C-5 of Glc2), 76.6
(C-4 of Glc1), 76.1 (C-5 of Glc1), 73.5 (C-2 of Glc2), 73.3 (C-2

Polyphenolic Glycosides from the Roots of Polygala
J ournal of Natural Products, 2002, Vol. 65, No. 3 327
of Glc1), 70.1 (C-4 of Glc2), 69.7 (C-3 of Glc1), 68.4 (C-6 of Glc1),
61.0 (C-6 of Glc2); FABMS m/z 563 [M + H]+.
300 (4.64), 319 (4.73); 1H NMR, see Table 4; 13C NMR, see
Table 5; FABMS m/z 1291 [M + Na]+.
Da lm a isiose A (5): amorphous powder, [R]23D -5.8° (c 0.77,
MeOH); UV (MeOH) λmax (log ꢀ) 224 (4.43), 284 (4.46), 317
(4.29); 1H NMR (CD3OD) δ 7.73 (1H, d, J ) 16 Hz, Hγ of
p-cou.), 7.55 (1H, d, J ) 16 Hz, Hγ of cin.), 7.52 (2H, d, J ) 8
Hz, H-2, 6 of p-cou.), 7.34 (overlapped, H-2, 3, 4, 5, 6 of cin.),
6.74 (2H, d, J ) 8 Hz, H-3, 5 of p-cou.), 6.41 (1H, d, J ) 16 Hz,
Hâ of p-cou.), 6.07 (1H, d, J ) 16 Hz, Hâ of cin.), 5.77 (1H, d,
J ) 5 Hz, H-3 of Fru), 5.42 (1H, d, J ) 3.5 Hz, H-1 of Glc1),
4.85 (1H, dd, J ) 10, 10 Hz, H-4 of Glc1), 4.44 (1H, dd, J ) 5,
5 Hz, H-4 of Fru), 4.43 (1H, d, J ) 7.5 Hz, H-1 of Glc2), 4.31
(1H, m, H-5 of Glc1), 4.18 (1H, m, H-5 of Fru), 4.14 (1H, dd, J
) 12.5, 3 Hz, H-6 of Glc1), 4.09 (1H, dd, J ) 12.5, 5 Hz, H-6
of Glc1), 3.81 (overlapped, H-6 of Fru), 3.78 (overlapped, H-6
of Glc2), 3.75 (1H, dd, J ) 10, 10 Hz, H-3 of Glc1), 3.73
(overlapped, H-6 of Fru), 3.67 (2H, br s, H-1 of Fru), 3.62 (1H,
dd, J ) 12.5, 5 Hz, H-6 of Glc2), 3.52 (1H, dd, J ) 10, 3.5 Hz,
H-2 of Glc1), 3.31 (overlapped, H-3 of Glc2), 3.31 (overlapped,
H-4 of Glc2), 3.24 (1H, m, H-5 of Glc2), 3.18 (1H, dd, J ) 9,
7.5 Hz, H-2 of Glc2), 2.04 (3H, s, Ac); 13C NMR (CD3OD) δ
172.7 (Ac), 167.8 (CR of cin.), 167.5 (CR of p-cou.), 161.6 (C-4 of
p-cou.), 147.6 (Cγ of p-cou.), 147.1 (Cγ of cin.), 135.4 (C-1 of
cin.), 131.6 (C-4 of cin.), 131.4 (C-2, 6 of p-cou.), 130.1 (C-3, 5
of cin.), 129.3 (C-2, 6 of cin.), 127.1 (C-1 of p-cou.), 118.0 (Câ of
cin.), 117.1 (C-3, 5 of p-cou.), 114.7 (Câ of p-cou.), 107.2 (C-2 of
Fru), 104.5 (C-1 of Glc2), 93.5 (C-1 of Glc1), 85.0 (C-5 of Fru),
84.4 (C-4 of Fru), 78.6 (C-3 of Fru), 77.9 (C-4 of Glc2), 77.9
(C-5 of Glc2), 75.0 (C-2 of Glc2), 73.0 (C-2 of Glc1), 72.8 (C-4
of Glc1), 72.5 (C-3 of Glc1), 71.2 (C-3 of Glc2), 70.0 (C-5 of Glc1),
65.1 (C-1 of Fru), 64.6 (C-6 of Glc-1), 63.2 (C-6 of Fru), 62.4
(C-6 of Glc2), 20.9 (Ac); FABMS m/z 823 [M + H]+, 845 [M +
Na]+.
Da lm a isiose M (18): amorphous powder, [R]23 -10.7° (c
D
0.29, MeOH); UV (MeOH) λmax (log ꢀ) 220 (4.62), 231 (4.63),
300 (4.65), 320 (4.74); 1H NMR, see Table 4; 13C NMR, see
Table 5; FABMS m/z 1311 [M + H]+, 1333 [M + Na]+.
Da lm a isiose N (19): amorphous powder, [R]23 -67.9° (c
D
0.40, MeOH); UV (MeOH) λmax (log ꢀ) 231 (4.58), 300 (4.57),
318 (4.63); 1H NMR, see Table 4; 13C NMR, see Table 5;
FABMS m/z 1479 [M + Na]+.
Da lm a isiose O (20): amorphous powder, [R]23 -61.5° (c
D
1.17, MeOH); UV (MeOH) λmax (log ꢀ) 219 (4.68), 234 (4.69),
297 (4.65), 325 (4.75); 1H NMR, see Table 4; 13C NMR, see
Table 5; FABMS m/z 1487 [M + H]+, 1509 [M + Na]+.
Da lm a isiose P (21): amorphous powder, [R]23 -61.8° (c
D
0.33, MeOH); UV (MeOH) λmax (log ꢀ) 219 (4.63), 232 (4.63),
298 (4.59), 323 (4.67); 1H NMR, see Table 4; 13C NMR, see
Table 5; FABMS m/z 1467 [M + Na]+.
Acid Hyd r olysis of Com p ou n d s 1-4. Compound 1 (30
mg) was stirred with 2 N aqueous HCl (3 mL) and dioxane (3
mL) at 100 °C for 1 h. The reaction mixture was extracted
with diethyl ether. From the H2O layer, D-glucose (tR 12.1 min)
and L-rhamnose (tR 7.6 min) were detected by GC (Supelco
SPB-1, 0.25 × 30 m, 215 °C). The diethyl ether layer afforded
compound 1a (7 mg) as an alycone. 1a was dissolved in
methanol and methylated by diazomethane-diethyl ether
solution to afford methyl ether 1b (7 mg). Compounds 2 (2 mg),
3 (2 mg), and 4 (10 mg) were hydrolyzed in the same way.
The sugar residues from 1-4 were identified in the same way
as described for the oligosaccharide mentioned below. Com-
pound 2 afforded 1a (1 mg), D-glucose, L-rhamnose, and
sinapinic acid, and 1a was identified by HPLC [ODS, 4.6 mm
× 25 cm, CH3CN-H2O (32.5:67.5), 1.0 mL/min, UV 260 nm]
and sinapinic acid (tR 8.4 min) was identified by HPLC [ODS,
4.6 mm × 25 cm, CH3CN-H2O (22.5:77.5), 1.0 mL/min, UV
260 nm]. Compound 3 afforded D-glucose and L-rhamnose.
Compound 4 afforded 4a (4 mg) as an aglycone and D-glucose.
1a : amorphous powder, 1H NMR (DMSO-d6) δ 8.13 (1H, dd, J
) 8, 1.5 Hz, H-5), 7.91 (1H, td, J ) 8, 1.5 Hz, H-7), 7.86 (1H,
dd, J ) 8, 1.5 Hz, H-5′), 7.72 (1H, dd, J ) 8, 1.5 Hz, H-8), 7.58
(1H, td, J ) 8, 1.5 Hz, H-6), 7.33 (1H, t, J ) 8 Hz, H-4′), 7.31
(1H, overlapped, H-3′); 13C NMR (DMSO-d6) δ 172.5 (C-4),
165.1 (C-2), 155.3 (C-11), 154.0 (C-9), 144.9 (C-1′), 142.7 (C-
2′), 134.9 (C-7), 126.4 (C-6), 125.4 (C-5), 124.7 (C-4′), 123.9 (C-
10), 121.2 (C-3′), 118.3 (C-8), 114.0 (C-6′), 113.8 (C-5′), 104.6
Da lm a isiose B (7): amorphous powder, [R]23 -46.0° (c
D
0.49, MeOH); UV (MeOH) λmax (log ꢀ) 210 (4.43), 228 (4.55),
300 (4.64), 309 (4.66); 1H NMR, see Table 2; 13C NMR, see
Table 5; FABMS m/z 1293 [M + H]+, 1315 [M + Na]+.
Da lm a isiose C (8): amorphous powder, [R]23 -67.8° (c
D
0.20, MeOH); UV (MeOH) λmax (log ꢀ) 220 (4.53), 232 (4.54),
298 (4.48), 323 (4.54); 1H NMR, see Table 2; 13C NMR, see
Table 5; FABMS m/z 1375 [M + Na]+.
Da lm a isiose D (9): amorphous powder, [R]23 -41.3° (c
D
0.99, MeOH); UV (MeOH) λmax (log ꢀ) 228 (4.50), 300 (4.59),
309 (4.62); 1H NMR, see Table 2; 13C NMR, see Table 5;
FABMS m/z 1273 [M + Na]+.
Da lm a isiose E (10): amorphous powder, [R]23 -69.0° (c
D
1
(C-3); FABMS m/z 281 [M + H]+. 4a : amorphous powder, H
0.29, MeOH); UV (MeOH) λmax (log ꢀ) 230 (4.61), 299 (4.64),
313 (4.68); 1H NMR, see Table 2; 13C NMR, see Table 5;
FABMS m/z 1303 [M + Na]+.
NMR (DMSO-d6) δ 8.05 (1H, dd, J ) 8, 2 Hz, H-7), 7.92 (1H,
dd, J ) 8, 2 Hz, H-6′), 7.82 (1H, td, J ) 8, 2 Hz, H-5), 7.74
(1H, dd, J ) 8, 2 Hz, H-4), 7.49 (1H, td, J ) 8, 2 Hz, H-6), 7.40
(1H, td, J ) 8, 2 Hz, H-4′), 7.13 (1H, s, H-10), 7.07 (1H, dd, J
) 8, 2 Hz, H-3′), 7.02 (1H, td, J ) 8, 2 Hz, H-5′); 13C NMR
(DMSO-d6) δ 177.1 (C-3), 160.7 (C-2), 156.5 (C-9), 155.8 (C-
2′), 134.0 (C-5), 132.5 (C-4′), 128.5 (C-6′), 125.1 (C-6), 124.6
(C-7), 123.1 (C-8), 119.4 (C-5′), 118.3 (C-4), 117.7 (C-1′), 117.0
(C-3′), 111.0 (C-10); FABMS m/z 239 [M + H]+.
Da lm a isiose F (11): amorphous powder, [R]23 -45.7° (c
D
0.44, MeOH); UV (MeOH) λmax (log ꢀ) 229 (4.58), 298 (4.62),
310 (4.64); 1H NMR, see Table 2; 13C NMR, see Table 5;
FABMS m/z 1281 [M + H]+, 1303 [M + Na]+.
Da lm a isiose G (12): amorphous powder, [R]23 -41.4° (c
D
0.89, MeOH); UV (MeOH) λmax (log ꢀ) 219 (4.46), 233 (4.48),
297 (4.43), 323 (4.51); 1H NMR, see Table 3; 13C NMR, see
Table 5; FABMS m/z 1311 [M + H]+, 1333 [M + Na]+.
Hyd r olysis of Oligosa cch a r id es 5 a n d 7-21. Each
sample (2 mg) was stirred with 1 N aqueous NaOH (50 µL) at
65 °C for 1 h. After acidification with 1 N HCl (100 µL), the
reaction mixture was diluted with H2O (500 µL) and extracted
with EtOAc (100 µL). The EtOAc layer was washed with H2O
(500 µL). To the EtOAc layer was added O-(4-nitrobenzyl)-
N,N′-diisopropylisourea (NBDI) (3 mg), and the reaction
mixture was stirred for 1 h at 65 °C. From the reaction
mixture, acetic acid (tR 9.4 min) was detected from 5, 7-14,
16, and 18-20, p-coumaric acid (tR 14.9 min) was detected
from 5, 7, 9, 10, 11, and 13-19, ferulic acid (tR 15.8 min) was
detected from 8, 10, 11, 12, and 14-21, benzoic acid (tR 22.8
min) was detected from 7-21, and cinnamic acid (tR 31.9 min)
was detected from 5 by HPLC [Develosil Ph-A, 4.6 mm × 25
cm, CH3CN-H2O (45:55), 1.0 mL/min, UV 273 nm]. The H2O
layer was passed through a column equipped with Amberlite
IRA-60E + IR-120B (1:1) (7 mm × 7 cm) and eluted with H2O.
The H2O elute was concentrated and dried in a small sample
tube, 0.5 N HCl (50 µL) was added, and the mixture was
Da lm a isiose H (13): amorphous powder, [R]23 -11.5° (c
D
0.86, MeOH); UV (MeOH) λmax (log ꢀ) 211 (4.64), 229 (4.74),
300 (4.85), 313 (4.91); 1H NMR, see Table 3; 13C NMR, see
Table 5; FABMS m/z 1273 [M + Na]+.
Da lm a isiose I (14): amorphous powder, [R]23 -66.2° (c
D
1.07, MeOH); UV (MeOH) λmax (log ꢀ) 230 (4.73), 300 (4.80),
315 (4.84); 1H NMR, see Table 3; 13C NMR, see Table 5;
FABMS m/z 1427 [M + H]+, 1449 [M + Na]+.
Da lm a isiose J (15): amorphous powder, [R]23 -14.5° (c
D
0.37, MeOH); UV (MeOH) λmax (log ꢀ) 230 (4.70), 301 (4.77),
316 (4.83); 1H NMR, see Table 3; 13C NMR, see Table 5;
FABMS m/z 1239 [M + H]+, 1261 [M + Na]+.
Da lm a isiose K (16): amorphous powder, [R]23 -60.3° (c
D
0.68, MeOH); UV (MeOH) λmax (log ꢀ) 220 (4.64), 231 (4.67),
300 (4.69), 318 (4.73); 1H NMR, see Table 3; 13C NMR, see
Table 5; FABMS m/z 1457 [M + H]+, 1479 [M + Na]+.
Da lm a isiose L (17): amorphous powder, [R]23 -19.9° (c
D
0.47, MeOH); UV (MeOH) λmax (log ꢀ) 220 (4.60), 231 (4.62),

328 J ournal of Natural Products, 2002, Vol. 65, No. 3
Kobayashi et al.
warmed for 10 min at 100 °C. The reaction mixture was passed
though a column equipped with Amberlite IRA-60E (7 mm ×
5 cm), eluted with H2O, and concentrated and dried in a small
tube. D-Cysteine methyl ester in pyridine (3 mg/25 µL) was
added, and the mixture was stirred for 1.5 h at 65 °C. To the
reaction mixture were added hexamethyldisilazane (15 µL) and
trimethylsilyl chloride (15 µL), and the mixture was stirred
for 30 min at 65 °C. The reaction mixture was diluted with
H2O and extracted with hexane. From the hexane extract,
D-glucose (tR 12.1 min) and D-fructose (tR 8.9 min) were
detected from 5 and 7-21, and L-rhamnose (tR 7.4 min) was
detected from 7-12, 14, 16, 19, 20, and 21 by GC (Supelco
SPB-1, 0.25 mm × 30 m, 215 °C). The tR of L-glucose, l-fructose,
and D-rhamnose was 11.3, 9.8, and 7.6 min, respectively.7
Refer en ces a n d Notes
(1) Kobayashi, W.; Miyase, T.; Suzuki, S.; Noguchi, H.; Chen, X.-M. J .
Nat. Prod. 2000, 63, 1066-1069, and references therein.
(2) Saitoh, H.; Miyase, T.; Ueno, A. Chem. Pharm. Bull. 1994, 42, 1879-
1885.
(3) Hara, S.; Okabe, H.; Mihashi, K. Chem. Pharm. Bull. 1986, 34, 1843-
1845.
(4) Paolo, E. R. D.; Hamburger, M. O.; Stoeckii-Evans, H.; Rogers, C.;
Hostettmann, K. Helv. Chim. Acta 1989, 72, 1455-1462.
(5) Hansen, P. E. Org. Magn. Reson. 1979, 12, 109-142.
(6) Pourfarzam, M.; Bartlett, K. J . Chromatogr. 1991, 570, 253-276.
(7) Retention times for D-rhamnose and L-fructose were obtained from
their enantiomers (L-rhamnose + L-cysteine methylester, D-fructose
+ L-cysteine methyl ester).
NP010434N