全 文 ：灰毛豆叶片甲醇提取物中黄酮
类物质的研究
裴晓丽 1，丁文兵 1，2，黄 蕊 1，李冠华 1，李有志 1，2*
（1.植物病虫害生物学与防控湖南省重点实验
室， 湖南农业大学植物保护学院， 湖南长沙
410128；2.国家植物功能成分利用工程技术研
究中心，湖南农业大学，湖南长沙 410128）
摘 要 [目的] 研究灰毛豆叶片甲醇提取物
中黄酮类化合物，并测试它们对斜纹夜蛾卵巢
细胞(SL 细胞)的细胞毒性。 [方法]对甲醇提取
物的分离采用了多种色谱技术分离纯化，并结
合各种光谱分析（包括 UV，1D，2D NMR 分析
以及 HR-ESR-MS）进行结构鉴定。细胞毒性的
测试采用的是 MTT 法。 [结果] 从灰毛豆叶片
甲醇提取物中分离得到 6 个黄酮类化合物 ：
6-methoxykaempferol (1), 6-methoxykaempferol
7-O-α-rhamnopyranoside (2), 6-methoxykaempferol
3-O-α-rhamnopyranosyl(1→2)[α-rhamnopyranos-
yl(1→6)]-β-galactopyranoside (3), 6-methoxyka-
empferol 3-O-α-rhamnopyranosyl (1→2) [α-rha-
mnopyranosyl(1→6)]-β-galactopyranoside-7-O-α-
rhamnopyranoside (4), pongachin (5), 5,7-dim-
ethoxy-8-(3-hydroxy-3-methylbut-1Z-enyl)flavan-
one (6)。除了化合物 5 之外其余化合物均为首
次从灰毛豆中分离得到。 细胞毒性测试发现
pongachin (5) 具有明显的细胞毒性，其 IC50为
4.4 mg/L，化合物 1，3 和 5 的细胞毒性均大于
阳性对照鱼藤酮 。 [结论 ] 灰毛豆叶片中 6-
methoxykaempferol 类化合物（1-4）含量相对较
大， 化合物 1，3 和 5 在对 SL 细胞的活性实验
中细胞毒性均大于阳性对照鱼藤酮，值得进一
步研究。
关键词 灰毛豆；黄酮类化合物；斜纹夜蛾；细
胞毒性
基金项目 国家自然科学基金（31071715）。
作者简介 裴晓丽 （1987- ），女 ，山西运城
人， 硕士研究生， 研究方向 : 植物源农药，E-
mail：530949467@qq.com。 * 通讯作者， 博士，
教授，主要从事杀虫植物成分与昆虫毒理方面
的研究。
收稿日期 2013-07-05
修回日期 2013-08-01
Study on Flavonoids from the Methanol Extracts
of Tephrosia purpurea Leaves
Xiaoli PEI1, Wenbing DING1,2, Rui HUANG1, Guanhua LI1, Youzhi LI1,2*
1. Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, College of Plant Protection, Hu-
nan Agricultural University, Changsha 410128, China;
2. National Research Center of Engineering & Technology for Utilization of Botanical Functional Ingredients, Hunan Agricultural
University, Changsha 410128, China
Supported by the National Natural Science Foundation of China (31071715).
*Corresponding author. E-mail: liyouzhi2008@sina.com
Received: July 5, 2013 Accepted: August 1, 2013A
Agricultural Science & Technology, 2013, 14(8): 1204-1208
Copyright訫 2013, Information Institute of HAAS. All rights reserved Storage and Processing
Abstract [Objective] The aim was to determine flavonoids from the MeOH extracts
of Tephrosia purpurea leaves and their cytotoxicitives against the ovarian cells from
Sprodenia litura (SL cells). [Method] The compounds were isolated with column
chromatography and their structures were established on the basis of various spec-
troscopic analysis (including UV, 1D and 2D NMR analyses as well as HR-ESR-
MS). The cytotoxicity against the SL cells was evaluated by using MTT method.
[Result] Six known flavonoids, 6-methoxykaempferol (1), 6-methoxykaempferol 7-O-
α-rhamnopyranoside (2), 6-methoxykaempferol 3-O-α-rhamnopyranosyl (1→2) [α-rham-
no-pyranosyl (1→6)]-β-galactopyranoside (3), 6-methoxykaempferol 3-O-α-rhamnopyra-
nosyl(1→2)[α-rhamnopyranosyl(1→6)]-β-galactopyranoside-7-O-α-rhamnopyranoside(4),
pongachin (5), 5,7-dimethoxy-8-(3-hydroxy-3-methylbut-1Z-enyl) flavanone (6) were is-
olated and determined. Except compound 5, the others were isolated from T. pur-
purea for the first time. For the cytotoxicity compound 5 had significant activity with
the IC50 value of 4.4 mg/L while compound 1 and 3, whose cytotoxicity exceeded
rotenone, also showed moderate activity. [Conclusion] Of all the compounds from
T. purpurea leaves, the content of 6-methoxykaempferol compounds was consider-
able. The profiles of these compounds against SL cells suggested that compounds
1, 3 and 5, whose cytotoxicity exceeded rotenone, were worth further research.
Key words Tephrosia purpurea; Flavonoids; Sprodenia litura; Cytotoxic activity
T ephrosia (family: Leguminosae;sub-family: Papilionaceae) is atropical and sub-tropical genus
of more than 400 species mostly found
in India and Africa. As the main natural
source of rotenoids, some species of
the Tephrosia, such as T. apollinea,
T. falciformis, and T. vogelii have been
widely studied. Moreover, some
species are also used as green ma-
nure and soil-keeping plants as well as
medicine[1-3].
Tephrosia purpurea, commonly
known as Pila in Sinhala, Kolinji in
Tamil, is a copiously branched xylo-
phyta and distributed in subtropical ar-
eas including Indian, the south of Chi-
na, Hawaii and so on. The whole plant
is toxic especially the root[1]. As a tra-
ditional medicine, T. purpurea is used
as a remedy for the treatment of impo-
tency, asthma, diarrhea, rheumatism,
and urinary disorders [4] and is said to
have the property of healing all types
of wounds[5]. The extracts and some
compounds of this plant have shown
various biological activities including
antibacterial , antidiabetic and an-
tioxidant, anti- inflammatory and can-
cer chemopreventive activities[6-9]. The
main chemical ingredients of T. pur-
purea revealed the presence of
rotenoids, flavanoids, sterols, sapo-
nins, tannins and their glycoside
derivatives[10-14], but little was reported
on the insecticida activity. Our previ-
ous studies had researched insectici-
dal activity of the methanol extracts
from each part of T. purpurea and re-
vealed all the parts except the xylem
of trunk possessed insecticidal prop-
erty [15]. The extracts from the seeds
DOI:10.16175/j.cnki.1009-4229.2013.08.011
Agricultural Science & Technology2013
Fig.1 Structures of compounds 1-6
and stem bark parts showed remark-
able insecticidal activity, while the ex-
tract from leaves also displayed some
insecticidal activities [15]. Further phy-
tochemistry studies were done on the
bark and seeds of T. purpurea, which
obtained some insecticidal compounds
such as isolonchocarpin, tephrosin,
12α-hydroxyrotenone and rotenone[16].
This report focused on chemical
investigation of T. purpurea leaves, re-
sulting in the isolation of six flavonoids:
6-methoxykaempferol (3, 4′ , 5, 7-te-
trahydroxy 6-methoxy flavone) (1) [17],
6-methoxykaempferol 7-O-α-rhamno-
pyranoside (2), 6-methoxykaempferol
3-O-α-rhamnopyranosyl (1→2) [α-rh-
amnopyranosyl (1→6)]-β-galactopyra-
noside (3), 6-methoxykaempferol 3-O-
α-rhamnopyranosyl (1→2) [α-rhamno-
pyranosyl(1→6)]-β-galactopyranoside-
7-O-α-rhamnopyranoside (4) [18], pon-
gachin (5) [19], 5, 7-dimethoxy-8-(3-hyd-
roxy-3-methylbut-1Z-enyl) flavanone
(6) (Fig.1) [18]. Meanwhile compounds
(1-6) were evaluated for the cytotoxi-
city against the ovarian cells from
Sprodenia litura (SL cells).
Materials and Methods
Materials
General experimental procedures
UV spectra were measured on Shi-
madzu UVmini-1240 spectrometer in-
strument (Shimadzu Corporation,
Tokyo, Japan). NMR spectra were
recorded on a Bruker DRX-400 spec-
trometer (400 MHz for 1H and 100
MHz for 13C with TMS as the internal
standard). HR-ESIvMS and ESI-MS
were recorded on an API QSTAR
mass spectrometer. The enzymevla-
beled detector was from Bio-Rad Lab-
oratories (Shanghai) Ltd. Sephadex
LH-20 (GE Healthcare, Uppsala, Swe-
den) was used for column chromatog-
raphy. Chromatography was per-
formed on silica gel (200-300 mesh;
Qingdao Marine Chemical Group, Co.
Qingdao, China), octadecylsilyl silica
gel (ODS) (30-50 μm; YMC CO. Ltd,
Kyoto, Japan) and semi-preparative
HPLC [Waters 1 525 pump, Waters
2 489 UV detector at 254 and 365 nm,
with a YMC-Pack ODS-A column (5
μm, 250×20 mm)].
Plant material The leaves of T. pur-
purea were collected from Jiangyong
County of Hunan Province, southern
China, in Oct. 2010, and identified by
Professor Yecong Zhong from Guang-
xi Academy of Forestry. An authenti-
cated voucher specimen (No. 200607)
of this plant was deposited at College
of Plant Protection, Hunan Agricultural
University．
Methods
Extraction and isolation Air dried
and ground leaves of T. purpurea
(10.5 kg) were extracted with MeOH
(3 ×50 L) at room temperature, and
then the solvent was concentrated un-
der reduced pressure. The residue
(214.7 g) was subjected to silica gel
column chromatography (CC), elutd
with a systemof CHCl3-MeOHmixtures
(100:0 -0:100) to obtain 24 fractions
(Fr.1-24). Fraction 22 (6.7 g) (CHCl3-
MeOH, 60:40) was further separated
by ODS column, using MeOH -H2O
(30:70-90:10) as eluents, to afford
subfractions (A-M). The subfraction I
(0.87 g) (MeOH-H2O, 50:50) was frac-
tionated over a Sephadex LH-20
(MeOH isocratic elution), and then was
further purified by silica gel CC, with
gradient mixtures of CHCl3 -MeOH
(80:20-60:40) to yield compound 1 (24
mg) and 2 (47 mg). The subfraction G
(1.76 g) (MeOH-H2O, 40:60) was car-
ried out over Sephadex LH-20, eluted
with MeOH and yielded three subfrac-
tions (G1-G3), G1 (100 mg) was sub-
jected to semi-preparative HPLC
(MeOH-H2O, 38:62, 5.0 ml/min) to af-
ford compound 3 (25 mg, tR=19.5 min).
G2 (130 mg) was subjected to semi-
preparative HPLC (MeOH-H2O, 38:62,
5.0 ml/min) to obtain compound 4 (25
mg, tR = 23 min). Fraction 11 (3.7 g)
(eluted from CHCl3), which was
passed over silica gel CC with CHCl3-
MeOH gradient elution (100:0-90:10),
resulted in the crystallization of com-
pound 5 (100 mg). Fraction 16 (4.5 g)
was passed over silica gel CC, with
the elution of CHCl3-MeOH (100:0-
90:10) to obtain fractions 16-A20-30 and
then purified by Sephadex LH-20 elut-
ing with MeOH to provide pure com-
pound 6 (30 mg).
Cytotoxicity assays
Cell culture SL cells were obtained
from the Key Laboratory of Pesticide
and Chemical Biology of Ministry of
Education, South China Agricultural
University, China, and cultured with
Grace’s insect cell culture medium
(Gibco, America) containing 9% new
born calf serum.
Solution preparation Each com-
pound dissolving in DMSO was sus-
pended in distilled water, and then was
diluted into a series of five different
concentrations, with gentle shaking to
ensure a homogeneous test solution.
The final concentration of DMSO was
5 g/L.
Groups for experiment Cytotoxicity
of compounds 1-6 was performed by
MTT assay as described by Wang RJ
against SL cells [20]. Cells in the loga-
rithmic growth phase were plated onto
1205
Agricultural Science & Technology 2013
Table 1 13C NMR Spectroscopic data of
compounds 1-4
Position
1 2 3 4
δC δC δC δC
2 147.5 148.1 159.3 159.8
3 135.8 136.2 134.6 134.5
4 176.6 176.7 180.2 179.9
5 151.9 151.5 152.2 154.3
6 131.4 132.4 133.1 134.3
7 157.7 155.4 159.2 156.8
8 94.2 94.4 95.5 95.6
9 152.1 152.0 154.3 153.5
10 103.9 105.5 106.7 108.2
1′ 122.2 122.0 123.5 122.1
2′/6′ 130.0 130.2 132.7 132.6
3′/5′ 115.9 115.9 116.7 117.0
4′ 159.7 159.9 161.8 163.2
6-OMe 60.5 60.8 61.0 61.6
Rha-1 99.4 100.7
2 70.5 71.9
3 70.8 72.4
4 72.0 73.7
5 70.8 71.7
6 18.4 18.4
Glc-1 101.3 101.0
2 77.7 77.7
3 75.8 75.9
4 70.8 70.9
5 75.5 75.7
6 67.3 67.5
Rha′-1 102.3 102.0
2 72.2 72.3
3 72.4 72.4
4 74.0 74.0
5 69.7 69.8
6 18.5 18.2
Rha″-1 103.1 102.8
2 72.5 72.6
3 72.4 72.5
4 74.2 74.2
5 69.9 70.0
6 18.0 17.8
(100 MHz, 1 -2 in DMSO -d6, 3 -4 in
MeOD-d4).
96 wells cell culture plate and divided
into the different groups for each com-
pound as follows: (1) experimental
groups with each compound for five
concentrations, (2) control group with-
out DMSO, (3) negative control group
with DMSO (the concentrations of
DMSO was 5 g/L). These groups were
cultured in the incubator for 48 h at
27.5 ℃. The values of OD575 were read
using an enzyme-labeled detector.
Rotenone (98% purity) was used as
positive control. All treatments and
controls were replicated five times.
Data statistic and analysis
IC50 (Median inhibitory concentra-
tion) value of each compound against
SL cells was calculated by Probit Anal-
ysis (DPS software, version 9.5). The
95% confidence interval, IC50 values
and degrees of freedom of the x2
goodness of fit tests, and regression
equations were recorded in Table 2.
Whenever the goodness of x2 was
found to be significant (P<0.05), a het-
erogeneity correction factor was used
in calculation of the confidence inter-
val.
Results and Analysis
Structure identification of com-
pounds
All structures of these compounds
(1-6) were established on the basis of
various spectroscopic analysis (in-
cluding UV, 1D and 2D NMR analyses
as well as HR-ESR-MS) and the result
was the following:
Compound 1 was obtained as
light yellow crystals (MeOH), mp 268-
271℃. The ESI-MS and NMR spectral
data of 1 agreed well with those given
in the literature for 6-methoxykaem-
pferol. The HMBC experiment of 1 was
employed to provide further confirma-
tion of the structure, which led to as-
signing the structure of 1 as 6-meth-
oxykaempferol (3, 4′, 5, 7-tetrahydroxy
6-methoxy flavone) (Fig.1). Its molecu-
lar formula was determined as
C16H12O7. 1H NMR (400 MHz, DMSO):
6.55 (1H, s, H-8), 8.05 (2H, d, J=8.9
Hz, H-2′,6′), 6.93 (2H, d, J=8.9 Hz, H-
3′,5′), 3.77 (3H, s, 6-OMe), 9.43 (1H,
br s, 3-OH), 10.13 (1H, br s, 4′-OH),
12.57 (1H, br s, 5-OH), 10.71 (1H, br
s, 7-OH). The 13C NMR Spectroscopic
data was shown in Table 1.
The 1H and 13C NMR spectra sug-
gested that 2 -4 contained the 6-me-
thoxykaempferol (1) played the role as
their aglycone, but difference in sugar
residues.
Compound 2 was obtained as a
yellow powder. The UV spectrum
showed absorption maxima at 365 nm
(log 1.07) and 268 nm (log 1.40). Its
molecular formula was determined as
C22H22O11 from ESI-MS ions at m/z 461
[M-H]-, 497 [M+Cl]- as well as the HR-
ESI-MS ion at m/z 461.106 5 [M-H]-.
1H NMR (400 MHz, DMSO): 6.99 (1H,
s, H-8), 8.11 (2H, d, J=8.9 Hz, H-2′,
6′ ), 6.94 (2H, d, J=8.9 Hz, H-3′ ,5′ ),
3.77 (3H, s, 6-OMe), 9.58 (1H, br s,
3-OH), 10.18 (1H, br s, 3-OH), 12.49
(1H, br s, 3-OH). Rha: 5.58 (1H, d, J=
1.6 Hz, H-1), 3.91 (1H, br s, H-2), 3.70
(1H, m, H-3), 3.36 (1H, m, H-4), 3.48
(1H, m, H-5), 1.15 (3H, d, J=6.1 Hz, H-
6). The 13C NMR Spectroscopic data
was shown in Table 1. The structure of
2 was identified as 6-methoxykaemp-
ferol 7-O-α-rhamnopyranoside (Fig.1).
Compound 3 was obtained as a
yellow powder. Its molecular formula
was determined as C34H42O20. 1H NMR
(400 MHz, MeOD): 6.51 (1H, s, H-8),
8.06 (2H, d, J=8.9 Hz, H-2′,6′), 6.92
(2H, d, J=8.9 Hz, H-3′,5′), 3.88 (3H, s,
6-OMe). Glc: 5.60 (1H, d, J=8.0 Hz, H-
1), 3.92 (1H, dd, J=9.6, 8.0 Hz, H-2),
3.69 (1H, dd, J=9.6, 3.5 Hz, H-3), 3.76
(1H, dd, J=3.5, 1.1 Hz, H-4), 3.62 (1H,
ddd, J=6.9, 5.6, 1.1 Hz, H-5), 3.71 (1H,
dd, J=10.5, 5.6 Hz, H-6), 3.45 (1H, dd,
J=10.5, 6.9 Hz, H-6). Rha′ : 4.51 (1H,
d, J=1.8 Hz, H-1), 3.54 (1H, dd, J=3.4,
1.8 Hz, H-2), 3.49 (1H, dd, J=9.5, 3.4
Hz, H-3), 3.26 (1H, t, J=9.5 Hz, H-4),
3.52 (1H, dd, J=9.5, 6.2 Hz, H-5), 1.18
(1H, d, J=6.2 Hz, H-6). Rha″: 5.23 (1H,
d, J=1.7 Hz, H-1), 4.00 (1H, dd, J=3.4,
1.8 Hz, H-2), 3.80 (1H, dd, J=9.5, 3.4
Hz, H-3), 3.34 (1H, t, J=9.6 Hz, H-4),
4.06 (1H, dd, J=9.6, 6.2 Hz, H-5), 0.98
(1H, d, J=6.2 Hz, H-6). The 13C NMR
Spectroscopic data was shown in
Table 1. The structure of 3 was identi-
fied as 6-methoxykaempferol 3-O-α-
rhamnopyranosyl (1→2)[α-rhamnopy-
ranosyl (1→6)]-β-galactopyranoside
(Fig.1).
Compound 4 was obtained as a
yellow powder. Its molecular formula
was determined as C40H52O24. 1H NMR
(400 MHz, MeOD): 6.89 (1H, s, H-8),
8.10 (2H, d, J=8.9 Hz, H-2′ ,6′), 6.91
(2H, d, J=8.9 Hz, H-3′,5′), 3.87 (3H, s,
6-OMe). Rha: 5.64 (1H, d, J=1.8 Hz,
H-1), 4.10 (1H, dd, J=3.5, 1.8 Hz, H-2),
3.91 (1H, dd, J=9.6, 3.5 Hz, H-3), 3.50
(1H, t, J=9.5 Hz, H-4), 3.68 (1H, dd, J=
9.5, 6.2 Hz, H-5), 1.30 (1H, d, J=6.2
Hz, H-6). Glc: 5.66 (1H, d, J=7.8 Hz,
H-1), 3.94 (1H, dd, J=9.5, 7.8 Hz, H-2),
1206
Agricultural Science & Technology2013
Table 2 Cytotoxicity of compounds 1-6 and rotenone against SL cells of Sprodenia litura
after treatment for 48 h
Compounds Toxicity regressive equation IC50 and the 95% confidenceinterval∥mg/L x
2
1 y=2.54+1.70x 27.8 (20.5-38.7) 1.160 2
2 y=1.45+1.67x 134.1 (89.1-182.4) 0.331 1
3 y=2.13+1.51x 78.8 (52.9-109.7) 0.244 2
4 y=0.33+2.05x 189.7 (144.4-246.9) 1.922 3
5 y=3.92+1.69x 4.4 (2.9-7.7) 0.990 9
6 y=0.21+(1.99±0.28)x 250.0 (190.3-333.5) 0.507 8
Rotenone y=1.77+1.586x 108.0 (47.4-372.2) 0.993 6
3.70 (1H, dd, J=9.5, 3.5 Hz, H-3), 3.77
(1H, dd, J=3.5, 1.0 Hz, H-4), 3.63 (1H,
ddd, J=6.9, 5.6, 1.0 Hz, H-5), 3.71 (1H,
dd, J=10.6, 5.6 Hz, H-6), 3.46 (1H, dd,
J=10.6, 6.9 Hz, H-6). Rha′ : 4.51 (1H,
d, J=1.7 Hz, H-1), 3.52 (1H, dd, J=3.4,
1.7 Hz, H-2), 3.47 (1H, dd, J=9.5, 3.4
Hz, H-3), 3.25 (1H, t, J=9.5 Hz, H-4),
3.51 (1H, dd, J=9.5, 6.2 Hz, H-5), 1.19
(1H, d, J =6.2 Hz, H-6). Rha: 5.22
(1H, d, J=7.8 Hz, H-1), 4.00 (1H, dd,
J=3.4, 1.8 Hz, H-2), 3.79 (1H, dd, J=
9.6, 3.4 Hz, H-3), 3.34 (1H, t, J=9.6
Hz, H-4), 4.04 (1H, dd, J=9.6, 6.2 Hz,
H-5), 1.01 (1H, d, J=6.2 Hz, H-6). The
13C NMR Spectroscopic data was
shown in Table 1. The structure of 4
was identified as 6-methoxykaem-
pferol 3-O-α-rhamnopyranosyl (1→2)
[α-rhamnopyranosyl (1→6)]-β-galac-
topyranoside-7-O-α-rhamnopyranosi-
de (Fig.1).
Compound 5 was obtained as a
white crystal. Its molecular formula
was determined as C21H20O4. 1H NMR
(500 MHz, CDCl3): 1.42 (3 H, s, -CH3),
1.44 (3 H, s, -CH3), 2.69 (1 H, dd, J=
16.3 Hz, 3.0 Hz, 3-H), 2.95 (1 H, dd,
J=16.3 Hz, 12.7 Hz, 3-H), 3.82 (3 H, s,
-OMe), 5.53 (1 H, dd, J=10.2 Hz, 3.0
Hz, H-2), 5.57 (1 H, d, J=8.0 Hz, H-
3″), 6.10 (1 H, s, H-6), 6.56 (1 H, d, J=
8.0 Hz, H-10), 7.37 (1 H,t, J=5.8 Hz, H-
4′), 7.44 (2 H, t, J=6.2 Hz, H-2′, H-6′),
7.56 (2 H, d, J=5.8 Hz, H-3′, H-5′); 13C
NMR (125 MHz, CDCl3): 29.4 (-CH3),
29.6 (-CH3), 47.3 (C-3), 57.3 (-OMe),
79.5 (C-2″ ), 80.9 (C-2), 95.6 (C-6),
104.5 (C-10), 107.6 (C-8), 117.7 (C-
4″), 128.0 (C-2′ , 6′), 128.4 (C-4′),
130.2 (C-3″), 130.5 (C-3′, 5′), 141.5
(C-1′), 160.5 (C-9), 161.4 (C-7), 164.1
(C-5), 188.8 (C-4). The structure of 5
was identified as pongachin (Fig.1).
Compound 6 was obtained as a
white powder. Its molecular formula
was determined as C22H24O5. 1H NMR
(400 MHz, CDCl3): 5.45 (1H, dd, J=
12.5 Hz , 3.2 Hz, H-2), 2.91 (2H, dd,
J =16.6 Hz , 12.5 Hz H-3), 2.89 (1H,
dd, J =16.6 Hz , 3.2 Hz, H-3 ), 6.20
(1H, s, H-6), 7.47 (2H, m, H-2′ ,6′ ),
7.42 (2H, m, H-5′,3′ ), 7.30 (1H, m,
H-4′), 3.02 (3H, s, 5-OMe), 3.10 (3H,
s, 7-OMe), 5.98 (1H, d, J=12.4 Hz, H-
1″ ), 5.90 (1H, d, J=12.4 Hz, H-2″ ),
1.34 (3H, s, 3″-CH3); 13C NMR (100
MHz, CDCl3): 78.9 (C-2), 45.5 (C-3),
189.5 (C-4), 162.0 (C-5), 89.0 (C-6),
161.9 (C-7), 108.0 (C-8), 160.0 (C-9),
106.3 (C-10), 138.7 (C-1′ ), 126.1 (C-
2′), 128.8 (C-3′), 126.1 (C-6′), 128.8 (C-
5′), 128.6 (C-4′), 117.1 (C-1″), 141.5
(C-2″), 71.5 (C-3″), 55.8 (5-OMe), 56.2
(7-OMe). The structure of 6 was iden-
tified as 5,7-dimethoxy-8-(3-hydroxy-
3-methylbut-1Z-enyl)flavan-one(Fig.1).
Cytotoxicity of compounds
Compounds (1-6) were evaluated
for the cytotoxicity against ovarian
cells from Sprodenia litura (SL cells),
with rotenone as positive standard.
The cytotoxicities were evaluated by
the conventional MTT method [21]. The
results listed in Table 2 showed that
the compound 5 exhibited significant
cytotoxicity with the IC50 value of 4.4
(mg/L) in our 2-day MTT test while
compound 1, whose cytotoxicity ex-
ceeded rotenone, also showed mod-
erate activity in the tested compounds.
As shown in Table 2, compounds 1-4
against SL cells were arranged in the
following order from high to low: 1>3>
rotenone >2>4 with the IC50 values of
27.8, 78.8, 108.0, 134.1, 189.7 (mg/L),
respectively.
Conclusion and Discussion
In this study, four known 6-
methoxykaempferol chemotype com-
pounds (1-4 ) together with two preny-
lated flavonoids (5-6 ) were isolated
and identified from T. purpurea with
considerable content. Except com-
pound 5, the others were isolated from
T. purpurea for the first time.
The chemotype of 6-methoxy-
kaempferol and its derivatives were re-
ported to have medical value including
anti-inflammatory, analgesic property
and relaxant activity [ 22-24 ] , while little
was reported on the insecticidal activi-
ty. In the biological activity test, the
aglycon (1 ) showed the best biological
activity among the four, while glycosy-
lation at either 3-OH or 7-OH position,
as shown by 3 and 2, significantly re-
duced the cytotoxicity. The bisdesmo-
side 4, with sugar residues at 3-OH
and 7-OH, was found to be inactive in
the investigation. The activity profile of
compounds 1 - 4 suggested that the
presence of the sugar groups at 3-OH
and 7-OH in 6-methoxykaempferol
might affect the cytotoxicity against SL
cells.
Rotenoid was usually known as
the main insecticidal components of
Tephrosia. However, various com-
pounds contributed to bioefficacy
such as insecticidal, ovicidal, repel-
lent, and anti-feeding activities against
various insect species jointly or inde-
pendently [25]. According to this study,
compounds 1, 3 and 5, whose cyto-
toxicity exceeded rotenone, deserve
to be further researched.
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common grounding system (>10 kV,
1.2/50 s).
M-structure As for electric appli-
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