全 文 ： 370 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 2012 年 9 月 第 10 卷 第 5 期

Chinese Journal of Natural Medicines 2012, 10(5): 0370−0373
doi: 10.3724/SP.J.1009.2012.00370
Chinese
Journal of
Natural
Medicines







Two new phenylpropanoids isolated from the
rhizomes of Alpinia galanga
ZHAO Ling1, CHEN Lv-Yi2, LIANG Jing-Yu3 *
1School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, 430023, China;
2School of Pharmacy, South-Central University for Nationalities, Wuhan, 430074, China;
3Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009, China
Available online Sep. 2012
[ABSTRACT] AIM: To study the chemical constituents and bioactivity of the rhizomes of Alpinia galangal Willd. (Zingiberaceae).
METHODS: The chemical constituents were isolated using various chromatographic methods and crystallization, and the chemical
structures were elucidated on the basis of spectral analysis. In addition, cytotoxic activities toward four tumor cell lines (HepG2,
EC109, A549 and Vero) were tested by MTT method. RESULTS: Six phenylpropanoids were obtained and their structures were iden-
tified as (S)-1’-ethoxy chavicol acetate (1), (E)-4-acetoxy cinnamyl ethyl ether (2), (E)-4-hydroxycinnamaldehyde (3), (E)-4-acetoxy
cinnamyl alcohol (4), 4-acetoxy cinnamyl acetate (5), and 4, 4’[(2E, 2E)-bis(prop-2-ene)-1, 1-oxy]-diphenyl-7, 7-diacetate (6).
CONCLUSION: Compounds 1 and 2 were two new phenylpropanoids. Compound 5 shows selective cytotoxic activity on human lung
adenocarcinoma cell A549 (IC50 19.35 μmol·L−1).
[KEY WORDS] Alpinia galanga; Zingiberaceae; Phenylpropanoids; Chemical constituent; Cytotoxic activity
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2012)05-0370-04

1 Introduction
The Alpinia galanga Willd. (Zingiberaceae) is widely
cultivated in China, India, and Southeast Asian countries,
such as Thailand, Malaysia, and the Philippines. The rhi-
zomes of this plant are extensively used as a flavoring in
traditional foods, and it is also used in traditional Chinese
medicine as a stomachic [1]. Recent studies showed that the
rhizomes of the plant have antitumor [2-3], antibacterial [4-6],
anti-inflammatory [7], and anti-ulcer effects [8]. In order to
determine the biologically active compounds, two new
phenylpropanoids (S)-1-ethoxy chavicol acetate (1) and
(E)-4-acetoxy cinnamyl ethyl ether (2), along with four
known phenylpropanoids (E)-4-hydroxylcinnamaldehyde (3),
(E)-4-acetoxy cinnamyl alcohol (4), 4-acetoxy cinnamyl ace-
tate (5) and 4, 4[(2E, 2E)-bis(prop-2-ene)-1, 1-oxy]-di-
phenyl-7, 7-diacetate (6) were isolated. This paper describes
the isolation and structural elucidation of the two new com-
pounds. Cytotoxic evaluations showed that compound 5 ex-

[Received on] 23-Mar.-2012
[*Corresponding author] LIANG Jing-Yu: Prof., jyliang08@126.
com
These authors have no any conflict of interest to declare.
hibited selective activity against human lung adenocarcinoma
cell A549.
2 Experimental
2.1 General experimental procedures
NMR spectra were recorded on a Bruker ACF-500/300
NMR instrument (1H: 500/300 MHz, 13C: 125/75 MHz) with
TMS as internal standard. Mass spectra were obtained on a
MS Agilent 1100 Series LC/MSD Trap mass spectrometer
(ESI-MS) and a Micro Q-TOF MS (HRESIMS), respectively.
All solvents used were of analytical grade (Tianjin Chemical
Plant). Column chromatography was performed on silica gel
(100−200 mesh, 200−300 mesh, Qingdao Marine Chemical
Co., Ltd.) and Sephadex LH-20 (Pharmacia).
2.2 Plant material
The rhizomes of A. galanga were collected in Bobai
Prefecture, Guangxi Province and identified by SONG Xue-
Hua, the curator of the China Pharmaceutical University
(China). A voucher specimen is deposited in the Department
of Natural Medicinal Chemistry, China Pharmaceutical Uni-
versity.
2.3 Extraction and isolation
The rhizomes of A. galanga (16 kg) were crushed into
small pieces and extracted twice with 95% ethanol. The
combined extracts were evaporated in vacuo, and the result-
ZHAO Ling, et al. /Chinese Journal of Natural Medicines 2012, 10(5): 370−373
2012 年 9 月 第 10 卷 第 5 期 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 371

ing residue was suspended in water and subsequently ex-
tracted with petroleum ether and EtOAc. A portion (161.5 g)
of the EtOAc extract (478 g) was sequentially submitted to
column chromatography (silica gel, petroleum ether−EtOAc
1 : 0−0 : 1) to afford several fractions. After repeated column
chromatography and recrystallisation, 1 (200.5 mg), 2 (300.1
mg), 3 (0.9 g), 4 (3.2 g), 5 (2.2 g) and 6 (0.5 g) were ob-
tained.
3 Structural Identification
Compound 1 was obtained as a pale yellow oil, and
showed [α]31.0 D −30.1 (c 1.42, EtOH). It gave prunosus color
with vanillin-sulfuric acid. Its molecular formula was deter-
mined as C13H16O3 [(M + Na)+ m/z 243.099 3; calcd. 243.099
7] by high resolution ESI-MS. The 1H NMR spectral data
(Table 1) showed an AA′BB′ spin system at δH 7.05 and 7.34,
typical of a para-substituted benzene ring, and confirmed by
the 13C NMR data (Table 1) of two resonances equivalent
resonances [δ 121.4 (C-3, C-5), 127.8 (C-2, C-6)]. From the
1H NMR spectral data, the signals at δH 5.25 (1H, ddd, J =
17.0, 1.5, 1.0 Hz), 5.18 (1H, ddd, J = 10.0, 1.5, 1.0 Hz),
5.91(1H, ddd, J = 17.0, 10.0, 6.5 Hz) and 4.73 (1H, br d, J =
6.5 Hz) indicated the presence of an allyl group. Furthermore,
the downfield chemical shift of δH 4.73 revealed the allyl
group might have an oxygen atom on the other side. The
protons signal at δH 1.22 (3H, t, J = 7.0 Hz), 3.52 (1H, dq, J =
9.0, 7.0 Hz) and 3.43 (1H, dq, J = 9.0, 7.0 Hz) belong to the
ethoxy group. Meanwhile, the methyl group at δH 2.28 (3H, s),
along with resonances at δC 21.1 and 169.4 indicated the
presence of an acetyl group. These data were similar to the
known compound 1′S-acetoxychavicol acetate [10], except that
the acetoxyl group at C-1′ was replaced by the ethoxy group.
This was supported by the HMBC (Fig. 1) correlations of
H-1′ with C-1′′ and H-1′′ with C-1′. The configuration of C-1′
was determined as S ([α]31.0 D −30.1) by comparison of the
optical rotation with 1′S-acetoxychavicol acetate ([α]25 D －60.1)
[9]. Thus, compound 1 was elucidated as (S)-1-ethoxy chav-
icol acetate.
Compound 2 was obtained as a pale yellow grease and
showed prunosus color with vanillin-sulfuric acid. Its mo-
lecular formula was determined as C13H16O3 [(M + Na)+ m/z
243.098 9; calcd. 243.099 7] by high resolution ESI-MS. The
1H NMR and 13C NMR spectral data (Table 1) revealed the
presence of one para-substituted benzene ring [7.37 (2H, d, J
= 8.6 Hz), 7.03 (2H, d, J = 8.6 Hz)], two olefinic protons
[6.58 (1H, d, J = 15.9 Hz), 6.26 (1H, dt, J = 15.9, 6.0 Hz)],
one oxygen-bearing methylene [4.12 (2H, d, J = 6.0 Hz)] and
one ethoxyl group [3.49 (2H, q, J = 7.0 Hz), 1.22 (3H, t, J =
7.0 Hz)]. Meantime, the 1H NMR signals at δH 2.28 (3H, s)
and the 13C NMR signals at δC 21.1, 169.4 indicated the
presence of an acetyl group. These data were similar to those
of trans-p-coumaryl diacetate [10], except that the acetoxyl
group at C-1′ was replaced by the ethoxyl group. The struc-
ture was further confirmed by long-range correlations from
H-1′ to C-1, C-2; from H-2′ to C-1′, C-3′; and from H-1′′to
C-3′, C-2′′ in the HMBC spectrum (Fig. 2). On the basis of
the above evidence, compound 2 was elucidated to be
(E)-4-acetoxy cinnamyl ethyl ether.
Compound 3 White needle crystals (MeOH), mp
139−141 °C, ESI-MS m/z 147 [M − H]−, 1H NMR (DMSO-d6,
300 MHz) δ: 9.59 (1H, d, J = 7.9 Hz, H-3′), 6.65 (1H, dd, J =
15.7, 7.9 Hz, H-2′), 7.61 (1H, d, J = 15.7 Hz, H-1′), 7.60 (2H,
Table 1 1H (500 MHz) and 13C (125 MHz) NMR data of 1 and 1H (300 MHz) and 13C (75 MHz) NMR data of 2 (CDCl3, J in Hz)
Compound 1 Compound 2
Position
δC δH HMBC (position) δC δH HMBC (position)
1 138.9 134.5
2 127.8 7.34 (m) 3, 4 127.6 7.37 (d, J = 8.6) 1, 3, 4
3 121.4 7.05 (m) 1, 4 121.5 7.03 (d, J = 8.6) 1, 4, 5
4 150.0 150.0
5 121.4 7.05 (m) 1, 4 121.5 7.03 (d, J = 8.6) 1, 4, 5
6 127.8 7.34 (m) 4, 5 127.6 7.37 (d, J = 8.6) 1, 4, 5, 1′
7 169.4 169.4
8 21.1 2.28 (s) 7 21.0 2.29 (s) 7
1′ 82.2 4.73 (br d, J = 6.5) 1, 2, 2′, 3′, 1′′ 131.1 6.58 (d, J = 15.9) 1, 2, 2′, 3′
2′ 140.0 5.91 (ddd, J = 17.0, 10.0, 6.5) 1, 1
′ 126.4 6.26 (dt, J = 15.9, 6.0) 1, 1′
3′ 116.1
3′a, 5.25 (ddd, J = 17.0,
1.5, 1.0)
3′b, 5.18 (ddd, J = 10.0,
1.5, 1.0)
1′, 2′ 70.9 4.12 (d, J = 6.0) 1′, 2′
1′′ 64.0 1
′′a, 3.52 (dq, J = 9.0, 7.0)
1′′b, 3.43 (dq, J = 9.0, 7.0) 1
′, 2′′ 65.6 3.49 (q, J = 7.0) 3′, 2′′
2′′ 15.2 1.22 (t, J = 7.0) 1′′ 15.0 1.22 (t, J = 7.0) 1′′

ZHAO Ling, et al. /Chinese Journal of Natural Medicines 2012, 10(5): 370−373
372 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 2012 年 9 月 第 10 卷 第 5 期



Fig. 1 Key correlations in the HMBC spectrum of com-
pound 1

Fig. 2 Key correlations in the HMBC spectrum of com-
pound 2
d, J = 8.8 Hz, H-2, 6), 6.85 (2H, d, J = 8.8 Hz, H-3, 5), 10.19
(1H, brs, 4-OH). Compound 3 was characterized as
trans-p-hydroxy-cinnamaldehyde by comparison of the
physical and spectral data with the literature [11].
Compound 4 White needle crystals (MeOH), mp
74−75 °C, ESI-MS m/z 193 [M + H]+, 1H NMR (CDCl3, 300
MHz) δ: 4.31 (2H, d, J = 5.7 Hz, H-3′), 6.31 (1H, dt, J = 15.9,
5.7 Hz, H-2′), 6.60 (1H, d, J = 15.9 Hz, H-1′), 7.38 (2H, d, J
= 8.6 Hz, H-2, 6), 7.05 (2H, d, J = 8.6 Hz, H-3, 5), 2.29 (3H,
s, 4-OCOCH3). Compound 4 was characterized as (E)-4-
acetoxy cinnamyl alcohol by comparison of the physical and
spectral data with the literature [11].
Compound 5 White lamellar crystals (petroleum
ether−EtOAc), mp 40−46 °C，ESI-MS m/z 257 [M + Na]+, 1H
NMR (CDCl3, 300 MHz) δ: 4.71 (2H, dd, J = 6.0, 1.5 Hz,
H-3′), 6.23 (1H, dt, J = 16.0, 6.0 Hz, H-2′), 6.62 (1H, d, J =
16.0 Hz, H-1′), 7.38 (2H, d, J = 7.0 Hz, H-2, 6), 7.05 (2H, d,
J = 7.0 Hz, H-3, 5), 2.29 (3H, s, 4-OCOCH3), 2.09 (3H, s,
3′-OCOCH3). Compound 5 was characterized as 4-acetoxy
cinnamyl acetate by comparison of the physical and spectral
data with the literature [10].
Compound 6 White needle crystals (petroleum
ether−EtOAc), mp 108−109 °C，ESI-MS m/z 384 [M + NH4]+,
1H NMR (CDCl3, 500 MHz) δ: 4.19 (4H, dd, J = 6.0, 1.5 Hz,
H-1, 1′), 6.26 (2H, dt, J = 16.0, 6.0 Hz, H-2, 2′), 6.61 (2H, d,
J = 16.0 Hz, H-3, 3′), 7.39 (4H, d, J = 8.6 Hz, H-5, 5′, 9, 9′),
7.05 (4H, d, J = 8.6 Hz, H-6, 6′, 8, 8′), 2.29 (6H, s, 7,
7′-OCOCH3). Compound 6 was characterized as 4, 4 [(2E,
2E)-bis (prop-2-ene)-1, 1-oxy]-diphenyl-7, 7-diacetate by
comparison of the spectral data with the literature [12].
4 Cytotoxicity Assay
All compounds were assayed for cytotoxic activities to-
ward four tumor cell lines (HepG2, EC109, A549, and Vero)
by the MTT test in vitro. Among them, compound 5 shows
selective cytotoxic activity on the human lung adenocarci-
noma cell line A549 (IC50 19.35 μmol·L−1). Other compounds
showed no such activities (IC50 > 20 μmol·L−1).
References
[1] Flora Republicae Popularis Sinicae [M]. Vol. 16. Beijing:
Science Press, 1981: 71.
[2] Muangnoi P, Lu M, Lee J, et al. Cytotoxicity, apoptosis and
DNA damage induced by Alpinia galanga rhizome extract [J].
Planta Med, 2007, 73(8): 748-754.
[3] Panich U, Kongtaphan K, Onkoksoong T, et al. Modulation of
antioxidant defense by Alpinia galanga and Curcuma aro-
matica extracts correlates with their inhibition of UVA-induced
melanogenesis [J]. Cell Biol Toxicol, 2010, 26(2): 103-116.
[4] Kiranmayee R, Bhuvaneswari Ch, Lakshmi MN, et al. Anti-
bacterial activity of Alpinia galanga (L.) Willd crude extracts
[J]. Appl Biochem Biotechnol, 2010, 162(3): 871-884.
[5] Oonmetta J, Suzuki T, Gasaluck P, et al. Current pharmacol-
ogical and phytochemical studies of the plant Alpinia galanga
[J]. LWT Food Sci Technol, 2006, 39(10): 1214-1220.
[6] Weerakkody NS, Caffin N, Turner MS, et al. In vitro antim-
icrobial activity of less-utilized spice and herb extracts against
selected food-borne bacteria [J]. Food Control, 2010, 21(10):
1408-1411.
[7] Yu ES, Min HJ, Lee K, et al. Anti-inflammatory activity of
p-coumaryl alcohol-γ-O-methyl ether is mediated through
modulation of interferon-γ production in Th cells [J]. Br J
Pharmacol, 2009,156(7): 1107-1114.
[8] Matsuda H, Pongpiriyadacha Y, Morikawa T, et al.
Gastroprotective effects of phenylpropanoids from the rhi-
zomes of Alpinia galanga in rats: structural requirements and
mode of action [J]. Eur J Pharmacol, 2003, 471(1): 59-67.
[9] Azuma H, Miyasaka K, Yokotani T, et al. Lipase-catalyzed
preparation of optically active 1′-acetoxychavicol acetates and
their structure–activity relationships in apoptotic activity
against human leukemia HL-60 cells [J]. Bioorg Med Chem,
2006, 14(6): 1811-1818.
[10] Noro T, Sekiya T, Katoh M, et al. Inhibitors of xanthine oxi-
dase from Alpinia galanga [J]. Chem Pharm Bull, 1988, 36(1):
244-248.
[11] Zhu XL, Dou YH, Huang XF, et al. Studies on the chemical
constituents of Alpinina galangal Willd. [J]. Mod Chin Med,
2008, 10(11): 13-15.
[12] Zhu XL, Yang MH, Luo JG, et al. A new phenylpropanoid
from Alpinia galanga [J]. Chin J Nat Med, 2009, 7(1): 19-21.
ZHAO Ling, et al. /Chinese Journal of Natural Medicines 2012, 10(5): 370−373
2012 年 9 月 第 10 卷 第 5 期 Chin J Nat Med Sep. 2012 Vol. 10 No. 5 373


大高良姜根茎中的两个新苯丙素化合物
赵 玲 1, 陈旅翼 2, 梁敬钰 3*
1武汉工业学院生物与制药工程学院 武汉 430023;
2中南民族大学药学院 武汉 430074;
3中国药科大学天然药物化学教研室 南京 210009
【摘 要】 目的：研究大高良姜的化学成分及活性。方法：运用各种柱色谱方法和重结晶手段分离纯化化合物, 并通过波
谱数据鉴定化合物结构。此外, 运用 MTT 法测定了化合物对四种肿瘤细胞株(HepG2, EC109, A549 和 Vero)的细胞毒活性。结
果：从大高良姜中分离得到 6 个苯丙素类化合物, 其结构分别鉴定为(S)-1-ethoxy chavicol acetate (1), (E)-4-acetoxy cinnamyl ethyl
ether (2), (E)-4-hydroxy-cinnamaldehyde (3), (E)-4-acetoxy cinnamyl alcohol (4), 4-acetoxy cinnamyl acetate (5) 和 4, 4[(2E, 2E)-bis
(prop-2-ene)-1, 1-oxy]-diphenyl-7, 7-diacetate (6). 结论：化合物 1 和 2 为新化合物。化合物 5 对 A549 肿瘤细胞有一定的细胞毒
活性, IC50为 19.35 μmol·L−1。
【关键词】 大高良姜; 姜科; 苯丙素; 化学成分; 细胞毒