A novel hexanordammarane glycoside from the leaves and stems of Panax quinquefolium L.

Article abstract of DOI:10.1080/14786419.2010.551768

One novel hexanordammarane glycoside, ginsenoside R₁₀, was isolated from the leaves and stems of Panax quinquefolium L. as a minor constituent. It is the first time that a hexanordammarane glycoside isolated from the plant of Panax quinquefolium L. Its structure was elucidated as 3-O-β-D-glucopyranosyl-3β, 12β-dihydroxy-22, 23, 24, 25, 26, 27-hexanordammarane-20-one (1), by the combination analysis of one-dimensional NMR, two-dimensional NMR and mass spectrometry.

Full text of DOI:10.1080/14786419.2010.551768

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Natural Product Research: Formerly
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A novel hexanordammarane glycoside
from the leaves and stems of Panax
quinquefolium L.
Jin-Ping Liu a , Dan Lu a & Ping-Ya Li a
a Institute of Frontier Medical Science, Jilin University ,
Changchun 130021 , China
Published online: 20 Oct 2011.
To cite this article: Jin-Ping Liu , Dan Lu & Ping-Ya Li (2012) A novel hexanordammarane glycoside
from the leaves and stems of Panax quinquefolium L., Natural Product Research: Formerly Natural
Product Letters, 26:8, 744-748, DOI: 10.1080/14786419.2010.551768
To link to this article: http://dx.doi.org/10.1080/14786419.2010.551768
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Natural Product Research
Vol. 26, No. 8, April 2012, 744–748
A novel hexanordammarane glycoside from the leaves and stems
of Panax quinquefolium L.
*
Jin-Ping Liu, Dan Lu and Ping-Ya Li
Institute of Frontier Medical Science, Jilin University, Changchun 130021, China
(Received 23 August 2010; final version received 30 December 2010)
One novel hexanordammarane glycoside, ginsenoside R10, was isolated
from the leaves and stems of Panax quinquefolium L. as a minor
constituent. It is the first time that a hexanordammarane glycoside isolated
from the plant of Panax quinquefolium L. Its structure was elucidated as
3-O-ꢀ-D-glucopyranosyl-3ꢀ, 12ꢀ-dihydroxy-22, 23, 24, 25, 26, 27-hexanor-
dammarane-20-one (1), by the combination analysis of one-dimensional
NMR, two-dimensional NMR and mass spectrometry.
Keywords: Panax quinquefolium L.; hexanordammarane; ginsenoside R10
1. Introduction
It was reported that the leaves and stems of Panax quinquefolium L. showed similar
medical effects as the roots of Panax quinquefolium L. such as anti-hemorrhagic
shock, protective effect against liver injury and anticancer (Mo, 2001; G. Y. Li, Zeng,
Meng, X. Li, & Wang, 2009). The chemical compositions of the leaves and stems of
P. quinquefolium were also investigated in the recent research such as quinquenoside
L3 (Wang, W. Li, & X. Li, 1998), quinquenoside L9 (Wang et al., 2001),
quinquenosides L10, L14 and L16 (Chen et al., 2009). As part of an ongoing research
on the chemical constituents of the leaves and stems of Panax quinquefolium L., this
article describes the isolation and the structural elucidation of a new hexanordam-
marane minor constituent, ginsenoside R10 (1), by the chemical and spectroscopic
methods (1D and 2D NMR, MS). The structure of 1 has been determined as 3-O-ꢀ-
D-glucopyranosyl-3ꢀ, 12ꢀ-dihydroxy-22, 23, 24, 25, 26, 27-hexanordammarane-20-
one. It is the first time that a hexanordammarane glycoside is isolated from the plant
of Panax quinquefolium L.
2. Results and discussion
Repeated CC of the n-BuOH extract partition of the water extract of the leaves and
stems of Panax quinquefolium L. led to the isolation of a new hexanordammarane
glycoside (1).
*Corresponding author. Email: lipy@jlu.edu.cn
ISSN 1478–6419 print/ISSN 1478–6427 online
ß 2012 Taylor & Francis
http://dx.doi.org/10.1080/14786419.2010.551768
http://www.tandfonline.com

Natural Product Research
745
Figure 1. The structure and HMBC correlations of glycoside 1.
Compound 1 was obtained as white amorphous solid (MeOH) that gave
positive reaction to the Libermann–Burchard test. HR-ESI-MS: m/z 537.3447
[M ꢀ H]ꢀ (calcd for 537.3506), indicated the molecular formula of 1 to be
C30H50O8. Acid hydrolysis of 1 with 1.0 M aqueous HCl yielded D-glucose as the
only sugar component. The 1H NMR spectra of 1 displayed six methyl singlet
signals at ꢁ 0.76 (3H, s), ꢁ 0.92 (3H, s), ꢁ 0.95 (3H, s), ꢁ 0.98 (3H, s), ꢁ 1.31 (3H, s),
ꢁ 2.40 (3H, s). The sugar was determined as ꢀ-glucose by the proton (ꢁ 4.94, d,
1
J ¼ 7.5 Hz) in the H NMR spectrum and an anomeric carbon signal at ꢁ 107.1,
respectively. Except for the signals of a glucopyranosyl unit, the aglycone of
compound 1 revealed the presence of five tert-methyls, seven methylenes, six
methines (two of them were oxygen substituted), four quaternary carbons as well
as one methyl ketone. In comparison with a normal dammarane saponin
ginsenoside Rh2 (Figure 1), isolated from the roots of Panax quinquefolium L. and
ginseng whose aglycone was protopanaxadiol, the chemical shifts of 1 showed
close resemblance with those of ginsenoside Rh2 (Kitagawa, Yoshikawa, &
Yoshihara, 1983), except the signals caused by the side-chain. In the case of
glycoside 1, the presence of a methyl ketone (ꢁ 213.5 and ꢁ 30.7) was observed,
instead of the carbon signals of the side-chain (C-20, 21, 22, 23, 24, 25, 26 and
27). The side-chain’s chemical shifts of 1 showed close resemblance with those of
notoginsenoside R10 (Li, Teng, & Yang, 2001) and 3ꢀ,6ꢂ,12ꢀ-triol-22, 23, 24, 25,
26, 27-hexanordammaran-20-one (Wu et al., 2007). The 13C NMR data of 1
compared with ginsenoside Rh2 and notoginsenoside R10 suggested the presence
of a methyl ketone as notoginsenoside R10.
In the HMBC spectrum, the long-range correlation between the methyl proton
signal at ꢁ 2.40 (H-21) and the carbon signals at ꢁ 213.5 (C-20) and ꢁ 52.9 (C-17)
could be identified. Moreover, the HMBC spectrum also showed the long-range
correlation between the proton signal at ꢁ 2.37 (H-13) and the carbon signals at ꢁ 71.4
(C-12), ꢁ 51.6 (C-14) and ꢁ 213.5 (C-20). In addition, the long-range correlation
between the proton signal at ꢁ 0.92 (3 H, s, H-30) and the carbons at ꢁ 33.0 (C-15),
40.3 (C-8), ꢁ 0.95 (3 H, s, H-18) and ꢁ 40.3 (C-8), 35.6 (C-7), 51.6 (C-14) could also be
identified in the HMBC correlations. So, the aglycone of 1 was considered to have a
hexanor- dammarane skeleton with carbonyl oxygen linked on C-20. The location of
the O-ꢀ-D-glucopyranosyl unit was determined to be on C-3 of the aglycone by
2DNMR spectra (Figure 2), and by analysis of two-dimensional NMR spectra, the
proton and carbon signals of 1 were assigned based on the above evidence. The
structure of 1 could be characterised as 3-O-(ꢀ-D-glucopyranosyl)-3ꢀ, 12ꢀ-
dihydroxy-22, 23, 24, 25, 26, 27-hexanordammarane-20-one. Compound 1 is a

746
J.-P. Liu et al.
Figure 2. The structures of ginsenoside Rh2 and notoginsenoside R10.
minor glycoside in the leaves and stems of Panax quinquefolium L. It is the
first time that a hexanordammarane glycoside is isolated from the plant of
Panax quinquefolium L. From the viewpoint of biogenetic pathway, this
special nortriterpenoid derivative may correspond to a pregnane skeleton
(Li et al., 2001).
3. Experimental
3.1. General experimental procedures
HR-ESI-MS spectrum was recorded by using Ionspec 7.0 TFT-ICR-MS (IonSpec
Corporation, Lake Forest, CA, USA). NMR spectra were measured at 500 MHz for
1H NMR, 125.8 MHz for 13C NMR, and 500 MHz for HMBC and HMQC on a
Bruker Avance-500 spectrometer (Karlsruhe, Germany). NMR spectra were
measured in pyridine-d5 by using TMS as the internal standard (Cambridge
Isotope Laboratories, Inc., Andover, MA, USA). Chemical shifts (ꢁ) are expressed in
ppm. Preparative HPLC was carried out on a 2998 Photodiode Array Detector and
SunFire Prep C18 Column (10 mm, 10 ꢁ 150 mm), 1525 BINARY HPLC PUMP
(Waters), the mobile phase was MeOH–H2O (73 : 27), flow rate: 1.0 mL minꢀ1
,
detection wavelength: 203 nm. Silica gel H (200–300 mesh; Qingdao Marine
Chemical, Inc., Qingdao, China) was used in column chromatography. Also, silica
gel G plates (Qingdao Marine Chemical, Inc.) were used in thin layer
chromatography.
3.2. Plant material
The leaves and stems of P. quinquefolium L. were collected in the Jingyu County of
Jilin Province (China) in September 2007, and identified by Prof Jin-Min Zhang.
A voucher specimen (No. 20070908) has been deposited at the Institute of Frontier
Medical Science, Jilin University, China.

Natural Product Research
747
3.3. Extraction and isolation
Air-dried leaves and stems of P. quinquefolium L. (2.5 kg) were extracted with hot
water (25 L ꢁ 3) and the water soluble fraction was extracted with CHCl3 and
n-BuOH in turn. The n-BuOH extract part was subjected to macro-reticular
absorption resin (D101), and was eluted with H2O (50 L) and 95% EtOH (50 L), and
then afforded the EtOH fraction (388 g). A part of the EtOH fraction (150 g) was
subjected to Si-gel CC eluting with CHCl3–MeOH mixture to give 120 fractions.
Fractions No. 78–83 was then subjected to preparative HPLC MeOH–H2O (65 : 35)
to obtain compound 1 (11 mg, 0.007%).
3.4. NMR data of compound 1
1
Compound 1: H NMR (500 MHz, pry-d5): ꢁ 1.49 (1H, m, H-1e), ꢁ 0.79 (1H, m, H-
1a), ꢁ 2.23 (1H, m, H-2e), ꢁ 1.82 (1H, m, H-2a), ꢁ 3.37 (1H, dd, J ¼ 11.5, 4.5 Hz, H-3),
ꢁ 0.72 (1H, m, H-5), ꢁ 1.51 (1H, m, H-6e), ꢁ 1.35 (1H, m, H-6a), ꢁ 1.45 (1H, m, H-7e),
ꢁ 1.19 (1H, m, H-7a),ꢁ 1.40 (1H, m, H-9), ꢁ 1.37 (1H, m, H-11e), ꢁ 1.24 (1H, m,
H-11a), ꢁ 3.83 (1H, m, H-12), ꢁ 2.37 (1H, d, J ¼ 9.0 Hz, H-13), ꢁ 1.96(1H, m, H-15e),
ꢁ 1.40 (1H, m, H-15a), ꢁ 1.97 (1H, m, H-16e), ꢁ 1.78 (1H, m, H-16a), ꢁ 3.08 (1H, m,
H-17), ꢁ 0.95 (3H, s, H-18), ꢁ 0.76 (3H, s, H-19), ꢁ 2.40 (3H, s, H-21), ꢁ 1.31 (3H, s,
H-28), ꢁ 0.98 (3H, s, H-29), ꢁ 0.92 (3H, s, H-30), ꢁ 4.94 (1H, d, J ¼ 7.5 Hz, H-10), ꢁ 4.04
(1H, m, H-20), ꢁ 4.24 (1H, m, H-30), ꢁ 4.22 (1H, m, H-40), ꢁ 4.02 (1H, m, H-50), ꢁ 4.59
(1H, m, H-60), ꢁ 4.41 (1H, m, H-60).
13C NMR (125.8 MHz, pyr-d5): ꢁ 39.3 (C-1), ꢁ 26.8 (C-2), ꢁ 88.8 (C-3), ꢁ 39.8 (C-4),
ꢁ 56.5 (C-5), ꢁ 18.5 (C-6), ꢁ 35.6 (C-7), ꢁ 40.3 (C-8), ꢁ 51.1 (C-9), ꢁ 37.2 (C-10), ꢁ 30.1
(C-11), ꢁ 71.4 (C-12), ꢁ 55.0 (C-13), ꢁ 51.6 (C-14), ꢁ 33.0 (C-15), ꢁ 27.7 (C-16), ꢁ 52.9
(C-17), ꢁ 15.8 (C-18), ꢁ 16.6 (C-19), ꢁ 213.5 (C-20), ꢁ 30.7 (C-21), ꢁ 28.2 (C-28), ꢁ 16.8
(C-29), ꢁ 17.1 (C-30), ꢁ 107.1 (C-10), ꢁ 75.9 (C-20), ꢁ 78.9 (C-30), ꢁ 72.0 (C-40), ꢁ 78.5 (C-
50), ꢁ 63.2(C-60).
Acknowledgements
This work was financially supported by the Supporting Program of the ‘Eleventh Five-year
Plan’ for Sci & Tech Research of China (No. 2007BAI38B04). We are grateful to Professor
Zhang Jing-min for identification of the plant material. We are also very grateful to Chun-Yu
Wang (Key Laboratory for Supramolecular Structure and Materials of Ministry of Education,
Jilin University) for the measurement of NMR.
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