Flavonoid and benzophenone glycosides from Coleogyne ramosissima

Article abstract of DOI:10.1016/S0031-9422(00)00192-8

A benzophenone glucoside and two flavonol glycosides were isolated together with 27 known polyphenols from the aerial parts of Coleogyne ramosissima, and their structures were elucidated by spectroscopic and chemical methods as iriflophenone 2-O-β-glucopyranoside, isorhamnetin 3-O-2(G)-rhamnopyranosylrutinoside-7-O-α-rhamnopyranoside and limocitrin 3-O-rutinoside-7-O-β-glucopyranoside, respectively. (C) 2000 Elsevier Science Ltd.

Full text of DOI:10.1016/S0031-9422(00)00192-8

Phytochemistry 54 (2000) 695±700
www.elsevier.com/locate/phytochem
Flavonoid and benzophenone glycosides from Coleogyne
ramosissima
Hideyuki Itoa, Eisei Nishitania, Takao Konoshimab, Midori Takasakib,
Mutsuo Kozukab, Takashi Yoshidaa,*
aFaculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama 700-8530, Japan
bKyoto Pharmaceutical University, Yamashina-ku, Kyoto 607-8414, Japan
Received 21 January 2000; received in revised form 11 April 2000
Abstract
A benzophenone glucoside and two ¯avonol glycosides were isolated together with 27 known polyphenols from the aerial
parts of Coleogyne ramosissima, and their structures were elucidated by spectroscopic and chemical methods as iri¯ophenone 2-
O-b-glucopyranoside, isorhamnetin 3-O-2G-rhamnopyranosylrutinoside-7-O-a-rhamnopyranoside and limocitrin 3-O-rutinoside-
7-O-b-glucopyranoside, respectively. 7 2000 Elsevier Science Ltd. All rights reserved.
Keywords: Coleogyne ramosissima; Rosaceae; Iri¯ophenone 2-O-glucopyranoside; Isorhamnetin 3-O-2G-rhamnopyranosylrutinoside-7-O-rhamno-
pyranoside; Limocitrin 3-O-rutinoside-7-O-glucopyranoside
1. Introduction
compounds in large amount. In the present study,
three new polyphenols, a benzophenone and two ¯avo-
nol glycosides along with 27 known polyphenols were
isolated from the aerial parts of the plant. We describe
herein the structure elucidation of these new polyphe-
nols.
Coleogyne ramosissima Torr. (Rosaceae) is a densely
branched shrub widely distributed in western North
America. As for the chemical constituents of this
plant, the presence of condensed tannins (Provenza
and Malechek, 1983) and cyanogenic glycosides
(Fikenscher et al., 1981) in the leaves was reported,
and lignans (Tachimoto et al., 1993) and phenylpropa-
noid glycosides (Tachimoto et al., 1994) were isolated
from the stem extract as inhibitors of Epstein±Barr
virus early antigen activation induced by 12-O-tertade-
canoylphorbol-13-acetate. The polar polyphenolic con-
stituents in the plant have, however, been investigated
little, although reversed-phase HPLC of the aqueous
acetone extract with diode array detection at 280 nm
suggested the occurrence of unidenti®ed UV-sensitive
2. Results and discussion
A concentrated solution of an aq. acetone homogen-
ate of the dried aerial parts of C. ramosissima was
extracted with Et2O, AcOEt and n-BuOH, successively.
A new benzophenone glucoside, iri¯ophenone 2-O-b-
glucopyranoside (1), and two new ¯avonoid glycosides,
isorhamnetin 3-O-2G-rhamnopyranosylrutinoside-7-O-
a-rhamnopyranoside (2) and limocitrin 3-O-rutinoside-
7-O-b-glucopyranoside (3) were obtained by repeated
column chromatography of the AcOEt and the n-
BuOH extracts, respectively. Twenty-seven known
polyphenols were also isolated from the AcOEt and n-
BuOH extracts as described in Section 3, and were
identi®ed as (+)-catechin, ( )-epicatechin, procyani-
* Corresponding author. Tel.: +81-86-251-7936; fax: +81-86-251-
7936.
E-mail address: yoshida@pheasant.pharm.okayama-u.ac.jp (T.
Yoshida).
0031-9422/00/$ - see front matter 7 2000 Elsevier Science Ltd. All rights reserved.
PII: S0031-9422(00)00192-8

696
H. Ito et al. / Phytochemistry 54 (2000) 695±700
Chart 1.
dins B-1, B-2, B-3, B-4, B-5, C-1, isoquercitrin (Mark-
ham et al., 1978), quercetin 3-O-b-xylopyranoside
(Kadota et al., 1990), 8-methoxykaempferol 3-O-b-
sophoroside1) (Ferreres et al., 1989), (2)-eriodictyol 7-
O-b-glucopyranoside (Cui et al., 1990), (+)-taxifolin 7-
O-b-glucopyranoside (Foo and Karchesy, 1989), (2)-
naringenin 8-C-b-glucopyranoside (Ge et al., 1991), 1-
O-(E )-cinnamyl-b-glucopyranose (Tachimoto et al.,
1994), (E )-coniferyl alcohol 9-O-b-glucopyranoside
(Andersson and Lundgren, 1988), picein (Perry et al.,
1996), liriodendrin (Deyama, 1983) and N 1-, N 5-, N 10
-
tri-p-(E)-coumaroylspermidine (Strack et al., 1990;
Bokern et al., 1995; Werner et al., 1995), and eight iso-
rhamnetin glycosides, isorhamnetin 3-O-2G-rhamno-
pyranosylrutinoside (4) (Masterova et al., 1991),
isorhamnetin 3-O-b-glucopyranoside (5) (Ishida et al.,
1988), narcissin (6) (Markham et al., 1978), limocitrin
3-O-b-glucopyranoside (7) (Ulubelin et al., 1984), limo-
1 This compound was erroneously reported to be 6-methoxykaemp-
ferol 3-O-b-sophoroside in our study on the anti-tumor promoting
activity of the rosaceous plants (Ito et al., 1999).

H. Ito et al. / Phytochemistry 54 (2000) 695±700
697
citrin 3-O-rutinoside (8) (Harborne, 1981), isorhamne-
tin 3-O-b-glucuronopyranoside (9) (Merfort and Wen-
disch, 1988), isorhamnetin 3-O-rutinoside-7-O-b-
glucopyranoside (10) (Dembinska-Migas et al., 1985)
and spinacetin 3-O-b-glucopyranoside (11) (Merfort
and Wendisch, 1987), by comparison of the physico-
chemical data with those reported (see Chart 1).
region, and a methoxyl signal ꢀd 3.85), which were
similar to the aglycone signals of isorhamnetin glyco-
sides (4±6). Further, the signals due to four anomeric
proton signals ‰d 5.63 (d, J ˆ 7 Hz), 5.54, 5.02, 4.37
(each br s )] and three methyl groups ‰d 1.11, 0.93, 0.70
(each 3H, d, J ˆ 6 Hz)] revealed the presence of three
rhamnosyl residues in 2. In fact, the 13C NMR spec-
trum of 2 (Table 1) was very similar to that of iso-
rhamnetin 3-O-2G-rhamnopyranosylrutinoside (4),
except for the presence of signals ascribable to an
extra rhamnose core and a remarkable up®eld shift
ꢀD d 2.7 ppm) of the aglycone C-7. The linkage pos-
Compound 1 was obtained as a pale yellow amor-
phous powder. The molecular formula was established
as C19H20O10 by HRESIMS (m/z 409.1186 [M + H]+,
calculated for C19H20O10 + H, 409.1135). The 1H
NMR spectrum of 1 indicated the presence of a 1-, 4-
disubstituted ‰d 7.64 (2H, d, J ˆ 8 Hz) and d 6.84 (2H,
d, J ˆ 8 Hz)] and 1-, 2-, 4-, 6-tetrasubstituted benzene
nucleus ‰d 6.33 (1H, d, J ˆ 2 Hz) and d 6.12 (1H, d,
Table 1
13C NMR spectral data for compounds 2, 3, 4 and 8 (126 MHz in
DMSO-d6)
4
J ˆ 2 Hz)]. A seven-spin system characteristic of a C1
glucopyranose residue was observed in the aliphatic
region. The 13C NMR spectrum showed a ketonic car-
bon resonance besides 12 sp2 and glucose carbon sig-
nals, suggesting that 1 is a glucoside of iri¯ophenone.
The HMBC spectrum of 1 revealed that the anomeric
proton at d 4.80 (H-10) was correlated through three-
bond coupling with the signal at d 158.7 (C-2) which
also showed two-bond coupling with one ꢀd 6.33, H-3)
of the two meta-coupled proton signals in the phloro-
glucinol unit (Fig. 1). The location of the glucose resi-
due was thus established to be C-2 of the aglycone. A
b-glucosidic linkage was suggested from the coupling
constant ꢀJ ˆ 8 Hz) of the anomeric proton signal.
Based on these data, compound 1 was iri¯ophenone 2-
O-b-glucopyranoside.
Compound 2, a yellow powder, showed an [M +
H]+ ion peak at m/z 917 in ESIMS. The 1H NMR
spectrum of 2 exhibited 1,3,4-trisubstituted benzene
signals ‰d 7.85 (d, J ˆ 2 Hz), 7.54 (dd, J ˆ 2, 8 Hz)
and 6.90 (d, J ˆ 8 Hz)], meta-coupled signals ‰d 6.78
and 6.43 (each d, J ˆ 2 Hz)] in the aromatic proton
2
4
3
8
Aglycone moiety
C-2
C-3
157.1
132.9
177.5
161.1
99.5
157.0
133.0
177.7
161.4
99.2
C-2
C-3
155.9 156.1
133.4 133.3
177.9 177.7
157.0 156.5
C-4
C-5
C-4
C-5
C-6
C-7
C-6
C-7
98.8
99.2
161.8
95.0
164.5
94.4
156.2 157.2
129.2 127.7
149.9 149.7
105.5 104.1
121.2 121.4
113.3 113.3
147.1 147.1
148.3 148.9
115.6 115.6
122.5 122.4
C-8
C-9
C-8
C-9
156.2
105.8
120.9
113.4
149.9
147.1
115.4
122.5
55.8
157.0
104.6
121.7
113.7
149.6
147.3
115.6
122.6
56.2
C-10
C-1'
C-2'
C-3'
C-4'
C-5'
C-6'
OMe
C-10
C-1'
C-2'
C-3'
C-4'
C-5'
C-6'
8-OMe
3'-OMe
60.8
55.7
61.2
55.8
Sugar moieties
Glucose (Glc)
C-10
C-20
C-30
C-40
C-50
C-60
Rutinose (Rut)
Glucose
98.8
77.8
77.2
70.5
76.0
66.5
99.1
78.2
77.3
70.8
76.3
67.4
C-10
C-20
C-30
C-40
C-50
C-60
101.4 101.5a
74.5
76.8
70.3
76.1
67.2
74.5
76.6
70.3
76.2
67.1
Rhamnose (Rha-1, 2, 3)
C-11,12,12' 101.04
100.98
Rhamnose
C-11
C-21
C-31
101.4
101.3
101.1 101.1a
70.5
70.8
71.9
68.4
17.9
70.6
70.8
72.0
68.5
17.9
98.6
C-21,22,22'
C-31,32,32'
C-41,42,42'
70.7 (3C)
70.5 (3C)
71.9 (2C)
71.8
70.7 (2C) C-41
70.9 (2C) C-51
72.1 (2C) C-61
C-51,52,52'
C-61,62,62'
68.5
68.8 (2C) Glucose (Glc)
68.4 (2C)
18.1
17.8
C-12
C-22
C-32
C-42
C-52
C-62
100.4
18.1
17.5
73.4
77.4
69.8
76.5
61.5
17.3
a These values may be interchanged.
Fig. 1. HMBC correlations of 1.

698
H. Ito et al. / Phytochemistry 54 (2000) 695±700
itions of sugar units with the aglycone in 2 were sub-
stantiated by HMBC spectrum ꢀJCH ˆ 6 Hz), in which
H-10 ꢀd 5.63) of glucose and H-12' (d 5.54) of rham-
nose (Rha-3) were correlated with C-3 ꢀd 132.9) and
C-7 ꢀd 161.8) of the aglycone through three-bond
couplings, respectively. Similarly, anomeric proton sig-
nals of the other rhamnoses (Rha-1 H-11 at d 5.02 and
Rha-2 H-12 at d 4.37) exhibited three-bond cross
peaks with C-20 at d 77.8 and C-60 at d 66.5 carbon
resonances of glucose, respectively (Fig. 2). These spec-
tral data indicated the presence of the 2G-rhamnopyra-
nosylrutinosyl and a rhamnosyl residue, at positions 3
and 7 of the aglycone, respectively. Enzymatic hy-
drolysis of 2 with hesperidinase (Ishida et al., 1988)
a€orded isorhamnetin 3-O-b-glucopyranoside (5), nar-
cissin (6) and 4 to establish that the 2G-rhamnopyrano-
sylrutinose unit is located at O-3. On the basis of these
data, the structure of 2 was determined as isorhamne-
tin 3-O-2G-rhamnopyranosylrutinoside-7-O-a-L-rham-
nopyranoside.
responded to those of 8 (Harborne, 1981) and querce-
tin 7-O-glucoside (Markham et al., 1978). A signi®cant
di€erence in the 13C NMR spectrum is in the chemical
shifts of C-7, C-8 and C-10 ꢀD d 1.0, +1.5 and +1.4
ppm). This suggests that 3 is a 7-O-glucoside of 8. The
proposed structure 3 was substantiated by the HMBC
spectrum which showed correlation of H-11 ꢀd 5.43)
with C-7 ꢀd 156.2) of the aglycone through three-bond
coupling. Further, the anomeric proton of the glucose
core exhibited an NOE correlation with H-6 at d 6.65,
and 8-methoxy signal at d 3.84 displayed NOE with
H-6' signal at d 7.56. Upon enzymatic hydrolysis with
b-glucosidase, 3 yielded 8 as judged by reversed-phase
HPLC. Consequently, compound 3 was established as
limocitrin 3-O-rutinoside-7-O-b-glucopyranoside.
3. Experimental
3.1. General
Compound 3 was obtained as a yellow powder. The
UV and ESIMS (m/z 817 [M + H]+) suggested that 3
1
is also a ¯avonoid glycoside. The H NMR spectrum
Optical rotations were recorded on a JASCO DIP-
1
1000 polarimeter. H (500 MHz) and 13C NMR (126
of 3 showed signals attributable to an aglycone at d
7.88 (d, J ˆ 2 Hz), 7.56 (dd, J ˆ 2, 8 Hz) and 6.94 (d,
J ˆ 9 Hz) (B-ring), d 6.65 (s) and two methoxyl signals
ꢀd 3.84 and 3.83), which were similar to those of limo-
citrin 3-O-rutinoside (8) except for a down®eld shift of
H-6 (D 0.37 ppm). The spectrum also exhibited three
anomeric proton signals at d 5.43 (d, J ˆ 8 Hz), d 5.06
(d, J ˆ 7:5 Hz) and d 4.39 (br s ) and a methyl signal
at d 0.95 (d, J ˆ 6 Hz), indicating the presence of a
rhamnose and two hexose units in 3. The sugar moiety
carbon resonances in the 13C NMR spectrum of 3 cor-
MHz) spectra were measured on a Varian VXR-500
instrument, and chemical shifts are given in d-values
from tetramethylsilane. Average J-values for one-bond
coupling and two- or three-bond coupling in HMQC
and HMBC were set at 140 Hz and 6 or 8 Hz. ESI-
MS were measured with a Micromass Auto Spec OA-
Tof mass spectrometer (solvent: 50% MeOH contain-
ing 0.1% AcONH4, ¯ow rate: 0.02 ml min 1). Normal
phase HPLC was conducted on a YMC-Pack SIL A-
003 (YMC Co., Ltd.), column ꢀ4:6 Â 250 mm) devel-
(55:33:11:1) containing oxalic acid (450 mg l 1) (¯ow
rate, 1.5 ml min 1; detection 280 nm) at room tem-
perature. Reversed-phase HPLC was performed on a
YMC-Pack AQ-302 (ODS) (YMC Co., Ltd.) column
ꢀ4:6 Â 150 mm) developed with 10 mM H3PO4±10 mM
KH2PO4±EtOH±AcOEt (42.5:42.5:10:5) (¯ow rate, 1.0
ml min 1; detection 280 nm) at 408C.
oped
with
n-hexane±MeOH±THF±HCO2H
3.2. Plant material
The aerial parts of C. ramosissima Torr. were col-
lected at Beaver Dam Mts, Utah, USA in May 1992,
and the plant was identi®ed by Dr. R.J. Kass, Brigham
Young University, Utah. A voucher specimen is depos-
ited at the Herbal Garden of Kyoto Pharmaceutical
University.
3.3. Extraction and isolation
The dried aerial parts (1 kg) were homogenized (3Â)
in Me2CO±H2O (7:3) ꢀ3 l  3† and the homogenate
was ®ltered. The ®ltrate was concentrated and
Fig. 2. HMBC Correlations Of 2.

H. Ito et al. / Phytochemistry 54 (2000) 695±700
699
extracted with Et2O ꢀ1 l  4), EtOAc ꢀ1 l  6† and n-
BuOH saturated with H2O ꢀ1 l  6). The EtOAc
extract (16.5 g) was subjected to chromatography over
Toyopearl HW-40 (coarse) (2.2 cm i.d. Â 66 cm) with
MeOH±H2O (4:6 4 5:5 4 6:4 4 7:3) 4 Me2CO±H2O
(7:3) in a stepwise gradient mode. Fractions showing
similar HPLC patterns were combined and further
puri®ed by a combination of CC over Toyopearl HW-
40 (®ne), MCI-gel CHP-20P and/or YMC GEL ODS-
AQ 120-S50 with aq. MeOH to a€ord (+)-catechin
(35 mg), ( )-epicatechin (270 mg), procyanidins B-2
(203 mg), B-3 (47 mg), B-4 (383 mg), B-5 (9 mg), C-1
(16 mg), isoquercitrin (47 mg), quercetin 3-O-b-xylop-
pyranoside (9 mg), (2)-eriodictyol 7-O-b-glucopyrano-
side (119 mg), (2)-naringenin 8-C-b-glucopyranoside
(11 mg), 1-O-cinnamyl-b-glucopyranose (125 mg), (E )-
coniferyl alcohol 9-O-b-glucopyranoside (5 mg), picein
(308 mg), N 1-, N 5-, N 10-tri-p-(E )-coumaroylspermi-
dine (59 mg), iri¯ophenone 2-O-b-glucopyranoside (1)
(6 mg), isorhamnetin 3-O-b-glucopyranoside (5) (36
mg), narcissin (6) (85 mg), limocitrin 3-O-b-glucopyra-
noside (7) (10 mg), limocitrin 3-O-rutinoside (8) (22
mg) and spinacetin 3-O-b-glucopyranoside (11) (12
mg).
The n-BuOH extract (56.3 g) was subjected to col-
umn chromatography over Diaion HP-20 (6.0 cm i.d.
 40 cm) and developed with H2O 4 MeOH±H2O (1:9
4 2:8 4 3:7 4 4:6 4 6:4) 4 MeOH in a stepwise gra-
dient mode. The eluate with MeOH±H2O (2:8) was
further subjected to chromatography over MCI-gel
CHP-20P to give picein (145 mg). The eluate with
MeOH±H2O (3:7) was similarly subjected to repeated
column chromatographies to give ( )-epicatechin (66
mg), procyanidins B-1 (40 mg), B-2 (94 mg), C-1 (29
mg), isorhamnetin 3-O-rutinoside-7-O-b-glucopyrano-
side (10) (23 mg), (+)-taxifolin 7-O-b-glucopyranoside
(8 mg), isorhamnetin 3-O-2G-rhamnopyranosylrutino-
side-7-O-a-rhamnopyranoside (2) (39 mg) and limoci-
trin 3-O-rutinoside-7-O-b-glucopyranoside (3) (12 mg).
From the eluate with MeOH±H2O (4:6), procyanidins
B-2 (52 mg), C-1 (10 mg), liriodendrin (9 mg), 8-meth-
oxykaempferol 3-O-sophoroside (63 mg) and isorham-
netin 3-O-2G-rhamnopyranosylrutinoside (5) (120 mg)
were obtained after repeated column chromatography.
The eluate with MeOH±H2O (6:4) was similarly puri-
®ed by a combination of column chromatographies to
yield 6 (63 mg) and 8 (17 mg). The known polyphenols
were identi®ed by comparison of their physical data
with the reported values.
D2O): d 7.64 (2H, d, J ˆ 8 Hz, H-2',6'), 6.84 (2H, d,
J ˆ 8 Hz, H-3', 5'), 6.33 (1H, d, J ˆ 2 Hz, H-3), 6.12
(1H, d, J ˆ 2 Hz, H-5), 4.80 (1H, d, J ˆ 8 Hz, H-10),
3.86 (1H, dd, J ˆ 2:5, 12 Hz, H-60), 3.64 (1H, dd,
J ˆ 6, 12 Hz, H-60), 3.44 (1H, ddd, J ˆ 2:5, 6, 9 Hz,
H-50), 3.41 (1H, t, J ˆ 9 Hz, H-30), 3.2±3.4 (over-
lapped with HDO, H-40), 3.02 (1H, dd, J ˆ 8, 9 Hz,
H-20). 13C NMR (MeOH-d4): d 197.5 (C-7'), 163.6 (C-
4'), 162.4 (C-4), 159.7 (C-6), 158.7 (C-2), 133.5 (2C)
(C-2',6'), 124.4 (C-1'), 115.8 (2C) (C-3',5'), 102.4 (C-
10), 99.8 (C-1), 98.1 (C-5), 95.9 (C-3), 78.2 (C-50), 77.8
(C-30), 74.7 (C-20), 71.1 (C-40), 62.5 (C-60).
3.3.2. Isorhamnetin 3-O-2G-rhamnosylrutinoside-7-O-a-
L-rhamnopyranoside (2)
A yellow powder, ‰aŠ
358 (MeOH; c 1.0). UV lmax
D
MeOH nm ꢀlog e): 248 sh (4.01), 254 (4.17), 265 sh
(4.07), 354 (4.07). ESIMS m/z: 917 [M + H]+, 939 [M
+ Na]+. HRESIMS m/z: 917.2965 [M + H]+, calcu-
lated for C40H52O24
+
H, 917.2927. 1H NMR
(DMSO-d6): d 7.85 (1H, d, J ˆ 2 Hz, H-2'), 7.54 (1H,
dd, J ˆ 2, 8 Hz, H-6'), 6.90 (1H, d, J ˆ 8 Hz, H-5'),
6.78 (1H, br s, H-8), 6.43 (1H, br s, H-6), 5.63 [1H, d,
J ˆ 7 Hz, glucose (Glc) H-10], 5.54 (1H, br s, Rha-3
H-12'), 5.02, 4.37 (each 1H, br s, Rha-1, 2 H-11,12),
3.85 (3H, s, 3'-OCH3), 1.11 (3H, d, J ˆ 6 Hz, Rha-3
H-62'), 0.93, 0.70 (each 3H, d, J ˆ 6 Hz, Rha-1, 2 H-
61,62). 13C NMR: Table 1.
3.3.3. Enzymatic hydrolysis of 2 with hesperidinase
A solution of 2 (1 mg) in 0.05 M McIlvaine bu€er
(pH 3.8) (1 ml) and 4% glucose-DMSO (0.1 ml) was
incubated with hesperidinase (1 mg) at 378C for 21 h.
The reaction mixture was analyzed by reversed-phase
HPLC to show peaks identical with those of the auth-
entic 4±6.
3.3.4. Limocitrin 3-O-rutinoside-7-O-b-glucopyranoside
(3)
A yellow powder, ‰aŠ
238 (MeOH; c 1.0). UV lmax
D
MeOH nm ꢀlog e): 257 (4.13), 270 sh (4.05), 360 (4.01);
+ AlCl3 276, 305 sh, 377, 418. ESIMS m/z: 817 [M +
H]+. HRESIMS m/z: 817.2423 [M + H]+, calculated
for C35H44O22 + H, 817.2402. 1H NMR (DMSO-d6
+ D2O): d 7.88 (1H, d, J ˆ 2 Hz, H-2'), 7.56 (1H, dd,
J ˆ 2, 8 Hz, H-6'), 6.94 (1H, d, J ˆ 8 Hz, H-5'), 6.65
(1H, s, H-6), 5.43 (1H, d, J ˆ 8 Hz, Glc H-10), 5.06
(1H, d, J ˆ 7:5 Hz, Glc H-11), 4.39 (1H, br s, Rha H-
12), 3.84 (3H, s, 8-OCH3), 3.83 (3H, s, 3'-OCH3), 0.95
(3H, d, J ˆ 6 Hz, Rha H-62) 13C NMR: Table 1.
3.3.1. Iri¯ophenone 2-O-b-glucopyranoside (1)
A
pale yellow amorphous powder, ‰aŠ
248
3.3.5. Enzymatic hydrolysis of 3 with b-glucosidase
A solution of 3 (1 mg) in 0.1 M acetate bu€er (pH
5) (1 ml) was incubated with b-glucosidase (1 mg) at
378C for 1 h. Reversed-phase HPLC of the reaction
mixture showed a peak identical with that of 8.
D
(MeOH; c 1.0). UV lmax MeOH nm ꢀlog e): 233 sh
(4.08), 289 (4.12). ESIMS m/z: 409 [M + H]+. HR-
ESIMS m/z: 409.1186 [M + H]+, calculated for
C19H20O10 + H, 409.1135. 1H NMR (Me2CO-d6
+

700
H. Ito et al. / Phytochemistry 54 (2000) 695±700
Chinese medicine. IX.: On the antihemorrhagic principles in
Acknowledgements
Typha lactifolia L. Chemical & Pharmaceutical Bulletin 36 (11),
4414±4420.
The authors are grateful to the SC-NMR Labora-
tory of Okayama University for NMR experiments.
This work was supported in part by a Grant-in-Aid
for Scienti®c Research (No. 10,557,207) from the Min-
istry of Education, Science, Sports and Culture of
Japan.
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