From the roots of Codonopsis pilosula (Franch) Nannf (Campanulaceae) five triterpenoids: taraxerol, taraxeryl acetate, 14-a-taraxeran-3-one and D:B-friedoolean-5-ene-3-b-ol as well as a-spinasterone were isolated. Their structures have been identified by MS, 1H-, 13C-NMR spectroscopy and comparison with reported data

Sung et al, Tran Van

Vietnam Academy of Science and Technology: Journals Online

Journal of Chemistry, Vol. 46 (4), P. 515 - 520, 2008 
 
TRITERPENES FROM THE ROOTS OF CODONOPSIS PILOSULA 
Received 30 October 2007 
Trinh Thi Thuy1, Tran Van Sung1, Katrin Frank2 and  Ludger Wessjohann2
1Institute of Chemistry, Vietnam Academy of Science and Technology 
2Leibniz-Institute of Plant Biochemistry,Weinberg 3, D-06120 Halle/Saale, Germany 
 
Summary 
 From the roots of Codonopsis pilosula (Franch) Nannf (Campanulaceae) five triterpenoids: 
taraxerol, taraxeryl acetate, 14-α-taraxeran-3-one and D:B-friedoolean-5-ene-3-β-ol as well as 
α-spinasterone were isolated. Their structures have been identified by MS, 1H-, 13C-NMR 
spectroscopy and comparison with reported data.  
 
I - Introduction 
Codonopsis pilosula (Franch) Nannf is one 
of the most famous traditional medicine and 
sometimes as cheaper substitute like ginseng in 
Chinese and Vietnamese drugs. It has been used 
for a long time and still being used widely today 
as a remedy for appetite, psychoneurosis, 
fatigue, dyspepsia and possessing adaptogenic, 
anti-stress properties [1]. Our previous study on 
the roots of C. pilosula (Franch) Nannf has led 
to the isolation and structural identification of 
lobetyol, 5-hydroxymethyl-2-furandehyde as 
well as bis-(2-ethylhexyl)-phthalate [2]. In this 
paper we report the isolation and structural 
elucidation of four triterpenoids, taraxerol (1), 
taraxeryl acetate (2), 14-α-taraxeran-3-one (3-
taraxeranone, 3), D:B-friedoolean-5-en-3-α-ol 
(4) and α-spinasterone (5).  
II - Experiment 
1. General  
Optical rotation [α]D: Digital Polarimeter 
Jasco DIP 1000. EI-MS: ADM 402, 70 eV, 
Finnigan TSQ 700. NMR: VARIAN 300 
spectrometer at 300 MHz (1H) and 75.5 MHz 
(13C, 13C-APT). Chemical shifts were referenced 
to internal TMS (δ = 0, 1H) and CDCl3 (δ = 77.0, 
13C). CC: Silica gel 60, 0.06 - 0.20 mm (Merck) 
for the first column, silica gel 60, 40-63 μm 
(Merck) for the following columns. TLC: Silica 
gel 60 F-254 (Merck). 
2. Plant material  
The roots of C. pilosula were bought in 
Hanoi market, Vietnam in May 2005. The 
species was identified by Dr. Ngo Van Trai, 
Institute of Materia Medica, Hanoi. A voucher 
specimen (Nr. 1) is deposited in the Institute of 
Chemistry, VAST, Hanoi.  
3. Extraction and isolation 
The ground and dried roots of C. pilosula 
(2.4 kg) were extracted four time with  MeOH 
(95%) at room temperature. MeOH was 
evaporated in vacuo, and the aq. solution (1.25 
kg) was partitioned with n-hexane followed by 
EtOAc and n-BuOH (each four time), giving 
45.0 g, 21.0 g and 29.3 g extracts, respectively. 
The n-hexane extract was separated on silica gel 
using n-hexane-CHCl3 (20:80→90:10) and then 
CHCl3-MeOH (98:2 → 90:10) to afford 24 
fractions (F1-F24, 150 ml/Fr.). Compounds 1-5 
were further purified by CC on silica gel or
 515
 crystallization. 
Table 1: 1H-NMR data of taraxeryl acetate (2) and α-spinasterone (5)  
(300 MHz, δppm, CDCl3, J in Hz) 
H 2 5 5 [8] 
1 1.39 m 1.92 dd (14.6; 3.1) 1.46 m; 2.13 ddd (6.1; 
14.6; 14.6) 
1.47 m; 2.12 ddd (6.1; 
14.6; 14.6) 
2 1.66 m; 2.02 m 2.28 br d (14.5) 2.28 br d (15) 
3 4.45 dd (10.3; 6.2) - - 
4 - 2.24  m 2.23  m 
5 1.33 m 1.80  m 1.81  m 
6 1.43 m, 1.46 m 1.83  m 1.82  m 
7 1.29 m,1.62 m 5.18  m 5.18  m 
9 1.54 m 1.77  m 1.76  m 
11 1.42-1.46 m 1.56, 1.75 m 1.55, 1.75 m 
12 0.93 - 1.33 m 1.27, 2.04 1.27, 2.04 m 
14 - 1.83 m 1.83 m 
15 5.53 dd (7.0; 3.2) 1.39 m, 1.50 m 1.40 m, 1.52 m 
16 1.65 m, 2.01 m 1.29 m, 1.77 m 1.29 m, 1.67 m 
17 - 1.30 m 1.30 m 
18 1.03 m 0.58 s 0.58 s 
19 1.43 m, 2.05 m 1.02 s 1.02 s 
20 - 2.04 m 2.05 m 
21 1.32 - 1.42 m 1.03 d (6.8) 1.03 d (6.7) 
22 1.32 - 1.42 m 5.13 dd (8.4; 15.0) 5.16 dd (8.5; 15.2) 
23 0.95 s 5.02 dd (8.5; 15.0) 5.02 dd (8.8; 15.3) 
24 0.82 s 1.55 m 1.56 m 
25 0.91 s 1.56 m 1.57 m 
26 1.09 s 0.82 d (6.2) 0.82 d (6.1) 
27 0.88 s 0.84 d (6.4) 0.84 d (6.7) 
28 0.86 s 1.19 m;  1.41 m 1.18 m;  1.41 m 
29 0.95 s 0.81 t (6.4) 0.81 t (7.3) 
30 0.91 s - - 
COCH3 2.04 s - - 
 
 
 516
Table 2: 13C-NMR data of  compounds 1-5 (CDCl3, 75.5 MHz, δppm) 
C 1* 2 3 4 5
1 38.56 37.73 38.31 18.30 38.79
2 26.60 28.86 28.22 27.88 38.15
3 78.65 80.98 212.96 76.32 211.68
4 38.83 39.01 42.16 40.86 44.26
5 55.39 55.63 53.08 141.45 42.88
6 18.68 18.76 18.30 121.96 30.10
7 35.63 35.15 35.36 23.71 116.88
8 38.55 37.92 39.28 47.45 139.33
9 48.56 48.76 42.79 34.88 48.85
10 37.81 37.72 37.45 49.70 34.44
11 17.39 17.59 18.30 34.65 21.76
12 36.51 35.83 35.65 30.41 39.35
13 37.54 37.41 36.03 39.33 43.37
14 157.76 157.78 58.19 37.87 55.02
15 116.56 116.84 30.03 32.12 23.02
16 37.40 36.70 32.79 36.07 28.55
17 37.62 37.58 39.70 30.15 55.85
18 49.68 49.20 59.44 43.08 11.98
19 41.17 41.23 41.55 35.12 12.34
20 28.64 28.86 28.22 28.32 40.86
21 34.95 33.72 32.44 33.16 21.76
22 33.55 33.13 30.54 38.99 137.92
23 27.75 28.04 18.74 29.02 129.37
24 15.33 15.59 6.93 25.53 51.25
25 15.33 16.67 14.73 16.30 31.91
26 29.63 29.99 18.74 19.70 19.06
27 25.76 25.99 32.13 18.52 21.45
28 29.73 29.89 31.83 32.10 25.45
29 33.13 33.41 35.07 34.59 12.34
30 21.15 21.42 20.33 32.47 -
C=O - 170.82 - - -
COCH3 - 21.36 - - -
* CDCl3:CD3OD (95:5) 
 
a) Taraxerol (1)  
Fractions 12-13 (Fr.12-14, 1.35g) were 
separated on silica gel, eluted with hexane-
EtOAc (90:10) and then crystallization to give 
360 mg (0.0150 %). [α]24D -22O (c 1.0, CHCl3). 
EI-MS 70 eV, m/z (rel. int.): 426 [M]+ (16), 411 
 517
[M-15]+  (14), 302 (52), 287 (40), 218 (37), 205 
(38), 204 (100), 189 (30), 135 (19), 121 (24), 
107 (22), 95 (23); ). 1H-NMR (CDCl3, 300 MHz, δ ppm): 0.97 (Me-23), 0.82 (Me-24), 0.95 (Me-
25), 1.09 (Me-26), 0.91 (Me-27), 0.80 (Me-28), 
0.93 (Me-29), 0.91 (Me-30). 13C-NMR (125 
MHz, CDCl3+CD3OD): data see table 2.  
b) Taraxeryl acetate (2)  
Compound 2 was isolated from Fr. 4+5 by 
CC (silica gel, n-hexane-EtOAC 98:2). White 
needles from EtOAc, yield 60 mg (0.0032 %). 
ESI-MS (m/z): 491 [M+Na]+ (C32H52O2). EI-MS, 
70 eV, m/z (rel. int.): 468 [M]+ (41), 453 (24), 
408 (20), 344 (60), 329 (26), 269 (24), 218 (40), 
204 (100), 189 (30), 135 (18), 121 (17), 109 
(21) 107 (10), 95 (24). 1H- and 13C-NMR data 
see Table 1 and 2.  
c) 3-Taraxeranone (14-α-taraxeran-3-one, 3) 
Compound 3 was isolated from Fr.10 by CC 
(silica gel, n-hexane-EtOAC-CHCl3, 90:10:1). 
White needles from EtOAc, yield 56mg (0.0023 
%). [α]24D – 45 (c 2, CHCl3, lit. [5] + 31O). ESI-
MS, m/z: 449 [M+Na]+ (C30H50O).
 EI-MS, 70 
eV, m/z (rel. int.): 426 [M]+ (87), 411 (29), 341 
(14), 302 (32), 273 (63), 205 (54), 191 (17), 179 
(46), 163 (51), 123 (100), 109 (77), 95 (82), 69 
(47);  1H-NMR (CDCl3, 300 MHz, δ ppm): 1.05 
(Me-23), 1.00 (9H, Me-24, Me-25, Me-29), 
1.18 (Me-26), 0.87 (Me-27), 0.88 (Me-28), 0.95 
(Me-30). 13C-NMR data see table 2.  
d) D:B-friedoolean-5-ene-3-β-ol (4)  
Compound 4 was isolated from Fr.10 and 
purified by CC [silica gel, n-hexane-EtOAc-
CHCl3 (90:10:1)]. Powder from EtOAc, yield 28 
mg (0.0012 %). EI-MS, 70 eV, m/z (rel. int.): 
426 [M]+ (18), 409 (30), 274 (C20H34, 100), 259 
(92), 205 (C14H21O, 47), 137 (C10H17, 30), 134 
(45), 109 (46), 95 (42), 81 (23); 1H-, 13C-NMR 
data see Table 1 and 2. 
e) α-Spinasterone [(22E)-5α-stigmasta-7,22-
diene-3-one (5)]  
Compound 5 was isolated from Fr.10 and 
purified by CC [silica gel, n-hexane-EtOAC-
CHCl3 (90:10:5)]. White needles from EtOAc, 
yield 45 mg (0.0019 %). EI-MS 70 eV, m/z (rel. 
int.): 410 [M]+ (56), 397 (32), 395 [M-Me]+, 367 
[M-C3H7]
+ (52), 298 [M-C8H16]
+ (32), 271, 269 
(loose of the side chain, 75), 257 (13), 244 (38), 
229 (77), 95 (40), 83 (22), 55 (24). 13C-NMR 
data see table 2. 
III - Results and Discussion 
The residue of an ethanol extract of C. 
pilosula was partitioned with n-hexane, EtOAc, 
n-BuOH, successively. The n-hexane extract, 
after evaporation of solvent was 
chromatographed on column over silica gel and 
then crystallization to afford compounds 1-5. 
Compounds 1-5 showed no fluorescence under 
UV light with λmax 254 and 366 nm. 
The EI-MS spectrum of 1 gave a mol peak at 
m/z 426 [M]+, corresponding to the molecular 
formula C30H50O. The APT
 and 13C-NMR spectra 
showed the presence of 30 carbons (CH3x8, 
CH2x10, CHx5, Cqx7), suggested that 1 has a 
triterpene skeleton. This was further confirmed 
by the signals of 8 tertiary methyl signals in the 
1H-NMR spectrum. One double bond was 
confirmed by olephinic methine signal at δH 
5.53 (dd, J = 7.0; 3.2 Hz) and δC  157.76 (C-14), 
116.56 (C-15). The structure of 1 was identified 
as taraxerol (14-taraxeren-3β-ol) by comparison 
of its 1H- and 13C-NMR spectra (Table 1) with 
reported data [3, 4]. Taraxerol showed antiulcer 
activity and is a cancer chemopreventive agent. 
It was isolated for the first time from 
Taraxacum officinale and then frequently found 
in other plants (Rhododendron spec., Euphorbia 
spec.) [3, 4]. 
Compound 2 was obtained as white needles 
from n-hexane extract using column 
chromatography on silica gel. The EI-MS 
spectrum gave a mol peak at m/z 468 [M]+ (41), 
corresponding to the molecular formula 
C32H52O2. The 
1H- and 13C-NMR spectral data 
were similar to those of 1, except the presence 
of an acetyl group (δH 2.04 and δC 170.28, 
21.36), therefore 2 was identified as taraxeryl 
acetate. Its 1H- and 13C-NMR spectral data were 
identical to reported data [4]. 
Compound 3 was obtained as white needles. 
The molecular formula of 3 was established as 
 518
C30H50O by combination of 
13C-NMR and mol 
peak at m/z 426 [M]+(87). The 13C-NMR 
spectrum showed the presence of a keton group 
(δC 212.96) and an absence of olephinic signals, 
suggesting that 3 is taraxeranone. In 
combination of its MS and NMR spectra, the 
structure of 3 was determined as 14α-taraxeran-
3-one (3-taraxeranone) [5]. 
  
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19 20 21
22
23 24
H
H
25 26
27
28
29 30
RO
1: R = H   Taraxerol
2: R = Ac Taraxeryl acetate
H
 
H
H
H
3:14-α-Taraxeran-3-one
H
O
 
1
2
3
4 5
6
7
8
9
10
11
12
13
14
15
16
17
18
19 20 21
22
23 24
25 26
27
29 30
HO
H
HH
4: D:B-fiedoolean-5-ene-3-β-ol
28
 
5: α-Spinasterone
H
O
3 5
17
18
22
7
23
24
25
26
27
28
21
19
20
29
2
4 6
89
10
11
12
13
14 15
16
1
 
  
  
The EI-MS spectrum of 4 gave a mol peak at 
m/z 426 [M]+(18), indicating a same molecular 
formula (C30H50O) as 1. The APT
 and 13C-NMR 
spectra showed the presence of 30 carbons 
(CH3x8, CH2x10, CHx5, Cqx7). In comparison 
of the 1H- and 13C-NMR spectral data (table 2) 
with reported data [6], the structure of 4 was 
identified as D: B-friedoolean-5-ene-3-β-ol, 
which was isolated for the first time from 
Securinega tinctorena and its isomer (D:B-
friedoolean-5-ene-3-α-ol) was found in 
Euphorbia royleana (Euphorbiaceae) [6, 7]. 
D:B-friedoolean-5-ene-3-β-ol is a relatively rare 
triterpene alcohol, which can be an intermediate 
in the biosynthesis of friedeline.  
The molecular formula of 5 (C29H46O) was 
determined by combination of molecular ion 
peak at m/z 410 [M]+ in EI-MS as well as its 13C-
NMR spectra. The 1H-NMR spectrum displayed 
two methyl doublets at δH 0.82, 0.84 (each 3H, 
d, J = 6.4 Hz), corresponding to one isopropyl 
group. The mass spectrum showed a fragment at 
m/z 269 (loss of the side chain) and fragments 
were common to related steroids, suggested that 
5 is a stigmasta-diene skeleton. This was further 
confirmed by the presence of three olephinic 
methine carbons at δC 116.88 (C-7), 137.92 (C-
22), 129.37 (C-23) and quaternary carbon at δC 
139.33 (C-8) in the 13C-NMR spectrum. The 1H-
NMR spectrum showed the presence of two 
olephinic protons at δH 5.13, 5.02 with trans 
configuration (J = 15.0 Hz). Combination of the 
MS, 1H- and 13C-NMR spectra (tables 1&2), the 
structure of 5 was identified as (22E)-5α-
stigmasta-7,22-diene-3-one (α-spinasterone), 
which was isolated from the heartwood of 
Albizzia julibrissin and the bark of Acacia 
concinna [8].  
 519
The presence of taraxerol as main component 
and its derivatives is a chemical support for the 
taxonomy of Codonopsis species used in 
Vietnamese traditional medicine. 
Acknowledgements: We thank BMBF, 
Germany, for financial support in form of a 
project. We are indebted Dr. Ngo Van Trai, 
Institute of Materia Medica, Hanoi for the 
identification of the plant material. 
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1. Heidrun Noerr, H. Wagner. Planta Medica, 
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3. Atta UR-Rahman, Nighat Sultana, Parzana 
Akhter, Parzana Nighat, M. Iqbal Choudary.  
Nat. Prod. Lett. 10, 249-256 (1997).  
4. Damon R. Billodeaux, Gloria A. Benavides. 
Acta Cryst. 55, 2129 - 2131 (1999). 
5. Yueh-Hsiung Kuo, Pei-Hung Chu. Journal 
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6. L. Carvalho, J. Seita. Nat. Prod. Lett. 2 (1), 
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Triterpene from the roots of Codonopsis pilosula.

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Triterpene from the roots of Codonopsis pilosula.

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