New dammarane-type saponins from the roots of panax notoginseng

Article abstract of DOI:10.1002/hlca.201300155

Three new dammarane-type triterpenoid saponins, 1-3, were isolated and identified as (20S)-20-O-[β-D-xylopyranosyl-(1→6)-β-D- glucopyranosyl-(1→6)-β-D-glucopyranosyl]dammar-24-ene-3β, 6α,12β, 20-tetrol (1), (20S)-6-O-[(E)-but-2-enoyl-(1→6)-β- D-glucopyranosyl]dammar-24-ene-3β,6α,12β,20-tetrol (2), and (20S)-6-O-[β-D-xylopyranosyl-(1→2)-β-D-xylopyranosyl] dammar-24-ene-3β,6α,12β,20-tetrol (3) from the roots of Panax notoginseng (Burkill) F.H.Chen (Araliaceae). Their structures were elucidated on the basis of spectroscopic analyses, including 1D- and 2D-NMR techniques and HR-ESI-MS, as well as by acidic hydrolysis. Copyright

Full text of DOI:10.1002/hlca.201300155

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New Dammarane-Type Saponins from the Roots of Panax notoginseng
by Li Qiua)b)c), Yang Jiaoc), Gui-Kun Huangc), Ji-Zhao Xiec), Jian-Hua Miaob),
and Xin-Sheng Yao*a)
a) Institute of Traditional Chinese Medicine & Natural Products, Jinan University, Guangzhou 510632,
Guangdong, P. R. China (phone/fax: þ 86-20-85225849; e-mail: tyaoxs@jnu.edu.cn)
b) Institute of Guangxi Medicinal Plant, Nanning 530023, Guangxi, P. R. China
c) School of Pharmaceutical Science, Guangxi Medical University, Nanning 530021, Guangxi, P. R. China
Three new dammarane-type triterpenoid saponins, 1 – 3, were isolated and identified as (20S)-20-O-
[b-d-xylopyranosyl-(1 ! 6)-b-d-glucopyranosyl-(1 ! 6)-b-d-glucopyranosyl]dammar-24-ene-3b,6a,12b,
20-tetrol (1), (20S)-6-O-[(E)-but-2-enoyl-(1 ! 6)-b-d-glucopyranosyl]dammar-24-ene-3b,6a,12b,20-te-
trol (2), and (20S)-6-O-[b-d-xylopyranosyl-(1 ! 2)-b-d-xylopyranosyl]dammar-24-ene-3b,6a,12b,20-te-
trol (3) from the roots of Panax notoginseng (Burkill) F.H.Chen (Araliaceae). Their structures were
elucidated on the basis of spectroscopic analyses, including 1D- and 2D-NMR techniques and HR-ESI-
MS, as well as by acidic hydrolysis.
Introduction. – All saponins present in the roots of Panax notoginseng, as well as
their derivatives, have been approved by the State Food and Drug Administration in
China as clinical drugs, which are widely used in the prevention and treatment of
cardiovascular diseases. Recent pharmaceutical studies have disclosed diverse bio-
activities of the saponins from Panax notoginseng, such as anti-inflammatory [1][2],
antitumor [3][4], antioxidant [5], hepatoprotective [6], immunomodulative, and
immune-adjunctive activities [7]. A detailed phytochemical investigation of the root of
Panax notoginseng was carried out in the present work. As a result, three new
dammarane-type saponins, 1 – 3, one natural compound, 7, and other 20 known
dammarane-type saponins, 4 – 6 and 8 – 24, were isolated and identified, of which 14 and
15 were isolated for the first time.
Results and Discussion. – The 80% EtOH extract of the air-dried root of Panax
notoginseng was chromatographed repeatedly to afford compounds 1 – 24 (Figs. 1 and
2). Three new minor saponins, 1 – 3, and one new natural compound, 6’-O-
acetylginsenoside Rh1 (7) [8], along with 20 known compounds, 6-O-(b-d-glucopyr-
anosyl)-20-O-(b-d-xylopyranosyl)-3b,6a,12b,20(S)-tetrahydroxydammar-24-ene (4)
[9], (20S)-ginsenoside Rh1 (5) [10], (20R)-ginsenoside Rh1 (6) [11], (20S)-ginsenoside
Rg2 (8) [12], notoginsenoside-R2 (9) [10], ginsenoside-F1 (10) [13], ginsenoside Rg1
(11) [14], notoginsenoside-R1 (12) [10], ginsenoside Re (13) [15], (20S)-protopanaxa-
triol-20-O-b-d-glucopyranosyl-(1 ! 6)-b-d-glucopyranoside (14) [16], ginsenoside-Rh4
(15) [17], vinaginsenoside R4 (16) [18], (20R)-ginsenoside Rg3 (17) [19], ginsenoside-
Rd (18) [18], ginsenoside Rb1 (19) [14], ginsenoside Ra3 (20) [20][21], notoginsenoside
ꢀ 2014 Verlag Helvetica Chimica Acta AG, Zꢁrich

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Fig. 1. New compounds 1 – 3 isolated from the root of Panax notoginseng
Fig. 2. Chemical structures of 4 – 24 isolated from the root of Panax notoginseng
Fa (21) [19], notoginsenoside R4 (22) [14], notoginsenoside D (23) [22], and
notoginsenoside-G (24) [23], were also isolated and identified by comparison of their

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spectroscopic data with those reported in the literature. Among them, two known
compounds, 14 and 15, were isolated for the first time from Panax notoginseng.
Compound 1 was obtained as a white, amorphous powder. Its molecular formula
was determined as C47H80O18 by HR-ESI-MS (m/z 955.5229 ([M þ Na]þ,
1
C47H80NaO18þ; calc. 955.5242)). The H- and 13C-NMR data (Table) of 1 were very
similar to those of ginsenoside F1 (10), except for two sets of signals due to a glucose
1
and a xylose units. In the H-NMR ((D5)pyridine) spectrum of 1, diagnostic signals
were found for a sapogenin moiety with those of eight Me groups at d(H) 0.97, 1.01,
1.09, 1.44, 1.60, 1.64, 1.64, 1.97 (each s, Me(30), Me(18), Me(19), Me(29), Me(26),
Me(21), Me(27), Me(28), resp.), and of an olefinic H-atom at d(H) 5.31 (t, J ¼ 6.2,
HꢀC(24)). In the 13C-NMR spectrum of 1, 47 C-atom signals were detected, including
two olefinic C-atom signals of C(24) (d(C) 126.0) and C(25) (d(C) 131.0). The signals
of C(5) and C(20) were shifted downfield to d(C) 61.7 [24] and 83.4, respectively. In
1
addition, the H-NMR spectrum showed signals for three anomeric H-atoms at d(H)
4.94 (d, J ¼ 7.5, HꢀC(1’’’)), 5.01 (d, J ¼ 7.7, HꢀC(1’’)), and 5.11 (d, J ¼ 7.8, HꢀC(1’)),
which showed HSQCs to anomeric C-atom signals at d(C) 105.9 (C(1’’’)), 105.5 (C(1’’))
and 98.0 (C(1’)), respectively. Based on the coupling constants of the anomeric H-
atoms, all sugar substituents were identified as b-configured.
Acid hydrolysis of 1 revealed the presence of xylose and glucose moieties, which
were in relative proportions of 1:2, as determined by GC/MS analysis [25]. All these
data suggested that compound 1 was a protopanaxatriol-type ginsenoside, and that the
trisaccharide unit was attached to C(20). The exact oligoglycoside structure at C(20) in
1 was determined from the HMBC spectrum, which showed HMBCs from HꢀC(1’)
(d(H) 5.11) to C(20) (d(C) 83.4), from HꢀC(1’’) (d(H) 5.01) to C(6’) (d(C) 70.3), and
from HꢀC(1’’’) (d(H) 4.94) to C(6’’) (d(C) 69.9) (Fig. 3). The relative configurations at
the ring junctions were confirmed by a NOESY spectrum, which revealed the
correlations from HꢀC(3) (d(H) 3.48 – 3.53) to HꢀC(5) (d(H) 1.20) and Me(28) (d(H)
1.97), from HꢀC(6) (d(H) 4.37 – 4.39) to Me(18) (d(H) 1.01) and Me(19) (d(H) 1.09),
from HꢀC(12) (d(H) 4.19 – 4.23) to HꢀC(9) (d(H) 1.52 – 1.56) and Me(30) (d(H) 0.97)
(Fig. 4). The NOE correlation from Me(17) (d(H) 2.57) to HꢀC(12) (d(H) 4.19 –
4.23), Me(21) (d(H) 1.64), and Me(30) (d(H) 0.97) suggested that the configuration
at C(20) should be (S). Furthermore, the HMBCs from Me(26) (d(H) 1.60) and
Me(27) (d(H) 1.64) to C(24) (d(C) 126.0) and from HꢀC(24) (d(H) 5.31) to C(26)
(d(C) 25.8) and C(27) (d(C) 17.9) confirmed that the C¼C bond was located at the side
1
train. The H- and 13C-NMR assignments for 1 were accomplished unambiguously
based on HSQC, HMBC, and TOCSY data. Thus, the structure of 1 was determined as
(20S)-20-O-[b-d-xylopyranosyl-(1 ! 6)-b-d-glucopyranosyl-(1 ! 6)-b-d-glucopyrano-
syl]dammar-24-ene-3b,6a,12b,20-tetrol.
Compound 2 was obtained as an amorphous powder. Its molecular formula was
determined as C40H66O10 by HR-ESI-MS (m/z 1413.9362 ([2 M þ H]þ, C80H133O2þ0 ; calc.
1413.9390)) and NMR data. The 1H- and 13C-NMR data (Table) of 2 were very similar
to those of ginsenoside Rh1 (5), except for a set of signals arising from the presence of a
butenoyl unit. In the 1H-NMR ((D5)pyridine) spectrum of 2, signals of eight Me groups
at d(H) 0.91, 1.04, 1.23, 1.41, 1.55, 1.61, 1.65, 2.01 (each s, Me(30), Me(19), Me(18),
Me(21), Me(29), Me(26), Me(27), Me(28), resp.) and of an olefinic H-atom at d(H)
5.31 (t, J ¼ 7.2, HꢀC(24)) were displayed, and in the 13C-NMR spectrum of 2, 40 C-atom

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Fig. 3. Key HMBCs of 1 – 3
signals were displayed, including those of two olefinic C-atom signals of C(24) (d(C)
126.2) and C(25) (d(C) 130.8). The signals of C(5) and C(6) were shifted downfield to
d(C) 61.4 and 80.0, respectively. In addition, the 1H-NMR spectrum showed an
anomeric H-atom signal at d(H) 5.06 (d, J ¼ 7.8, HꢀC(1’)), which showed HSQC to the
C-atom signal at d(C) 106.2 (C(1’)). Based on the coupling constant of the anomeric H-
atom, the sugar substituent was identified as b-configurated, and acid hydrolysis of 2
revealed the presence of a glucose moiety, identified by GC/MS analysis [25]. The
above data suggested that compound 2 was also a protopanaxatriol-type ginsenoside,
and that the sugar unit was at C(6). The location of the sugar unit at C(6) (d(C) 80.0)
was established by the HMBC experiment (Fig. 3). The anomeric H-atom at d(H) 5.06
(d, J ¼ 7.8, HꢀC(1’)) was correlated through a three-bond coupling with C(6) (d(C)
80.0), and the HꢀC(6) signal (d(H) 4.38) correlated, in turn, with the anomeric C-atom
signal at d(C) 106.2 (C(1’)). The presence of a butenoyl group was confirmed on the
basis of HSQC and HMBC data (HMBCs from HꢀC(3’’) (d(H) 7.09) to C(1’’) (d(C)
166.5), and from HꢀC(4’’) (d(H) 1.74) to C(2’’) (d(C) 123.1) and C(3’’) (d(C) 144.8)).

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The HMBCs from HꢀC(6’) (d(H) 5.11) to C(1’’) (d(C) 166.5), coupled with the C-atom
signal of C(6’) downfield-shifted (d(C) 63.0 to 65.1) by comparing it with that of
ginsenoside Rh1, revealed that the butenoyl group was at C(6’) of the glucose unit. The
relative configurations at the ring junctions were confirmed by NOESY spectrum,
which have revealed the correlations from HꢀC(3) (d(H) 3.50) to HꢀC(5) (d(H)
1.41 – 1.43) and Me(28) (d(H) 2.01), from HꢀC(6) (d(H) 4.38) to Me(18) (d(H) 1.23)
and Me(19) (d(H) 1.04), and from HꢀC(12) (d(H) 3.90 – 3.94) to HꢀC(9) (d(H) 1.57)
and Me(30) (d(H) 0.91) (Fig. 4). The NOE correlations from HꢀC(17) (d(H) 2.30 –
2.34) to HꢀC(12) (d(H) 3.90 – 3.94), Me(21) (d(H) 1.41), and Me(30) (d(H) 0.91)
suggested that the configuration at C(20) should be (S). Furthermore, the HMBCs
from Me(26) (d(H) 1.61) and Me(27) (d(H) 1.65) to C(24) (d(C) 126.2), from
HꢀC(24) (d(H) 5.31) to C(26) (d(C) 25.8) and C(27) (d(C) 17.9) confirmed that the
1
C¼C bond is located in the side chain. The H- and 13C-NMR assignments for 2 were
accomplished unambiguously based on HSQC and HMBC data. Thus, the structure of
2 was determined as (20S)-(6-O-[(E)-but-2-enoyl-(1 ! 6)-b-d-glucopyranosyl]dam-
mar-24-ene-3b,6a,12b,20-tetrol.
Fig. 4. Selected NOESY correlations of 1 – 3
Compound 3 was isolated as a white amorphous powder. Its molecular formula was
determined as C40H68O12 by HR-ESI-MS (m/z 763.4605 [M þ Na]þ, C40H68NaO1þ2 ;
calc.763.4608) and NMR data. Comparing the 1H- and 13C-NMR data of 3 with those of
1 and 2, all these three compounds turned out to possess the same aglycone moiety. In
1
the H-NMR ((D5)pyridine) spectrum of 3, signals of eight Me groups at d(H) 0.87,
0.96, 1.12, 1.39 (2ꢁ), 1.60, 1.63, 2.03 (each s, Me(30), Me(19), Me(18), Me(21),
Me(29), Me(26), Me(27), Me(28), resp.) and of an olefinic H-atom at d(H) 5.30 (t, J ¼
7.1, HꢀC(24)) were displayed, and in the 13C-NMR spectrum of 3, 40 C-atom signals
were detected, including those of two olefinic C-atom signals of C(24) (d(C) 126.3) and
C(25) (d(C) 130.7). The signals of C(5) and C(6) were shifted downfield to d(C) 61.2,
1
and 78.8, respectively. In addition, the H-NMR spectrum exhibited two anomeric H-
atom signals at d(H) 5.09 (d, J ¼ 6.2, HꢀC(1’)) and 5.65 (d, J ¼ 7.4, HꢀC(1’’), which
showed HSQCs to C-atom signal at d(C) 104.2 (C(1’)) and 104.9 (C(1’’)). Based on the
coupling constant of the anomeric H-atom, the sugar substituent was identified as b-
configurated, and acid hydrolysis of 3 revealed the presence of a xylcose moiety,
identified by GC/MS analysis [25]. All these data suggested that compound 3 was also a
protopanaxatriol-type ginsenoside, and the sugar unit was at C(6). The location of the
sugar unit at C(6) (d(C) 78.8) was established by the HMBC experiment (Fig. 3). The
signal of the anomeric H-atom at d(H) 5.09 (d, J ¼ 6.2, HꢀC(1’)) correlated through a

Helvetica Chimica Acta – Vol. 97 (2014)
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three-bond coupling with that of C(6) (d(C) 78.8), and the HꢀC(6) signal (d(H) 4.32)
correlated, in turn, with that of the anomeric C-atom at d(C) 104.2 (C(1’)). The relative
configurations at the ring junctions were confirmed by NOESY spectrum, which
exhibited correlations from HꢀC(3) (d(H) 3.45 – 4.49) to HꢀC(5) (d(H) 1.38) and
Me(28) (d(H) 2.03), from HꢀC(6) (d(H) 4.32) to Me(18) (d(H) 1.12) and Me(19)
(d(H) 0.96), and from HꢀC(12) (d(H) 3.90) to HꢀC(9) (d(H) 1.52) and Me(30) (d(H)
0.87) (Fig. 4). The NOE correlation from HꢀC(17) (d(H) 2.29) to HꢀC(12) (d(H)
3.90), Me(21) (d(H) 1.39), and Me(30) (d(H) 0.87), suggested that the configuration at
C(20) should be (S). Furthermore, the HMBCs from Me(26) (d(H) 1.60) and Me(27)
(d(H) 1.63) to C(24) (d(C) 126.3), from HꢀC(24) (d(H) 5.30) to C(26) (d(C) 25.8) and
C(27) (d(C) 17.6) confirmed that the C¼C bond was located in the side train. The 1H-
and 13C-NMR assignments for 3 were achieved unambiguously based on HSQC and
HMBC data. Thus, the structure of 3 was determined as (20S)-6-O-[b-d-xylopyranosyl-
(1 ! 2)-b-d-xylopyranosyl]dammar-24-ene-3b,6a,12b,20-tetrol.
Financial supports by Youth Science Foundation of Guangxi Botanical Garden of Medicinal Plants
(Grant No. 201101) and Guangxi Natural Science Foundation (Grant No. 2013GXNSFBB053001) are
gratefully acknowledged. The authors are grateful to Dr. Ying-Hui Duan of Institute of Traditional
Chinese Medicine & Natural Products of Jinan University for his kindly help in recording HR-ESI mass
spectra. The authors also thank Ms. Ying-Liang Wei of Guangxi Institute of Analysis for recording the
NMR and mass spectra.
Experimental Part
General. Column chromatography (CC): silica gel G (SiO2; 100 – 200 and 200 – 300 mesh, Qingdao
Sea Chemical Factory, P. R. China), D101 macroporous resin (Cangzhou Bon Adsorber Technology Co.,
Ltd., P. R. China), and Sephadex LH-20 (Amersham Biosciences, Germany). Medium-pressure liquid-
chromatography (MPLC): MCI gel (CHP20P, 75 – 150 mm; Mitsubishi Chemical Corporation, Japan) and
reversed-phase C18 silica gel (40 – 63 mm; Merck, Germany). TLC: precoated silica gel G plates (Qingdao
Sea Chemical Factory, P. R. China); visualization with 10% H2SO4 in alcohol, followed by heating.
MPLC: Eyela Ceramic VSP 3050 pump, Eyela glass column (300 ꢁ 30 mm). Prep. HPLC: Shimadzu LC-
6AD pump, Shimadzu SPD-20A UV detector, YMC ODS-A (20 ꢁ 250 mm, 10 mm). M.p.: X-4 micro-
melting-point apparatus (Shanghai, P. R. China); uncorrected. IR Spectra: PerkinElmer Spectrum One
FT-IR spectrometer. NMR Spectra: Bruker ARX-600 spectrometer in (D5)pyridine with TMS as an
internal standard. ESI-MS: Agilent 1100-LC/MS DTrap SL. HR-ESI-MS: Waters API QSTAR Pular-1
mass spectrometer and a Waters Synapt G2 MS mass spectrometer. GC/MS: Agilent 7000B Triple Quad
GC/MS-7890 GC system.
Plant Material. The roots of Panax notoginseng were purchased in Yi Xin Pharmaceutical Co. Ltd.
(Nanning, Guangxi) in October, 2011. The sample was authenticated by TCM-Pharmacist Jia-Fu Wei
from Guangxi Zhuang Autonomous Region Food and Drug Administration. A voucher specimen
No. SQ20111019 was deposited with the Laboratory of Natural Products of the College of Pharmacy,
Guangxi Medical University.
Extraction and Isolation. Dried roots (8.9 kg) of Panax notoginseng were extracted with tenfold 80%
EtOH under reflux, and ca. 1,800 g of extract were obtained. The extract was suspended in H2O and then
partitioned with CH2Cl2 and BuOH successively.
The BuOH (1517 g) extract was subjected to CC (D101 macroporous resin; H2O/EtOH 100 :0 !
0 :100): Frs. B1 – B5. Fr. B3 (256 g) was subjected to CC (D101 macroporous resin; H2O/EtOH
90 :10 ! 70 :30): Frs. B31 – B32. Fr. B32 (56 g) was subjected to CC (SiO2; CH2Cl2/MeOH 100 :1 !
0 :100): Frs. B321 – B327. Fr. B326 (10 g) was recrystallized from MeOH/H2O to yield 11 (5536 mg).
The mother soln. from Fr. B326 was subjected to MPLC (RP C18 silica gel; H2O/MeOH 100: 0 ! 0 :100):
Frs. B3261 – B3266. Fr. B3264 (0.21 g) and Fr. B3265 (0.53 g) were submitted to prep. HPLC (MeOH/

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Helvetica Chimica Acta – Vol. 97 (2014)
H2O 70 :30) to afford 1 (51 mg), 24 (141 mg), and 16 (145 mg). Fr. B325 (37 g) was subjected to MPLC
(RP C18 silica gel; H2O/MeOH 100: 0 ! 0 :100): Frs. B3251 – B3255. Fr. B3251 (0.2 g) was submitted to
prep. HPLC (MeOH/H2O 50 :50) to furnish 12 (17 mg). Fr. B3254 (1.4 g) was purified by CC (Sephadex
LH-20; CHCl3/MeOH, 1:1) and prep. HPLC (MeOH/H2O 53 :47) to afford 4 (79 mg), 11 (105 mg), and
14 (53 mg). Fr. B327 (8.8 g) was subjected to MPLC (RP C18 silica gel; H2O/MeOH 100: 0 ! 0 :100):
Frs. B3271 – B3277. Fr. B3275 (7.7 g) was separated by CC (Sephadex LH-20; CHCl3/MeOH 1:1) and
prep. HPLC (MeOH/H2O 62 :38) to give 19 (5536 mg), 20 (87 mg), 21 (422 mg), 22 (320 mg), and 23
(68 mg). Fr. B5 (150 g) was subjected to CC (SiO2; CHCl3/MeOH/H2O 100 :0 :0 ! 6 :4 :0.4): Frs. B51 –
B55. Fr. B55 (80 g) was purified by CC (SiO2; CHCl3/MeOH 10 :1 ! 1:1): Frs. B521 – B524. Fr. B523
(8.1 g) was separated by MPLC (MCI gel; H2O/MeOH, 30 :70 ! 0 :100) and prep. HPLC (MeOH/H2O
70 :30) to yield 6 (129 mg) and 10 (30 mg). Fr. B524 (2.1 g) was separated by MPLC (RP C18 silica gel;
H2O/MeOH 70: 30 ! 0 :100), MPLC (MCI gel; H2O/MeOH 100 :0 ! 0 :100), and prep. HPLC (MeOH/
H2O 50 :50) to furnish 3 (9 mg), 5 (100 mg), 8 (100 mg), and 9 (50 mg). Fr. B54 (100 g) was subjected to
CC (SiO2; CHCl3/MeOH 8 :2 ! 7:3): Frs. B541 – B543. Fr. B543 (92 g) was re-subjected to CC (SiO2;
AcOEt/EtOH/H2O 5 :1:0 ! 6 :3 :0.3) to afford 17 (200 mg) and 18 (531 mg).
The CH2Cl2 extract (72 g) was subjected to CC (SiO2; CH2Cl2/MeOH 1:0 ! 5 :1): Frs. C1 – C7.
Fr. C6 (8.2 g) was separated by CC (SiO2; CHCl3/MeOH 20 :1 ! 5 :1): Frs. C61 – C65. Fr. C64 (2.8 g) was
submitted to MPLC (MCI gel; H2O/MeOH, 50 :50 ! 0 :100): Frs. C641 – C644. Fr. C641 (1.1 g) was
subjected to MPLC (RP C18 silica gel; H2O/MeOH 100: 0 ! 0 :100), and prep. HPLC (MeOH/H2O
60 :40) to give 2 (15 mg), 7 (54 mg), 15 (15 mg), and 16 (135 mg).
(20S)-20-O-[b-d-Xylopyranosyl-(1 ! 6)-b-d-glucopyranosyl-(1 ! 6)-b-d-glucopyranosyl]dammar-
24-ene-3b,6a,12b,20-tetrol (¼(3b,6a,12b)-3,6,12-Trihydroxydammar-24-en-20-yl O-b-d-Xylopyranosyl-
(1 ! 6)-O-b-d-glucopyranosyl-(1 ! 6)-b-d-glucopyranoside; 1). White amorphous power (MeOH).
1
M.p. 187 – 1888. IR (KBr): 3399, 2927, 1638, 1384, 1042. H- and 13C-NMR: see the Table. HR-ESI-MS
(pos.): 955.5229 ([M þ Na]þ, C47H80NaO1þ8 ; calc. 955.5242).
(20S)-6-O-[(E)-But-2-enoyl-(1 ! 6)-b-d-glucopyranosyl]dammar-24-ene-3b,6a,12b,20-tetrol
(¼(3b,6a,12b)-3,12,20-Trihydroxydammar-24-en-6-yl 6-O-[(2E)-1-Oxobut-2-en-1-yl]-b-d-glucopyrano-
side; 2). White amorphous power (MeOH). M.p. 159 – 1608. IR (KBr): 3399, 2933, 2962, 1712, 1655,
1
1384, 1029. H- and 13C-NMR: see the Table. HR-ESI-MS (pos.): 1413.9362 ([2 M þ H]þ, C80H133Oþ20
;
calc. 1413.9390).
(20S)-6-O-[b-d-Xylopyranosyl-(1 ! 2)-b-d-xylopyranosyl]dammar-24-ene-3b,6a,12b,20-tetrol
(¼(3b,6a,12b)-3,12,20-Trihydroxydammar-24-en-6-yl 2-O-b-d-Xylopyranosyl-b-d-xylopyranoside; 3).
1
White amorphous power (MeOH). M.p. 171 – 1728. IR (KBr): 3391, 2932, 1642, 1384, 1033. H- and
13C-NMR: see the Table. HR-ESI-MS (pos.): 763.4605 ([M þ Na]þ, C40H68NaO1þ2 ; calc. 763.4608).
Acid Hydrolysis of 1 – 3. Each compound (1.5 mg) was hydrolyzed with 1.5 ml of 1m HCl at 1008 for
4 h. The mixture was extracted with CHCl3 (3ꢁ), and the aq. residue was evaporated under reduced
pressure. Then, 1 ml of pyridine and 2 mg of NH2OH · HCl were added to the residue, and the mixture
was heated at 908 for 1 h. After cooling, Ac2O (0.5 ml) was added, and the mixture was heated at 908 for
1 h. The mixtures were evaporated under reduced pressure, and the resulting aldononitrile peracetates
were analyzed by GC/MS using standard aldononitrile peracetates as reference samples.
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Received April 12, 2013