全 文 ：
201 Journal of Chinese Pharmaceutical Sciences http://www.jcps.ac.cn
Chemical constituents from Murraya tetramera Huang
Ying Zhou, Wenguang Wang, Pengfei Tu, Yong Jiang*
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science
Center, Beijing 100191, China
Abstract: The aim of this study is to perform a phytochemical investigation of the aerial parts of Murraya tetramera Huang.
Nine compounds were isolated by silica gel and Sephadex LH-20 chromatography, together with preparative TLC and HPLC methods.
By analysis of the MS and NMR spectroscopic data and comparison with those in literature, these nine compounds were identified as
3,3-[oxybis(methylene)]bis(9-methoxy-9H-carbazole) (1), murrastifoline B (2), murrayaquinone A (3), 2,4-dimethoxyphenol (4),
1,2-dimethoxy-4-nitrobenzene (5), 3-methylanisole (6), (–)-syringaresinol-4-O-β-D-glucopyranoside (7), β-sitosterol (8), and
octadecanyl-3-methoxy-4-hydroxylbenzeneacrylate (9), respectively. Among them, compounds 4–6 and 9 were described from
the genus Murraya for the first time, and compounds 2–9 were isolated from M. tetramera for the first time.
Keywords: Rutaceae, Murraya tetramera, Chemical constituents
CLC number: R284 Document code: A Article ID: 1003–1057(2016)3–201–05
Received: 2015-10-25, Revised: 2015-11-18, Accepted: 2015-12-15.
Foundation items: National Science Fund for Excellent Young Scholars
(Grant No. 81222051), and National Key Technology R&D Program
“New Drug Innovation” of China (Grant Nos. 2012ZX09301002-002-002
and 2012ZX09304-005).
*Corresponding author. Tel.: 86-10-82802719,
E-mail: yongjiang@bjmu.edu.cn
http://dx.doi.org/10.5246/jcps.2016.03.024
1. Introduction
The genus Murraya consists of about 14 species and
2 varieties all over the world and 9 species and 1 variety
in China, mainly distributed in the west of Guangxi
Province and the southeast of Yunnan Province[1]. This
genus plants grow mainly in the sunny limestone
mountain, and have the efficacies of dispelling wind,
relieving the exterior syndromes, promoting qi flow,
activating blood circulation, removing blood stasis,
anti-inflammation, analgesia, as well as antipyresis[2].
Murraya tetramera Huang is a traditional folk medicine
for the local resident in Guangxi Province, however, its
bioactive ingredients have not been clarified. Previous
reports revealed the existence of alkaloids, flavonoids,
anthraquinones, and coumarins from this plant[3–5]. In order
to further specify the chemical composition of M. tetramera,
a phytochemical investigation was performed, which led to







the isolation and structural identification of 9 compounds
(Fig. 1), including 3,3-[oxybis(methylene)]bis(9-methoxy-
9H-carbazole) (1), murrastifoline B (2), murrayaquinone
A (3), 2,4-dimethoxyphenol (4), 1,2-dimethoxy-4-
nitrobenzene (5), 3-methylanisole (6), (–)-syringaresinol-4-
O-β-D-glucopyranoside (7), β-sitosterol (8), and octadecanyl-
3-methoxy-4-hydroxylbenzeneacrylate (9). Among them,
compounds 4–6 and 9 were obtained from the genus
Murraya for the first time, and compounds 2–9 were
isolated from M. tetramera for the first time.
2. Experimental
2.1. General procedures
NMR data were acquired with a Varian INOVA-500
NMR spectrometer (Varian Inc. Palo Alto, CA), using
TMS as internal standard. EI-MS was obtained on an
AEI-MS-50 mass spectrometer. HPLC separations were
performed on a Waters semi-preparative instrument
equipped with a Waters Nova-pak C18 column (7.8 mm×
300 mm, 5 μm), detected at UV 254 and 285 nm. Silica
gel (Qingdao Haiyang Chemical Factory), Sephadex
LH-20 (Phamacia Company), macroporous D-101 resin
(Bao’en Chemical Co. Ltd, Hebei), and ODS (40–63 µm,

202 Zhou, Y. et al. / J. Chin. Pharm. Sci. 2016, 25 (3), 201–205
Merck ＆ Co., Inc.) were applied for column chroma-
tography. All the other analytical reagents are of
products of Beijing Chemical Factory.
2.2. Plant materials
The aerial parts of M. tetramera were collected in
2011 from Guangxi Province and authenticated by
Prof. Pengfei Tu. A voucher specimen (No. MT-YZ-
201108) is deposited in the Herbarium of Peking
University Modern Research Center for Traditional
Chinese Medicine.
2.3. Extraction and isolation
The leaves and stems of M. tetramera (8.7 kg) were
extracted with 95% aqueous EtOH at 80 °C twice, 3 h
each time. After removal of solvent under reduced
pressure, the residue was suspended in H2O (1.5 L) and
extracted with CHCl3 to yield CHCl3–soluble extract
(88 g) and aqueous-soluble part (14.9 g). The CHCl3
extract (87.5 g) was chromatographed on a silica gel
column chromatography (CC) using a gradient of
petroleum ether (PE)–EtOAc (9:1→8:2→7:3→6:4→
5:5→4:6→0:100, v/v) to give 11 fractions (A–K). Fr. A
was fractionated on a silica gel CC and eluted with
CHCl3–EtOAc (1:0→8:1→7:1→0:1, v/v) to afford
four fractions (Aa–Ad). Fr. Aa and Fr. Ac were purified
by preparative TLC to yield compounds 1 (2.3 mg) and 3
(1.1 mg), respectively. Fr. Ab was purified by a silica
gel CC (eluted with CH2Cl2) and Sephadex LH-20
(eluted with PE–CHCl3–MeOH, 5:5:1, v/v/v), and then
purified by preparative TLC, using PE–acetone (8:2,
v/v) as eluent to yield compound 6 (2.4 mg). Fr. Ad
was separated by a silica gel CC (200–300 mesh)
and eluted with n-hexane–acetone (40:1→20:1→
10:1→1:1, v/v) and was purified by preparative TLC
(developed by CH2Cl2) to yield compound 9 (3.2 mg).
Fr. C was crystallized to afford compound 8 (39.0 mg)
and the residue was separated by Sephadex LH-20
and eluted with PE–CHCl3–MeOH (5:5:1, v/v/v) to
afford 3 subfractions (Fc-1–Fc-3). Subfractions Fc-1
and Fc-2 were purified by preparative TLC using
CHCl3 as eluent to yield compounds 4 (1.9 mg) and
5 (2.6 mg), respectively. Subfraction Fc-3 was purified
by preparative TLC, using CHCl3 as eluent to yield
compound 2 (2.2 mg).
The aqueous-soluble fraction (14.0 g) was subjected
to a D101 macroporous resin CC and eluted with
water, 40% aqueous EtOH, 60% aqueous EtOH, and
95% aqueous EtOH, successively. The resulting eluents
were concentrated in vacuo to yield 4 residues. The
40% eluent (4 g) was chromatographed on a silica gel
OH
OCH3H3CO
OCH3
OCH3O2N CH3H3CO
O
O
OMe
MeO
OMe
OMe7
8
2
1
2
1
4
24
1
3
O
OH
O
OH
OH
HO
OH
H
H
HO
H
1
4
7
8
9
N
HN
H3CO
CH3
OCH3
1
36
4a4b
8a 8b
N
H
O
O
CH3
1
3
4
6
4a
4b
8a 8b
N
N
OCH3
OCH3
O
1
35
8
1
3
6
8
4a
4a
4b
4b
8a 8b
8b
8a
1
3 4
5
4
1
H3CO
HO
O
O 9
1
3
4a
4b
5
78a
8b
6
Figure 1. Structures of compounds 1–9.

203 Zhou, Y. et al. / J. Chin. Pharm. Sci. 2016, 25 (3), 201–205
CC and eluted with CHCl3–MeOH (5:1→1:1, v/v) to
obtain 3 fractions (Fr. A–Fr. C). Fr. B was separated by
semi-preparative HPLC (Waters Nova-pak C18 column,
7.8 mm×300 mm, 5 μm; 50%–100% MeOH for 30 min;
2 mL/min; 254 and 285 nm) to obtain compound 7
(1.9 mg).
3. Identification
3.1. 3,3-[Oxybis(methylene)]bis(9-methoxy-9H-
carbazole) (1)
Brown gum solid. EI-MS m/z 436 [M]+, 226, 210.
1H NMR (CDCl3, 500 MHz) δ: 8.07 (2H, s, H-4, 4),
8.03 (2H, d, J 7.5 Hz, H-5, 5), 7.53 (6H, overlapped,
H-1, 1, 2, 2, 8, 8), 7.48 (2H, t, J 7.5 Hz, H-7, 7), 7.26
(2H, t, J 7.5 Hz, H-6, 6), 4.77 (4H, s, 2×-CH2), 4.12
(6H, s, 2×-OCH3).
13C NMR (CDCl3, 125 MHz) δ: 138.1
(C-8a, 8a), 137.4 (C-8b, 8b), 130.3 (C-3, 3), 126.8
(C-2, 2), 126.3 (C-7, 7), 120.6 (C-5, 5), 120.5 (C-4, 4),
120.2 (C-6, 6), 120.1 (C-4b, 4b), 120.1 (C-4a, 4a),
108.5 (C-8, 8), 108.4 (C-1, 1), 72.6 (2×-CH2), 63.5
(2×-OCH3). All above data are in good agreement
with those of 3,3-[oxybis(methylene)]bis(9-methoxy-
9H-carbazole)[6].
3.2. Murrastifoline B (2)
Yellow gum solid. 1H NMR (CDCl3, 500 MHz) δ:
10.05 (br s, N-H), 8.04 (1H, d, J 7.8 Hz, H-5), 7.93
(1H, d, J 7.8 Hz, H-5), 7.55 (1H, s, H-4), 7.51 (1H, s,
H-4), 7.45 (1H, d, J 7.9 Hz, H-8), 7.41 (1H, d, J 7.9 Hz,
H-7), 7.36 (1H, t, J 7.9 Hz, H-7), 7.22 (1H, d, J 7.9 Hz,
H-8), 7.18 (1H, t, J 7.7 Hz, H-6), 7.17 (1H, t, J 7.7 Hz,
H-6), 6.79 (1H, s, H-2), 6.78 (1H, s, H-2), 3.93 (3H, s,
1-OCH3), 3.83 (3H, s, 1-OCH3), 2.54 (3H, s, 3-CH3).
13C NMR (CDCl3, 125 MHz) δ: 146.7 (C-1), 145.6 (C-1),
141.4 (C-8a), 139.4 (C-8a), 131.4 (C-8b), 129.1 (C-8b),
125.6 (C-7), 125.5 (C-7), 125.0 (C-4b), 124.0 (C-4b),
123.5 (C-3), 123.2 (C-3), 120.6 (C-4a), 120.1 (C-4a),
119.2 (C-6), 118.8 (C-6), 112.8 (C-4), 112.5 (C-4),
110.9 (C-5), 110.8 (C-5), 109.7 (C-8), 109.2 (C-8),
107.7 (C-2), 105.0 (C-2), 55.8 (1-OCH3), 55.4 (1-OCH3),
21.7 (3-CH3). All above data are in good agreement with
those of murrastifoline B[7].
3.3. Murrayaquinone A (3)
Brown gum solid. 1H NMR (acetone-d6, 500 MHz) δ:
8.18 (1H, d, J 7.6 Hz, H-5), 7.65 (1H, d, J 7.6 Hz, H-8),
7.43 (1H, t, J 7.6 Hz, H-7), 7.36 (1H, t, J 7.6 Hz, H-6),
6.58 (1H, s, H-2), 2.15 (3H, s, -CH3).
13C NMR (acetone-d6,
125 MHz) δ: 183.1 (C-4), 179.9 (C-1), 148.3 (C-8a),
137.3 (C-8b), 135.8 (C-3), 131.5 (C-7), 126.4 (C-8),
124.1 (C-4b), 123.7 (C-6), 122.1 (C-5), 116.2 (C-4a),
113.5 (C-2), 15.0 (-CH3). All above data are in good
agreement with those of murrayaquinone A[8].
3.4. 2,4-Dimethoxyphenol (4)
Pale yellow gum solid. 1H NMR (CDCl3, 500 MHz)
δ: 7.94 (1H, dd, J1 6.6 Hz, J2 2.6 Hz, H-5), 7.76 (1H, d,
J 6.6 Hz, H-6), 6.91 (1H, d, J 2.6 Hz, H-3), 3.99 (3H,
s, 3-OCH3), 3.97 (3H, s, 1-OCH3);
13C NMR (CDCl3,
125 MHz) δ: 155.5 (C-1), 149.8 (C-4), 142.5 (C-2),
118.8 (C-5), 110.8 (C-6), 107.4 (C-3), 57.4 (3-OCH3),
57.3 (1-OCH3). All above data are in good agreement
with those of 2,4-dimethoxyphenol[9].
3.5. 1,2-Dimethoxy-4-nitrobenzene (5)
Brown gum solid. 1H NMR (CDCl3, 500 MHz) δ: 7.94
(1H, dd, J1 8.9 Hz, J2 2.6 Hz, H-5), 7.77 (1H, d, J 2.6 Hz,
H-3), 6.93 (1H, d, J 8.9 Hz, H-6), 4.00 (3H, s, 1-OCH3),
3.99 (3H, s, 2-OCH3);
13C NMR (CDCl3, 125 MHz)
δ: 154.6 (C-1), 149.0 (C-2), 141.2 (C-4), 118.0 (C-5),
110.0 (C-6), 106.6 (C-3), 56.6 (1-OCH3), 56.5 (2-OCH3).
All above data are in good agreement with those of
1,2-dimethoxy-4-nitrobenzene[10].
3.6. 3-Methylanisole (6)
Colorless liquid. 1H NMR (CDCl3, 500 MHz) δ: 7.64
(1H, t, J 8.7 Hz, H-5), 7.51 (1H, d, J 8.7 Hz, H-4), 6.96
(1H, d, J 8.7 Hz, H-6), 6.02 (1H, s, H-2), 4.01 (3H,
s, -OCH3), 2.97 (3H, s, -CH3);
13C NMR (CDCl3,
125MHz) δ: 151.7 (C-1), 133.3 (C-4), 124.2 (C-3),
114.7 (C-5), 114.6 (C-6), 110.7 (C-2), 57.0 (-OCH3),

204 Zhou, Y. et al. / J. Chin. Pharm. Sci. 2016, 25 (3), 201–205
32.2 (-CH3). All above data are in good agreement
with those of 3-methylanisole[11].
3.7. (–)-Syringaresinol-4-O-β-D-glucopyranoside (7)
Pale yellow powder. –6.37 (c 0.11, CH3OH).
1H NMR (CD3OD, 500 MHz) δ: 6.70 (2H, s, H-2, 6),
6.64 (2H, s, H-2, 6), 4.88 (d, J 8.5 Hz, Glc-H-1), 4.87
(1H, overlapped with H2O peak, H-7), 4.74 (d, J 4.0 Hz,
H-7), 4.28, 3.90 (4H, m, H-9, 9), 3.89 (6H, s, 3, 5-OMe),
3.88 (6H, s, 3, 5-OMe), 3.69 (1H, m, Glc-H-6), 3.51
(1H, dd, J1 8.5 Hz, J2 8.0 Hz, Glc-H-2), 3.47 (1H, m,
Glc-H-5), 3.45 (1H, m, Glc-H-4), 3.23 (1H, m, Glc-H-3),
3.24 (1H, m, H-8), 3.10 (1H, m, H-8); 13C NMR
(CD3OD, 125 MHz) δ: 154.2 (C-3, 5), 149.2 (C-3, 5),
139.6 (C-1), 136.0 (C-4), 135.5 (C-4), 133.0 (C-1),
105.3 (Glc-C-1), 104.7 (C-2, 6), 104.4 (C-2, 6), 87.4
(C-7), 87.1 (C-7), 78.1 (Glc-C-3), 77.7 (Glc-C-5), 75.6
(Glc-C-2), 72.8 (C-9), 72.7 (C-9), 71.2 (Glc-C-4), 62.4
(Glc-C-6), 57.0 (3, 5-CH3), 56.7 (3, 5-OCH3), 55.5 (C-8),
55.3 (C-8). All above data are in good agreement with
those of (–)-syringaresinol-4-O-β-D-glucopyranoside[12].
3.8. β-Sitosterol (8)
White crystal (in CHCl3), mp: 139–142 °C.
1H NMR
(CDCl3, 500 MHz) δ: 5.30 (1H, m, H-6), 3.52 (1H, m,
H-3), 1.00 (3H, s, H-19), 0.92 (3H, d, J 6.5 Hz, H-21),
0.85 (3H, d, J 7.0 Hz), 0.83 (3H, d, J 6.9 Hz), 0.81 (3H,
d, J 6.8 Hz), 0.68 (3H, s, H-18); 13C NMR (CDCl3,
125 MHz) δ: 140.9 (C-5), 121.9 (C-6), 72.0 (C-3), 56.9
(C-14), 56.2 (C-17), 50.3 (C-9), 46.0 (C-13), 42.5 (C-24),
42.4 (C-4), 40.0 (C-16), 36.3 (C-10), 36.2 (C-20), 34.0
(C-23), 32.1 (C-1), 32.0 (C-2), 32.0 (C-7), 31.9 (C-8),
29.4 (C-27), 28.4 (C-12), 26.0 (C-22), 24.6 (C-15),
23.2 (C-25), 21.4 (C-11), 19.9 (C-28), 19.7 (C-21),
19.5 (C-29), 19.3 (C-19), 12.1 (C-26), 12.0 (C-18).
All above data are in good agreement with those of
β-sitosterol[13] .
3.9. Octadecanyl-3-methoxy-4-hydroxylbenzenea-
crylate (9)
White crystal (in the mixture of CHCl3–MeOH).
1H NMR
(CDCl3, 500 MHz) δ: 7.61 (1H, d, J 15.9 Hz, H-7),
7.08 (1H, dd, J1 8.1 Hz, J2 1.5 Hz, H-6), 7.04 (1H, d,
J 1.5 Hz, H-2), 6.92 (1H, d, J 8.1 Hz, H-5), 6.29 (1H,
d, J 15.9 Hz, H-8), 4.20 (2H, t, J 6.7 Hz, -OCH2-), 3.92
(3H, s, OCH3), 1.28 (32H, br s, 16×-CH2-), 0.89 (3H, t,
J 6.8 Hz, -CH3);
13C NMR (CDCl3, 125 MHz) δ: 167.4
(C-9), 148.2 (C-3), 147.1 (C-4), 144.7 (C-7), 127.5
(C-1), 123.2 (C-6), 116.2 (C-8), 115.0 (C-5), 109.8
(C-2), 64.8 (-OCH2-), 56.2 (-OCH3), 32.1 (-CH2), 29.9
(11×-CH2-), 29.5 (-CH2-), 29.1 (-CH2-), 26.2 (-CH2-),
22.8 (-CH2-), 14.2 (-CH3). All above data are in good
agreement with those of octadecanyl-3-methoxy-4-
hydroxylbenzeneacrylate[14].
4. Discussion
So far, there have been few reports about the chemical
constituents from M. tetramera, and most previous
pharmacological works focused on the crude extract
of M. tetramera[15–17]. In this study, a phytochemical
investigation of M. tetramera led to the isolation and
structural identification of nine compounds. This is the
first report for compounds 4–6 and 9 from this genus
plants, and compounds 2–9 were described from this
species for the first time. These data will be useful
for the further research and development of this
traditional medicinal plant.
Acknowledgements
This work was financially supported by National
Science Fund for Excellent Young Scholars (Grant
No. 81222051), and National Key Technology R&D
Program “New Drug Innovation” of China (Grant Nos.
2012ZX09301002-002-002 and 2012ZX09304-005).
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205 Zhou, Y. et al. / J. Chin. Pharm. Sci. 2016, 25 (3), 201–205
四数九里香的化学成分研究
周颖, 王文光, 屠鹏飞, 姜勇*
北京大学医学部 药学院 天然药物及仿生药物国家重点实验室, 北京 100191
摘要: 对四数九里香的地上部位进行了化学成分研究。运用硅胶和Sephadex LH-20柱色谱及制备HPLC和TLC等多
种分离方法进行分离纯化, 共分离得到9个化学成分, 通过NMR和MS数据分析, 并结合文献对比, 鉴定其结构分别为
3,3-[oxybis(methylene)]bis(9-methoxy-9H-carbazole) (1), murrastifoline B (2), murrayaquinone A (3), 2,4-二甲氧基苯酚 (4),
1,2-二甲氧基-4-硝基苯 (5), 间甲基苯甲醚 (6), 丁香树脂酚葡萄糖苷 (7), β-谷甾醇 (8), 3-甲氧基-4-羟基反式苯丙烯酸正十
八醇酯 (9)。化合物4–6和9为首次从九里香属植物中分离得到, 化合物2–9为该种植物中首次分离得到。
关键词: 芸香科; 四数九里香; 化学成分