Flavonoids from Ziziphus jujuba Mill var. Spinosa

Article abstract of DOI:10.1016/S0040-4020(00)00842-5

Eight flavonoid compounds were isolated from the seeds of Ziziphus jujuba Mill var. Spinosa. On the basis of chemical and spectral analyses their structures were elucidated as swertish (1), puerarin (2), 6'''-feruloylspinosin (3), apigenin-6-C-β-D-glucopy

Full text of DOI:10.1016/S0040-4020(00)00842-5

TETRAHEDRON
Pergamon
Tetrahedron 56 (2000) 8915±8920
Flavonoids from Ziziphus jujuba Mill var. spinosa
a
a
b
b
Gong Cheng,a Yanjing Bai,a Yuying Zhao,a, Jing Tao, Yi Liu, Guangzhong Tu, Libin Ma,
*
Ning Liaoc and Xiaojie Xuc
aDivision of Natural Medicinal Chemistry, Beijing Medical University, Beijing 100083, People's Republic of China
bBeijing Institute of Microchemistry, Beijing 100091, People's Republic of China
cCollege of Chemistry and Molecular Engineering, Peking University, Beijing 100871, People's Republic of China
Received 28 January 2000; revised 30 August 2000; accepted 14 September 2000
AbstractÐEight ¯avonoid compounds were isolated from the seeds of Ziziphus jujuba Mill var. Spinosa. On the basis of chemical and
spectral analyses their structures were elucidated as swertish (1), puerarin (2), 6000-feruloylspinosin (3), apigenin-6-C-b-d-glucopyranoside
(4), spinosin (5), 6000-feruloylisospinosin (6), isospinosin (7), and isovitexin-200-O-b-d-glucopyranoside (8). Flavonoids 6 and 7 are novel
compounds. Rotamers exist for compounds 1, 3 and 5, which are reported for the ®rst time. Compounds 2, 4 and 8 were isolated from this
plant for the ®rst time. Spinosin and swertish possess signi®cant sedative activity. q 2000 Elsevier Science Ltd. All rights reserved.
Introduction
extracted with 95% aqueous EtOH. The extract was
dissolved in 50% aqueous EtOH and extracted with petro-
leum ether. The 50% aqueous EtOH layer was concentrated
and the residue obtained was extracted with n-BuOH. The
butanolic extract was isolated using silica gel column,
Sephadex LH-20 column and HPLC to obtain compounds
1±8 (Fig. 1).
The seeds of Ziziphus jujuba Mill var. spinosa (Bunge)
Huex. H.F. Chou are used as a sedative medicine in
China. In our recent research, a 95% aqueous EtOH extract
of the seeds were separated by repeated chromatagraphy to
give eight ¯avonoid compouds. On the basis of chemical
and spectral analyses their structures were elucidated as
swertish (1), puerarin (2), 6000-feruloylspinosin (3), api-
genin-6-C-b-d-glucopyranoside (5, 7, 40- trihydroxy-
¯avone-6-C-b-d-glucopyranoside, 4), spinosin (5),
6000-feruloylisospinosin (6), isospinosin (7), isovitexin-200-
O-b-d-glucopyranoside (5, 7, 40-trihydroxy¯avone-6-C-b-
d-glucopyranosyl-(1!2)-b-d-glucopyranoside, 8) (Fig.1).
Flavonoids 6 and 7 are novel compounds named 6000-feru-
loylisospinosin and isospinosin. Rotamers exist for
compounds 1, 3 and 5, which are reported for the ®rst
time. Compounds 2, 4 and 8 were isolated from this plant
for the ®rst time. Spinosin and swertish possess signi®cant
sedative activity. The oral administration of spinosin and
swertish (4£1025 mol/Kg) prolonged pentobarbital induced
sleeping time by 29±31% compared to the control group.
Swertish used to study the sedative activity was afforded by
the acidic hydrolysis of 5.
Compound 5, yellow powder, mp 237±2408C. UV lmax
(nm) MeOH: 271, 334; NaOMe: 271, 389; AlCl3: 281,
301, 352, 383; AlCl3/HCl: 281, 302, 350, 382; NaOAc:
271, 390. The UV spectrum showed 5 had the structure of
5,40-dihydroxy¯avonoid. NFAB-MS gave a quasi molecular
ion peak at m/z: 607[M21]2. 1H NMR and 13C NMR spec-
tra possess separate signals (Experimental and Table 1),
which were almost superimposable on those of the mixture
of spinosin and zivulgarin reported by Zeng Lu1 the
structure of spinosin is 5,40-dihydroxy-7-methyoxy-
¯avone-6-C-b-d-glucopyranosyl (1!2)-b-d-glucopyrano-
side, zivulgarin is 5,40-dihydroxy-7-methyoxy¯avone-6-C-
b-d-glucopyranosyl (1!4)-b-d-glucopyranoside. Com-
pound 5 also had the same Rf value as the mixture of spino-
1
sin and zirulgorin on TLC. In a comparison of the H and
13C NMR spectra of 5 with those of spinosin,2 except for the
phenomenon of separate signals all of the signals of 5 were
almost superimposable on those previously reported for
spinosin. One- and two-dimensional NMR techniques (1H
NMR, 13C NMR, COSY, HMBC) permitted assignments of
all the 1H and 13C signals of 5. HMBC experiments showed
correlation of the H-1 of the terminal glucose with C-2 of
the internal glucose, and H-1 of the internal glucose with C-
6 of the aglycone (Fig. 1, spinosin). These results permitted
us to conclude two glucose linked together at C-2 and the
sugar chain bound to C-6 of the aglycone of 5.
Results and Discussion
The seeds of Ziziphus jujuba Mill var. spinosa were
Keywords: Ziziphus jujuba Mill var. spinosa, ¯avonoid, ¯avone-C-glyco-
side; conformers; spinosin; 6000-feruloylisospinosin; isospinosin; sedative
activity.
* Corresponding author. Tel.: 110-62091592; fax: 110-62015584;
e-mail: nmecham@mail.bjmu.edu.cn
0040±4020/00/$ - see front matter q 2000 Elsevier Science Ltd. All rights reserved.
PII: S0040-4020(00)00842-5

8916
G. Cheng et al. / Tetrahedron 56 (2000) 8915±8920
Figure 1. The structure of compounds 1±8.
Table 1. 13C NMR data of compounds 1, 3, 5 (125 Mz, DMSO-d6) (pBˆspinosin)
No.
pB[2]
5 (298 K).
5 (393 K)
1 (298 K)
3(298 K)
Aglycone 2
3
4
5
6
163.7
102.8
181.6
159.9
108.7
164.3
90.2
163.66
163.66
102.99
182.15
160.42
108.58
164.94
90.67
163.59
102.77
181.48
159.68
109.00
164.11
90.30
163.7
103.0
181.8
159.5
109.6
164.9
90.1
163.8
103.0
182.2
160.2
109.6
164.9
91.0
163.6
102.9
181.7
159.4
108.7
165.1
89.9
163.8
103.1
182.2
160.7
108.7
164.1
90.6
102.90
181.82
159.57
108.58
163.72
90.20
7
8
9
10
156.9
104.1
120.9
127.9
115.6
161.1
156.85
104.09
120.92
128.34
115.59
160.78
55.99
70.64
80.58
78.19
70.41
81.42
61.40
105.08
74.48
76.25
69.23
76.25
60.10
156.96
104.36
120.92
128.34
115.89
161.22
56.42
70.99
81.06
78.55
70.41
81.67
61.40
105.22
74.62
76.25
69.52
76.25
60.59
156.58
104.11
120.89
127.73
115.59
160.78
55.89
70.72
80.06
78.09
70.45
80.60
61.39
104.26
74.22
76.04
69.85
75.73
60.68
156.7
104.1
121.0
128.4
115.8
161.2
56.2
70.8
72.5
79.0
69.6
156.8
104.5
121.0
128.4
115.9
161.2
56.4
70.9
72.8
79.0
70.3
156.8
103.9
121.1
128.4
115.70
161.1
56.3
70.62
80.1
78.5
70.2
81.9
61.4
105.1
74.3
76.2
68.2
73.1
157.0
104.4
121.2
128.5
115.75
161.1
56.0
70.96
81.5
78.7
70.3
81.9
61.4
105.6
74.3
76.2
68.9
73.3
10
20 60
30 50
40
OCH3
Inner glc100
70.7
80.3
78.2
70.3
81.0
61.3
104.6
74.3
76.1
69.6
75.9
60.5
200
300
400
500
81.5
61.7
81.7
61.7
600
Outer glc1000
2000
3000
4000
5000
6000
62.0
62.4
Feruloyl 1000 0
2000 0
125.3
110.8
147.77
149.2
115.3
122.9
144.6
113.6
166.1
55.6
125.5
110.9
147.85
149.2
115.4
129.0
144.7
114.0
166.2
55.6
3000 0
4000 0
5000 0
6000 0
7000 0
8000 0
9000 0
OCH3

G. Cheng et al. / Tetrahedron 56 (2000) 8915±8920
8917
Figure 2. Two conformers of spinosin from CS Chem 3D Pro.
When 1H and 13C NMR spectra of 5 were measured at high
temperature (393 K, 1208C in DMSO-d6), the multiplicity
collapsed to a ®rst order spectrum, and on decreasing to
room temperature multiplicity was observed again. On the
basis of these observations, the structure of 5 was estab-
lished as spinosin and it is proposed that rotational isomers
(Fig. 2) must exist in spinosin. Thus the report of the
mixture of spinosin and zivulgarin1 is incorrect. The
compound is not a mixture but instead rotamers of
spinosin. A very interesting phenomenon was observed.
Like spinosin, swertish (1) and 6000-ferulloylspinosin (3)
also had the same NMR phenomenon -serial separate
signals. All these compounds have 5-OH and 7-OCH3,
6-C-glycose, but in compounds 2, 4 and 6±8 the separate
signals were not observed. Obviously, two conformers are
produced by the rotational barrierd 7-OCH3 in ¯avone-6-C-
glycoside.
power term for the repulsive part rather than the more
customary 12th-power term.
Grid Scan was used as the method of conformer search. This
procedure enables systematic exploration of conformations
by stepwise alteration of selected torsion angles over speci-
®ed range of values. The spatial hindrance of the methoxyl
group on the aglycone moiety to the free rotation of the link
bond between the aglycone part and the saccharide part
seems to be the main reason of the existence of two similar
conformers. So the torsion 1 and torsion 2 were consideed in
the confomational search. Torsion 3 was added in order to
compare compounds A, B and C to show the difference
between the effects of methoxyl group and hydroxyl group
on the whole molecular conformation in total. All 3 torsions
were set to rotate from 0 to 3508 and once per 308, thus 1728
conformations in total were generated in the Grid Scan. To
avoid the irrational interatom distance generated by the
rotation, which will cause system energy rise to an
unreasonable level, each conformer obtained in the rotation
was minimized from its current point to a reasonable con-
former. Thus, the energies of those conformers are appro-
priate and comparable.
As mentioned above, compound 5 showed two stable
conformers in NMR spectroscopy. In order to uncover the
factors stabilizing these two conformers at room tempera-
ture, conformational analysis was performed on compound
A (5), B (8) and C (See Fig. 3).
The conformational analysis was performed using the MSI
insight ii software package.3 The consistent Force Field
(CFF91)4 was chosen to assign parameters to the
compounds. CFF91 is optimised to suit the prediction of
gas-phase geometries, vibrational frequencies, conforma-
tional energies, torsion barriers and crystal structures of
small models. The CFF91 force®eld employs quartic poly-
nomials for bond stretching and angle bending and a three-
term Fourier expansion for torsions. The out-of-plane (also
called inversion) coordinate is de®ned according to Wilson
et al.5 The van der Waals interactions use an inverse 9th-
For each compound, an energy vs. conformer graph was
generated after rotation of each torsion and minimization
of each conformer. For compound A, the lowest energy is
202.67 kcal/mol (conformer 679), and the highest energy is
294.53 kcal/mol (conformer 1055). For compound B, the
lowest energy is 193.59 kcal/mol (conformer 1520), and
the highest energy is 247.54 kcal/mol (conformer 291).
For compound C, the lowest energy is 234.41 kcal/mol
(conformer 1552), and the highest energy is 319.30 kcal/
mol (conformer 1326). It must be mentioned here that
since the structures of these three compounds are different,
Figure 3. The structures of the compounds used in the conformational analysis. (The torsions used in conformation analysis are indicated on the compound
A(5) and are similar for the other two compounds.)

8918
G. Cheng et al. / Tetrahedron 56 (2000) 8915±8920
Figure 4. The conformational analysis results of Compounds A, B and C.
the absolute energy can not useful for comparison. Only the
energy difference between conformers of the same
compound is meaningful.
2018C, UV lmax (nm): 271, 325; NaOMe: 248, 381;
NaOAc: 269, 331 (sh); AlCl3: 278, 304, 334, 380; AlCl3/
HCl: 278, 304, 334, 381, the UV spectum showed 6 had a
structure of 5,40-dihydroxy¯avonoid. FAB-MS gave a
pseudo-molecular ion peak at (m/z) 785[M11]1. HR-MS
showed a molecular formula C38H40O18, calcd 784.7232,
obs. 807.2135 [M1Na]1. In the 13C NMR the signals of 6
were in good agreement with those of 6000-feruloylspinosin6
except for the signals at 109.4 and 90.6 for C-6 and C-8. The
two 13C NMR signals of 6 were observed at 94.9 and 104.8,
which were identical with the signals at 94.5 and 105.2 due
to C-6 and C-8 of 5, 40-dihydroxy-7-methoxy¯avone-8-C-
b-d-glucosid.7 Based on these observation the structure of 6
was determined to be 5,40-dihydroxy-7-methyoxy¯avone-8-
C-6000-feruloyl-b-d-glucopyranosyl(1!2)-b-d-glucopyrano-
side. Compound 6 is a novel compound named 6000-feruloyl-
isospinosin.
In Fig. 4.1, there is an obvious energy barrier between two
energy minima, which agrees with the existence of the two
stable conformers of Compound A in NMR spectroscopy.
As for compound B, the substitution of a methoxyl group
with a hydroxyl group reduced the spatial hindrance to the
rotation of the link bond (Torsion-1). In the conformational
analysis Fig. 4.2, the energy barrier is much lower than for
compound A. Thus compound B dose not show iso-
conformers in NMR spectroscopy at room temperature
with so smooth a barrier between the two potential minima.
When the two hydroxyl groups in the aglycone moiety are
both methylated to form compound C, the conformation
analysis also re¯ects the enhanced hindrance caused by
two methoxyl group. The energy barrier in Fig. 4.3 is deeper
when compared to Fig. 4.1, which con®rms that two
conformers of compound C can exist.
Compound 7 was obtained as yellow powder. UV lmax
(nm) MeOH: 269, 329; NaOMe: 272, 380; NaOAc: 269,
358; AlCl3: 276, 304, 342, 385; AlCl3/HCl: 277, 303, 339,
382, the UV spectrometer showed 7 had the structure of 5,
Compound 6 was obtained as a yellow powder, mp 200±

G. Cheng et al. / Tetrahedron 56 (2000) 8915±8920
8919
40-dihydroxy¯avonoid. TOF-MS and FAB-MS gave the
same quasimolecular ion peak at m/z 609[M11]1 as spino-
sin (5). Negative HR-MS showed a molecular formula
C28H31O15, calcd 607.1668, obs. 607.1670 [M21]2. In the
13C NMR of 7 the signals for the aglycone were in good
agreement with those of compound 6, the signals for the
sugars were identical with those of 5(1208C). The structure
of 7 was identi®ed as 5,40-dihydroxy-7-methoxy¯avone-8-
C-b-d-glucopyranosyl(1!2) -b-d-glucopyranoside. Com-
pound 7 is a novel compound named isospinosin.
Pharmacognosy and deposited in Division of Natural
Medicinal Chemistry, Beijing Medical University.
Extraction and isolation
The seeds of the plant were extracted with 95% aqueous
EtOH. The extract was dissoved in 50% aqueous EtOH
and was extracted with petrol. The 50% aqueous EtOH
layer was concentrated and the residue obtained was
extracted with n-BuOH. The n-BuOH extract was fractioned
by silica gel column eluting with gradient CHCl3±MeOH±
H2O to give fractions A±C. Fraction A was puri®ed using
Sephadex LH-20 and separated by HPLC (43% aqueous
MeOH) to yield 1 (10 mg).Fraction B was fractioned by
Sephadex LH-20(MeOH) to afford fractions 1±3. Fraction
1 was subjected to CC on silica gel eluting with EtOAc±
EtOH±H2Oˆ10:1:5, to give 2 (13 mg) and 3 (40 mg).
Fraction 2 and 3 were each separated by HPLC (43%
aqueous MeOH), to give 4 (15 mg) and 6 (18 mg). Fraction
C was crystallised (CHCl3±EtOH) to give 5 (2.4 g). The
mother-liquor was subjected to HPLC (52% aqueous
MeOH) to afford 7 (12 mg) and 8 (8 mg).
In the 13C NMR spectra of compounds 1, 3, 4 and 8, signals
were found to be identical with those of known compounds
swertish,2 6000-feruloylspinosin,6 apigenin-6-C-b-d-gluco-
pyranoside8 and isovitexin-200-O-b-d-glucopyranoside,
respectively,9 but the multiplicity caused by rotatrenal
isomeism in compounds 1 and 3 was also observed in
their NMR spectra.
Experimental
General experimental procedures
Acidic hydrolysis of 5
Melting points were determined on an X4 apparatus and are
uncorrected. UV spectra were taken in MeOH on a
A solution of 5 in 2 M HCl/H2O was re¯uxed at 1008C for
6 h. The reaction mixture was extracted with n-BuOH. The
n-BuOH layer was washed with H2O and evaporated to
dryness to obtain a yellow solid, its mp, Rf and 13C NMR
data were identical with an authentic sample of swertish.
1
Shimadzu UV 260 spectrometer. H NMR and 13C NMR,
H±H COSY, HMQC and HMBC spectra were recorded
with a Bruker AM-500 instrument. FAB-MS was taken on
a KYKY-ZHP-5# spectrometer, TOF-MS was recorded on
MALDI-TOF instrument. Silica gel was purchased
from Marine Chemical Factory in Qingdao. Sephadex
LH-20, RP-18(Chemical Reagent Factory, Tian Jin) were
used. TLC was performed on a RP-18 precoated layer
(Merck).
Identi®cation
Compound 1 swertish. Yellow powder mp 225±2288C,
UV lmax (nm): MeOH: 270, 329, NaOMe: 270, 390,
AlCl3: 278, 302, 343, 373. HRFAB MS m/z:
Plant material
445.1131[calcd
for
C22H21O10(M2H)2,445.1140].1H
NMR (500 MHz DMSO-d6), (splitting caused by rotational
isomerism): 7.97 (2H, d, Jˆ7.9 Hz, 20, 60-H); 6.93 (2H, d,
Jˆ7.9 Hz, 30, 50-H); 6.87,6.85 (1H, s, 8-H); 6.83, 6.82 (s,
3-H). 13C NMR data see Table 1.
The seeds of Ziziphus jujuba Mill var. spinosa were
collected in Baoji, Shaanxi Province of China and were
identi®ed by Professor Zheng Junhua in Division of
Table 2. C NMR data of compounds 4, 6±8 (125 MHz,DMSO-d6) (pA: 5, -40-dihydroxy-7-methoxy¯avon-8-C-b-d-glycopyranoside)
13
No.
pA[7]
4
6
7
8
No.
6
7
8
2
3
4
5
6
7
8
9
164.0
102.6
181.8
161.0
95.4
163.1
105.2
155.3
104.6
123.1
128.8
115.8
161.0
163.0
102.6
181.9
160.5
108.9
163.0
93.9
156.3
102.6
121.1
128.2
115.9
161.1
163.3
102.5
182.0
161.1
94.9
164.0
104.8
155.2
104.2
121.5
128.9
115.8
161.4
56.3
71.4
79.8
78.2
70.2
81.8
60.9
163.5
102.7
182.3
161.2
95.1
164.3
104.9
155.3
104.3
121.7
129.1
115.9
161.5
56.0
71.4
80.9
78.2
70.1
81.9
60.9
163.5
102.8
182.0
161.2
108.0
163.0
93.0
156.4
103.9
121.2
128.5
116.0
181.2
1000
104.2
74.4
75.8
68.9
73.6
104.9
74.5
76.2
69.3
76.2
60.2
105.3
74.7
76.4
69.4
76.5
60.5
2000
3000
4000
5000
6000
62.4
1000 0
2000 0
3000 0
4000 0
5000 0
6000 0
7000 0
8000 0
9000 0
OCH3
125.6
111.1
147.9
149.2
115.5
123.0
144.7
114.3
166.3
56.6
10
10
20 60
30 50
40
OCH3
100
200
300
400
500
600
73.1
70.7
78.7
70.4
81.7
61.2
73.2
70.5
78.9
70.2
81.3
61.3
70.5
80.5
78.3
70.4
81.7
61.4

8920
G. Cheng et al. / Tetrahedron 56 (2000) 8915±8920
Compound 2 puerarin. White powder, mp 203±2058C,
UV lmax (nm): MeOH: 250, 303(sh), NaOAc: 258, 342,
AlCl3: 250, 305(sh)8 H NMR, (500 MHz DMSO-d6): 8.3
mp 200±2018C, FAB-MS, m/z: 785[M11]1 HRFAB MS
m/z: 807.2135 [calcd for C38H40O18Na [M1Na]1,
807.2112]. IR (KBr) n
1
cm21: 3316, 2918, 1669, 1642,
max
(1H, s, 2-H), 7.9 (1H, d, Jˆ8.7 Hz, 5-H), 7.4 (2H, d,
Jˆ8.4 Hz, 20, 60-H), 7.0 (1H, d, Jˆ8.7 Hz, 6-H),6.8 (2H,
d, Jˆ8.4 Hz, 30,50-H), 4.8 (1H, d, Jˆ9.8 Hz 100-H). FAB-
MS, m/z: 439, 417, 386, 328, 307, 296, 288, 273, 258,
245, 225. HRFAB MS m/z: 415.1036 [calcd for
C21H19O9(M2H)2, 415.1035].13C NMR (dppm, C-2±10;
10 ±60; 100 ±600): 152.5, 120.3, 174.8, 126.1, 114.9, 161.0,
112.6, 156.1, 116.7; 122.6, 129.9, 114.9, 157.1, 114.9,
129.9; 73.4, 70.8, 78.6, 70.4, 81.7, 61.3. On the basis of
the spectral analysis the structure of compound 2 was
identi®ed as puerarin (7,40-dihydroxy-iso¯avone-8-C-b-d-
glucopyranoside).
1605, 1589, 1507, 1486, 1446, 1431, 1396, 1348, 1276,
1202, 1171, 1083, 1057, 1025. UV lmax (nm): MeOH:
271, 325; NaOAc: 269, 331(sh); AlCl3: 278, 304, 334,
380. 1H NMR (500 MHz DMSO-d6): 7.98 (2H, d,
Jˆ8.6 Hz, 20, 60-H), 7.48 (1H, d, Jˆ15.9 Hz, 7000 0-H), 7.30
(1H, d, Jˆ1.5 Hz, 2000 0-H), 7.08 (1H, dd, Jˆ8, 1.5 Hz, 6000 0
-
H), 6.85 (2H, d, Jˆ8.6 Hz, 30, 50-H), 6.81 (1H, d, Jˆ8.2 Hz,
5
000 0-H), 6.70 (1H, s, 6-H), 6.43 (1H, s, 3-H), 6.29 (1H, d,
Jˆ15.9 Hz, 8000 0-H), 4.82 (1H, d, Jˆ9.6 Hz, glc 100-H), 4.03
(1H, d, Jˆ7.6 Hz, glc 1000-H), 3.84 (3H, s, 7-OCH3),3.82
(3H, s, feruloyl-OCH3). 13C NMR data see Table 2.
Compound 7 (isospinosin). Yellow powder, mp 215±
2168C IR (KBr) nmaxcm21: 3330, 2923, 1643, 1601, 1512,
1480, 1447, 1410, 1379, 1345, 1299, 1252, 1195, 1178,
1167, 1115, 1069, 1018. UV lmax (nm) MeOH: 269, 329;
NaOMe: 272, 380; NaOAc: 269, 358; AlCl3: 276, 304, 342,
385; AlCl3/HCl: 277, 303, 339, 382, TOF-MS and FAB-
MS: 609[M11]1. NHR-MS showed a molecular formula
C28H31O15, calcd 607.1663, obs. 607.1670 [M21]2.1H
NMR (500 MHz DMSO-d6): 13.3 (s, 5-OH), 8.01 (2H, d,
Jˆ8.6 Hz, 30, 50-H), 6.89 (2H, d, Jˆ8.6 Hz, 20, 60-H), 6.77
(1H, S, 6-H), 6.46 (1H, S 3-H), 4.82 (1H, d, Jˆ10.0 Hz, glc
100-H), 4.03 (1H, d, Jˆ7.5 Hz, glc 1000-H).3.84 (3H, s,
OCH3), 13C NMR data see Table 2.
Compound 3 (6000-feruloylspinosin). Yellow powder, mp
2248C (dec.), UV lmax (nm) MeOH: 274, 328, NaOMe:
269, 380, AlCl3: 284, 302, 336, 385, NaOAc: 273, 3338
1H NMR (500 MHz DMSO-d6), (splitting caused by
rotational isomerism): 7.81 (d, Jˆ7.9 Hz 20,60-H), 7.18,
7.06, (d, Jˆ15.7 Hz, 7000 0-H), 7.17, 7.04 (d,Jˆ2.0 Hz,
feruloyl- 2000 0-H), 6.89 (d, Jˆ7.9 Hz, 30,50-H), 6.85, 6.83
(s, 3-H), 6.77, 6.67 (s, 8-H), 6.72 (dd, Jˆ8.1,2.0 Hz, 6000 0
-
H),6.67 (d, Jˆ8.1 Hz, 5000 0-H), 6.23, 6.15 (d, Jˆ15.7 Hz,
8
000 0-H),4.67 (t, Jˆ9.3 Hz, glc100-H), 4.27, 4.23 (t,
Jˆ7.9 Hz, glc1000-H) FAB-MS, m/z: 783 [M21]2, 612,
607, 459, 443, 409, 379, 352, 339. HRFAB MS m/z:
783.2135 [calcd for C38H39O18(M2H)2,783.2142]. 13C
NMR data see Table 1.
Compound
8
(isovitexin-200-O-b-d-glucopyranoside).
Yellow powder, mp 206±2088C TOF-MS: 617 [M1Na]1,
633 [M1K]1. 1H NMR (500 MHz DMSO-d6) 13.6 (s,
5-OH), 7.91 (2H, d, Jˆ8.8 Hz, 20, 60-H), 6.92 (2H, d,
Jˆ8.8 Hz, 30, 50-H), 6.75 (1H, s, 8-H), 6.48 (1H, s, 3-H),
4.64 (1H, d, Jˆ9 Hz, glc 100-H), 4.15 (1H, d, Jˆ7.6 Hz, glc
1000-H). 13C NMR data see Table 2.
Compound 4 (5, 7, 40-trihydroxy¯avone-6-C-b-d-gluco-
side). Yellow powder, mp 242±2448C, UV lmax (nm):
MeOH: 271, 329; NaOMe: 279(sh), 380; AlCl3: 277, 301,
1
343, 380; AlCl3/HClˆAlCl3; NaOAc; 278, 329, 395. H
NMR (500 MHz DMSO-d6): 13.5 (1H, brs, 5-OH), 7.88
(2H, d, Jˆ8.6 Hz, 30, 50-H), 6.91 (2H, d, Jˆ8.6 Hz, 20, 60-
H), 6.69 (1H, s, 3-H), 6.41 (1H, s, 8-H), 4.57 (1H, d, 9.9 Hz,
100-H). 13C NMR data see Table 2.
References
Compound 5 (spinosin). Yellow powder, mp 237±2408C,
UV lmax (nm) MeOH: 271, 334; NaOMe: 271, 389; AlCl3:
281, 301, 352, 383; AlCl3±HCl: 281, 302, 350, 383;
NaOAc: 271, 390. NFAB-MS m/z: 607[M21]2. HRFAB
MS m/z: 607.1657 [calcd for C28H31O15(M2H)2,
607.1663]. 1H NMR (500 MHz DMSO-d6) (at room
temperature): 7.97 (2H, d, Jˆ8.7 Hz, 20, 60-H), 6.95 (2H,
d, Jˆ8.7 Hz, 30, 50-H), 6.83, 6.84 (1H, s, 3-H), 6.67, 6.80
(1H, s, 8-H), 4.67, 4.69 (1H, d, Jˆ9.8 Hz, glc 100-H), 4.15,
4.17(1H, d, Jˆ8.5 Hz, glc 1000-H). 1H NMR (at 1208C):7.88
(2H, d, Jˆ8.7 Hz, 20, 60-H), 6.98 (2H, d, Jˆ8.7 Hz, 30,
50-H), 6.73 (1H, s, 3-H), 6.67 (1H, s, 8-H), 4.77 (1H, d,
Jˆ9.7 Hz, glc 100-H), 4.25 (1H, d, Jˆ7.7 Hz, glc 1000-H),
3.92 (3H, s, OCH3),3.89 (3H, s, OCH3), 13C NMR data
see Table 1.
1. Zeng, L.; Zhang, R. Y.; Wang, X. Acta Pharmaceutica 1987, 22
(2), 114.
2. Woo, W. S.; Kang, S. S.; Shim, S. H.; Wangner, H.; Chari,
V. M.; Seligmann, O.; Obrmeier, G. Phytochemistry 1979, 18, 353.
3. InsightII Version 97.5 software; Molecular Simulations Inc.:
Burlington, MA, 1999.
4. Maple, J. R.; Hwang, M. J.; Stock®sch, T. P.; Dinur, U.;
Waldman, M.; Bwing, C. S.; Hagler, A. T. J. Comput. Chem.
1994, 15, 162±182.
5. Wilson, E. B.; Decius, J. C.; Cross, P. C. Molecular Vibrations;
Dover: New York, 1980.
6. Woo, W. S.; Kang, S. S.; Wangner, H.; Seligmann, O.; Chari,
V. M. Phytochemistry 1980, 19, 2791.
7. Fujita, M.; Inoue, T. Chem. Pharm. Bull. 1982, 30, 2342.
8. Osterdahl, B. G. Acta. Chem. Scand. 1978, B32, 93.
9. Zeribun, B.; Lockwood, G. B.; Waigh, R. D. J. Nat. Prod. 1987,
50, 322.
Compound 6 (6000-feruloylisospinosin). Yellow powder,