Compound 7 had a molecular formula
C51H82O22, as deduced by the HRESIMS at m/z
1081.4988 [M+Cl]- (calcd. for C51H82ClO22,
1081.4986) and the 13C NMR data. The 1H and 13C
NMR spectra, in combination with DEPT, HSQC,
COSY, and HMBC spectra showed the presence of
two tertiary methyls at δH 0.83 (s, H3-18) and 1.07
(s, H3-19), one secondary methyl group at H 1.00
(d, J = 7.0 Hz, H3-21), a trisubstituted olefinic
proton at δH 5.40 (d, J = 3.0 Hz, H-6), and an
acetalic carbon signal at C 110.8 (C-22), implying
that 7 possesses the 5,6-spirostane skeleton [21].
The 1H NMR spectrum of 7 contained signals for
four anomeric protons at H 4.52 (d, J = 8.0 Hz, H-
1 ), 5.22 (br s, H-1 ), 4.86 (H-1 ), and 4.22 (d, J =
7.5 Hz, H-1 ), that showed HSQC correlations with
anormeric carbons at C 100.4 (C-1 ), 102.3 (C-1 ),
102.9 (C-1 ), and 104.7 (C-1 ), respectively,
indicating that 7 possesses four sugar units.
Comparison of the 13C NMR data of the sugar units
with those reported previously suggested the
presence of two glucopyranoses and two
rhamnopyranoses. The relatively large couplingVJC, 55(2)
constant (J > 7.5 Hz) of the anomeric proton of the
glucose revealed the -configuration, whereas the -
oriented anomeric form of the rhamnose was defined
based on the chemical shift values of its C-3 and C-5
positions [22]. In the HMBC spectrum, the
correlation between the anomeric proton of a
glucose unit at H 4.22 (H-1 ) and C 72.8 (C-27)
indicated that this glucose was located at C-27
position of the aglycone (figure 2). The sugar
sequence at C-3 of the aglycone was identified as -
L-rhamnopyranosyl-(1 4)[ -L-rhamnopyranosyl-
(1 2)]- -D-glucopyranoside by the HMBC
correlations between H 4.86 (H-1 ) and C 79.9
(Glc C-4), between H 5.22 (H-1 ) and C 79.3 (Glc
C-2), and between H 4.52 (H-1 ) and C 79.3 (C-3).
This assignment was also supported by the ROESY
correlations between H 4.86 (H-1 ) and H 3.54 (H-
4 ), between H 5.22 (H-1 ) and H 3.42 (H-2 ), and
between H 4.52 (H-1 ) and H 3.61 (H-3) (figure 3).
On the basis of the above analysis, along with
comparison of the NMR data of 7 with those of the
very recently reported spirostane-type saponin, the
structure of compound 7 was established as shown
in Figure 1, namely dioscoroside D [20].
In summary, our phytochemical study on the
acidic methanol extract of D. metel resulted in the
isolation and identification of seven compounds,
including pterodontriol B (1), disciferitriol (2),
scopolamine (3), adenosine (4), thymidine (5),
ilekudinoside C (6), and dioscoroside D (7). Among
the isolated compounds, pterodontriol B,
disciferitriol, ilekudinoside C, and dioscoroside D
were reported for the first time from the Datura
genus
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Vietnam Journal of Chemistry, International Edition, 55(2): 188-195, 2017
DOI: 10.15625/2525-2321.2017-00442
188
Chemical constituents of Datura metel L.
Nguyen Thi Mai
1
, Nguyen Thi Cuc
2
, Tran Hong Quang
2*
1
Hanoi University of Transport and Communications
2
Institue of Marine Biochemistry, Vietnam Academy of Science and Technology (VAST)
Received 17 October 2016; Accepted for publication 11 April 2017
Abstract
Chemical investigation of an acidic methanol extract of the whole plant of Datura metel resulted in the isolation of
seven compounds, including pterodontriol B (1), disciferitriol (2), scopolamine (3), adenosine (4), thymidine (5),
ilekudinoside C (6), and dioscoroside D (7). Their structures were elucidated by extensive spectroscopic methods,
including 1D and 2D NMR and mass spectra and in comparison with reported data in the literature. Among the isolated
compounds, pterodontriol B, disciferitriol, ilekudinoside C, and dioscoroside D were reported for the first time from the
Datura genus.
Keywords. Datura metel, Solanaceae, sesquiterpene, triterpenoid saponin, steroidal saponin
1. INTRODUCTION
Datura metel L. is an annual herb that belongs to
the Solanaceae family. It has tropical American
origin and is widely cultivated in many tropical and
temperate regions. In the Vietnamese traditional
medicine, D. metel has been used for the treatment
of coughs, bronchial asthma, and rheumatism [1]. Its
leaves have been used as anesthetics in surgery, a
fumigant in bronchial asthma, and anti-contractive
agents in the stomach ulcers [1]. The flowers of D.
metel have been used widely in the Chinese
traditional medicine for the treatment of asthma,
convulsions, pain, and rheumatism for centuries [2].
Previous studies of the pharmacological effects have
shown that D. metel seeds exhibits a hypoglycemic
activity in normal and alloxan-induced diabetic rats
[3], the chloroform extract of D. metel displays an
antifungal effect toward several pathogenic species
of Aspergillus [4], and the seeds and fruit pulps of
D. metel have a high antioxidant activity [5].
Chemical studies have demonstrated that the major
chemical components of D. metel are withanolide-
type steroids [6-12], which have been shown to
suppress NO production in lipopolysaccharide
(LPS)-stimulated RAW264.7 cells [11, 12], and
exhibit cytotoxicity against HCT-116, A549, DLD-
1, BGC-823, and K562 cancer cell lines [6, 7, 10]. In
addition, the isolation of some megastigmane
sesquiterpenes and amide alkaloids from D. metel
was also reported [13, 14]. In the present study, we
report the isolation and structural elucidation of
seven compounds from the acidic methanol extract
of the whole plants of D. metel.
2. MATERIAL AND METHODS
2.1. Plant material
The whole plants of D. metel were collected in
Thai Binh province, Vietnam during May 2015, and
identified by Dr. Bui Van Thanh, Institute of
Ecology and Biological Resources. A voucher
specimen (NCCT-CDM-5.2015) was deposited at
the Herbarium of the Institute of Marine
Biochemistry, VAST.
2.2. General experimental procedures
All NMR spectra were recorded on a Bruker
AM500 FT-NMR spectrometer (500 MHz for
1
H
and 125 MHz for
13
C-NMR), chemical shifts are
reported in ppm using TMS as an internal standard.
ESIMS spectra were recorded on Agilent 1100.
Column chromatography (CC) was performed on
silica gel 230-400 mesh or RP-18 resins (150 μm,
Fuji Silysia Chemical Ltd.). Compounds were
visualized by spraying with aqueous 10% H2SO4 and
heating for 5 minutes.
2.3. Extraction and isolation
VJC, 55(2) 2017 Tran Hong Quang et al.
189
The whole plant of D. metel was dried (5 kg),
ground, and extracted with MeOH/acetic acid (pH =
5.0) under sonication at room temperature. After
concentration under reduced pressure, the MeOH
extract (300 g) was suspended in water and
partitioned with CHCl3 to give CHCl3 and aqueous
fractions. The aqueous fraction was alkalinized by
adding NH4OH until pH = 9.0, and then partitioned
successively with CH2Cl2 and EtOAc to provide
CH2Cl2, EtOAc, and aqueous fractions, respectively.
The CH2Cl2 and EtOAc fractions were combined
and subjected to a reversed phase (RP) C18 column
chromatography (CC), eluted with MeOH-H2O
(10:17, v/v) to provide three subfractions (DME1-
DME3). Subfraction DME1 was fractionated using a
silica gel CC, being eluted with EtOAc-MeOH-H2O
(20:1:0.01, v/v/v) to give 3 (15 mg). The aqueous
fraction was neutralized and subjected to
fractionation through a Diaion HP-20 column, being
eluted with a stepwise gradient of MeOH in water
(25–100 %) to give four fractions (DMW1–DMW4).
Fraction DMW2 was fractionated using reversed
phase (RP) C18 column chromatography (CC), being
eluted with MeOH-H2O (1:3, v/v) to yield
subfractions DMW21–DMW25. Subfraction
DMW22 was then fractionated on a silica gel
column, being eluting with CH2Cl2-MeOH-H2O
(5:1:0.05, v/v/v) to give 4 (6 mg) and 5 (8 mg).
Fraction DMW4 was subjected to a RP C18 CC,
being eluted with a stepwise gradient of MeOH-H2O
(1:2–4:1, v/v) to yield five subfractions (DMW41–
DMW5). Fraction DMW42 was separated using a
silica gel CC, being eluted with CH2Cl2-MeOH-H2O
(6:1:0.05, v/v/v) to provide four subfractions
(DMW421–DMW424). Subfraction DMW421 was
fractionated using a silica gel CC, being eluted with
EtOAc-MeOH-H2O (13:1:0.05, v/v/v) and further
purified by a silica gel CC, being eluted with
CH2Cl2-MeOH-H2O (6:1:0.1, v/v/v) to give 2 (7
mg). Subfraction DMW424 was fractionated by a
RP C18 CC, eluted with acetone-H2O (1:3, v/v) and
further purified by a silica gel CC, being eluted with
CH2Cl2-MeOH-H2O (6.5:1:0.05, v/v/v) to give 1 (7
mg). Subfraction DMW44 was fractionated using a
silica gel CC, being eluted with EtOAc-MeOH-H2O
(2.5:1:0.1, v/v/v) to provide five subfractions
(DMW441–DMW445). Subfraction DMW443 was
fractionated by a RP C18 CC, being eluted with
MeOH-H2O (2:1, v/v) and further purified by a silica
gel CC, being eluted with CH2Cl2-MeOH-H2O
(3:1:0.1, v/v/v) to yield 7 (12 mg), and 6 (10 mg).
Pterodontriol B (1): white, amorphous powder;
C15H28O3, M = 256; ESIMS: m/z 255 [M H] ;
1
H
(CD3OD, 500 MHz) and
13
C NMR data (CD3OD,
125 MHz), see table 1.
Disciferitriol (2): white, amorphous powder;
C15H28O3, M = 256; ESIMS: m/z 255 [M H] ;
1
H
(CD3OD, 500 MHz) and
13
C NMR data (CD3OD,
125 MHz), see table 1.
Scopolamine (3): white, amorphous powder;
C17H21NO4, M = 303; ESIMS: m/z 304 [M+H]
+
;
1
H
(CD3OD, 500 MHz) and
13
C NMR data (CD3OD,
125 MHz), see table 2.
Adenosine (4): white, amorphous powder;
C10H13N5O4, M = 267; ESIMS: m/z 268 [M+H]
+
;
1
H
(CD3OD, 500 MHz) and
13
C NMR data (CD3OD,
125 MHz), see table 2.
Thymidine (5): white, amorphous powder;
C10H14N2O5, M = 242; ESIMS: m/z 241 [M H] ;
1
H
(CD3OD, 500 MHz) and
13
C NMR data (CD3OD,
125 MHz), see table 2.
Ilekudinoside C (6): white, amorphous powder;
C41H66O14, M = 782; ESIMS: m/z 805 [M+Na]
+
;
1
H
(CD3OD, 500 MHz) and
13
C NMR data (CD3OD,
125 MHz), see table 3.
Dioscoroside D (7): white, amorphous powder;
C51H82O22, M = 1046; HRESIMS: m/z 1081.4988
[M+Cl] (calcd. for C51H82ClO22, 1081.4986);
1
H
(CD3OD, 500 MHz) and
13
C NMR data (CD3OD,
125 MHz), see table 3.
3. RESULTS AND DISCUSSION
Compound 1 was obtained as a white,
amorphous powder. Its molecular formula was
identified as C15H28O3 by an ESIMS ion peak at m/z
255 [M H] ,
along with the
13
C NMR data. The
1
H
NMR of 1 showed the signals of four tertiary methyl
groups at δH 1.26 (6H, s, H3-12 and H3-13), 1.11
(3H, s, H3-14), and 0.93 (3H, s, H-15), and one
oxymethine proton at δH 3.26 (1H, m, H-1) (Table
1). The
13
C NMR and DEPT spectra contained
signals for 15 carbons, including three non-
protonated carbons (two of which were oxygenated),
three methines (of which one was oxygenated), five
methylenes, and four methyl carbons. Comparison
of the
1
H and
13
C NMR data of 1 with those of the
reported ent-eudesmane sesquiterpene, pterodontriol
B revealed that the structures of these compounds
are similar [15]. The minor differences between the
13
C chemical shifts of these compounds observed at
C-3, C-9, and C-11 might be due to the different
solvents that these compounds were recorded in (1:
in CD3OD; pterodontriol B: in C5D5N). In the
HMBC spectrum, the HMBC correlations from δH
1.26 (H3-12 and H3-13) to δC 43.1 (C-7) and 75.2 (C-
11), from δH 1.11 (H3-14) to δC 42.0 (C-3), 72.8 (C-
4), and 48.4 (C-5), and from δH 0.93 (H3-15) to δC
80.8 (C-1), 48.4 (C-5), 39.2 (C-9), and 39.8 (C-10)
indicated that four methyl groups and three hydroxyl
VJC, 55(2) 2017 Chemical constituents of Datura metel L.
190
groups are located at C-11, C-4, and C-10 positions
(figure 2). Thus, compound 1 was determined to be
pterodontriol B.
Compound 2 was isolated as a white, amorphous
powder and its molecular formula was established as
C15H28O3 by the presence of an ion [M H] at m/z
255 in the ESIMS. The
1
H and
13
C NMR spectra of 2
were found nearly identical with those of 1, except
for significant difference of the chemical shift of C-7
(2: δH 1.33/ δC 50.7 vs 1: δH 1.70/ δC 43.1) (table 1),
suggesting that these compounds have different
configuration at C-7. This was supported by a good
agreement when comparing the
1
H and
13
C NMR
data of 2 with those reported for the 7-epimer of 1,
disciferitriol (table 1) [15]. The different
13
C
chemical shift between these compounds at C-11
could be explained by influence of the different
solvents used (2: in CD3OD; disciferitriol: in
C5D5N). Therefore, compound 2 was identified as
disciferitriol.
The molecular formula of compound 3 was
determined to be C17H21NO4 by the observation of
an ion [M+H]
+
at m/z 304 in the ESIMS and
13
C
NMR spectroscopic analysis. The
1
H NMR spectrum
contained signals for five aromatic protons at H
7.32 (3H) and 7.37 (2H) which were characteristic
of a phenyl ring. The
1
H NMR spectrum further
exhibited signals for oxygenated proton signals at δH
Table 1:
1
H and
13
C NMR data for
compounds 1 and 2
C C
#1
1 C
#2
2
C
a,b
H
a,c
C
a,b
H
a,c
1 80.1 80.8 3.26 (m) 79.5 80.3 3.24 (m)
2 30.1 29.4 1.63
*
30.4 29.3 1.66
*
3 39.0 42.0 1.75 (m)
1.53
*
42.8 41.9 1.75 (m)
1.53 (m)
4 71.7 72.8 71.7 72.5
5 48.2 48.4 1.64
*
54.0 54.0 1.26
*
6 21.8 21.7 2.03 (br d,
13.5)/1.53
*
22.3 23.1 1.67 (m)
1.29 (m)
7 42.8 43.1 1.70 (m) 50.6 50.7 1.33 (m)
8 21.9 21.6 1.80 (m)
1.63 (m)
23.1 22.6 1.95 (m)
1.15 (m)
9 42.4 39.2 1.60
*
1.53
*
42.4 42.0 1.95
*
1.09
*
10 39.6 39.8 40.1 40.1
11 74.0 75.2 71.0 73.4
12 29.8 28.7
d
1.26 (s)
d
27.9 26.8
d
1.19 (s)
d
13 30.4 29.5
d
1.26 (s)
d
28.0 27.4
d
1.20 (s)
d
14 23.1 22.0 1.11 (s) 23.4 22.5 1.11 (s)
15 14.4 14.1 0.93 (s) 14.0 13.7 0.88 (s)
a
Recorded in CD3OD,
b
125 MHz,
c
500MHz,
d
Signals are
interchangeable;
*
Overlapped signal;
#1
C of pterodontriol
B [15] in C5D5N;
#2
C of disciferitriol [15] in C5D5N
Figure 1: Chemical structures of compounds 1-7 from D. metel
VJC, 55(2) 2017 Tran Hong Quang et al.
191
Table 2:
1
H and
13
C NMR data for compounds 3-5
C C
#1
3
C
#2
4
C
#3
5
C
a,b
H
a,c
C
d,b
H
d,c
C
a,b
H
a,c
1 57.3 58.6 3.17 (m)
2 55.8 56.4 3.57 (d, 3.5) 152.4 152.3 8.13 (s) 152.37 152.4
4 55.4 56.1 3.03 (d, 3.5) 149.0 149.0 138.16 138.1 7.83 (d, 1.0)
5 59.2 58.7 3.28 (m) 119.3 119.3 111.51 111.5
6 30.4 29.4 1.50 (d, 16.0)
2.13 (m)
156.2 156.1 166.42 166.5
7 66.3 67.1 4.99 (t, 5.5)
8 30.2 29.6 1.71 (d, 16.0)
2.22 (m)
139.9 139.8 8.34 (s)
10 41.6 39.1 2.52 (s)
1′ 171.2 172.8 87.9 87.9 5.87 (d, 6.5) 86.25 86.2 6.30 (t, 7.0)
2′
53.8 55.9 3.79 (m) 73.4 73.4 4.59 (dd, 5.5,
6.5)
41.22 41.1 2.26 (m)
3′
63.4 64.4 3.77 (m)
4.15 (t, 11.0)
70.6 70.6 4.14 (dd, 3.0,
5.5)
72.21 72.2 4.42 (m)
4 85.9 85.8 3.96 (m) 88.82 88.8 3.93 (m)
5
61.6 61.6 3.53 (br d, 12.0)
3.67 (br d, 12.0)
62.84 62.8 3.82 (dd, 12.0,
3.0)
3.76 (dd, 12.0,
3.5)
1′′ 135.4 137.5
2′′, 6′′ 128.3 129.9 7.32*
4′′ 127.6 128.8 7.32*
3′′, 5′′ 127.3 129.2 7.37*
5-CH3 12.44 12.4 1.90 (d, 1.0)
a
Recorded in CD3OD,
b
125 MHz,
c
500MHz,
d
in DMSO-d6;
*
Overlapped signal;
#1
C of scopolamine [16] in CDCl3;
#2
C of adenosine [17] in DMSO-d6;
#3
C of thymidine [18] in CD3OD.
3.57 (d, J = 3.5 Hz, H-2), 3.03 (d, J = 3.5 Hz, H-4),
4.99 (t, J = 5.5 Hz, H-7), 3.77 (m, H-3 a), and 4.15
(t, J = 11.0 Hz, H-3 b), and two protons bearing
nitrogens at δH 3.17 (m, H-1) and 3.28 (m, H-5), and
the down-filed signal of a methyl group bearing
nitrogen at δH 2.52 (H3-10). Analysis of the
13
C
NMR and HSQC indicated the presence of one
carbonyl carbon at δC 172.8 (C-1 ), five aromatic
methines at δC 137.5 (C-1 ), 129.9 (C-2 and 6 ),
128.8 (C-4 ), and 129.2 (C-3 and 5 ), one epoxy
group at δC 56.4 (C-2) and 56.1 (C-4), one
oxymethine at δC 67.1 (C-7), one oxymethylene at δC
64.4 (C-3 ), two methines bearing nitrogen at δC 58.6
(C-1) and 58.7 (C-5), two methylenes, and one
methyl group.
The
1
H and
13
C NMR data of 3 (in CD3OD)
showed a similarity with those of scopolamine (in
CDCl3), suggesting that the structures of both
compounds are identical (table 2) [16]. By the
HMBC correlations observed between δH 2.52 (H3-
10) and δC 58.6 (C-1) and between δH 3.57 (H-2) and
δH 3.03 (H-4) and δC 58.7 (C-5), the positions of the
methyl and epoxy groups were assigned to C-10 and
C-4/C-5, respectively (figure 2). The overall
structure of 3 was subsequently assigned by the
HMBC correlations between δH 3.79 (H-2 ) and δC
137.5 (C-1 ), 129.9 (C-2 and C-6 ), between δH
3.77 and 4.15 (H2-3 ) and δC 172.8 (C-1 ), 55.9 (C-
2 ), and 137.5 (C-1 ), and between δH 4.99 (H-7)
and δC 172.8 (C-1 ). Based on the above analysis,
compound 3 was determined to be scopolamine.
The ESIMS of compound 4 exhibited an ion
[M+H]
+
at m/z 268, corresponding with the
molecular formula C10H13N5O4. The
1
H NMR
spectrum showed signals of two down-field aromatic
protons at δH 8.13 (s, H-2) and 8.34 (s, H-8),
suggesting that these protons are connecting with
nitrogen atoms. The
1
H NMR further displayed a
signal for one anomeric proton at δH 5.87 (d, J = 6.5
Hz) revealing that 4 has one sugar moiety. The
13
C
NMR contained 10 carbon signals, including two
down-field aromatic methines at δC 152.3 (C-2) and
139.8 (C-8), three non-protonated carbons at δC
149.0, 119.3 (C-5), and 156.1, suggesting that 4
VJC, 55(2) 2017 Chemical constituents of Datura metel L.
192
possesses the purine nucleus. The five remaining
carbon signals, including four oxymethines and one
oxymethylene group were assigned to a -
ribofuranose by comparing with the data reported in
the literature (table 2) [17]. Thus, compound 4 was
identified as adenosine.
Table 3:
1
H and
13
C NMR data for compounds 6 and 7
C C
#1
6 C
#2
7 C C
#1
6 C
#2
7
C
a,b
H
a,c
C
a,b
H
a,c
C
a,b
H
a,c
C
a,b
Ha,c
1 47.3 47.3 0.88
*
2.05
*
37.5 38.5 1.10
*
/1.90
* 1′ 106.6 106.2 4.31 (d, 7.5) 100.3 100.4 4.52 (d,
8.0)
2 66.9 68.0 3.81
*
30.1 30.7 1.62
*
/1.94
* 2′ 73.1 72.9 3.60* 77.9 79.3 3.42 (dd,
7.5, 9.5)
3 88.4 88.5 3.48 (d, 9.5) 78.1 79.3 3.61 (m) 3′ 74.9 74.6 3.53* 77.8 77.9 3.61*
4 44.7 45.2 39.0 39.5 2.31 (t,
12.0)/2.48 (dd,
3.0, 13.0)
4′ 69.7 70.0 3.84* 78.5 79.9 3.54
(t, 9.5)
5 47.7 47.7 1.22
*
140.8 141.8 5′ 67.8 67.8 3.65*
3.92 (d,
10.5)
76.9 76.5 3.34
*
6 18.2 18.7 1.39
*
/1.49
*
121.8 122.6 5.40 (d, 3.0) 6 61.3 61.9 3.67
*
3.82 (br
d, 11.0)
7 33.2 33.6 1.33
*
/1.68
*
32.3 33.1 1.58
*
/2.01
*
1″ 95.7 95.7 5.37 (d, 8.0) 102.0 102.3 5.22 (br
s)
8 40.8 41.0 31.7 32.7 1.68
* 2″ 74.1 73.9 3.33* 72.5 72.1 3.95*
9 48.2 48.8 1.64
*
50.3 51.6 0.99
* 3″ 78.9 78.2 3.42* 72.8 72.4 3.68*
10 37.8 38.5 37.1 38.0 4″ 71.4 71.2 3.38* 74.1 73.9 3.43 (dd,
9.0, 9.5)
11 23.9 24.5 1.98
*
21.1 21.9 1.56
* 5″ 79.2 78.5 3.36* 69.5 69.7 4.14*
12 126.2 127.0 5.27 (br s) 39.8 40.9 1.23
*
/1.79
*
6″ 62.5 62.5 3.82*/3.71* 18.6 17.8 1.27
(d, 6.5)
13 138.4 139.3 40.4 41.4 1″ 102.9 102.9 4.86*
14 42.6 43.4 56.6 57.7 1.18
*
2″ 72.5 72.1 3.86*
15 28.7 29.2 1.11
*
/1.96
*
32.2 32.7 1.31
*
/2.01
*
3″ 72.7 72.3 3.65*
16 24.7 25.2 1.78
*
/2.09
*
81.1 82.2 4.42 (q, 7.5)
4″
73.9 73.7 3.43 (dd,
9.0, 9.5)
17 48.4 48.4 62.8 63.7 1.78
5″ 70.4 70.6 3.95*
18 53.4 54.1 2.27 (d, 12.0) 16.3 16.7 0.83 (s)
6″
18.5 17.9 1.28
(d, 6.0)
19 39.2 40.2 1.00
*
19.4 19.8 1.07 (s)
1′″
105.0 104.7 4.22 (d,
7.5)
20 39.4 40.3 1.41
*
42.0 42.9 1.93
* 2′″ 75.1 75.1 3.19*
21 30.8 30.6 1.31
*
15.0 14.8 1.00 (d, 7.0) 3′″ 78.6 78.0 3.36*
22 36.8 37.5 1.67
*
/
1.79 (br d,
13.5)
109.5 110.8
4′″
71.6 71.6 3.29
*
23 63.8 64.0 3.28 (d,
11.5)/3.71
*
31.3 31.9 1.63
*
1.73
*
5′″ 78.5 78.0 3.28*
24 14.7 14.4 0.76 (s) 23.9 24.2 1.57
*
/1.68
*
6′″ 62.8 62.7 3.69*/
3.88
*
25 17.4 17.8 1.07 (s)
36.7 37.1 1.91
*
26 17.6 17.9 0.86 (s) 63.6 64.4 3.52 (br d,
11.0)
3.76
*
27 23.8 24.0 1.15 (s) 72.0 72.8 3.33
*
/3.80
*
28 176.2 177.9
29 17.8 17.6 0.92 (d, 6.5)
30 21.3 21.5 1.30 (d, 6.5)
a Recorded in CD3OD,
b125 MHz, c500MHz; *Overlapped signal; #1 C of ilekudinoside C [19] in C5D5N;
#2 C of dioscoroside D in
[20] C5D5N
VJC, 54(5) 2016 Chemical constituents of Datura metel
193
The molecular formula of thymidine (5),
C10H14N2O5 was deduced by its ESIMS ion at m/z
241 [M H] and
1
H and
13
C NMR spectra. The
1
H
NMR spectrum showed signals for one aromatic
proton at δH 7.83 (d, J = 1.0 Hz, H-4), one methyl
group at δH 1.90 (d, J = 1.0 Hz, 5-CH3), and one
anomeric proton at δH 6.30 (t, J = 7.0 Hz, H-1 ). The
13
C NMR spectrum displayed 10 carbon signals,
including two carbonyl carbons at δC 152.4 (C-2)
and 166.5 (C-6), one aromatic methine at δC 138.1
(C-4), one non-protonated aromatic carbon at
δC111.5 (C-5), and one methyl at δC 12.4 (5-CH3),
suggesting the presence of a methyl-pyrimidinedione
structural moiety. The remaining carbon signals,
including three oxymethines, one oxymethylene, and
one methylene carbons were assigned to a deoxy-β-
D-ribofuranoside by comparing with the reported
values (table 2) [18]. So the structure of thymidine
(5) was established as shown in figure 1.
Ilekudinoside C (6) was isolated as a white,
amorphous powder. Its molecular formula was
C53H86O21, as deduced by ESIMS at m/z 805
[M+Na]
+
and its
13
C NMR spectrum. The
13
C NMR
spectrum exhibited 41 carcbon signals, of which 30
were assigned to a triterpenoid alglycone and 11 to a
saccharide moiety. The
1
H NMR spectrum of 6
contained signals for six methyl groups at δH 0.76 (s,
H3-24), 1.07 (s, H3-25), 0.86 (s, H3-26), 1.15 (s, H3-
27), 0.92 (d, 6.5, H3-29), and 1.30 (d, 6.5, H3-30), a
trisubstituted olefinic proton at δH 5.27 (br s), and
two anomeric protons at δH 4.31 (d, 7.5, H-1 ) and
5.37 (d, 8.0, H-1 ). The signals at δC 127.0 and
139.3 in the
13
C NMR spectrum, assignable to C-12
and C-13, suggested the presence of a
12
-ursane-
type triterpene. Signals at δC 88.5 (C-4) and 177.9
(C-28) in the
13
C NMR spectrum suggested that 6 is
a bisdesmosidic ursane-type saponin. The sugar
units were identified as one glucopyranose and one
arabinopyranose based on comparing the
13
C NMR
data of 6 with those reported previously in the
literature [19]. The relatively large spin couplings of
the anomeric protons (J > 7.5 Hz) were indicative of
the -arabinopyranose and -glucopyranose. In the
HMBC spectrum, the HMBC correlation from H
3.48 (H-3) to C 68.0 (C-2) suggested that a
hydroxyl group is attached to C-2 position (Figure
2). The HMBC correlations between δH 4.31 (H-1 )
and δC 88.5 (C-3) and between δH 5.37 (H-1 ) and δC
177.9 (C-28) indicated that the arabinose and
glucose were located at C-3 and C-28, respectively.
Based on the data obtained and comparing with
those of the reported compound (table 3) [19], the
structure of ilekudinoside C (6) was elucidated as
shown in figure 1.
Figure 2: Selected HMBC correlations of
compounds 1-3, 6, and 7 and COSY correlations of
compound 7
Figure 3: Selected ROESY correlations of
compound 7
Compound 7 had a molecular formula
C51H82O22, as deduced by the HRESIMS at m/z
1081.4988 [M+Cl]
-
(calcd. for C51H82ClO22,
1081.4986) and the
13
C NMR data. The
1
H and
13
C
NMR spectra, in combination with DEPT, HSQC,
COSY, and HMBC spectra showed the presence of
two tertiary methyls at δH 0.83 (s, H3-18) and 1.07
(s, H3-19), one secondary methyl group at H 1.00
(d, J = 7.0 Hz, H3-21), a trisubstituted olefinic
proton at δH 5.40 (d, J = 3.0 Hz, H-6), and an
acetalic carbon signal at C 110.8 (C-22), implying
that 7 possesses the
5,6
-spirostane skeleton [21].
The
1
H NMR spectrum of 7 contained signals for
four anomeric protons at H 4.52 (d, J = 8.0 Hz, H-
1 ), 5.22 (br s, H-1 ), 4.86 (H-1 ), and 4.22 (d, J =
7.5 Hz, H-1 ), that showed HSQC correlations with
anormeric carbons at C 100.4 (C-1 ), 102.3 (C-1 ),
102.9 (C-1 ), and 104.7 (C-1 ), respectively,
indicating that 7 possesses four sugar units.
Comparison of the
13
C NMR data of the sugar units
with those reported previously suggested the
presence of two glucopyranoses and two
rhamnopyranoses. The relatively large coupling
VJC, 55(2) 2017 Chemical constituents of Datura metel L.
194
constant (J > 7.5 Hz) of the anomeric proton of the
glucose revealed the -configuration, whereas the -
oriented anomeric form of the rhamnose was defined
based on the chemical shift values of its C-3 and C-5
positions [22]. In the HMBC spectrum, the
correlation between the anomeric proton of a
glucose unit at H 4.22 (H-1 ) and C 72.8 (C-27)
indicated that this glucose was located at C-27
position of the aglycone (figure 2). The sugar
sequence at C-3 of the aglycone was identified as -
L-rhamnopyranosyl-(1 4)[ -L-rhamnopyranosyl-
(1 2)]- -D-glucopyranoside by the HMBC
correlations between H 4.86 (H-1 ) and C 79.9
(Glc C-4), between H 5.22 (H-1 ) and C 79.3 (Glc
C-2), and between H 4.52 (H-1 ) and C 79.3 (C-3).
This assignment was also supported by the ROESY
correlations between H 4.86 (H-1 ) and H 3.54 (H-
4 ), between H 5.22 (H-1 ) and H 3.42 (H-2 ), and
between H 4.52 (H-1 ) and H 3.61 (H-3) (figure 3).
On the basis of the above analysis, along with
comparison of the NMR data of 7 with those of the
very recently reported spirostane-type saponin, the
structure of compound 7 was established as shown
in Figure 1, namely dioscoroside D [20].
In summary, our phytochemical study on the
acidic methanol extract of D. metel resulted in the
isolation and identification of seven compounds,
including pterodontriol B (1), disciferitriol (2),
scopolamine (3), adenosine (4), thymidine (5),
ilekudinoside C (6), and dioscoroside D (7). Among
the isolated compounds, pterodontriol B,
disciferitriol, ilekudinoside C, and dioscoroside D
were reported for the first time from the Datura
genus.
Acknowledgment. This research was supported by
NAFOSTED under grant number 104.01-2014.69.
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Corresponding author: Tran Hong Quang
Institute of Marine Biochemistry
Vietnam Academy of Science and Technology
No. 18, Hoang Quoc Viet, Cau Giay District, Hanoi
E-mail: quangth2004@yahoo.com.
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