Two new compounds from Aconitum tanguticum

Article abstract of DOI:10.1080/10286020.2014.904292

A phytochemical investigation on the whole plant of Aconitum tanguticum (Ranunculaceae) resulted in the isolation and characterization of two new phenylpropanoid glycosides (1 and 2). Their structures were elucidated as 4-hydroxyphenethoxy-8-O-β-d-[6-O-(4-O-β-d-glucopyranosyl)-sinapoyl] -glucopyranoside (1) and 3,4-dimethoxy-trans-cinnamic acid-9-O-β-d- allopyranoside (2) on the basis of spectroscopic data (1D NMR, 2D NMR, and MS) and comparison with the literature data. 2014

Full text of DOI:10.1080/10286020.2014.904292

This article was downloaded by: [Selcuk Universitesi]
On: 20 December 2014, At: 15:47
Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered
office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK
Journal of Asian Natural Products
Research
Publication details, including instructions for authors and
subscription information:
http://www.tandfonline.com/loi/ganp20
Two new compounds from Aconitum
tanguticum
Yan-Rong Liac, Lu Xua, Chun Liab, Zhi-Min Wangabc & Li-Xin Yanga
a Institute of Chinese Materia Medica, China Academy of Chinese
Medicine Sciences, Beijing 100700, China
b Shaanxi University of Chinese Medicine, Xi'an 712046, China
c National Engineering Laboratory for Quality Control Technology
of Chinese Herbal Medicines, Beijing 100700, China
Published online: 07 Apr 2014.
Click for updates
To cite this article: Yan-Rong Li, Lu Xu, Chun Li, Zhi-Min Wang & Li-Xin Yang (2014) Two new
compounds from Aconitum tanguticum, Journal of Asian Natural Products Research, 16:7, 730-734,
DOI: 10.1080/10286020.2014.904292
To link to this article: http://dx.doi.org/10.1080/10286020.2014.904292
PLEASE SCROLL DOWN FOR ARTICLE
Taylor & Francis makes every effort to ensure the accuracy of all the information (the
“Content”) contained in the publications on our platform. However, Taylor & Francis,
our agents, and our licensors make no representations or warranties whatsoever as to
the accuracy, completeness, or suitability for any purpose of the Content. Any opinions
and views expressed in this publication are the opinions and views of the authors,
and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content
should not be relied upon and should be independently verified with primary sources
of information. Taylor and Francis shall not be liable for any losses, actions, claims,
proceedings, demands, costs, expenses, damages, and other liabilities whatsoever
or howsoever caused arising directly or indirectly in connection with, in relation to or
arising out of the use of the Content.
This article may be used for research, teaching, and private study purposes. Any
substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing,
systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &

Conditions of access and use can be found at http://www.tandfonline.com/page/terms-
and-conditions

Journal of Asian Natural Products Research, 2014
Vol. 16, No. 7, 730–734, http://dx.doi.org/10.1080/10286020.2014.904292
Two new compounds from Aconitum tanguticum
Yan-Rong Lia,b, Lu Xua, Chun Lia,c*, Zhi-Min Wanga,b,c and Li-Xin Yanga
aInstitute of Chinese Materia Medica, China Academy of Chinese Medicine Sciences, Beijing
100700, China; Shaanxi University of Chinese Medicine, Xi’an 712046, China;
b
cNational Engineering Laboratory for Quality Control Technology of Chinese Herbal Medicines,
Beijing 100700, China
(Received 3 December 2013; final version received 11 March 2014)
A phytochemical investigation on the whole plant of Aconitum tanguticum
(Ranunculaceae) resulted in the isolation and characterization of two new
phenylpropanoid glycosides (1 and 2). Their structures were elucidated as 4-
hydroxyphenethoxy-8-O-b-D-[6-O-(4-O-b-D-glucopyranosyl)-sinapoyl]-glucopyrano-
side (1) and 3,4-dimethoxy-trans-cinnamic acid-9-O-b-D-allopyranoside (2) on the
basis of spectroscopic data (1D NMR, 2D NMR, and MS) and comparison with the
literature data.
Keywords: Ranunculaceae; Aconitum tanguticum; chemical composition; phenylpro-
panoid glycosides
1. Introduction
matory fractions of Ponka (the pharmaco-
logical screening results of different
fractions from Ponka have not yet been
published). As the continuation of our
work, two new phenylpropanoid glyco-
sides were isolated from this plant. This
paper deals with the determination of the
structures of new compounds (Figure 1).
Aconitum tanguticum (Maxim.) Stapf.
(Ranunculaceae), a perennial herbaceous
plant, mainly distributes in Qinghai,
Gansu, Sichuan and Yunnan provinces in
China [1]. In Tibetan medicine, the whole
plant is named “Ponka” and has been used
as a traditional Tibetan folk medicine for
the treatment of hepatitis, influenza,
gastricism, nephritis, and other diseases
for thousands of years [2]. Now, Ponka is
used in a large number of Tibetan patent
drugs [3,4]. Recently, a lot of compounds
such as diterpenoid alkaloids [5–8],
flavonoid glycosides [9–10], and phenolic
acid derivatives [11–12] have been
isolated from this plant, but there has
been no activity reports of these monomer
compounds. So, until now, the effective
material base of Ponka is not clear, due to
the low level of the quality control and
formulation of relative drugs. In order to
clarify the medicinal material base of
Ponka, we have continuously studied the
chemical compositions of the anti-inflam-
2. Results and discussion
The 30% and 60% ethanol eluent of EtOH
extract of A. tanguticum was subjected to
silica gel, ODS, Sephadex LH-20 column
chromatography (CC), and preparative
HPLC to give compounds 1 and 2. The
structures of new compounds were eluci-
dated on the basis of spectroscopic data
(1D NMR, 2D NMR, and MS) and
comparison with the literature data.
Compound 1 was obtained as an
amorphous powder, whose molecular
formula, C31H40O16, was confirmed by
the HRTOFMS at m/z 691.2203
1
[M þ Na]þ. The H NMR spectroscopic
*Corresponding author. Email: lichun@icmm.ac.cn
q 2014 Taylor & Francis

Journal of Asian Natural Products Research
731
O
7´´
2´´
H3CO
6´
9´´
O
OH2C
1´´
6´´
8´´
3´´
4´´
6´
2
5
O
7
HO
HOH2C
4´
7
2
4´
HO
OCH3
OCH3
1
O
6´´´
HOH2C
1´
5´
2´
3
4
5´
2´
3
1
O
9
O
5´´
8
3´
O
8
OH
HO
O
6
HO
4´´´
4
5´´´
2´´´
1´
OH
3´
1´´´
HO
OCH3
6
OH
3´´´
5
OH
OH
1
2
Figure 1. Structures of compounds 1 and 2.
data showed two groups of aromatic
protons, including one AA0BB0 system
attributed to a 1,4-disubstituted aromatic
ring at dH 7.00 (2H, d, J ¼ 7.8 Hz) and
6.60 (2H, d, J ¼ 7.8 Hz), and the other
group attributed to a 1,3,4,5-tetrasubsti-
tuted aromatic ring at dH 7.03 (2H, s); two
aromatic methoxy singles at dH 3.78 (6H,
s, 300, 500-OCH3), and two anomeric protons
at dH 4.24 (1H, d, J ¼ 7.8 Hz) and 5.02
(1H, d, J ¼ 7.8 Hz). The 13C NMR
spectrum showed a group of characteristic
signals due to a sinapoyl group at dC 129.9
(C-100), 107.0 (C-200, 600), 153.2 (C-300, 500),
136.7 (C-400), 145.3 (C-700), 117.5 (C-800),
and 166.9 (C-900), and a group of charac-
teristic signals due to a p-hydroxyphenyl
ethanol at dC 129.0 (C-1), 130.1 (C-2, 6),
115.4 (C-3, 5), 156.0 (C-4), 35.3 (C-7), and
70.3 (C-8). The 1H and 13C NMR spectra of
1 were closely similar to those of 4-
hydroxyphenethoxy-8-O-b-D-[6-O-(4-O-
b-D-glucopyranosyl)-feruloyl]-glucopyra-
noside, which has been isolated from the
same plant before [11], except that the
NMR signals of the feruloyl unit were
replaced by those attributed to a sinapoyl
unit in 1 (Table 1). The HMBC spectrum of
1 showed correlations of H-1000 with C-400,
H-200 and H-600 with C-700, H-60 with C-900,
and H-10 with C-8. Therefore, compound 1
was elucidated to be 4-hydroxyphe-
nethoxy-8-O-b-D-[6-O-(4-O-b-D-gluco-
pyranosyl)-sinapoyl]-glucopyranoside.
Compound 2 was isolated as a white
amorphous powder, whose molecular
formula C17H22O9, was deduced by the
HRESIMS at m/z 393.1154 [M þ Na]þ. In
the 1H and 13C NMR spectra, compound 2
showed proton signals corresponding to a
1,3,4-trisubstituted benzene ring at dH 7.25
(1H, d, J ¼ 1.8 Hz, H-2), 7.20 (1H, d,
J ¼ 8.4 Hz, H-5), and 7.17 (1H, dd,
J ¼ 8.4, 1.8 Hz, H-6), with a trans-double
bond at dH 7.65 (1H, d, J ¼ 18.0 Hz, H-7),
6.45 (1H, d, J ¼ 18.0 Hz, H-8), and two
methoxy signals at dH 3.79 (3H, s, 3-
OCH3) and 3.90 (3H, s, 4-OCH3), as well
as a carbonyl carbon at dC 168.0 (C-9).
These data indicated that 2 had a 3,4-
dimethoxy cinnamic acid moiety. In
addition, seven oxygen-bearing proton
signals assignable to a glycosidic moiety
were observed at dH 5.34 (1H, d,
J ¼ 7.8 Hz), 3.71 (1H, dd, J ¼ 7.8,
3.0 Hz), 4.18 (1H, t, J ¼ 3.0 Hz), 3.63
(1H, dd, J ¼ 12.0, 3.0 Hz), 3.87–3.89 (1H,
m), 3.86 (1H, dd, J ¼ 12.0, 2.4 Hz), and
3.69 (1H, dd, J ¼ 12.0, 6.0 Hz), with the
corresponding carbon signals at dC 98.8,
70.6, 71.5, 67.2, 74.5, and 61.4. This
glycosidic moiety was identified as D-
allopyranoside, as acid hydrolysis of 2
liberated a D-allose which was determined
by gas chromatography (GC) analysis
[13]. The large coupling constant of the
anomeic proton at dH 5.34 (1H, d,
J ¼ 7.8 Hz) indicated the D-allopyranoside
was in a b-configuration. Furthermore, the
NMR data of b-D-allopyranosyl group
were in good agreement with those of cis-
ferulic acid 4-O-b-D-allopyranoside [14].
HMBC correlations of H-10 with C-9
confirmed that the allopyranosyl group is
linked to C-9. Thus, the structure of 2 was
characterized as 3,4-dimethoxy-trans-cin-
namic acid-9-O-b-D-allopyranoside.

732
Y. Li et al.
land). HRTOFMS and HRESIMS data
3. Experimental
were recorded on an Agilent Technologies
6250 Accurate-Mass Q-TOF LC/MS spec-
trometer (Santa Clara, CA, USA). ESI-MS
were measured on Agilent 6130SQ-MSD
(Agilent, USA). Preparative HPLC was
performed on a LC 3000 instrument
(Beijing Chuangxintongheng Science and
Technology Co., Ltd., Beijing, China)
connected to a UV 3000 detector, using an
Intersil-ODS column (250 £ 20 mm,
10 mm; GL Sciences Inc., Tokyo, Japan).
TLC plate precoated with silica gel GF254
3.1 General experimental procedures
Melting point was determined on an X-4
micromelting point apparatus (Cany Pre-
cision Instruments Co., Ltd., Shanghai,
China). IR spectra were recorded on a
Nicolet 5700 FT-IR spectrometer (Thermo
Nicolet Corporation, Madison, WI, USA).
UV spectra were measured on T6 New
Century UV–vis spectrophotometer
(Pgeneral, Beijing, China). NMR spectra
were acquired with Bruker Advance 600
spectrometer (Bruker, Fallanden, Switzer-
Table 1. 1H NMR (600 MHz) and 13C NMR (150 MHz) spectroscopic data of compounds 1 and 2.
Position
1
2
dC
dH
dC
dH
1
2
3
4
5
6
7
8
129.0
130.1
115.4
156.0
115.4
130.1
35.3
128.9
111.0
149.0
149.6
115.8
122.1
144.7
115.5
7.00 (1H, d, 7.8 Hz)
6.60 (1H, d, 7.8 Hz)
7.25 (1H, d, 1.8 Hz)
6.60 (1H, d, 7.8 Hz)
7.00 (1H, d, 7.8 Hz)
2.71–2.73 (2H, m)
3.51–2.53 (1H, m)
3.77–3.78 (1H, m)
7.20 (1H, d, 8.4 Hz)
7.17 (1H, dd, 8.4, 1.8 Hz)
7.65 (1H, d, 18.0 Hz)
6.45 (1H, d, 18.0 Hz)
70.3
9
3-OCH3
4-OCH3
10
20
30
40
50
60
168.0
55.4
55.4
98.8
70.6
71.5
67.2
74.5
61.4
3.79 (3H, s)
3.90 (3H, s)
4.24 (1H, d, 7.8 Hz)
2.99–3.01 (1H, m)
3.16–3.18 (1H, m)
3.16–3.18 (1H, m)
3.31–3.33 (1H, m)
103.5
74.1
77.0
70.4
77.7
64.1
5.34 (1H, d, 7.8 Hz)
3.71 (1H, dd, 7.8, 3.0 Hz)
4.18 (1H, t, 3.0 Hz)
3.63 (1H, dd, 12.0, 3.0 Hz)
3.87–3.89 (1H, m)
4.23 (1H, dd, 12.0, 6.0 Hz)
4.36 (1H, dd, 12.0, 1.8 Hz)
3.69 (1H, dd, 12.0, 6.0 Hz)
3.86 (1H, dd, 12.0, 2.4 Hz)
100
129.9
107.0
153.2
136.7
153.2
107.0
145.3
117.5
166.9
102.7
73.8
200
7.03 (1H, s)
300
400
500
600
7.03 (1H, s)
7.58 (1H, d, 18 Hz)
6.65 (1H, d, 18 Hz)
700
800
900
1000
2000
3000
4000
5000
6000
5.02 (1H, d, 7.8 Hz)
3.25–3.27 (1H, m)
3.25–3.27 (1H, m)
3.16–3.18 (1H, m)
3.31–3.33 (1H, m)
3.40–3.42 (1H, m)
3.51–3.53 (1H, m)
3.78 (3H, s)
76.9
70.2
76.9
61.1
300-OMe
500-OMe
56.5
56.5
3.78 (3H, s)

Journal of Asian Natural Products Research
733
(20£ 20 cm) was purchased from Merck
(Darmstadt, Germany). ODS (octadecyl-
silyl, 50 mm; YMC Co., Ltd., Kyoto,
Shimogyo-ku, Japan), silica gel (200–
300 mesh; Qingdao Marine Chemical
Industry, Qingdao, China), and AB-8
macroporous adsorption resin (The
Chemical Plant of Nankai University,
Tianjin, China) were used for CC. Spots
were visualized under UV light or by
spraying with vanillic aldehyde–10%
concentrated sulfuric acid ethanol solution
(5:95, v/v) followed by heating. Solvents
[petroleum ether (60–908C), CHCl3,
EtOAc, MeOH, CH2Cl2, and EtOH] were
of analytical grade and purchased from
Beijing Chemical Company, Beijing,
China.
(8:2:0.125, 7:3:0.5, 6:4:1, 5:5:1) to give
10 fractions (fractions 1–10). Fraction 4
(3.0 g) was applied to silica gel CC and
gradiently eluted with CHCl3–MeOH–
H2O (9:1:0.05, 8:2:0.125, 7:3:0.5) to give
three fractions Fr.4-1–Fr.4-3. Fr.4-2
(110 mg) was further purified by prepara-
tive HPLC (CH3OH/H2O 35:65,
10 ml min– 1
)
to give compound
1
(tR ¼ 27.8 min, 30 mg). The fraction
(107 g) eluted with H2O–EtOH (4:6 v/v)
was subjected to silica gel CC and
gradiently eluted with CHCl3 –MeOH
(9:1, 8:2, 7:3, 6:4, 5:5), to give 12 fractions
(fractions 1–12). Fraction 4 (15 g) was
subjected to silica gel CC and gradiently
eluted
with
CHCl3 –MeOH–H2O
(8:2:0.125, 7:3:0.5, 6:4:1) to give Fr.4-1.
Fr.4-1 (83 mg) was further purified by
preparative HPLC (CH3OH/H2O 35:65,
10 ml min– 1
)
to give compound
2
3.2 Plant material
The whole plant of A. tanguticum (col-
lected in Qinghai province, China, in
August 2010) was purchased from Xining
Sanjiangbao Trade Co., Ltd., Xining,
China and identified by Researcher Xue-
Feng Feng of Institute of Chinese Materia
Medica, China Academy of Chinese
Medical Sciences. The specimen (No.
20100813) was deposited in the Institute
of Chinese Materia Medica, China Acad-
emy of Chinese Medical Sciences.
(tR ¼ 39 min, 11 mg).
3.3.1 4-Hydroxyphenethoxy-8-O-b-D-
[6-O- (4-O-b-D-glucopyranosyl)-
sinapoyl]-glucopyranoside
20
White amorphous powder, ½aꢀD –12.0 (c
0.02, MeOH); UV (MeOH) lmax (nm):
304.4; IR (KBr) nmax (cm–1): 3369, 1722,
1639, 1599; for 1H NMR (DMSO-d6,
600 MHz) and 13C NMR (DMSO-d6,
150 MHz) spectroscopic data, see
Table 1; (þ) ESI-MS: m/z 691
[M þ Na]þ; HRTOFMS: m/z 691.2203
[M þ Na]þ (calcd for C31H40O16Na
691.2200).
3.3 Extraction and isolation
Air-dried plants (10 kg) of A. tanguticum
were extracted with 95% EtOH. After
evaporation of the solvent under reduced
pressure, the residue (1.8 kg) was sus-
pended in H2O and extracted successively
with petroleum ether and EtOAc. The
residual aqueous solution was subjected to
CC on macroporous resin AB-8 and eluted
with H2O, H2O–EtOH (7:3 v/v), H2O–
EtOH (4:6 v/v), and H2O–EtOH (0.5:9.5
v/v), successively. The fraction (407.9 g)
eluted with H2O–EtOH (7:3 v/v) was
subjected to silica gel CC and gradiently
3.3.2 3,4-Dimethoxy-trans-cinnamic
acid-9-O-b-D-allopyranoside
20
White amorphous powder, ½aꢀD –20.0 (c
0.02, MeOH); UV (MeOH) lmax (nm):
1
318.0; for H NMR (CD3OD, 600 MHz)
and 13C NMR (CD3OD, 150 MHz) spec-
troscopic data, see Table 1. ESI-MS: m/z
393 [M þ Na]þ, 409 [M þ K]þ, 405
[M þ Cl]–, 739 [2M–H]–; HRESIMS:
m/z 393.1154 [M þ Na]þ (calcd for
eluted
with
CHCl3 –MeOH–H2O

734
Y. Li et al.
409.0892 References
C17H22O9Na
[M þ K]þ, 763.2415 [2M þ Na]þ.
393.1156),
[1] D.S. Luo, Chinese Tibetan Herbal Medi-
cine, (Nationalities Publishing House,
Beijing, 1997), p. 65.
[2] Nanjing University of Traditional Chinese
Medicine, Dictionary of Chinese medi-
cine, (Shanghai Science and Technology
Press, Shanghai, 2005), p. 3564.
[3] Pharmacopoeia Commission of People’s
Republic of China, Pharmacopoeia of
People’s Republic of China. 2010 ed.,
(Chinese Medical Science and Technol-
ogy Press, Beijing, 2010), p. 406, 416,
447.
[4] Pharmacopoeia Commission of People’s
Republic of China, Tibetan medicine
standards of Ministry of health of
People’s Republic of China, (Ministry of
Health of People’s Republic of China,
Beijing, 1995), pp. 110–447.
[5] B.S. Josh, Y.L. Baia, D.H. Chen, and S.W.
Pelletiera, Tetrahedron Lett. 34, 7525
(1993).
[6] S.J. Qu, C.H. Tan, Z.L. Liu, S.H. Jiang, L.
Yu, and D.Y. Zhu, Phytochem. Lett. 4,
144 (2011).
[7] H.Q. Wang, S.H. Jiang, P.M. Yang, M.H.
Ying, S.J. Lin, and D.Y. Zhu, Nat. Prod.
Res. Dev. 14, 13 (2002).
[8] L. Lin, D.L. Chen, X.Y. Liu, Q.H. Chen,
F.P. Wang, and C.Y. Yang, Nat. Prod.
Commun. 4, 897 (2009).
[9] L. Xu, X. Zhang, L.M. Lin, C. Li, Z.M.
Wang, and Y.M. Luo, J. Asian Nat. Prod.
Res. 15, 737 (2013).
3.4 Determination of the absolute
configuration of the sugar moiety
According to the reported method [13],
2 mg of compound 2 was hydrolyzed by
2 M HCl–H2O (2 ml) at 908C for 4 h. After
removal of HCl by evaporation and
extraction with CHCl3, the H2O extract
was evaporated and dried in vacuum to
give the monosaccharide residue. The
residue was dissolved in pyridine (1 ml)
containing L-cysteine methyl ester hydro-
chloride (2 mg) and heated at 608C for 2 h,
then evaporated under N2 stream and dried
in vacuum. The residue was dissolved in
0.2 ml of N-trimethylsilylimidazole and
heated at 608C for 1 h. The reaction
mixture was partitioned between n-hexane
and H2O (2 ml), and the n-hexane extract
was analyzed by GC (Agilent 7890A)
under the following conditions: capillary
column HP-5 (30 m £ 0.32mm£ 0.25mm);
FID detector; carrier gas N2 (flow rate ¼ 1-
mlmin–1); detector temperature (2808C);
injection temperature (2508C), and oven
temperature
gradient:
100–2808C
(108C min–1), and 2808C for 5 min. The
same procedure was applied to authentic
samples. By comparison with the retention
[10] L. Xu, Y.R. Li, C. Li, L.M. Lin, Z.M.
Wang, and Y.M. Luo, Chin. Med. Mater.
38, 2818 (2013).
time of the authentic sample (tR- -allose,
20.110min; tR- -allose, 20.415 min), D-allose
[11] L. Xu, M. Luo, L.M. Lin, X. Zhang, C. Li,
Z.M. Wang, and Y.M. Luo, J. Asian Nat.
Prod. Res. 15, 743 (2013).
[12] M. Luo, L.M. Lin, C. Li, Z.M. Wang, and
W.B. Guo, Chin. Med. Mater 37, 1245
(2012).
D
L
(tR 20.109min) was identified in the acid
hydrolysate of compound 2.
[13] J. Bai, Z.F. Fang, H. Chen, S.S. Yu, D.
Zhang, H.L. Wei, S.G. Ma, Y. Li, J. Qu, S.
Xu, J.H. Ren, F. Zhao, and J.H. Liu,
Carbohydr. Res. 361, 206 (2012).
[14] S.H. Yim, H.J. Kim, N. Jeong, K.D. Park,
Y.J. Lee, S.D. Cho, and I.S. Lee, Bull.
Korean Chem. Soc. 33, 1253 (2012).
Acknowledgements
This project was financially supported by
Science
Beijing
Natural
Foundation
(7132152) and the Fundamental Research
Funds for the Central Public Welfare Research
Institutes (ZZ070828).