Chinese

Journal of
Natural
Chinese Journal of Natural Medicines 2012, 10(2): 0115−0118 Medicines
doi: 10.3724/SP.J.1009.2012.00115

Chemical constituents from the water-soluble fraction

of wild Sargentodoxa cuneata
TANG Jian1, 2, MA Rui-Li1, OUYANG Zhen1*, CHEN Hai-Sheng2
1
School of Pharmacy, Jiangsu University, Zhenjiang 212013, China;
2
School of Pharmacy, Second Military Medical University, Shanghai 200433, China
Available online 20 Mar. 2012

[ABSTRACT] AIM: To study the chemical constituents from the water-soluble fraction of wild Sargentodoxa cuneata. METHODS:
The compounds were isolated by silica gel and HW-40C column chromatography. Their structures were elucidated on the basis of
spectral analysis of ESI-MS and NMR. RESULTS: Eight known glycosides were isolated and identified as (−)-lyoniresinol-9-O-β-D-
glucoside (1), (+)-lyoniresinol-9-O-β-D-glucoside (2), (−)-lyoniresinol-9′-O-β-D-glucoside (3), (+)-syringaresinol-4-O-β-D-glucoside
(4), androsin (5), (+)-syringin (6), sargentol (7) and daucosterol (8). CONCLUSION: Compounds 1, 3 and 6 were isolated from this
plant for the first time, and constituents of the water-soluble parts of wild S. cuneata were studied systematically for the first time.
[KEY WORDS] Sargentodoxa cuneata; (−)-Lyoniresinol-9-O-β-D-glucoside; (+)-Syringin; Sargentol
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2012)02-0115-04

1 Introduction
Sargentodoxa cuneata (Sargentodoxaceae, previously at-
tributed to Lardizabalaceae), a plant endemic to China, is
used as a traditional Chinese medicine or ethnic medicine
(called as Hongteng or Xueteng) possessing exact effects for
the treatment of rheumatic arthritis, acute appendicitis,
amenorrhea and painful menstruation [1]. The pharmacologi-
cal activities of crude extracts or isolates from this plant in-
cluded anti-inflammatory, antioxidant antimicrobial, antiviral,
anti-hyposia, vasodilating and myocardial ischemia inhibitory
effects[2-3]. Recent investigations about this herbal meidicine
concerned to immunosuppressive effects of water decoction,
which could be the basis of anti-inflammatory and curing
rheumatic arthritis [4-5]. Previous phytochemical studies on
this plant described several kinds of constituents from the
acetyl acetate fraction, including anthraquinones, triterpe-
[Received on] 03-May-2011
[Research funding] This project was supported by the National
Natural Science Foundation of China (No. 81072985 ) and the Ji-
angsu University Starting Foundation (No. 10JDG050).
*
[ Corresponding author] OUYANG Zhen: Prof., Tel: 86-511-
88791564; Fax: 86-511-88791564, E-mail: zhenouyang@ujs.edu.cn
These authors have no any conflict of interest to declare.
Copyright © 2012, China Pharmaceutical University.
Published by Elsevier B.V. All rights reserved.
noids, lignans, flavonoids and organic acids [2, 6-8], and phe-
nolic glycosides and lignan glycosides from the n-BuOH and
water-soluble fractions [6-9].
The herbs in previous investigation mostly were pur-
chased from the herbal stores, and in our pharmacological
studies, the water-soluble fraction of wild S. cuneata showed
higher inhibiting effect than that of purchased materials. So,
this paper described the investigation of chemical composi-
tions from water-soluble fraction of wild S. cuneata to ex-
plain the material basis for the excellent anti-inflammatory
and immunosuppressive activity.
The water-soluble fraction from 80% alcohol extraction
of the stem of wild S. cuneata was subjected to resin, silical
gel and HW-40C column chromatography to give eight gly-
cosides: (−)-lyoniresinol-9-O-β-D-glucoside (1), (+)-lyoni-
resinol-9-O-β-D-glucoside (2), (−)-lyoniresinol-9′-O-β-D-glu-
coside (3), (+)-syringaresinol-4-O-β-D-glucoside (4), aceto-
vanillone-4-O-β-D-glucoside (androsin, 5), (+)-syringin (6),
sargentol (7) and daucosterol (8).
There were few reports about aryltetrahydronaphthalene
lignan glycosides and cinnamic alcohol glycosides from this
plant [7], while we reported the isolation of these two classes
of compounds 1, 3 and 6 from this plant for the first time.
2 Apparatus and Regents
NMR spectra were determined on Bruker Avance II 600
NMR, for 1H NMR at 600 MHz and 13C NMR at 150 MHz.
ESI-MS spectra were recorded on Varian MAT-212 mass
 TANG Jian, et al. /Chinese Journal of Natural Medicines 2012, 10(2): 115−118
spectrometer. TLC analysis was run on HSGF254 silica gel
plates (10−40 μm, Yantai, China). Column chromatography
was performed on Diaion HP-20 macroporous resin (Mitsu-
bishi Chemical Industries, Ltd.), silica gel (200−300 mesh,
Yantai, China), and Toyopearl HW-40C (Tosoh).
3 Plant materials
The plant was collected in Qin Mountain, Shaanxi Prov-
ince, China in September 2009, and authenticated as Sargen-
todoxa cuneata Rehd. Et Wils by Prof. OUYANG Zhen. A
voucher specimen has been deposited in the Herbarium of
Pharmacognosy Laboratory (DXT0909).
4 Extraction and Isolation
The air-dried and powdered stems (6.0 kg) were perco-
lated with 80% alcohol for 24 × 2 h. The extract was concen-
trated to an aqueous residue, and then extracted with EtOAc
to give two portions. The water-soluble portion (12 L) was
evaporated under reduced pressure to remove the residual
EtOAc, filtrated. The filtration (10 L) was subjected to
Diaion HP-20 resin column (2.5 L, 80 mm × 500 mm),
eluting with water and the gradient aqueous alcohol
Fig. 1 Structures of compounds 1-3 and 7
5 Results and Discussion
Compound 1 White yellow powders. ESI-MS m/z
581.3 [M - H]−, 419.2 [581.3 − glc]−; According to the 1H-
and 13C NMR spectral data (Table 1), it was determined as
(−)-lyoniresinol-9-O-β-D-glucopyranoside by comparison
with the literature [10].
Compound 2 Yellow powders. ESI-MS m/z 581.3 [M
− H]−, 419.2 [581.3 − glc]−; According to the 1H- and 13C
NMR spectral data (Table 1), it was determined as (+)- lyon-
iresinol-9-O-β-D-glucopyranoside by comparison with the
literature [10].
Compound 3 Yellow powders. ESI-MS m/z 581.3 [M
− H]−, 419.2 [581.3 − glc]−; According to the 1H- and 13C
NMR spectral data (Table 1), it was determined as (−)- lyon-
iresinol-9′-O-β-D-glucopyranoside by comparison with the
literature [10].
Compound 4 Yellow powders. ESI-MS m/z 579.2 [M
− H]−, 417.2 [579.2 − glc]−; 1H NMR (600 MHz, DMSO-d6)
δ: 8.27 (1H, s, 4′-OH), 6.66 (2H, s, H-2″, 6″), 6.60 (2H, s,
H-2′, 6′), 4.67 (1H, d, J= 4.8 Hz, H-6), 4.62 (1H, d, J = 4.2
(15%–50%– 95%). The fraction eluted with 50% alcohol was
evaporated to give about 90 g of residue. The partial residue
(70 g) was subjected to silica gel column chromatography
(100–200 mesh, 1.0 Kg) using the gradient CH2Cl2 : CH3OH
(10 : 1–5 : 1–1 : 1–0 : 1, 1% H2O, V/V) as eluent to obtain
five fractions:Ⅰ(7 g), ⅡA (11g), ⅡB (6 g), Ⅲ (12 g), Ⅳ (9 g).
Fraction ⅡA (11 g) was subjected to silica gel column
chromatography, eluting with CH2Cl2 : CH3OH (10 : 1–5 :
1–2 : 1, 1% H2O, V/V) to afford six subfractions (subfr. 2a-f).
Subfr. 2d (550 mg) was purified repeatedly over HW-40C
column chromatography using 85% MeOH to yield 1 (11 mg)
and 2 (7 mg). Subfr. 2c (1.2 g) was separated by silica gel
column chromatography and further purified by Toyopearl
HW-40C to give 3 (11 mg) and 4 (74 mg). Fraction ⅡB (6 g)
was separated by silica gel column chromatography with
CH2Cl2 : CH3OH (10 : 1–5 : 1, 2% H2O, V/V) solvent system
to four subfractions (subfr. 2g-j). Subfr. 2i (1.2 g) was puri-
fied repeatedly over silica gel and HW-40C column chroma-
tography to yield 5 (30 mg), 6 (6 mg) and 7 (196 mg). The
partial fraction Ⅰ (0.9 g) was purified over silica gel column
chromatography, eluting with CH2Cl2 : CH3OH (10 : 1), to
obtain 8 (26 mg).
Hz, H-2), 4.17-4.19 (2H, m, H-4a, 8a), 3.79-3.82 (2H, m,
H-4b, 8b), 3.17-3.20 (2H, m, H-1, 5), 3.71 (6H, br s, 2 ×
OCH3), 3.70 (6H, br s, 2 × OCH3), 4.88 (1H, d, J = 7.2 Hz,
H-1″′), 3.59 (1H, dm, H-6″′a), 3.40 (1H, dm, H-6″′b),
3.03-3.15 (4H, m, H-2″′, 3″′, 4″′, 5″′); 13C NMR (150 MHz,
DMSO-d6) δ: 53.5 (C-1), 85.2 (C-2), 71.2 (C-4), 53.6 (C-5),
85.0 (C-6), 71.1 (C-8), 131.3 (C-1′), 103.6 (C-2′, 6′), 147.8
(C-3′, 5′), 137.1 (C-4′), 133.6 (C-1″), 104.1 (C-2″, 6″), 152.5
(C-3″, 5″), 134.8 (C-4″), 55.9 (3′, 5′-OCH3), 56.3 (3″,
5″-OCH3), 102.6 (C-1″′), 74.1 (C-2″′), 76.4 (C-3″′), 69.8
(C-4″′), 77.1 (C-5″′), 60.8 (C-6″′). It was determined as
(+)-syringaresinol-4-O- β-D-glucopyranoside by comparison
with the literature [11].
Compound 5 White powders. ESI-MS m/z 327.1 [M −
H]−; 1H NMR (600 MHz, DMSO-d6) δ: 7.59 (1H, dd, J = 8.4,
2.4 Hz, H-6), 7.48 (1H, d, J=1.8 Hz, H-2), 7.19 (1H, d, J=8.4
Hz, H-5), 3.74 (3H, br s, 3-OCH3), 2.55 (3H, br s, COCH3),
4.78 (1H, d, J=7.8 Hz, H-1′), 3.67 (1H, dm, H-6′a), 3.46 (1H,
dd, J=12.0, 6.0 Hz, H-6′b), 3.18-3.30 (4H, m, H-2′, 3′, 4′, 5′);
13
C NMR (150 MHz, DMSO-d6) δ: 130.6 (C-1), 110.8 (C-2),
148.5 (C-3), 152.9 (C-4), 114.0 (C-5), 122.5 (C-6), 196.3
(COCH3), 26.2 (COCH3), 55.5 (3-OCH3), 99.2 (C-1′), 73.1
 TANG Jian, et al. /Chinese Journal of Natural Medicines 2012, 10(2): 115−118

1
Table 1 H NMR and 13C NMR spectra for compounds 1-3 (DMSO-d6, 600 MHz and 150 MHz)
1 2 3
Position
δH (J, Hz) δC δH (J, Hz) δC δH (J, Hz) δC
1 133.4 133.5 133.8
2 6.35 s 106.1 6.36 s 106.2 6.39 s 106.2
3 147.6 147.7 148.1
4 137.7 137.7 138.8
5 147.6 147.7 148.1
6 6.30 s 106.1 6.29 s 106.2 6.39 s 106.2
7 4.30 d (5.4) 40.7 4.21 d (5.4) 40.3 4.31 d (5.4) 41.9
8 2.04 m 43.5 1.48 m 43.2 1.90 m 49.0
3.41-3.45 m; 3.43-3.48 m;
9 70.2 69.2 3.52-3.61 dm 62.4
3.67 m 3.68 m
1′ 128.6 128.8 129.4
2′ 6.56 s 106.8 6.55 s 106.6 6.58 s 106.9
3′ 147.0 147.3 148.0
4′ 137.4 137.4 138.1
5′ 146.6 146.5 147.0
6′ 125.0 124.9 125.6
2.50 m; 2.48 m; 2.58 dd (15.6, 12.0);
7′ 32.5 32.4 33.2
2.64 dd (15.0, 4.2) 2.60 dd (15.0, 4.2) 2.79 dd (15.0, 4.2)
8′ 1.98 m 39.5 1.48 m 39.0 1.85 m 37.3
3.25-3.30 m; 3.23-3.27 m; 3.52-3.55 m;
9′ 64.1 64.2 74.0
3.42-3.45 m 3.48-3.52 m 3.93 dd (9.6, 4.8)
3, 5-OCH3 3.65 br s 56.1 3.65 br s 56.0 3.74 br s 56.0
3′-OCH3 3.78 br s 55.8 3.77 br s 55.8 3.86 br s 55.7
5′-OCH3 3.29 br s 59.1 3.30 br s 59.0 3.33 br s 59.1
1″ 4.17 d (7.8) 103.2 4.09 d (7.8) 103.5 4.25 d (7.8) 103.8
2″ 3.25-3.30 (2H, m) 74.3
73.7 73.6
3″ 77.0 76.9 77.1
3.05-3.18 m 3.01-3.14 m
4″ 70.1 70.0 3.18 t (8.4) 70.8
77.1 77.1
5″ 3.35 m 77.2
6″ 3.44 m; 3.46 m; 3.66 dd (11.4, 5.4);
61.2 61.1 61.9
3.70 m 3.72 dd (11.4, 2.4) 3.84 dd (11.4, 2.4)

(C-2′), 76.6 (C-3′), 69.4 (C-4′), 76.9 (C-5′), 60.4 (C-6′). It
was determined as acetovanillone-4-O-β-D-glucoside (an-
drosin) by comparison with the literature [12].
Compound 6 White powders. ESI-MS m/z 371.1 [M −
H]−, 209.2 [371.1 − glc]−; 1H NMR (600 MHz, DMSO-d6) δ:
6.75 (2H, br s, H-2, 6), 6.55 (1H, d, J= 16.2 Hz, H-7), 6.33
(1H, dt, J= 15.6, 5.4 Hz, H-8), 4.22 (2H, dd, J= 6.0, 1.8 Hz,
H-9), 3.86 (6H, br s, 3, 5-OCH3), 4.87 (1H, d, J= 7.8, H-1′),
3.78 (1H, dd, J= 12.0, 1.8 Hz, H-6′a), 3.67 (1H, dd, J= 12.0,
5.4 Hz, H-6′b), 3.41-3.49 (3H, m, H-2′, 3′, 4′), 3.21 (1H, ddd,
J=9.6, 5.4, 2.4 Hz, H-5′); 13C NMR (150 MHz, DMSO-d6) δ:
135.3 (C-1), 105.5 (C-2, 6), 154.4 (C-3, 5), 135.9 (C-4),
131.3 (C-7), 130.1 (C-8), 63.6 (C-9), 57.1 (3, 5-OCH3), 105.4
(C-1′), 75.8 (C-2′), 77.9 (C-3′), 71.4 (C-4′), 78.4 (C-5′), 62.6
(C-6′). It was determined as (+)-syringin by comparison with
the literature [13].
Compound 7 White powders. ESI-MS m/z 387.1 [M −
H]−, 389.1 [M + H]+; 1H NMR (600 MHz, DMSO-d6) δ: 6.66
(2H, s, H-2, 6), 4.89 (1H, d, J=7.8 Hz, H-1″), 4.68 (1H, d,
J=3.6 Hz, H-1′), 4.21 (1H, dd, J=9.0, 6.6 Hz, H-3′a), 3.83
(1H, dd, J=9.0, 3.6 Hz, H-3′b), 3.76 (6H, br s, 3, 5-OCH3),
3.60 (1H, dd, J=11.4, 1.8 Hz, H-6″a), 3.41 (1H, dd, J=12.0,
6.0 Hz, H-6″b), 3.21 (1H, m, H-5″), 3.10-3.18 (3H, m, H-3″,
2″, 2′), 3.02 (H, m, H-4″); 13C NMR (150 MHz, DMSO-d6):
133.7 (C-1), 104.2 (C-2, 6), 152.7 (C-3, 5), 137.2 (C-4), 85.0
(C-1′), 53.5 (C-2′), 71.3 (C-3′), 56.4 (OCH3), 102.6 (C-1″),
74.1 (C-2″), 76.4 (C-3″), 69.9 (C-4″), 77.1 (C-5″), 60.9
(C-6″). It was determined as sargentol by comparison with
the literature [8].
Compound 8 was identified as daucosterol by the au-
thentic sample.
These displayed that constituents of wild S. cuneata
were certain consistents with those of cultured herbs [2, 6-9].
6 Anti-inflammatory assay
Compound 7 and the fractions eluted with 50% alcohol
of wild and cultured S. cuneata were evaluated in vivo for
their anti-inflammatory action, using the method of xy-
 TANG Jian, et al. /Chinese Journal of Natural Medicines 2012, 10(2): 115−118

lene-induced ear edema in mice. Briefly, 30 min after irriga-
tion of the samples, 50 μL of xylene was applied to the ante-
rior and posterior surfaces of the right ear of each mouse. 30
min after xylene application, mice were sacrificed and both
ears were removed. The ear edema was measured and the
inhibition percentage was calculated by the following for-
mula: Inhibition percentage = [(Control group mean − Test
group mean)/Control group mean]] × 100%. Compound 7
exhibited remarkable anti-inflammatory action, with inhibi-
tion of 47.2% and 29.1% at the dosage of 100 and 50 mg·kg-1.
Fraction (50% alcohol) of wild S. cuneata showed distinct
inhibiting effect on ear swelling (inhibition rates, 41.5% and
32.1%) at the dosage of 300 and 100 mg/kg, superior to
38.4% and 30.5% of cultured S. cuneata.
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野生大血藤水溶性部位中的糖苷类化合物
汤 建 1, 2, 马瑞丽 1, 欧阳臻 1*, 陈海生 2
1
江苏大学药学院, 镇江 212013;
2
第二军医大学药学院, 上海 200433

【摘 要】 目的：对野生大血藤水溶性部位成分进行分离和鉴定。方法：运用硅胶柱和凝胶柱层析等方法分离纯化化合物,
通过质谱和核磁等波谱手段进行结构鉴定。结果：从大血藤水溶性部位中分得 8 个糖苷类化合物：(−)-南烛木树脂酚-9-O-β-D-
葡萄糖苷(1)，(+)-南烛木树脂酚-9-O-β-D-葡萄糖苷(2)，(−)-南烛木树脂酚-9′-O-β-D-葡萄糖苷(3)，(+)-丁香树脂酚-4-O-β-D-葡萄
糖苷(4)，草夹竹桃苷(5)，(+)-丁香苷(6)，sargentol (7)，胡萝卜苷(8)。结论：化合物 1, 3 和 6 为首次从该植物中分得。并且也
是首次对野生大血藤水溶性部位的化学成分进行系统研究。
【关键词】 大血藤; (−)-南烛木树脂酚-9-O-β-D-葡萄糖苷; (+)-丁香苷; Sargentol

【基金项目】 国家自然科学基金（No. 81072985）和江苏大学高级人才启动基金（No. 10JDG050）资助项目