Three new 18,19-seco-ursane glycosides from Elsholtzia bodinieri

Article abstract of DOI:10.1016/j.phytol.2015.04.028

Abstract Chromatographic separation of an extract of the aerial part of Elsholtzia bodinieri resulted in the isolation of three new 18,19-seco-ursane glycosides, bodiniosides E-G (1-3). Their structures were elucidated as 2α,12β,23-trihydroxy-3-(β-d-glucopyranosyl)-19-oxo-18,19-seco-urs-13(18)-en-28-O-β-d-glucopyranosyl ester (1), 3-β-d-glucopyranosyl-19-β-d-glucopyranosyl-12β,21-dihydroxy-18,19-seco-urs-13(18)-en-28-oic acid (2), and 2α,12β,21-trihydroxy-3-β-d-glucopyranosyl-19-β-d-glucopyranosyl-18,19-seco-urs-13(18)-en-28-oic acid (3), respectively, by extensive NMR techniques, including 1D- and 2D-NMR experiments, as well as comparing with spectral data with those of the known analogues.

Full text of DOI:10.1016/j.phytol.2015.04.028

G Model
PHYTOL 947 1–5
Phytochemistry Letters xxx (2015) xxx–xxx
Contents lists available at ScienceDirect
Phytochemistry Letters
journal homepage: www.elsevier.com/locate/phytol
1
2
Short communication
Three new 18,19-seco-ursane glycosides from Elsholtzia bodinieri
3
4
Q1
Xue-Wei Zhao, Jin-Dong Zhong, Hong-Mei Li, Rong-Tao Li *
Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, PR China
A R T I C L E I N F O
A B S T R A C T
Article history:
Chromatographic separation of an extract of the aerial part of Elsholtzia bodinieri resulted in the isolation
of three new 18,19-seco-ursane glycosides, bodiniosides E-G (1–3). Their structures were elucidated as
Received 2 March 2015
Received in revised form 23 April 2015
Accepted 29 April 2015
Available online xxx
2a
,12
nosyl ester (1), 3-
en-28-oic acid (2), and 2
b
,23-trihydroxy-3-(
-glucopyranosyl-19-
,12 ,21-trihydroxy-3-
b
-
D
-glucopyranosyl)-19-oxo-18,19-seco-urs-13(18)-en-28-O-
-glucopyranosyl-12 ,21-dihydroxy-18,19-seco-urs-13(18)-
-glucopyranosyl-19- -glucopyranosyl-18,19-seco-
b-D-glucopyra-
b
-D
b
-D
b
a
b
b
-D
b-D
urs-13(18)-en-28-oic acid (3), respectively, by extensive NMR techniques, including 1D- and 2D-NMR
experiments, as well as comparing with spectral data with those of the known analogues.
ã 2015 Published by Elsevier B.V. on behalf of Phytochemical Society of Europe.
Keywords:
Elsholtzia bodinieri
Lamiaceae
18,19-seco-ursane glycoside
5
ꢁ1
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
1
. Introduction
hydroxyl groups (3428 cm ), carbonyl groups (1734 and
1
ꢁ
1
ꢁ1
1
701 cm ), and olefinic functional groups (1636 cm ). The H
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
The species Elsholtzia bodinieri Vaniot, belonging to the genus
Elsholtzia (Labiatae), is an annual herbaceous plant distributed in
Yunnan and Guizhou Provinces in China. It is generally known as
NMR spectrum (Table 1) of 1 revealed six methyl signals at dH 1.35
(3H, s, Me-24), 0.74 (3H, s, Me-25), 1.03 (3H, s, Me-26), 1.17 (3H, s,
Me-27), 2.06 (3H, s, Me-29), and 0.96 (3H, d, J = 6.6 Hz, Me-30),
three oxygen-bearing methines at dH 4.09 (1H, m, H-2), 3.28 (1H, d,
J = 9.1 Hz, H-3) and 4.56 (1H, dd, J = 1.5, 10.0 Hz, H-12), as well as an
olefinic proton at dH 6.54 (1H, s, H-18). In addition, a series of
typical signals of two sugar residues were recognized, including
two anomeric proton signals at
.32 (1H, d, J = 8.2 Hz, H-1 ), revealing
both sugar residues on the basis of their JH1,H2 coupling constants
“
Dongzisu” and used as herbal tea or folk medicine for the
1
1
1
1
1
1
1
1
1
1
2
2
2
2
treatment of cough, headache, pharyngitis, fever and hepatitis (
Jiangsu New Medical College, 1985). Previous phytochemical
studies on E. bodinieri showed that this plant was a rich source
of triterpenoids, diterpenoids and flavonoids as well as their
corresponding glycosides, including oleanane triterpenoid sapo-
nins (Zhu et al., 2002; Hu et al., 2007; Li et al., 2012), 18,19-seco-
ursane monodesmoside saponins (Li et al., 2005), clerodane
diterpenoid glycosides (Hu et al., 2008), flavonoid and flavanone
glycosides (Li et al., 2008). With an aim to discover more new
compounds from E. bodinieri, we further systematically investigat-
ed the n-BuOH soluble fraction of this plant, which led to the
isolation of three new 18,19-seco-ursane glycosides, bodiniosides
E-G (1–3). We report herein the isolation and structural elucidation
of the isolates.
0
d
H
4.93 (1H, d, J = 7.8 Hz, H-1 ) and
Q2
00
6
b
-configuration present in
3
(
Zhang et al., 2012) Fig. 1. On acid hydrolysis of 1, only
Glc) was detected as sugar residues by GC chromatography with
-cysteine derivatives. Except
D-glucose
(
the corresponding trimethylsilylated
L
13
for the signals belonging to the glucosyl groups, the C NMR and
DEPT spectra (Table 2) of 1 exhibited resonances for 30 carbons,
including eight quaternary carbons (two are carbonyls at
11.8 and 175.3 and an olefinic carbon at 147.6), seven methines
containing three oxygenated ones ( 66.9, d, C-2; 95.4, d, C-3; 69.1,
d, C-12) and an olefinic one ( 120.9, d, C-18), nine methylenes
including an oxygenated one ( 63.9, t, C-23), and six methyls (
8.1, q, C-24; 17.8, q, C-25; 18.1, q, C-26; 21.6, q, C-27; 28.5, q, C-29;
d
C
2
d
C
Q3
Q4
d
C
5
0
d
C
2
4
2. Results and discussion
51
d
C
d
C
5
2
1
2
2
2
2
2
5
6
7
8
9
19:3
D
Compound 1, ½
aꢀ
ꢁ29.05 (c 0.19, MeOH), white amorphous
53
54
55
56
57
16.3, q, C-30), which were assigned to 18, 19-seco-ursane triterpene
skeleton (Li et al., 2005; Kakuno et al.,1991). Careful comparison of
the H and C NMR data of 1 with those of laevigin B (Yan et al.,
013) suggested their structural similarities, except for the
presence of an additional Glc moiety in 1. The glycosidic linkages
powder, gave rise to a quasi-molecular ion peak at m/z 867.4117
+
1
13
(
[M + Na] ) in the HR-ESI-MS, which corresponded to the molecular
13
formula C42
68
H O17, and was further confirmed by its C NMR and
2
DEPT analysis. The IR absorption bands indicated the existence of
5
8
at C-3 and C-28 of the aglycone were determined by the HMBC
0
0
59
60
correlations (Fig. 2) from H-1 to C-3, from H-3 to C-1 , and from H-
0
0
1
to C-28, respectively. Furthermore, the chemical shift (
C-1”) of the anomeric carbon appeared in upfield further
C
d 96.4, d,
*
Corresponding author. Tel.: +86 871 65920569; fax: +86 871 65920570.
6
1
E-mail address: rongtaolikm@163.com (R.-T. Li).
http://dx.doi.org/10.1016/j.phytol.2015.04.028
874-3900/ã 2015 Published by Elsevier B.V. on behalf of Phytochemical Society of Europe.
1
Please cite this article in press as: Zhao, X.-W., et al., Three new 18,19-seco-ursane glycosides from Elsholtzia bodinieri. Phytochem. Lett. (2015),
http://dx.doi.org/10.1016/j.phytol.2015.04.028

G Model
PHYTOL 947 1–5
2
X.-W. Zhao et al. / Phytochemistry Letters xxx (2015) xxx–xxx
Table 1
1
5 H
H NMR data of compounds 1–3 in pyridine-d (600 MHz, d , mult, J in Hz).
No.
1
1
2
3
1.18 (overlap)
1.75 (overlap)
0.95 (m)
2.03 (m)
1.17 (overlap)
2.40 (m)
4.07 (m)
–
2
.39 (m)
2
4.09 (m)
–
2.12 (m)
3
5
6
3.28 (d, 9.1)
0.95 (m)
1.47 (m)
3.40 (dd, 4.4, 11.7)
0.84 (m)
1.48 (m)
1.19 (m)
1.39 (m)
1.20 (overlap)
1.61(br s)
2.31(m)
3.29 (d, 9.2)
0.89 (m)
1.45 (m)
1.14 (m)
1
.27 (m)
1.43 (m)
.20 (overlap)
7
9
1.44 (m)
1.20 (overlap)
1.62 (br s)
2.33 (m)
1.60 (m)
4.67 (dd, 2.2, 10.7)
1.01 (m)
2.36 (m)
1.63 (m)
1.86 (m)
6.43 (s)
1
1.60 (d, 9.2)
2.28 (m)
11
1.62 (m)
1.61(m)
1
2
4.56 (dd, 1.5, 10.0)
1.10 (m)
4.47 (dd, 2.1, 11.7)
1.02 (m)
2.47 (m)
1.64 (m)
1.86 (m)
6.45 (s)
4.30 (m)
2.05 (m)
4.66 (m)
–
15
2
.28 (m)
1.63 (m)
.94 (m)
16
1
1
8
6.54 (s)
–
1
9
4.26 (m)
2.30 (m)
4.67 (m)
–
2
0
2.42 (m)
1.73 (m)
21
2
.20 (m)
1.70 (m)
.52 (d, 12.6)
3.35 (d, 9.0)
.45 (m)
2
2
3
1.72 (m)
2.35 (dd, 6.9, 13.3)
1.11(s)
1.70 (m)
2.33 (dd, 6.9, 13.0)
1.19 (s)
2
2
4
–
–
24
25
26
27
29
30
1.35 (s)
0.74 (s)
1.03 (s)
1.17 (s)
2.06 (s)
0.96 (d, 6.6)
1.31(s)
0.79 (s)
0.95 (s)
1.25 (s)
1.27 (d, 6.4)
0.93 (d, 6.9)
1.36 (s)
0.83 (s)
0,97 (s)
1.08 (s)
1.26 (d, 6.8)
0.91 (d, 6.3)
Fig. 1. Structures of compounds 1–3.
3
1
2
3
4
5
6
-Glc
0
4.93 (d, 7.8)
4.10 (overlap)
4.20 (m)
4.14 (m)
4.31 (m)
4.95 (d, 8.1)
4.05 (m)
4.40 (m)
4.38 (m)
4.31 (m)
4.94 (d, 7.8)
4.06 (m)
4.27 (m)
4.37 (m)
4.31 (m)
ꢁ
76
observed at m/z 811.4470 (HR-ESI-MS, [M–H
2
O–H] ), consistent
0
0
0
0
0
7
7
with a molecular formula of C42 16 assigned by a combina-
70
H O
1
13
1
78
79
80
tional analysis of H, C NMR and DEPT spectra. The H NMR
spectrum (Table 1) of 2 showed seven methyl signals at 1.11 (3H,
s, Me-23), 1.31 (3H, s, Me-24), 0.79 (3H, s, Me-25), 0.95 (3H, s, Me-
6),1.25 (3H, s, Me-27),1.27 (3H, d, J = 6.4 Hz, Me-29), and 0.93 (3H,
d, J = 6.9 Hz, Me-30), four oxygen-bearing methines at 3.40 (1H,
dd, J = 4.4, 11.7 Hz, H-3), 4.47 (1H, dd, J = 2.1,11.7 Hz, H-12), 4.30 (1H,
m, H-19) and 4.66 (1H, m, H-21), and an olefinic proton at 6.45
(1H, s, H-18). In addition, the presence of two anomeric signals at
4.95 (1H, d, J = 8.1 Hz, H-1 ) and 4.88 (1H, d, J = 7.8 Hz, H-1 )
revealed the existence of two sugar residues with -configuration
Zhang et al., 2012). Acid hydrolysis of 2 afforded -glucose (Glc) as
sugar residues, which was confirmed by GC analysis of their
corresponding trimethylsilylated -cysteine derivatives. In addi-
tion to the sugar moieties, the C NMR spectrum (Table 2) showed
the presences of one carbonyl ( 179.1, s, C-28), seven methyls (
d
H
4.32 (dd, 4.7, 12.1)
4.37 (dd, 4.8, 11.8)
4.65 (m)
4.33 (dd, 4.7, 11.8)
4.45 (m)
4
.50 (m)
8
1
2
19-Glc
0
0
0
0
0
0
0
0
0
0
0
0
d
82
83
84
H-1
H-2
H-3
H-4
H-5
H-6
4.88 (d, 7.8)
3.98 (m)
4.20 (m)
4.41 (m)
3.86 (ddd, 2.5, 4.7, 9.3)
4.24 (m)
4.84 (d, 7.7)
3.98 (m)
4.16 (m)
4.26 (m)
3.95 (m)
4.59 (m)
H
d
H
8
5
0
00
d
H
8
6
2
8-Glc
b
0
0
0
0
0
0
0
0
0
0
0
0
H-1
H-2
H-3
H-4
H-5
H-6
6.32 (d, 8.2)
4.21 (m)
4.18 (m)
4.39 (m)
4.00 (m)
4.01 (m)
87
(
D
8
8
8
9
L
13
90
91
92
d
C
d
C
4
.24 (m)
1
2
7.1, q, C-23; 28.2, q, C-24; 16.9, q, C-25; 18.5, q, C-26; 22.1, q, C-27;
2.0, q, C-29; 8.8, q, C-30), eight methylenes, eight methines (four
oxygenated ones at dC 88.8, 68.1, 76.6, 77.5, and one olefinic one at
116.9), seven quaternary carbons (one olefinic at dC 149.5). The
9
9
3
4
6
6
6
6
6
6
6
6
7
7
7
2
3
4
5
6
7
8
9
0
1
2
supported the existence of ester glycoside at C-28 (Laura et al.,
012).
The relative configuration of 1 was determined through
inspection of the ROESY spectrum. The NOE correlation between
Me-24 and Me-25 indicated the -orientation of the hydroxy-
methylene group at C-23. Additionally, the ROESY cross-peaks of
H-2/Me-25 and H-3/H-5 established the -orientation of H-2 and
-orientation of H-3, respectively. Based on the above evidence,
the structure of 1 was established as 2 ,12 ,23-trihydroxy-3-(
d
C
2
1
13
95
H- and
C NMR spectra closely matched with those of
9
6
cornutaoside B, a known 18, 19-seco-ursane triterpene glycoside
isolated from Ilex cornuta (Wu et al., 2008). The differences
between compound 2 and cornutaoside B were the nature of the
sugars, including the types and linkage of the sugar residues.
Q5 97
a
9
9
8
9
b
0
100
101
102
103
a
HMBC correlations observed between H-1 and C-3 (dC 88.8) of the
a
b
b
-glu-
-
D
-
aglycone, indicated that one Glc moiety was linked to C-3.
Moreover, elaborative comparison of the NMR data of the aglycone
of 2 with those of cornutaoside B exhibited that the chemical shift
of C-19 in compound 2 obviously moved to downfield (from
68.4 in cornutaoside B to 76.6 in 2), which suggested that the
other Glc moiety was located at C-19. Furthermore, the HMBC
glucopyranosyl)-19-oxo-18,19-seco-urs-13(18)-en-28-O-
copyranosyl ester, and named bodinioside E.
b-D
1
1
1
04
05
06
7
7
7
3
4
5
19:3
D
d
C
Compound 2, ½
a
ꢀ
ꢁ88.97 (c 0.09, MeOH + CHCl
3
2:1), was
d
C
obtained as white amorphous powder and its molecular formula
was determined as C42 16 on basis of the quasi-molecular ion
70
H O
Please cite this article in press as: Zhao, X.-W., et al., Three new 18,19-seco-ursane glycosides from Elsholtzia bodinieri. Phytochem. Lett. (2015),
http://dx.doi.org/10.1016/j.phytol.2015.04.028

G Model
PHYTOL 947 1–5
X.-W. Zhao et al. / Phytochemistry Letters xxx (2015) xxx–xxx
3
Table 2
123
124
assignment. The relative configuration of 3 was established by
analysis of the ROESY spectrum. NOE correlations of H-2/Me-
1
3C NMR data of compounds 1–3 in pyridine-d
5
(150 MHz).
2
1
1
1
25
26
27
No.
1
3
25 and H-3/H-5 determined the
of H-3, respectively. Therefore, the structure of bodinioside G (3)
was assigned as 2 ,12 ,21-trihydroxy-3- -glucopyranosyl-19-
-glucopyranosyl-18,19-seco-urs-13(18)-en-28-oic acid.
b-oriented of H-2 and a-oriented
d
C
d
C
d
C
1
2
3
4
5
6
7
8
9
47.6 t
66.9 d
95.4 d
40.8 s
55.8 d
18.4 t
34.8 t
41.3 s
49.6 d
38.1 s
33.0 t
69.1 d
147.6 s
41.3 s
27.9 t
28.0 t
43.9 s
120.9 d
211.8 s
47.4 d
29.3 t
39.5 t
63.9 t
18.1 q
17.8 q
18.1 q
21.6 q
175.3 s
28.5 q
16.3 q
39.1 t
26.7 t
88.8 d
39.6 s
56.0 d
18.3 t
33.1 t
41.9 s
49.6 d
37.2 s
34.9 t
68.1 d
149.5 s
41.5 s
26.7 t
28.2 t
44.0 s
116.9 d
76.6 d
42.7 d
77.5 d
41.9 t
17.1 q
28.2 q
16.9 q
18.5 q
22.1 q
179.1 s
22.0 q
8.8 q
47.3 t
66.7 d
95.2 d
40.5 s
55.6 d
18.1 t
32.9 t
41.2 s
49.3 d
37.9 s
34.5 t
68.7 d
149.4 s
41.6 s
27.5 t
29.1 t
43.8 s
116.7 d
76.5 d
43.0 d
77.3 d
42.5 t
17.8 q
28.3 q
17.0 q
18.3 q
22.1 q
179.0 s
21.8 q
8.6 q
a
b
b-D
128
29
130
b-D
1
3
. Experimental
.1. General experimental procedures
D and 2D NMR spectra were recorded using Bruker Avance III-
3
1
1
1
31
32
33
1
1
0
1
1
600 instrument with tetramethylsilane (TMS) as an internal
standard. ESI and HR-ESI-MS were taken on an API Qstar Pulsar
instrument. Semi-preparative HPLC was performed on an Agilent
1
2
1
3
134
1
4
1
35
1200 liquid chromatograph with a ZORBAX SB-C18 (5 mm, 9.4 mm
1
5
ꢂ 250 mm) column. Column chromatography (CC) was carried out
on silica gel (200–300 mesh,100–200 mesh, 80–100 mesh, Qingdao
Marine Chemical Factory, Qingdao, China), Diaion HP-20SS (63–
1
6
1
36
1
7
1
8
137
138
1
9
1
50
m
m Mitsubishi Fine Chemical Industries Co., Ltd., Tokyo, Japan),
m, YMC Co., Ltd., Japan) and Sephadex LH-20
2
0
1
1
1
39
40
41
ODS-C18 (75
m
2
1
22
23
24
25
26
27
28
29
30
(Amersham Biosciences AB, Uppsala, Sweden). Fractions were
monitored by TLC plates (Silica gel H, Qingdao Marine Chemical
Factory, Qingdao, China), and spots were visualized by heating silica
142
143
2 4
gel plates sprayed with 5% H SO –EtOH.
1
1
44
45
3
.2. Plant material
The sample of E. bodinieri was collected from Honghe, Yunnan
3
1
2
3
4
5
6
-Glc
146
147
Province, PR China, in May 2008, and was identified by Prof. Hai-
Zhou Li. A voucher specimen (KMUST 20080003) was deposited at
the Laboratory of Phytochemistry, Faculty of Life Science and
Technology, Kunming University of Science and Technology.
0
106.6 d
75.6 d
78.6 d
71.6 d
78.7 d
62.6 t
107.1 d
75.8 d
78.3 d
71.5 d
78.4 d
62.7 t
106.3 d
75.4 d
78.0 d
71.3 d
78.4 d
62.4 t
0
0
0
0
0
0
1
1
48
49
150
3
.3. Extraction and isolation
Glc
0
0
0
0
0
0
0
0
0
0
0
0
1
2
3
4
5
6
2
1
2
3
4
5
6
104.5 d
75.4 d
78.7 d
71.9 d
78.8 d
63.1 t
104.3 d
75.1 d
78.4 d
71.4 d
78.5 d
62.5 d
1
51
The aerial parts of E. bodinieri (15.0 kg) were powdered and
152
153
154
extracted with 75% Me
ture and filtered. The filtrate was concentrated in vacuo and the
2
CO (3 ꢂ 35 L, 48 h, each) at room tempera-
3
resulting residue was extracted successively with CHCl , EtOAc and
n-BuOH, respectivley.
1
1
55
56
8-Glc
0
0
0
0
0
0
0
0
0
0
0
0
The n-BuOH extract (300.0 g) was separated over macroporous
resin CC (110 ꢂ1200 mm) eluting with MeOH/H O (gradient 10, 30,
0, 90, and 100%, each 2.5 L) to afford fractions I–IV. Fr. II (50 g) was
subjected to Sephadex LH-20 gel CC, eluting with MeOH/H
(gradient 10, 30, 50, 70, 90, and 100%, each 1 L) and silica gel CC
chloroform/MeOH, gradient 20:1 ! 0:1) to obtain subfractions II-
–II-4. Fr. II-4 (7.9 g) was chromatographed successively over silica
gel CC (chloroform/MeOH/H
96.4 d
74.0 d
78.8 d
71.2 d
79.4 d
62.4 t
157
2
Q6 158
6
1
1
1
1
1
59
60
61
62
63
2
O
(
1
2
O, gradient 20:1:0.1 ! 5:5:0.5), ODS
1
1
1
1
07
08
09
10
00
164
165
166
167
correlations from Me-29 and H-1 to C-19 further confirmed the
above conclusion. Thus, compound 2 was established as 3-
glucopyranosyl-19- -glucopyranosyl-12 ,21-dihydroxy-18,19-
(MeOH/H O, gradient 10% ! 100%), and semi-preparative HPLC
2
b
-D
-
(25% CH CN, 3 mL/min, t = 11.5 min) to obtain compound 1
3
R
b-
D
b
(30.4 mg). Fr. II-3 (1.49 g) was purified over silica gel CC (chloro-
form/MeOH/H O,
O, gradient 20:1:0.1 ! 5:5:0.5), ODS (MeOH/H
seco-urs-13(18)-en-28-oic acid, and given the name bodinioside F.
2
2
1
1
1
1
1
1
1
1
68
69
70
71
72
73
74
75
1
1
1
1
1
1
1
1
1
1
1
1
11
12
13
14
15
16
17
18
19
20
21
22
19:3
D
gradient 10% ! 100%), and semi-preparative HPLC (29% CH CN, 3
Compound 3, ½
a
ꢀ
ꢁ45.71 (c 0.32, MeOH), the molecular
3
mL/min, t = 12.5 min) to afford compound 3 (7.4 mg). Fr. III (42.0 g)
R
formula was determined to be C42
at m/z 827.4424 ([M–H
unsaturation. Detailed comparison of the H- (Table 1) and
NMR (Table 2) data of 3 with those of 2 revealed that they showed
highly similarity except for the signals at C-1, C-2 and C-3, which
indicated that compound 3 possessed different substitute mode in
ring A. The chemical shift of C-2 evidently moved to low field
H
70
O
17 as deduced by HR-ESI-MS
ꢁ
was isolated by Sephadex LH-20 gel CC (eluted with 10%, 30%, 60%,
2
O–H] ), indicating eight degrees of
1
13
90%, and 100% MeOH/H O) to yield subfractions III-1–III-2.
C
2
Compound 2 (10.0 mg) was purified from Fr. III-2 (17.0 g) by
repeatedly ODS (MeOH–H O, gradient 10% ! 100%), silica gel CC
2
(chloroform/MeOH/H O, gradient 20:1:0.1 ! 5:5:0.5) and semi-
2
preparative HPLC (22% CH CN, 3 mL/min, t = 24.5 min).
3
R
suggested that the methylene at C-2 (
was substituted by a hydroxyl group in 3 (
Moreover, the HMBC correlations (Fig. 2) from H-1 (
H-3 ( 3.29, d, J = 9.2 Hz) to C-2 further confirmed the above
d
H
2.03, 2.12;
d
C
26.7, t) in 2
66.7, d).
2.40, m) and
1
76
3.3.1. Bodinioside E (1)
White amorphous powder; ½
(KBr)
d
H
4.07,
d
C
1
9:3
177
178
a
ꢀ
ꢁ29.05 (c 0.19, MeOH); IR
d
H
D
ꢁ1
1
13
max
y 3428, 1734, 1701, 1636 cm ; H- and C NMR data, see
d
H
Please cite this article in press as: Zhao, X.-W., et al., Three new 18,19-seco-ursane glycosides from Elsholtzia bodinieri. Phytochem. Lett. (2015),
http://dx.doi.org/10.1016/j.phytol.2015.04.028

G Model
PHYTOL 947 1–5
4
X.-W. Zhao et al. / Phytochemistry Letters xxx (2015) xxx–xxx
Fig. 2. Key HMBC correlations for compounds 1 and 3.
1
1
79
80
+
199
200
Tables 1 and 2; ESI-MS (pos.) m/z 867 ([M + Na] ); HRESIMS (pos.)
HMDS–TMCS (hexamethyldisilazane–trimethylchlorosilane, 2:1)
was added, and the mixture was heated at 60 C for another 30 min.
+
ꢃ
m/z 867.4117 ([M + Na] ) (calcd for C42
H
68
O
17Na, 867.4143).
2
01
After centrifugation, the supernatant was subjected to GC analysis
1
81
ꢃ
ꢃ
202
203
204
3
.3.2. Bodinioside F (2)
under the following conditions: column temp 150–280 C at 5 /
19:3
D
ꢃ
1
1
1
1
82
83
84
85
min, carrier gas N (1 mL/min), injector temp 230 C and detector
temp 250 C, split ratio 1:10. The standards were prepared
White amorphous powder; ½
a
ꢀ
ꢁ88.97 (c 0.09, MeOH + CHCl
3
2
ꢃ
1
13
2
8
:1); H- and C NMR data, see Tables 1 and 2; ESI-MS (neg.) m/z
2
2
2
2
05
06
07
08
ꢁ
ꢁ
following the same procedure. Under these conditions, the
retention time of D-glucoside derivative was 22.03 min. During
co-injection studies, identical retention times were observed
between the different hydrolysates and authentic standards.
11 ([M–H
2
O–H] ); HRESIMS (neg.) m/z 811.4470 ([M–H
15, 811.4480).
2
O–H] )
(
calcd for C42
67
H O
1
86
3
.3.3. Bodinioside G (3)
1
1
1
1
87
88
89
90
19:3
D
1
White amorphous powder; ½
a
ꢀ
ꢁ45.71 (c 0.32, MeOH); H-
209
Competing financial interest
13
NMR and C NMR data, see Tables 1 and 2; ESI-MS (neg.) m/z 827
ꢁ
ꢁ
(
(
[M–H
2
O–H] ); HRESIMS (neg.) m/z 827.4424 ([M–H
2
O–H] )
Q7 210
The authors declare no competing financial interest.
calcd for C42
67
H O
16, 827.4423).
2
11
Supplementary data
1
1
91
92
3
2
.4. Acid hydrolysis and GC analysis of compounds 1–3 (Zhao et al.,
007)
212
13
1
D and 2D NMR, IR, EIMS, HREIMS data of compounds 1–3 are
2
available free of charge via the internet.
1
1
1
1
1
1
93
94
95
96
97
98
Compounds 1–3 (each 5.0 mg) were hydrolyzed with 9% HCl at
ꢃ
9
2
0 C for 5 h. The reaction liquid was filtered after being cooled to
214
Acknowledgment
ꢃ
–4 C, and the filtered liquor was freeze-dried to obtain dry
residual. The dried material was dissolved in dry pyridine (100
then 0.1 M
added, and the mixture was heated at 60 C for 1 h. Then 150
m
L),
L) was
L of
215
Q8 216
217
We gratefully acknowledge the financial support for this
research work by the National Natural Science Foundation of
China (21262021).
L
-cysteine methyl ester hydrochloride (200
m
ꢃ
m
Please cite this article in press as: Zhao, X.-W., et al., Three new 18,19-seco-ursane glycosides from Elsholtzia bodinieri. Phytochem. Lett. (2015),
http://dx.doi.org/10.1016/j.phytol.2015.04.028

G Model
PHYTOL 947 1–5
X.-W. Zhao et al. / Phytochemistry Letters xxx (2015) xxx–xxx
5
2
18
233
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Please cite this article in press as: Zhao, X.-W., et al., Three new 18,19-seco-ursane glycosides from Elsholtzia bodinieri. Phytochem. Lett. (2015),
http://dx.doi.org/10.1016/j.phytol.2015.04.028