Terpenic and phenolic glycosides from leaves of Breynia officinalis Hemsl.

Article abstract of DOI:10.1248/cpb.52.1086

From the leaves of Breynia officinalis, collected on Okinawa Island, six terpenic glucosides and six phenolic glycosides were isolated. Two of the terpenic glucosides were found to be known, and they were identified as turpinionoside B and betulalbuside A

Full text of DOI:10.1248/cpb.52.1086

1
086
Chem. Pharm. Bull. 52(9) 1086—1090 (2004)
Vol. 52, No. 9
Terpenic and Phenolic Glycosides from Leaves of Breynia officinalis
HEMSL.
a
a
,a
b
c
Hiroyuki MORIKAWA, Ryoji KASAI, Hideaki OTSUKA,* Eiji HIRATA, Takakazu SHINZATO,
d
e
Mitsunori ARAMOTO, and Yoshio TAKEDA
a
Department of Pharmacognosy, Graduate School of Biomedical Sciences, Hiroshima University; 1–2–3 Kasumi, Minami-
b
ku, Hiroshima 734–8551, Japan: Faculty of Agriculture, University of the Ryukyus; 1 Senbaru, Nishihara-cho, Nakagami-
gun, Okinawa 903–0129, Japan: Yona Field, Subtropical Field Science Center, Faculty of Agriculture, University of
Ryukyus; 685 Aza Yona, Kunigami-son, Kunigami-gun, Okinawa 905–1427, Japan: Iriomote Station, Tropical Biosphere
c
d
Research Center, University of the Ryukyus; 870 Aza Uehara, Taketomi-cho, Yaeyama-gun, Okinawa 907–1541, Japan:
e
and Faculty of Integrated Art and Sciences, The University of Tokushima; 1–1 Minamijosanjima-cho, Tokushima
7
70–8502, Japan. Received April 5, 2004; accepted May 27, 2004
From the leaves of Breynia officinalis, collected on Okinawa Island, six terpenic glucosides and six phenolic
glycosides were isolated. Two of the terpenic glucosides were found to be known, and they were identified as
turpinionoside B and betulalbuside A. The structures of the remaining terpenic glucosides were elucidated to be
megastigmane glucosides, named breyniaionosides A—D, using spectroscopic analyses. The absolute structure of
breyniaionoside D was determined using the modifed Mosher’s method. The absolute structure of the known
compound betulalbuside A was determined for the first time in this study. Six phenolic glycosides were found to
be arbutin and its derivatives. Two known compounds were found to be robustaside A and eximine. New com-
pounds were named isorobustaside A, and breyniosides A and B, and their structures were elucidated from spec-
troscopic evidence.
Key words Breynia officinalis; Euphorbiaceae; megastigmane glucoside; breyniaionoside; betulalbuside A; breynioside
Breynia officinalis (Euphorbiaceae) is a poisonous peren- ture of 1 was determined to be (3S,5R,6S,9S)-megastigman-
2
)
nial shrub 1.5—4 m in height, and grows in Okinawa, Tai- 7-ene-3,6,9-triol 9-O-b-D-glucopyranoside.
wan, and southern China. However, it is used as a remedy for
Breyniaionoside A (2), [a] ꢀ48.8°, was isolated as an
healing wounds and edema, as an ointment, and for syphilis amorphous powder and its elemental composition was deter-
and intestinal hemorrhage due to overwork by oral adminis- mined to be C H O using negative-ion high-resolution
D
1
9
32
9
1)
tration. Continuing our survey of Okinawan resource plants, (HR) FAB-MS. The IR spectrum of 2 showed a strong ab-
ꢀ1
the constituents of B. officinalis were investigated.
sorption band at 3376 cm suggestive of a glycosidic struc-
From a n-BuOH-soluble fraction of the MeOH extract of ture. The H- and C-NMR spectra showed signals attribut-
the leaves of B. officinalis, 12 compounds were isolated. Five able to a glucopyranosyl unit, and 13 carbon signals, which
1
13
(1—5) were found to be megastigmane glucosides, one (1) of included two tertiary methyls at the geminal position, one
which was a known compound, identified as turpinionoside secondary methyl, two methylenes, one methine, one quar-
2)
B. The other known compound, betulalbuside A (6) is 3,7- ternary carbon, one primary, secondary and tertiary carbinol,
dimethylocta-1,6-dien-3,8-diol 8-O-b-D-glucopyranosiode, respectively, one disubstituted trans double bond, and a ke-
1
1
of which absolute configuration at the 3-position was deter- tone functional group. The H– H correlation spectroscopy
mined for the first time. The known phenolic glucosides (7— (COSY) spectrum showed series correlations from H-7 (dH
3
)
4)
9
) were identified as arbutin, robustaside A, and ex- 5.95), H-8 (d 5.75), and H-9 (d 4.47) to H -10 (d 3.62).
H
H
2
H
5,6)
imine,
respectively. New phenolic compounds 10—12 These data indicated that this aglycone is a derivative of
were also derivatives of arbutin, like compounds 8 and 9. megastigmane, of which the 10-methyl was oxidized to a pri-
This paper deals with the structural elucidation of the new mary carbinol. The secondary methyl on the ring had the
compounds.
equatorial orientation, since H-4 (dH 2.46) appeared as a
triplet signal (Jꢁ13 Hz), and the side chain at the 6-position
was in the equatorial orientation, as judged from the phase-
Results and Discussion
Compounds 1—12 were isolated from the n-BuOH-solu- sensitive nuclear Overhauser enhancement spectroscopy (Ph-
ble fraction of a MeOH extract of the leaves of B. officinalis NOESY) spectrum in which H-7 (dH 5.95) crossed both
1
3
by the combination of highly porous synthetic resin (Diaion geminal methyls (d 0.93, 0.97) at the 1-position. The C-
H
HP-20), and normal- and reversed-phase column chromatog- NMR spectrum and the sign of the Cotton effect in the CD
raphy (RPCC). Droplet counter-current chromatography spectrum of the ring part were essentially the same as those
DCCC) and ODS-HPLC were also performed for the purifi- reported for ampelopsisionoside. The position of the gluco-
cation of the compounds. The details and yields are given in sidic linkage was determined to be the hydroxyl group at the
Experimental. 9-postion, as judged from heteronuclear multiple bond corre-
7)
(
Turpinionoside B (1) was isolated as an amorphous pow- lation (HMBC) spectrum in which H-1ꢂ (d 4.36) crossed C-
H
der, and NMR spectra indicated the presence of a megastig- 9 (d 80.0) and H-9 (d 4.47) C-1ꢂ (d 101.1). The mode of
C
H
C
mane skeleton and a glucopyranose unit. From spectroscopic glucosidic linkage was deduced to be b from the coupling
1
13
data, including H- and C-NMR spectra, and [a] the struc- constant of the anomeric proton. Therefore the structure of
D
*
To whom correspondence should be addressed. e-mail: hotsuka@hiroshima-u.ac.jp
© 2004 Pharmaceutical Society of Japan

September 2004
1087
Chart
1
1
3
Table 1.
C-NMR Data for Breyniaionosides A—D (2—5) and Betulal- an amorphous powder and its elemental composition was the
buside A (6) (CD OD, 100 MHz)
1
1
3
same as that of 3. The H– H COSY spectrum showed that
the spin-spin coupling systems of the ring moiety and the
side chain were also the same as those of 3. In the C-NMR
C
2
3
4
5
6
13
1
2
3
4
5
6
7
8
9
0
1
2
3
43.8
46.2
214.8
52.5
37.9
78.3
138.9
128.9
80.0
66.4
25.2
25.1
16.8
101.1
75.0
78.2
71.7
78.2
62.8
40.4
45.9
67.5
39.9
35.5
78.8
140.8
127.7
80.1
66.4
25.3
25.9
16.9
100.8
75.0
78.1
71.7
78.1
62.8
40.5
46.0
67.5
40.0
35.5
78.5
136.5
130.2
72.1
75.2
25.2
25.9
16.5
104.5
75.2
78.0
71.8
78.0
62.9
44.8
40.9
66.9
42.8
85.0
60.1
126.0
140.0
77.3
21.2
21.7
78.8
25.0
102.6
75.3
78.2
71.6
77.9
62.8
112.2
146.3
73.8
43.0
23.5
130.2
132.9
76.0
27.7
14.1
spectrum, the C-10 signal was shifted downfield (d
C
66.4Æ75.2) and the C-9 one upfield (d 80.1Æ72.1) com-
C
pared with those of 3. Thus 4 was assumed to be a positional
isomer of 3 in terms of the sugar moiety. This assumption
was confirmed by the HMBC spectrum, in which dH 3.65
(dd, Jꢁ10, 4 Hz) and 3.78 (dd, Jꢁ10, 7 Hz) (H -10) crossed
2
d 104.5 (C-1ꢂ) and d 4.31 (d, Jꢁ8 Hz, H-1ꢂ) crossed d
C
H
C
75.2. Therefore the structure of brenyaionoside C (4) was
1
elucidated to be (3S*,5R*,6S*,9x)-megastigman-7-ene-
1
1
1
3
,6,9,10-tetrol 10-O-b-D-glucopyranoside.
Breyniaionoside D (5), [a] ꢀ1.50°, was isolated as an
D
1
2
3
4
5
6
ꢂ
102.7
75.1
78.2
71.8
77.9
62.9
amorphous powder and its elemental composition was deter-
mined to be C H O based on negative-ion FAB-MS. The
ꢂ
ꢂ
ꢂ
ꢂ
ꢂ
1
9
32
8
1
13
H- and C-NMR spectra showed signals attributable to a
glucopyranosyl unit, and 13 carbon signals, which included
two tertiary methyls, one secondary methyl, three methyl-
enes, one of which bears an electronegative substitutent, two
methines with an oxygen atom, two quarternary carbons, one
breyniaionoside A (2) was elucidated to be (5R,6S,9x)- of which bears an electronegative substitutent, and a trans
1
1
megastigman-7-ene-6,9,10-triol-3-one 9-O-b-D-glucopyra- double bond. The H– H COSY spectrum showed two series
noside. The absolute configuration at the 9-position remains of spin–spin couplings: from H-6 to H-10 and H -2 to H -4.
2
2
to be determined, since in the case of adjacent secondary and Furthermore, H-2ax (dH 1.52) was coupled with H-12a (dH
primary alcohols like 9,10-diol, the result of application of a 3.47) due to long-range coupling via a w-letter interaction in
8)
b-D-glucopyranosylation-induced shift-trend for determina- 2 Hz. The HMBC spectrum show a cross peak between dH
tion of the absolute configuration of a secondary alcohol is 3.72 (d, Jꢁ8 Hz, H-12b) and d 85.0 (C-5). These results in-
C
9
)
known to be ambiguous, and also the modified Mosher’s dicate the presence of one more ring system, which satisfies
10)
method probably leads to some ambiguity.
the degree of unsaturation, acquired by HR-FAB-MS. Thus
Breyniaionoside B (3), [a] ꢀ66.5°, was isolated as an the structure of 5 was assumed to have an epoxy ring be-
D
amorphous powder and its elemental composition was deter- tween C-12 and C-5, like ascleposide C (13), isolated from
1
13
11)
mined to be C H O . The H- and C-NMR spectral data Asclepias fruticosa.
Since the HMBC spectrum also
1
9
34
9
were similar to those of 2, except for the absence of a ketone showed a cross peak between H-1ꢂ (dH 4.37) and C-9 (dC
functional group and the presence of a secondary alcohol at 77.3), breyniaionoside D (5) was a positional isomer of as-
d
67.5 with d 3.79 (tt, Jꢁ12, 4 Hz). From the coupling cleposide C (13) in terms of the sugar moiety. On application
8)
C
H
pattern of ring protons, the relative structure of the six-mem- of a b-D-glucopyranosylation-induced shift-trend for deter-
bered ring was judged to be the same as that of co-occurring mination of the absolute configuration, the 9-position was
turpinionoside B (1). Therefore the structure of breyniaiono- determined to have the R configuration, which is the same as
side B (3) was elucidated to be (3S*,5R*,6S*,9x)-megastig- in the case of 13. Since there was no probe for elucidation of
man-7-ene-3,6,9,10-tetrol 9-O-b-D-glucopyranoside. The ab- the absolute configuration of other carbons, breyniaionoside
solute configuration of the 9-position and that on the ring re- D (5) was enzymatically hydrolyzed and the modified Mosh-
6)
main to be determined for the same reason.
er’s method was applied for the aglycone (5a). As a result,
Breyniaionoside C (4), [a] ꢀ24.3°, was also isolated as the structure of the aglycone moiety was found to be the
D

1
088
Vol. 52, No. 9
same as that of 13, including the absolute configuration. lated glucopyranose moiety as in robustaside A (8) and ex-
Therefore, the structure of breyniaionoside D (5) was eluci- imine (9). The acyl moiety comprises nine carbon atoms,
dated to be (1S,3S,5R,6R,9R)-megastigman-7-ene-3,9-diol- which include a 1,4-disubstituted benzene ring, a cis double
5
,12-epoxide 9-O-b-D-glucopyranoside.
Compound 6 was isolated as an amorphous powder and its carbonyl functional group. From these data, the structure of
elemental composition was C H O . The one- and two-di- compound 10 was concluded to be a geometrical isomer of
bond [dH 5.80 (d, Jꢁ12 Hz) and 6.89 (d, Jꢁ12 Hz)], and a
1
6
28
7
mensional NMR data showed that the planar structure of 6 the acyl moiety of robustaside A isolated from Grevillea ro-
was a known compound, betulalbuside A, isolated from Be- busta, and it was named isorobustaside A according to the
12)
tula alba. However, the absolute configuration of the 3-po- mother compound.
sition was not fully determined. Thus commercially available
Breynioside A (11), [a] ꢀ38.4°, was isolated as colorless
D
(
3
3R)-(ꢀ)-linalool was oxidized with SeO to give (3R)-(ꢀ)- needles (mp 244—246 °C) and its elemental composition
2
12)
1
13
,7-dimethylocta-1,6-dien-3,8-diol ([a]D ꢀ8.60°).
Since was determined to be C H O . The H- and C-NMR data
19 20 9
5,6)
the optical rotation value of the aglycone (6a) obtained on showed a close resemblance to those of eximine (9) iso-
enzymatic hydrolysis showed [a] ꢃ7.59°, the absolute con- lated from Protea eximia, except for the presence of 4-hy-
D
figuration of the 3-position was concluded to be S.
droxybenozoate instead of benzoate. Therefore the structure
Isorobustaside A (10), [a]D ꢀ51.4°, was isolated as an of breynioside A was elucidated to be 4ꢄ-hydroxyeximine.
amorphous powder and its elemental composition was deter- Breynioside B (12), [a] ꢀ67.1°, was isolated as an amor-
D
1
13
mined to be C H O . The H- and C-NMR spectra showed phous powder and its elemental composition was determined
the presence of a para-substituted benzene ring and a 6-acy- to be C H O . The H- and C-NMR data showed a close
2
1
22
9
1
13
2
4
28 12
resemblance to eximine (9) with one more sugar moiety.
From the characteristic carbon signals, the extra sugar moiety
was concluded to be b-apiofuranose. When compared with
reported data for seguinoside A (14) isolated from Myrsine
3)
seguinii, the structure of breynioside B was concluded to be
ꢂ-benzoylseguinoside A.
6
Experimental
General Experimental Procedures A highly porous synthetic resin
(Diaion HP-20) was purchased from Mitsubishi Kagaku (Tokyo, Japan). Sil-
ica gel CC was performed on silica gel 60 (E. Merck, Darmstadt, Germany)
and reversed-phase [octadecyl silica gel (ODS)] open CC (RPCC) on Cos-
mosil 75C -OPN (Nacalai Tesque, Kyoto) [Fꢁ50 mm, Lꢁ25 cm, linear
1
8
gradient: MeOH–H O (1 : 9, 1 l)Æ(7 : 3, 1 l), fractions of 10 g were col-
2
lected]. The DCCC (Tokyo Rikakikai, Tokyo, Japan) was equipped with 500
glass columns (Fꢁ2 mm, Lꢁ40 cm), and the lower and upper layers of a
solvent mixture of CHCl –MeOH–H O–n-PrOH (9 : 12 : 8 : 2) were used as
Chart
2
3
2
the stationary and mobile phases, respectively. Five-gram fractions were col-
lected and numbered according to their order of elution with the mobile
phase. HPLC was performed on ODS (Inertsil; GL Science, Tokyo, Japan;
Fꢁ20 mm, Lꢁ250 mm), and the eluate was monitored with a UV detector
at 254 nm and a reflective index monitor. b-D-Glucosidase (emulsin) was
purchased from Sigma Chemical Co. (St. Louis, MO, U.S.A.), and crude
hesperidinase was a gift from Tanabe Pharmaceutical Co. Ltd. (R)-(ꢃ)- and
1
3
Table 2.
(
C-NMR Data for Isorobustaside A (10), Breyniosides A and B
11, 12), and Seguinoside A (14) (CD OD, 100 MHz)
3
a)
C
10
11
12
14
1
154.2
119.8
116.3
152.4
103.9
75.0
152.3
119.7
116.7
153.9
103.8
75.0
152.2
119.5
116.7
153.9
102.4
78.6
152.4
119.2
116.7
153.8
102.3
78.8
(
S)-(ꢀ)-a-MTPAs were from Nacalai Tesque Co., Ltd. (Kyoto, Japan).
2
3
4
1
2
3
4
5
6
1
2
3
4
5
6
7
8
9
1
2
3
4
7
, 6
, 5
A melting point was determined with a Yanagimoto micro-melting point
apparatus and is uncorrected. Optical rotations were measured on a Union
Giken PM-101 digital polarimeter with a 1-cm cell or a JASCO DIP-360 po-
larimeter with a 10-cm cell under an accumulation of eight times. IR spectra
were measured on a Horiba FT-710 Fourier transform infrared spectropho-
tometer and UV spectra on a JASCO V-520 UV/VIS spectrophotometer. CD
ꢂ
ꢂ
ꢂ
ꢂ
ꢂ
ꢂ
ꢄ
ꢄ
ꢄ
ꢄ
ꢄ
ꢄ
ꢄ
ꢄ
ꢄ
ꢅ
b)
b)
78.0
78.1
78.7
78.2
71.9
72.1
72.2
71.5
1
13
spectrum was recorded on a JASCO J-720 spectropolarimeter. H- and C-
NMR spectra were taken on a JEOL JNM a-400 spectrometer at 400 MHz
and 100 MHz, respectively, with tetramethylsilane (TMS) as an internal stan-
dard. Negative- and positive-ion HR-FAB-MS spectra were taken on a JEOL
JMS SX-102 spectrometer.
78.2
75.6
75.4
78.7
64.4
65.1
65.5
62.8
127.6
133.7
116.7
160.1
116.7
133.7
145.4
115.9
168.1
122.3
133.0
116.3
163.6
116.3
133.0
168.0
110.9
78.2
80.8
75.5
66.2
110.8
77.9
80.8
75.5
66.1
Plant Material Leaves of B. officinalis HEMSL. (Euphorbiaceae) were
collected in Kunigami-son, Kunigami-gun, Okinawa, Japan, in August 2000,
and a voucher specimen was deposited in the Herbarium of the Department
of Pharmacognosy, Graduate School of Biomedical Sciences, Hiroshima
University (00-BO-Okinawa-0828). A small amount of B. officinalis was
also collected in Taketomi-cho, Yaeyama-gun, Okinawa, in November 2001
(01-BO-1104).
131.4
130.7
129.7
134.4
167.9
Extraction and Fractionation Air-dried leaves of B. officinalis
4.11 kg) were extracted three times with MeOH. The MeOH extract was
ꢅ, 6ꢅ
ꢅ, 5ꢅ
ꢅ
(
concentrated to 3.0 l and then 150 ml of H O was added. This solution was
2
washed with 3.0 l of n-hexane (159 g) and then the methanolic layer was
concentrated to a viscous gum. The gummy residue was suspended in 3.0 l
ꢅ
of H O, and then extracted successively with 3.0 l each of EtOAc and n-
a) Data from ref. 3. b) May be exchangeable.
2

September 2004
1089
13
BuOH to afford 111 g and 46.6 g of the EtOAc- and n-BuOH-soluble frac- 5.63 (1H, dd, Jꢁ16, 8 Hz, H-8), 5.81 (1H, d, Jꢁ16 Hz, H-7); C-NMR
tions, respectively. Evaporation of the H O layer gave 313 g of residue. The (CD OD): Table 1; HR-FAB-MS (negative-ion mode) m/z: 405.2116
2
3
ꢀ
n-BuOH-soluble fraction was subjected to highly porous synthetic resin (Di- [MꢀH] (Calcd for C H O : 405.2125).
1
9
33
9
2
7
aion HP-20) CC (Mitsubishi Chemical Co., Ltd.; Fꢁ80 mm, Lꢁ55 cm),
Breyniaionoside C (4): Amorphous powder, [a]D ꢀ24.3° (cꢁ0.37,
ꢀ1 1
using H O–MeOH (4 : 1, 6 l), (2 : 3, 6 l), (3 : 2, 6 l), and (1 : 4, 6 l), and MeOH
(
MeOH eluate was subjected to silica gel (150 g) CC with increasing
MeOH). IR nmax (film): 3369, 2931, 1639, 1076, 1033 cm ; H-NMR
2
6 l), and 2-l fractions were collected. The residue (3.42 g) of the 20% (CD OD, 400 MHz) d: 0.84 (3H, d, Jꢁ7 Hz, H -13), 0.87 (3H, s, H -12),
3
3
3
0.98 (3H, s, H -11), 1.39 (1H, q, Jꢁ12 Hz, H-4ax), 1.41 (1H, ddd, Jꢁ12, 4,
3
amounts of MeOH in CHCl3 [CHCl3 (1 l), CHCl –MeOH (99 : 1, 1.5 l),
2 Hz, H-2eq), 1.66 (1H, t, Jꢁ12 Hz, H-2ax), 1.69 (1H, m, H-4eq), 1.95 (1H,
m, H-5), 3.22 (1H, dd, Jꢁ9, 8 Hz, H-2ꢂ), 3.26—3.28 (3H, overlapped, H-4ꢂ,
5’), 3.37 (1H, t, Jꢁ9 Hz, H-3ꢂ), 3.63—3.68 (1H, overlapped, H-6ꢂa), 3.65
(1H, dd, Jꢁ10, 4 Hz, H-10a), 3.78 (1H, dd, Jꢁ10, 7 Hz, H-10b), 3.78—3.83
(1H, overlapped, H-3), 3.87 (1H, dd, Jꢁ12, 2 Hz, H-6ꢂb), 4.31 (1H, d,
3
(39 : 1, 1.5 l), (19 : 1, 1.5 l), (37 : 3, 1.5 l), (9 : 1, 3 l), (17 : 3, 3 l), (4 : 1, 3 l),
(
3 : 1, 3 l), and (7 : 3, 3 l)], and then CHCl –MeOH–H O (70 : 30 : 4, 3 l), and
3
2
250-ml fractions were collected]. The residue (0.92 g) of the 12.5—15%
MeOH eluate was subjected to RPCC, and then the residue of fractions 28—
5 (135 mg) was purified by DCCC to give 85.0 mg of 7 in fractions 21—26
1
3
3
Jꢁ8 Hz, H-1ꢂ), 4.38 (1H, m, H-9), 5.71 (2H, s, H-7, 8); C-NMR (CD OD):
3
ꢀ
as colorless needles. The residue (78 mg) of fractions 80—88 obtained on
RPCC was purified by DCCC (31 mg in fractions 34—39), followed by
Table 1; HR-FAB-MS (negative-ion mode) m/z: 405.2148 [MꢀH] (Calcd
for C H O : 405.2125).
1
9
33
9
2
7
HPLC with MeOH–H O (3 : 7) to give 18.1 mg of 2. The residue (0.34 g) of
Breyniaionoside D (5): Amorphous powder, [a]D ꢀ1.50° (cꢁ1.21,
2
ꢀ1 1
1
5—17.5% MeOH eluate obtained on silica gel CC was subjected to RPCC,
and then the residue of fractions 70—79 (64 mg) was purified by DCCC
43 mg in fractions 16—21), followed by HPLC with MeOH–H O (3 : 17) to
MeOH). IR nmax (film): 3363, 2927, 1646, 1072, 1035 cm ; H-NMR
(CD OD, 400 MHz) d: 0.95 (3H, s, H -11), 1.15 (3H, s, H -13), 1.31 (3H, d,
3
3
3
(
Jꢁ6 Hz, H -10), 1.48 (1H, t, Jꢁ13 Hz, H-4ax), 1.52 (1H, td, Jꢁ13, 2 Hz, H-
2
3
give 30.0 mg of 3 and 5.6 mg of 4.
2ax), 1.70 (1H, dd, Jꢁ13, 7 Hz, H-2eq), 1.82 (1H, dd, Jꢁ13, 7 Hz, H-4eq),
2.06 (1H, d, Jꢁ8 Hz, H-6), 3.19 (1H, dd, Jꢁ9, 8 Hz, H-2ꢂ), 3.23 (1H, ddd,
Jꢁ9, 6, 2 Hz, H-5ꢂ), 3.34 (1H, t, Jꢁ9 Hz, H-4ꢂ), 3.36 (1H, t, Jꢁ9 Hz, H-3ꢂ),
3.47 (1H, dd, Jꢁ8, 2 Hz, H-12a), 3.67 (1H, dd, Jꢁ12, 6 Hz, H-6ꢂa), 3.72
(1H, d, Jꢁ8 Hz, H-12b), 3.83 (1H, dd, Jꢁ12, 2 Hz, H-6ꢂb), 4.37 (1H, d,
Jꢁ8 Hz, H-1ꢂ), 4.00 (1H, tt, Jꢁ13, 7 Hz, H-3), 4.41 (1H, quintet, Jꢁ6 Hz,
The residue (7.57 g) of the 40% MeOH eluate obtained on Diaion HP-20
CC was subjected to silica gel (450 g) CC with increasing amounts of
MeOH in CHCl [CHCl (2 l), CHCl –MeOH (99 : 1, 3 l), (39 : 1, 3 l), (19 : 1,
3
3
3
3
l), (37 : 3, 3 l), (9 : 1, 6 l), (17 : 3, 6 l), (4 : 1, 6 l), (3 : 1, 6 l), and (7 : 3, 6 l)],
and then CHCl –MeOH–H O (70 : 30 : 4, 6 l), and 250-ml fractions were col-
3
2
1
3
lected]. The residue (1.10 g) of the 7.5—10% MeOH eluate was subjected to
RPCC, and then the residue of fractions 122—134 (124 mg) was purified by
DCCC to give 27.3 mg of 6 in fractions 69—78. The residue (1.21 g) of the
H-9), 5.78 (1H, dd, Jꢁ16, 8 Hz, H-7), 5.80 (1H, dd, Jꢁ16, 6 Hz, H-8); C-
NMR (CD OD): Table 1; HR-FAB-MS (negative-ion mode) m/z: 387.1989
3
ꢀ
[MꢀH] (Calcd for C H O : 387.2019).
1
9
31
8
2
3
1
0—12.5% MeOH eluate obtained on silica gel CC was subjected to RPCC,
and then the residue of fractions 87—97 (152 mg) was purified by DCCC
38 mg in fractions 46—57), followed by HPLC with MeOH–H O (1 : 4) to
Betulalbuside A (6): Amorphous powder, [a] ꢀ32.7° (cꢁ1.89, MeOH).
D
ꢀ1 1
IR nmax (film): 3431, 2925, 1650, 1076 cm ; H-NMR (CD OD, 400 MHz)
3
(
d: 1.26 (3H, s, H -9), 1.54 (2H, m, H -5), 1.67 (3H, br s, H -10), 2.09 (2H,
2
3
2
3
afford 4.0 mg of 5. The residue (226 mg) of fractions 105—120 obtained on
RPCC was purified by DCCC (87 mg in fractions 44—55), followed by
m, H -4), 3.20 (1H, dd, Jꢁ9, 8 Hz, H-2ꢂ), 3.22 (1H, m, H-5ꢂ), 3.29 (1H, t,
2
Jꢁ9 Hz, H-4ꢂ), 3.34 (1H, t, Jꢁ9 Hz, H-3ꢂ), 3.66 (1H, dd, Jꢁ12, 6 Hz, H-
6ꢂa), 3.85 (1H, dd, Jꢁ12, 2 Hz, H-6ꢂb), 4.03 (1H, d, Jꢁ12 Hz, H-8a), 4.19
(1H, d, Jꢁ12 Hz, H-8b), 4.25 (1H, d, Jꢁ8 Hz, H-1ꢂ), 5.03 (1H, dd, Jꢁ11,
2 Hz, H-1a), 5.20 (1H, dd, Jꢁ18, 2 Hz, H-1b), 5.48 (1H, tq, Jꢁ7, 1 Hz, H-6),
HPLC with MeOH–H O (7 : 13) to give 41.3 mg of 1.
2
The residue (9.25 g) of the 60% MeOH eluate obtained on Diaion HP-20
CC was subjected to silica gel (450 g) CC with increasing amounts of
MeOH in CHCl [CHCl (2 l), CHCl –MeOH (99 : 1, 3 l), (39 : 1, 3 l), (19 : 1,
1
3
3
3
3
5.91 (1H, dd, Jꢁ18, 11 Hz, H-2); C-NMR (CD OD, 100 MHz): Table 1.
3
ꢀ
3
l), (37 : 3, 3 l), (9 : 1, 6 l), (17 : 3, 6 l), (4 : 1, 6 l), (3 : 1, 6 l), and (7 : 3, 6 l)],
HR-FAB-MS (negative-ion mode) m/z: 331.1765 [MꢀH] (Calcd for
and then CHCl –MeOH–H O (70 : 30 : 4, 6 l), and 500-ml fractions were col-
C H O : 331.1757).
3
2
16 27
7
2
2
lected]. The residue (0.49 g) of the 10—12.5% MeOH eluate was subjected
to RPCC, and then the residue (184 mg) of fractions 140—156 was purified
by DCCC to give 95.0 mg of 9 in fractions 105—115. The residue (1.04 g)
of the 12.5—15% MeOH eluate obtained on silica gel CC was subjected to
RPCC, and then the residue (194 mg) of fractions 113—123 was purified by
DCCC to give 127 mg of 11 in fractions 52—58. The residue (435 mg) of
RPCC fractions 133—152 was subjected to DCCC to give 263 mg of a crys-
talline material in fractions 58—67. This was recrystallized from MeOH to
give 11.4 mg of 8. The residue of the mother liquor was purified by HPLC
Isorobustaside A (10): Amorphous powder, [a]D ꢀ51.4° (cꢁ0.24,
MeOH). IR nmax (film): 3375, 1692, 1629, 1604, 1510, 1449, 1212, 1171,
ꢀ1
1074, 832, 777 cm . UV lmax (MeOH): 225 (4.09), 300 (4.11), 311 (4.13)
1
nm (log e). H-NMR (CD OD, 400 MHz) d: 3.41—3.44 (3H, m, H-2ꢂ, 3ꢂ,
3
4ꢂ), 3.59 (1H, m, H-5ꢂ), 4.31 (1H, dd, Jꢁ12, 6, H-6ꢂa), 4.48 (1H, dd, Jꢁ12,
2 Hz, H-6ꢂb), 5.80 (1H, d, Jꢁ12 Hz, H-8ꢄ), 6.64 (2H, d, Jꢁ9 Hz, H-3, 5),
6.72 (2H, d, Jꢁ9 Hz, H-3ꢄ, 5ꢄ), 6.89 (1H, d, Jꢁ12 Hz, H-7ꢄ), 6.91 (2H, d,
Jꢁ9 Hz, H-2, 6), 7.60 (2H, d, Jꢁ9 Hz, H-2ꢄ, 6ꢄ), H-1ꢂ was overlapped by a
ꢀ
DHO signal. HR-FAB-MS (negative-ion mode) m/z: 417.1170 [MꢀH]
with MeOH–H O (2 : 3) to give 4.7 mg of 10. The residue (0.77 g) of the
(Calcd for C H O : 417.1186).
2
21 21
9
22
12.5—15% MeOH eluate obtained on silica gel CC was subjected to RPCC,
Breynioside A (11): Colorless needles (MeOH), mp 244—246 °C, [a]D
ꢀ38.4° (cꢁ1.12, MeOH). IR n (film): 3369, 1697, 1607, 1509, 1280,
and then the residue (142 mg) of fractions 154—166 was purified by DCCC
max
ꢀ1
to give 76 mg of 12 in fractions 70—82.
1211, 1047, 833, 769 cm . UV lmax (MeOH): 216 (3.94), 258 (4.04) nm
2
7
1
Breyniaionoside A (2): Amorphous powder, [a]D ꢀ48.8° (cꢁ1.21, (log e). H-NMR (CD OD, 400 MHz) d: 3.41—3.48 (3H, m, H-2ꢂ, 3ꢂ, 4ꢂ),
3
ꢀ1
1
MeOH). IR n
(film): 3376, 2931, 1639, 1076, 1037 cm
CD OD, 400 MHz) d: 0.93 (3H, s, H -12), 0.95 (3H, d, Jꢁ6 Hz, H -13),
;
H-NMR
3.71 (1H, ddd, Jꢁ10, 7, 2 Hz, H-5ꢂ), 4.35 (1H, dd, Jꢁ12, 7, H-6ꢂa), 4.47
(1H, d, Jꢁ8 Hz, H-1ꢂ), 4.72 (1H, dd, Jꢁ12, 2 Hz, H-6ꢂb), 6.60 (2H, d,
Jꢁ9 Hz, H-3, 5), 6.84 (2H, d, Jꢁ9 Hz, H-3ꢄ, 5ꢄ), 6.93 (2H, d, Jꢁ9 Hz, H-2,
max
(
3
3
3
0
2
.97 (3H, s, H -11), 1.82 (1H, dd, Jꢁ13, 2 Hz, H-2eq), 2.12 (1H, dq, Jꢁ13,
Hz, H-4eq), 2.31 (1H, m, H-5), 2.46 (1H, t, Jꢁ13 Hz, H-4ax), 2.86 (1H, d,
3
1
3
6), 7.91 (2H, d, Jꢁ9 Hz, H-2ꢄ, 6ꢄ). C-NMR (CD OD): Table 2. HR-FAB-
3
ꢀ
Jꢁ13 Hz, H-2ax), 3.20 (1H, ddd, Jꢁ9, 6, 2 Hz, H-5ꢂ), 3.26 (1H, dd, Jꢁ9, 7
MS (negative-ion mode) m/z: 391.1005 [MꢀH] (Calcd for C H O :
1
9
19
9
H, H-2ꢂ), 3.26—3.32 (2H, overlapped, H-3ꢂ, 4ꢂ), 3.62 (2H, m, H -10), 3.65
391.1029).
Breynioside B (12): Amorphous powder, [a]D ꢀ67.1° (cꢁ0.89, MeOH).
IR nmax (film): 3421, 1708, 1603, 1509, 1290, 1227, 1076, 825, 773 cm
2
2
3
(1H, dd, Jꢁ12, 6 Hz, H-6ꢂa), 3.86 (1H, dd, Jꢁ12, 2 Hz, H-6ꢂb), 4.36 (1H, d,
ꢀ
1
Jꢁ7 Hz, H-1ꢂ), 4.47 (1H, m, H-9), 5.75 (1H, dd, Jꢁ16, 8 Hz, H-8), 5.95
.
1
3
1
(
(
[
1H, d, Jꢁ16 Hz, H-7); C-NMR (CD OD): Table 1; CD: ꢃ153 (285) [q] UV lmax (MeOH): 226 (4.03), 282 (3.33) nm (log e). H-NMR (CD OD,
3
3
nm) (cꢁ0.011 M, MeOH); HR-FAB-MS (negative-ion mode) m/z: 403.1950
MꢀH] (Calcd for C H O : 403.1968).
400 MHz) d: 3.43 (1H, t, Jꢁ10 Hz, H-3ꢂ), 3.58 (1H, d, Jꢁ11 Hz, H-5ꢄa),
ꢀ
19
31
9
3.61 (1H, d, Jꢁ11 Hz, H-5ꢄb), 3.62 (2H, m, H-2ꢂ, 3ꢂ), 3.72 (1H, ddd, Jꢁ10,
8, 2 Hz, H-5ꢂ), 3.79 (1H, d, Jꢁ10 Hz, H-4ꢄa), 3.98 (1H, d, Jꢁ2 Hz, H-2ꢄ),
4.10 (1H, d, Jꢁ10 Hz, H-4ꢄb), 4.40 (1H, dd, Jꢁ12, 8 Hz, H-6ꢂa), 4.71 (1H,
dd, Jꢁ12, 2 Hz, H-6ꢂb), 4.81 (1H, d, Jꢁ8 Hz, H-1ꢂ), 5.48 (1H, d, Jꢁ2 Hz, H-
1ꢄ), 6.58 (2H, d, Jꢁ9 Hz, H-3, 5), 6.91 (2H, d, Jꢁ9 Hz, H-2, 6), 7.49 (2H, t,
Jꢁ8 Hz, H-3ꢅ, 5ꢅ), 7.61 (1H, tt, Jꢁ8, 1 Hz, H-4ꢅ), 8.02 (2H, dd, Jꢁ8, 1 Hz,
2
7
Breyniaionoside B (3): Amorphous powder, [a]D ꢀ66.5° (cꢁ2.00,
ꢀ1
1
MeOH). IR n
(
0
(film): 3369, 2929, 1641, 1078, 1027 cm
;
H-NMR
max
CD OD, 400 MHz) d: 0.87 (3H, d, Jꢁ6 Hz, H -13), 0.88 (3H, s, H -12),
.97 (3H, s, H -11), 1.40 (1H, q, Jꢁ12 Hz, H-4ax), 1.42 (1H, ddd, Jꢁ12, 4,
3
3
3
3
2
Hz, H-2eq), 2.31 (1H, m, H-5), 1.66 (1H, t, Jꢁ12 Hz, H-2ax), 1.69 (1H, m,
1
3
H-4eq), 1.98 (1H, m, H-5), 3.18 (1H, ddd, Jꢁ9, 6, 2 Hz, H-5ꢂ), 3.26 (1H, dd,
Jꢁ9, 7 Hz, H-2ꢂ), 3.29—3.31 (2H, overlapped, H-3ꢂ, 4ꢂ), 3.61 (2H, m, H2- m/z: 507.1524 [MꢀH] (Calcd for C H O : 507.1503).
H-2ꢅ, 6ꢅ). C-NMR (CD OD): Table 2. HR-FAB-MS (negative-ion mode)
3
ꢀ
24
27 12
1
0), 3.65 (1H, dd, Jꢁ12, 6 Hz, H-6ꢂa), 3.79 (1H, tt, Jꢁ12, 4 Hz, H-3), 3.85
Enzymatic Hydrolysis of Breyniaionoside D (5) Breyniaionoside D
(1H, dd, Jꢁ12, 2 Hz, H-6ꢂb), 4.37 (1H, d, Jꢁ7 Hz, H-1ꢂ), 4.43 (1H, m, H-9),
(5, 3.8 mg) in 2 ml of H O was hydrolyzed with crude hesperidinase (5 mg)
2

1
090
Vol. 52, No. 9
at 37 °C for 24 h. The reaction mixture was partitioned with 2 ml of EtOAc.
H -9), 1.54 (2H, m, H -4), 1.63 (1H, br s, H-10), 2.07 (2H, m, H -5), 3.90
3
2
2
Evaporation of the organic layer resulted in 1.8 mg (81%) of aglycone (5a). (2H, s, H -8), 5.03 (1H, dd, Jꢁ11, 2 Hz, H-1a), 5.19 (1H, dd, Jꢁ18, 2 Hz, H-
2
13
Evaporation of the aqueous layer and trituration with MeOH gave glucose,
1b), 5.38 (1H, tq, Jꢁ7, 1 Hz, H-6), 5.91 (1H, dd, Jꢁ18, 11 Hz, H-2). C-
1
which was identified on TLC. Aglycone (5a): Colorless syrup, H-NMR NMR (CD OD, 100 MHz) d: 13.7 (C-10), 23.4 (C-4), 27.7 (C-9), 43.1 (C-
3
(
CD OD, 400 MHz) d: 0.94 (3H, s, H -11), 1.14 (3H, s, H -13), 1.25 (3H, d, 5), 69.0 (C-1), 73.8 (C-6), 112.1 (C-8), 126.9 (C-3), 135.9 (C-2), 146.3 (C-
3 3 3
ꢀ
Jꢁ6 Hz, H -10), 1.49 (1H, dd, Jꢁ13, 10 Hz, H-4ax), 1.51 (1H, ddd, Jꢁ13,
7). HR-FAB-MS (negative-ion mode) m/z: 169.1222 [MꢀH] (Calcd for
3
1
1
0, 2 Hz, H-2ax), 1.69 (1H, dd, Jꢁ13, 10 Hz, H-2eq), 1.83 (1H, dd, Jꢁ13, C H O : 169.1229).
1
0
17
2
2
2
0 Hz, H-4eq), 2.03 (1H, m, H-6), 3.46 (1H, dd, Jꢁ8, 2 Hz, H-12a), 3.71
D-Glucose: [a]D ꢃ52.6° (cꢁ0.95, H O, 24 h after being dissolved in the
2
(1H, d, Jꢁ8 Hz, H-12b), 4.01 (1H, tt, Jꢁ13, 10 Hz, H-3), 4.28 (1H, quintet,
solvent).
Jꢁ6 Hz, H-9), 5.78 (1H, dd, Jꢁ16, 8 Hz, H-7), 5.80 (1H, dd, Jꢁ16, 6 Hz, H-
Oxidation of (R)-(ꢀ)-Linalool (R)-(ꢀ)-Linalool (1.0 g) was oxidized
13
13)
8
). C-NMR (CD OD, 100 MHz) d: 21.5 (C-11), 24.0 (C-10), 25.0 (C-13),
with SeO (0.87 g) in 40 ml of EtOH at 50 °C for 3 h. The reaction mixture
3
2
40.9 (C-2), 42.8 (C-4), 44.7 (C-1), 60.1 (C-6), 66.9 (C-3), 69.1 (C-9), 78.9
was diluted with H O and then extracted three times with EtOAc. The
2
(
C-12), 84.9 (C-5), 124.4 (C-7), 142.0 (C-8). HR-FAB-MS (negative-ion residue obtained on evaporation of the EtOAc layer was subjected to silica
ꢀ
mode) m/z: 225.1499 [MꢀH] (Calcd for C H O : 255.1491).
gel (100 g) CC with CHCl and CHCl : MeOH (95 : 5). From the latter elu-
3 3
1
3
11
3
(R)- and (S)-MTPA Diesters of 5a A solution of 5a (0.9 mg) in 1 ml of
ate, 228 mg of (R)-(ꢀ)-3,7-dimethylocta-1,6-diene-3,8-diol (6b) was ob-
dehydrated CH Cl was reacted with (R)-MTPA (24 mg) in the presence of tained.
2
2
2
2
1
13
1
-ethyl-3-(3-dimethyalminopropyl)carbodiimide hydrochloride (12 mg) and
N,N-dimethylaminopyridine (10 mg), and the mixture was occasionally NMR spectra were essentially the same as those of the natural compound
stirred at 25 °C for 1 h. After the addition of 1 ml of CH Cl , the solution (6a).
Known Compounds Isolated Turpinionoside B (1): Amorphous pow-
Amorphous powder, [a]D ꢀ8.60° (cꢁ0.76, MeOH). The H- and C-
2
2
was successively washed with H O (1 ml), 5% HCl (1 ml), NaHCO -satu-
2
3
2
2
2)
rated H O (1 ml), and brine (1 ml). The organic layer was dried over Na SO
and then evaporated under reduced pressure. The residue was purified by
preparative TLC [silica gel (0.25-mm thickness, Merck), applied for 18 cm,
der, [a]D ꢀ8.6° (cꢁ0.76, MeOH). Arbutin (7): Colorless needles
22 3)
2
2
4
(MeOH), mp 163—165 °C, [a]D ꢀ55.8° (cꢁ0.56, MeOH). Robustaside A
22
D
(8): Colorless needles (MeOH), mp 210—213 °C, [a] ꢀ67.9° (cꢁ0.56,
4)
22
D
developed with CHCl –acetone (19 : 1) for 9 cm, and then eluted with
MeOH). Eximine (9): Colorless needles (MeOH), mp 208—210 °C, [a]
5,6)
3
CHCl –MeOH (9 : 1)] to furnish the 3,9-di-O-(R)-MTPA ester, 5b (1.4 mg,
ꢀ38.6° (cꢁ0.47, MeOH).
3
53%). Through a similar procedure, 5c (1.3 mg, 50%) was prepared from 5a
(
0.9 mg) by use of (S)-MTPA (26 mg), EDC (13 mg), and DMAP (9 mg).
Acknowledgements The authors are grateful for access to the supercon-
ducting NMR instrument in the Analytical Center of Molecular Medicine of
1
3,9-Di-O-(R)-MTPA Ester (5b): Colorless syrup. H-NMR (CDCl3,
4
00 MHz) d: 0.91 (3H, s, H -11), 1.16 (3H, s, H -13), 1.37 (3H, d, Jꢁ6 Hz, the Graduate School of Biomedical Sciences, Hiroshima University. This
3
3
3
H -10), 1.49 (1H, dd, Jꢁ13, 10 Hz, H-4ax), 1.60 (1H, ddd, Jꢁ13, 10, 2 Hz, work was supported by a Grant-in-Aid from the Ministry of Education, Sci-
H-2ax), 1.91 (1H, dd, Jꢁ13, 10 Hz, H-2eq), 1.98 (1H, dd, Jꢁ13, 10 Hz. H- ence, Sports, Culture and Technology of Japan (No. 13672216). Thanks are
4
eq), 2.05 (1H, d, Jꢁ9 Hz. H-6), 3.52 (1H, dd, Jꢁ8, 2 Hz, H-12a), 3.52 (3H,
also due to the Okinawa Foundation for the financial support with an Oki-
q, Jꢁ1 Hz, –OCH ), 3.54 (3H, q, Jꢁ1 Hz, –COCH ), 3.85 (1H, d, Jꢁ8 Hz, nawa Research Promotion Award (H.O.).
3
3
H-12b), 5.39 (1H, tt, Jꢁ13, 10 Hz, H-3), 5.56 (1H, quintet, Jꢁ6 Hz, H-9),
.70 (1H, dd, Jꢁ15, 6 Hz, H-8), 5.81 (1H, dd, Jꢁ15, 9 Hz, H-7), 7.32—7.43 References and Notes
5
(
6H, m), 7.48—7.53 (4H, m) (aromatic protons). HR-FAB-MS (positive-ion
1) Chang P. C., Wu C. C., “Herbs of Taiwan in Color,” Vol. 1, Nan Cun
(South Villa) Trade Co., Ltd., Taipei, ROC, 1979, p. 99.
2) Yu Q., Otsuka H., Hirata E., Shinzato T., Takeda Y., Chem. Pharm.
Bull., 50, 640—644 (2002).
ꢃ
mode, m-nitrobenzyl alcohol as a matrix) m/z: 681.2258 [MꢃNa] (NaI)
(Calcd for C H O F Na: 681.2263).
33 36 5 3
1
3
,9-Di-O-(S)-MTPA Ester (5c): Colorless syrup. H-NMR (CDCl3,
4
00 MHz) d: 0.87 (3H, s, H -11), 1.15 (3H, s, H -13), 1.42 (3H, d, Jꢁ6 Hz,
3
3) Zhong X.-N., Otsuka H., Ide T., Hirata E., Takushi A., Takeda Y., Phy-
tochemistry, 49, 2149—2153 (1998).
3
3
H -10), 1.47 (1H, ddd, Jꢁ13, 10, 2 Hz, H-2ax), 1.56 (1H, dd, Jꢁ13, 10 Hz,
H-4ax), 1.86 (1H, dd, Jꢁ13, 10 Hz, H-2eq), 2.01 (1H, dd, Jꢁ13, 10 Hz, H-
4) Ahmed, A. S., Nakamura N., Meselhy R., Makhboul M. A., El-Emary
N., Hattori M., Phytochemistry, 53, 149—154 (2000).
5) Verotta L., Orsini F., Pelizzoni F., Torri G., Rogers C. B., J. Nat. Prod.,
62, 1526—1531 (1999).
6) Perold G. W., Rosenberg M. E. K., Howard A. S., Huddle P. A., J.
Chem. Soc., Perkin Trans. I, 1979, 239—243 (1979).
7) Inada A., Nakamura Y., Konishi M., Murata H., Kitamura F., Toya H.,
Nakanishi T., Chem. Pharm. Bull., 38, 2437—2439 (1991).
8) Kasai R., Suzuno M., Asakawa J., Tanaka O., Tetrahedron Lett., 1977,
175—178 (1977).
4
2
eq), 2.02 (1H, d, Jꢁ9 Hz. H-6), 3.50 (3H, br s, –OCH ), 3.53 (1H, dd, Jꢁ8,
3
Hz, H-12a), 3.53 (3H, br s, –COCH ), 3.83 (1H, d, Jꢁ8 Hz, H-12b), 5.41
3
(
1H, tt, Jꢁ13, 10 Hz, H-3), 5.56 (1H, quintet, Jꢁ6 Hz, H-9), 5.61 (1H, dd,
Jꢁ15, 6 Hz, H-8), 5.73 (1H, dd, Jꢁ15, 9 Hz, H-7), 7.34—7.40 (6H, m),
.49—7.51 (4H, m) (aromatic protons). HR-FAB-MS (positive-ion mode,
7
ꢃ
m-nitrobenzyl alcohol as a matrix) m/z: 681.2258 [MꢃNa] (NaI) (Calcd
for C H O F Na: 681.2263).
33
36
7 6
DdS–dR in Hz at 400 MHz: H-2ax, ꢀ52 Hz; H-2eq, ꢀ50 Hz; H-4ax,
ꢃ29 Hz; H-4eq, ꢃ46 Hz; H-7, ꢀ33 Hz; H-8, ꢀ32 Hz; H -10, ꢃ21 Hz; H -
3
3
1
1, ꢀ12 Hz; H-12a, ꢀ4 Hz; H -13, ꢀ3 Hz.
Enzymatic Hydrolysis of Betulalbuside
9) Kijima H., Otsuka H., Ide T., Ogimi C., Hirata E., Takushi A., Takeda
Y., Phytochemistry, 42, 723—727 (1996).
3
A (6) Betulalbuside (6,
2
7.3 mg) in 2 ml of H O was hydrolyzed with emulsin (15 mg) at 37 °C for 10) Ohtani I., Kusumi T., Kashman Y., Kakisawa H., J. Am. Chem. Soc.,
2
24 h. The resultant hydrolyzate was separated by silica gel CC to give (S)-
113, 4092—4096 (1991).
(ꢃ)-3,7-dimethylocta-1,6-dien-3,8-diol as an aglycone (6a) (11.4 mg, 82%) 11) Abe F., Yamauchi T., Chem. Pharm. Bull., 48, 1908—1911 (2000).
and 14.3 mg of D-glucose.
S)-(ꢃ)-3,7-Dimethylocta-1,6-diene-3,8-diol (6a): Amorphous powder,
12) Tschesche R., Ciper F., Breitmaier E., Chem. Ber., 110, 3111—3117
(1977).
(
23
[
a] ꢃ7.59° (cꢁ0.76, MeOH). IR n
(film): 3366, 2924, 1641, 1455, 13) Sekine T., Fukasawa N., Ikegami F., Saito K., Fujii Y., Murakoshi I.,
D
max
ꢀ1
1
1
410, 1331, 999, 919 cm . H-NMR (CD OD, 400 MHz) d: 1.25 (3H, s,
Chem. Pharm. Bull., 45, 148—151 (1997).
3