全 文 ： 42 Chin J Nat Med Jan. 2011 Vol. 9 No. 1 2011 年 1 月 第 9 卷 第 1 期

Chinese Journal of Natural Medicines 2011, 9(1): 0042−0045
doi: 10.3724/SP.J.1009.2011.00042
Chinese
Journal of
Natural
Medicines







Anthraquinone Glycosides from the Roots of
Prismatomeris connata
HAO Jing1, 3, FENG Shi-Xiu1, 3, QIU Sheng-Xiang (Samuel)1, CHEN Tao2*
1Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of
Sciences, Guangzhou 510650, China; 2Shenzhen Fairy Lake Botanical Garden, Chinese Academy of Sciences, Shenzhen 518004, China;
3Graduate School of Chinese Academy of Sciences, Beijing 100049, China
Available online 20 Jan. 2011
[ABSTRACT] AIM: To study the anthraquinone glycosides and their cytotoxicity from the roots of Prismatomeris connata.
METHODS: The chemical constituents were isolated by Diaion HP-20 resin, silica gel, Sephadex LH-20 and ODS column chroma-
tography and the structures were elucidated on the basis of spectral analysis. The cytotoxic activity was evaluated by MTT method.
RESULTS: Six known anthraquinone glycosides were obtained and their structures were identified as 1-O-methylrubiadin
3-O-β-primeveroside (1), damnacanthol 3-O-β-primeveroside (2), rubiadin 3-O-β-primerveroside (3), lucidin 3-O-β-primeveroside (4),
1, 3-dihydroxy-2-(methoxymethyl)anthraquinone 3-O-β-primerveroside (5), and digiferruginol ω-gentiobiose (6). CONCLUSION:
All the compounds were isolated from this genus for the first time.
[KEY WORDS] Prismatomeris connata; Anthraquinone glycosides; Cytotoxic activity
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2011)01-0042-04

1 Introduction
The genus Prismatomeris (Rubiaceae) consists of about
16 species, being distributed in the tropical and subtropical
areas in Southeast Asia with only 1 species and 2 subspecies
being native to China [1]. P. tetrandra ssp. multiflora is dis-
tributed in Yunnan, Southwest China. Plants from Guangxi,
Guangdong and Fujian are named as P. connata. P. connata
ssp. hainanense, which was treated as P. tetrandra, repre-
senting plants distributed in Hainan [2]. Its roots, called
“Huang-gen”, are a folk Chinese medicine used for the
treatment of hepatitis and pneumoconiosis. Several medica-
ments prepared from them (like Huanggen Tablets) have been
used in clinic in China. Pharmacological studies showed that
the roots possess antibacterial, anti-inflammatory, and anti-
tumor activities [3-4]. Previous chemical investigation on the
roots reported the isolation of anthraquinones [5-6], however,
there was no chemical and pharmacological investigation

[Received on] 04-Aug-2010
[Research Funding] This project was supported by the Shenzhen
Urban Management Bureau Grant (Chen Tao 2006).
[*Corresponding author] CHEN Tao: Prof., Tel: 86-755-25825277,
Fax: 86-755-25825277, E-mail: taochen.mobg@gmail.com
These authors have no any conflict of interest to declare.
reported on anthraquinone glycosides from this species. In
continuation of our phytochemical studies on South China
medicinal plants, the n-BuOH extract from the roots of this
plant was investigated. Herein, we report the isolation, char-
acterization, and cytotoxicity evaluation of six known an-
thraquinone glycosides.
Small pieces of the air-dried roots of P. connata were
extracted with 95% EtOH. The extract was suspended in H2O
and partitioned with petroleum ether, EtOAc, and n-BuOH
successively. The n-BuOH-soluble fraction was separated by
repeated column chromatography over Diaion HP-20, silica
gel, ODS, and Sephadex LH-20. As a result, six known an-
thraquinone glycosides, 1-O-methylrubiadin 3-O-β-prime-
veroside (1), damnacanthol 3-O-β-primeveroside (2), ru-
biadin 3-O-β-primerveroside (3), lucidin 3-O-β-primevero-
side (4), 1,3-dihydroxy-2-(methoxymethyl)anthraquinone
3-O-β-primerveroside (5) and digiferruginol ω-gentiobiose (6)
were isolated and their structures were determined by spec-
troscopic analysis and by comparison of their physiochemical
and spectroscopic data with those reported. All these com-
pounds were reported from the genus for the first time.
In evaluation for cytotoxicity against human lung cancer
(A549) and human hepatoma (HepG2) cell lines, the six anth-
raquinone glycosides (1−6) were found to be almost inac-
HAO Jing, et al. /Chinese Journal of Natural Medicines 2011, 9(1): 42−45
2011 年 1 月 第 9 卷 第 1 期 Chin J Nat Med Jan. 2011 Vol. 9 No. 1 43



R1 R2 R3
1 CH3 H O-prim
2 CH3 OH O-prim
3 H H O-prim
4 H OH O-prim
4a H OH OH
5 H OCH3 O-prim
6 CH3 O-gent H
Fig. 1 Structures of compounds 1-6
tive (IC50 > 100 µmol·L−1) while the aglycone (4a) of 4 ex-
hibited potent inhibitory effect on both A549 and HepG2 cell
lines with IC50 of 6.72 and 9.38 µmol·L−1, respectively. This
result suggested that glycosylation might greatly weaken the
cytotoxic activity of anthraquinones.
2 Apparatus and Reagents
The 1H NMR (400 MHz) and 13C NMR (100 MHz)
spectra were recorded on a Bruker DRX-400 instrument us-
ing TMS as an internal standard. ESI-MS were collected on
MDS SCIEX API 2000 LC/GC/MS instrument. Automatic
fast column was performed with DR Flash II (Shanghai Li
Sui Chemical Engineering Co., Ltd., Shanghai, China). For
column chromatography, silica gel 60 (75-150 µm) produced
by Qingdao Marine Chemical Ltd. (Qingdao, China), De-
velosil ODS (75 µm, Nomura Chemical Co. Ltd., Japan), and
Sephadex LH-20 were used. All solvents were purchased
from the Guangzhou Chemical Reagent Factory (Guangzhou,
China).
3 Plant Material
The roots of P. connata were collected from Nanning,
Guangxi Province, China, in July 2008 and identified by Prof.
Chen Tao (Shenzhen Fairy Lake Botanical Garden). An au-
thenticated voucher specimen (No. 320959) was deposited at
the herbarium of South China Botanical Garden, Chinese
Academy of Sciences, Guangzhou, China.
4 Extraction and Isolation
The air-dried roots of P. connata (20 kg) were smashed
into small pieces and extracted three times with 95% EtOH at
room temperature. The combined EtOH solution was concen-
trated under vacuum to obtain a crude residue (380 g), which
was suspended in water and partitioned with petroleum ether
(60-90 °C), EtOAc, and n-BuOH successively.
The n-BuOH-soluble portion (150 g) was chromato-
graphed on a Diaion HP-20 macroporous resin column and
eluted successively with H2O and 95% EtOH. The 95%
EtOH eluant was collected and concentrated under vacuum to
afford a residue (90 g) which was then subjected to a silica
gel column and eluted with CHCl3-MeOH (9:1 to 6:4) to give
seven fractions (Frs. 1-7). Fr. 3 (8.0 g) was separated by silica
gel column chromatography using CHCl3-MeOH (9:1 to 8:2)
to afford five subfractions (Fr.3a−e). Fr. 3c was recrystallized
in CHCl3-MeOH (85:15) to give 1 (58 mg). Fr. 4 (12 g) was
subjected to a silica gel column and eluted with CHCl3-
MeOH (85:15 to 80:20) to afford six subfractions (Fr. 4a−f).
Fr. 4b and 4c were separately purified by ODS column chro-
matography using 60% and 65% MeOH to give 2 (35 mg)
and 3 (20 mg), respectively. Sub Fr. 4c was further applied to
a Sephadex LH-20 column using CHCl3-MeOH (1:3) fol-
lowed by purification with ODS column chromatography
using 55% and 60% MeOH to yield 4 (35 mg) and 5 (15 mg),
respectively. Fr. 5 (6 g) was subjected to silica gel column
chromatography eluting with CHCl3-MeOH-H2O (100:18:2)
to afford six subfractions (Fr. 5a-f). Fr. 5e was purified by
ODS column chromatography eluting with 55% MeOH to
yield 6 (12 mg).
5 Structural Identification
Compound 1 Yellow powder, C27H30O13, ESI-MS m/z
563.3 [M + H] +, 585.5 [M + Na] +, 1147.4 [2M + Na] +,
561.5 [M − H] −, 597.2 [M + Cl]−, 266.9 [M − glc − xyl − H]−;
1H NMR and 13C NMR (400 MHz and 100 MHz, DMSO-d6):
see Tables 1 and 2. It was identified as 1-O- methylrubiadin
3-O-β-primeveroside by comparison of the physical and
spectral data with the literature [8].
Compound 2 Yellow powder, C27H30O14, ESI-MS m/z
601.4 [M + Na] +, 617.4 [M + K] +, 1179.1 [2M + Na] +, 447
[M − xyl + H] +, 285.1 [M − glc − xyl + H] +, 613.0 [M +
Cl]−, 283.1 [M − glc − xyl − H]−; 1H NMR and 13C NMR
(400 MHz and 100 MHz, DMSO-d6): see Tables 1 and 2. It
was identified as damnacanthol 3-O-β-primeveroside by
comparison of the physical and spectral data with the litera-
ture [9].
Compound 3 Yellow powder, C26H28O13, ESI-MS m/z
571.1 [M + Na] +, 587.2 [M + K] +, 1119.2 [2M + Na] +,
1135.1 [2M + K] +, 547.2 [M − H]−, 583.5 [M + Cl]−, 253.1
[M − glc − xyl − H]−; 1H NMR and 13C NMR (400 MHz and
100 MHz, DMSO-d6): see Tables 1 and 2. It was
identified as rubiadin 3-O-β-primerveroside by comparison of
the physical and spectral data with the literature [8].
Compound 4 Yellow powder, C26H28O14, ESI-MS m/z
587.2 [M + Na] +, 603.2 [M + K] +, 1151.2 [2M + Na] +,
563.4 [M − H] −, 599.2 [M + Cl]−, 269.2 [M − glc − xyl −
H]−; 1H NMR and 13C NMR (400 MHz and 100 MHz,
DMSO-d6): see Tables 1 and 2. It was identified as lucidin
3-O-β-primeveroside by comparison of the physical and
spectral data with the literature [8].
HAO Jing, et al. /Chinese Journal of Natural Medicines 2011, 9(1): 42−45
44 Chin J Nat Med Jan. 2011 Vol. 9 No. 1 2011 年 1 月 第 9 卷 第 1 期

Table 1 1H NMR spectral data of compounds 1−6
δH (J = Hz) Position
1 2 3 4 5 6
1-OH 13.0 13.06 13.01
3 8.10 d (8.0)
4 7.70 s 7.74 s 7.47 s 7.48 s 7.49 s 8.05 d (8.0)
5 8.15 dd (8.0, 2.0) 8.15 dd (8.0, 2.0) 8.24 dd (8.0, 2.0) 8.24 dd (8.0, 2.0) 8.24 dd (8.0, 2.0) 8.17 dd (8.0, 2.0)
6 7.92 m 7.92 m 8.18 m 8.19 m 7.95 m 7.93 m
7 7.92 m 7.92 m 8.18 m 8.19 m 7.95 m 7.93 m
8 8.17 dd (8.0, 2.0) 8.16 dd (8.0, 2.0) 8.25 dd (8.0, 2.0) 8.25 dd (8.0, 2.0) 8.26 dd (8.0, 2.0) 8.19 dd (8.0, 2.0)
2-CH3 2.26 s 2.16 s
2-CH2OH
4.60 m
4.66 m
4.61 m
4.67 m
4.81 d (13.8)
5.00 d (13.8)
2-CH2OCH3
4.69 d (10.0)
4.62 d (10.0)
2-CH2OCH3 3.43 s
1-OCH3 3.81 s 3.88 s 3.81 s
1 5.14 d (7.6) 5.12 d (7.2) 5.13 d (7.6) 5.14 d (7.6) 5.14 d (7.6) 4.33 d (7.6)
2 -5 2.99-3.60 m 2.98-3.68 m 2.97-3.64 m 2.99-3.69 m 2.98-3.71 m 2.99-3.63 m
6 3.95 d (9.2) 3.67 m
3.96 d (9.2)
3.68 m
3.92 d (9.2)
3.64 m
3.94 d (9.2)
3.68 m
3.95 d (9.2)
3.68 m
4.05 d (9.0)
3.69 m
1 4.14 d (7.2) 4.16 d (7.2) 4.14 d (7.2) 4.15 d (7.2) 4.15 d (7.2) 4.31 d (7.8)
2 -5(6) 2.99-3.60 m 2.98-3.68 m 2.97-3.64 m 2.99-3.69 m 2.98-3.71 m 2.99-3.63 m

Table 2 13C NMR spectral data of compounds 1−6
δC Position
1 2 3 4 5 6
1 160.2 160.7 161.8 162.4 162.0 157.9
2 129.2 131.6 121.2 123.9 123.7 140.4
3 160.3 160.6 161.6 162.3 161.9 134.3
4 108.6 109.1 106.3 106.8 106.4 122.7
5 126.5 126.3 126.9 127.0 126.7 126.2
6 134.8 133.6 135.1 135.2 134.9 133.9
7 135.0 134.7 135.2 135.3 134.7 134.6
8 127.0 126.7 127.3 127.3 127.0 126.8
9 182.7 180.0 187.6 187.6 187.2 181.6
10 181.1 182.1 182.0 182.0 181.5 182.4
11 134.2 132.1 133.5 133.4 132.8 134.4
12 134.4 134.4 133.2 133.2 133.0 132.2
13 120.3 120.5 111.4 111.8 111.4 125.0
14 132.5 135.7 132.4 134.2 133.8 134.4
2-CH3 9.5 8.9
2-CH2OCH3 51.9 51.4 50.9 64.6
2-CH2OCH3 69.1
1-OCH3 61.3 62.7 61.9
1 100.7 101.0 100.7 101.1 100.8 102.4
2 73.7 73.3 73.7 73.7 73.3 73.6
3 76.1 75.9 76.5 76.2 76.4 76.7
4 69.6 69.2 69.5 69.5 69.5 70.0
5 76.5 76.4 76.1 76.9 75.8 76.0
6 68.4 68.0 68.4 68.4 68.0 68.5
1 104.5 104.1 104.5 104.5 104.1 103.4
2 73.6 73.3 73.6 73.7 73.4 73.5
3 76.8 75.7 76.8 76.1 76.5 76.9
4 69.9 69.5 69.9 69.9 69.6 70.0
5 66.0 65.6 66.0 66.1 65.6 76.6
6 61.0

HAO Jing, et al. /Chinese Journal of Natural Medicines 2011, 9(1): 42−45
2011 年 1 月 第 9 卷 第 1 期 Chin J Nat Med Jan. 2011 Vol. 9 No. 1 45

Compound 5 yellow powder, C27H30O14, ESI-MS m/z
601.5 [M + Na] +, 617.5 [M + K] +, 447.0 [M − xyl + H] +,
577.6 [M − H] −; 1H NMR and 13C NMR (400 MHz and 100
MHz, DMSO-d6): see Tables 1 and 2. It was identified as 1,
3-dihydroxy-2-(methoxymethyl)anthraquinone 3-O-β-primer-
veroside by comparison of the physical and spectral data with
the literature[8,10].
Compound 6 yellow powder, C28H32O14, ESI-MS m/z
615.2 [M + Na] +, 631.2 [M + K] +, 1027.3 [2M + Na] +,
431.0 [M − glc + H] +, 269.1 [M − glc − glc + H]+,591.1 [M
− H] −, 627.4 [M + Cl] −, 267.2 [M − glc − glc − H] −; 1H
NMR and 13C NMR (400 MHz and 100 MHz, DMSO-d6):
see Tables 1 and 2. It was identified as rubiadin 3-O-β-
primerveroside by comparison of the physical and spectral
data with the literature[11].
Hydrolysis of compound 4 Compound 4 (20 mg) was
dissolved in 15 mL 2 mol·L−1 aqueous HCl and allowed to
stand at 50 °C for 6 h, and was then added with H2O and
EtOAc (each 30 mL). The EtOAc layer was washed with
H2O and then evaporated to dryness. The residue was sub-
jected to silica gel column chromatography using
CHCl3-MeOH-H2O (100:5:0.5) to afford compound lucidin
(4a) (5.0 mg)[12].
Lucidin (4a): yellow powder, C15H10O5, ESI-MS m/z
269.1 [M − H] −; 1H NMR (CDCl3, 400 MHz): δ 8.24 (1H, d,
J = 8.0 Hz, H-8), 8.21 (1H, d, J = 8.0 Hz, H-5), 8.15 (2H, m,
H-6 and H-7), 7.26 (1H, s, H-4), 4.54 (2H, s, CH2-2); 13C
NMR (DMSO-d6, 100 MHz): δ 162.2 (C-1), 126.7 (C-2),
162.3 (C-3), 109.6 (C-4), 126.6 (C-5), 133.9 (C-6), 134.3
(C-7), 127.2 (C-8), 180.8 (C-9), 183.2 (C-10), 132.7 (C-11),
135.0 (C-12), 118.4 (C-13), 135.1 (C-14), 51.3 (CH2-2).
6 Cytotoxicity assay
Cytotoxicity was determined by MTT method[7] using
human lung cancer (A549) and human hepatoma (HepG2)
cells grown in RPMI-1640 medium plus 10% heat-inac-
tivated fetal bovine serum. Briefly, exponentially growing
cells were seeded into a 96-well plate at a density of 1.0 × 104
cells/well. After 24 h incubation, the cells were treated in
triplicate with or without various concentrations of test sam-
ples. After 48 h of incubation at 37 °C, 5 % CO2, MTT re-
agent (5 mg·mL−1) was added to each well for 4 h. And the
resulting crystals were dissolved in DMSO (150 µL) and then
shaken for another 15 min. The absorbance was recorded on a
microplate reader (TECAN Genios) at a wavelength of 570
nm. The inhibition percentages were calculated from reduc-
tion of absorbance in the control assay which was treated
with 1% DMSO alone.
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黄根中的蒽醌苷成分
郝 静 1, 3, 冯世秀 1, 3, 邱声祥 1, 陈 涛 2*
1中国科学院华南植物园, 广州 510650; 2深圳市中国科学院仙湖植物园, 深圳 518004;
3中国科学院研究生院，北京 10049
【摘 要】 目的：对黄根中的蒽醌苷类化合物及其细胞毒活性进行研究。方法：采用大孔吸附树脂柱、硅胶柱、凝胶柱及
反相柱色谱法分离单体化合物, 根据波谱技术鉴定结构, 采用 MTT 法进行细胞毒活性测定。结果：从黄根乙醇提取物的正丁醇
部分离得到 6 个蒽醌苷类化合物, 1-O-methylrubiadin 3-O-β-primeveroside (1), damnacanthol 3-O-β-primeveroside (2), rubiadin
3-O-β-primerveroside (3), lucidin 3-O-β-primeveroside (4), 1, 3-dihydroxy-2-(methoxymethyl) anthraquinone 3-O-β-primerveroside (5)
and digiferruginol ω-gentiobiose (6)。结论：化合物 1−6 均为首次从该植物中分离得到, 蒽醌成苷后生物活性减弱。
【关键词】 黄根; 蒽醌苷; 细胞毒活性

【基金项目】 深圳市城市管理局科技项目(陈涛 2006)资助