Cyclic glycolipids from glandular trichome exudates of Cerastium glomeratum

Article abstract of DOI:10.1016/j.phytochem.2012.07.001

Fourteen cyclic glycolipids, named glomerasides A-N, have been isolated from the glandular trichome exudate of Cerastium glomeratum (Caryophyllaceae). Their structures were determined by spectroscopic analysis of the glycolipids, as well as by application of the Ohrui-Akasaka method to the fatty acid methyl esters derived from the glycolipids and GCMS studies of trimethylsilyl ether derivatives of the methyl esters. The various glomerasides have a glycosidic linkage between the anomeric hydroxy group of the glucose and the C-11, C-10 or C-9 positions of the docosanoyl moiety. They also contained an ester linkage between the C-6 hydroxy group of the glucose ring and the carboxyl group of the oxygenated fatty acid to form their macrocyclic structures. The glucose moiety was optionally acetylated and/or malonylated at the C-2 or C-3 hydroxy groups. Among these compounds, the 1,6′-cyclic ester of 11(R)-(2-O-acetyl-β- d-glucopyranosyloxy)docosanoic acid (glomeraside D) was the most abundant (25%).

Full text of DOI:10.1016/j.phytochem.2012.07.001

Phytochemistry 82 (2012) 149–157
Contents lists available at SciVerse ScienceDirect
Phytochemistry
journal homepage: www.elsevier.com/locate/phytochem
Cyclic glycolipids from glandular trichome exudates of Cerastium glomeratum
⇑
Teigo Asai, Yuki Nakamura, Yui Hirayama, Kiyoshi Ohyama, Yoshinori Fujimoto
Department of Chemistry and Materials Science, Tokyo Institute of Technology, Meguro, Tokyo 152-8551, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
Fourteen cyclic glycolipids, named glomerasides A–N, have been isolated from the glandular trichome
exudate of Cerastium glomeratum (Caryophyllaceae). Their structures were determined by spectroscopic
analysis of the glycolipids, as well as by application of the Ohrui–Akasaka method to the fatty acid methyl
esters derived from the glycolipids and GCMS studies of trimethylsilyl ether derivatives of the methyl
esters. The various glomerasides have a glycosidic linkage between the anomeric hydroxy group of the
glucose and the C-11, C-10 or C-9 positions of the docosanoyl moiety. They also contained an ester link-
age between the C-6 hydroxy group of the glucose ring and the carboxyl group of the oxygenated fatty
acid to form their macrocyclic structures. The glucose moiety was optionally acetylated and/or malony-
lated at the C-2 or C-3 hydroxy groups. Among these compounds, the 1,60-cyclic ester of 11(R)-(2-O-
Received 31 August 2010
Received in revised form 19 June 2012
Available online 30 July 2012
Keywords:
Cerastium glomeratum
Caryophyllaceae
Glandular trichome
Cyclic glycolipid
Ohrui–Akasaka method
acetyl-b-
D
-glucopyranosyloxy)docosanoic acid (glomeraside D) was the most abundant (25%).
Ó 2012 Elsevier Ltd. All rights reserved.
1. Introduction
In this paper, the isolation and structure elucidation of fourteen
cyclic glycolipids, named glomerasides A–N from the glandular tri-
Glandular trichomes produce secondary metabolites of diverse
classes such as terpenoids, phenylpropanoids, polyketides and
fatty acid derivatives (Schilmiller et al., 2008). The secondary
metabolites secreted by glandular trichomes are thought to have
diverse biological activities, such as protecting the aerial parts of
plants against herbivores and pathogens (Duke et al., 2000; Spring,
2000). As far as glycolipid structures isolated from glandular tri-
chome exudates are concerned, glycosyloxy-fatty acids have been
reported from Ibicella lutea and Proboscidea louisiana (Martynia-
ceae) (Asai et al., 2010), as well as a glyceride having a glycosyl-
oxy-fatty acyl moiety from Cerasus yedoensis (Rosaceae) (Asai and
Fujimoto, 2011) and a series of 1,20-cyclic ester derivatives of
chome exudate of the plants are described. All were 1,60-cyclic es-
ter derivatives of (b-D-glucopyranosyloxy)dodecanoic acids.
2. Results and discussion
The exudate sample was obtained by rinsing an upper portion
of the aerial part with Et2O. The extract showed an identical TLC
pattern as the sample that was obtained by gently wiping the sur-
face of the calyxes with oil-free cotton. The extract was subjected
to silica gel column chromatography to give Pools I–VI. Compounds
1, 2 and 3 were obtained in a 28:45:27 ratio by a reversed-phase
HPLC separation of Pool I. Compounds 4 and 5 were similarly ob-
tained in a 82:18 ratio from Pool II and compounds 6, 7 and 8 were
obtained from Pool III in a 26:43:31 ratio. NMR spectroscopic anal-
yses of Pool IV (a mixture of compounds 9 and 10), Pool V (a mix-
ture of compounds 11 and 12) and Pool VI (a mixture of
compounds 13m and 14m) indicated that all possessed malonyl
groups. For example, compound 9, the major constituent in Pool
V, exhibited signals at d 4.90 (H2-200) in the 1H NMR and at d 41.1
(C-200), 166.7 (C-100) and 170.1 (C-300) in the 13C NMR spectra. Pools
IV–VI were converted to the corresponding methyl esters and then
separated by HPLC. Compounds 9m and 10m were obtained from
Pool IV in a 79:21 ratio, compounds 11m and 12m were obtained
from Pool V in an 87:13 ratio and compounds 13m and 14m were
obtained from Pool VI in a 75:25 ratio.
(b-D-glucopyranosyloxy)fatty acids (gallicasides A–H) from Silene
gallica (Caryophyllaceae) (Asai and Fujimoto, 2010).
The characterization of the unique cyclic glycolipids from S. gal-
lica, described above, is the first study to analyze secondary metab-
olites from glandular trichome exudates of plants belonging to the
Caryophyllaceae family. These rare cyclic glycolipids structures
prompted us to further investigate other Caryophyllaceous plants.
To this regard, Cerastium glomeratum has glandular trichomes in an
upper portion of the aerial part including calyxes through field re-
search. C. glomeratum, a plant native to Europe, and is thought to
have migrated to Japan about 100 years ago and is generally found
in roadsides nowadays. This plant flowers in April and May
(Shimizu, 1995). A study on fatty acids of this plant has been
reported previously (Jamieson and Reid, 1971).
Compound 1, named glomeraside A, showed a pseudo-molecu-
lar ion at m/z 543.3894 [M+H]+ in the positive HRFABMS that cor-
responded to the molecular formula C30H54O8. The IR absorption
bands at 3590, 3440 and 1720 cm–1 suggested the presence of
⇑
Corresponding author. Tel./fax: +81 357342241.
E-mail address: fujimoto.y.aa@m.titech.ac.jp (Y. Fujimoto).
0031-9422/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.phytochem.2012.07.001

150
T. Asai et al. / Phytochemistry 82 (2012) 149–157
hydroxy and carbonyl groups. The 1H NMR spectrum (Table 1)
showed signals of seven protons assignable to a b-glucopyranosyl
moiety acylated at the 30-O and 60-O positions [d4.38 (d, J = 7.7
Hz, H-10), 3.48 (ddd, J = 9.1, 7.7, 2.5 Hz, H-20), 4.92 (t, J = 9.1 Hz,
H-30), 3.57 (td, J = 9.1, 4.8 Hz, H-40), 3.52 (m, H-50), 4.37 (dd,
J = 12.1, 1.8 Hz, Ha-60) and 4.30 (dd, J = 12.1, 4.7 Hz, Hb-60)]. These
resonances were unambiguously assigned through analysis of the
H–H COSY spectrum. Furthermore, an oxymethine proton at d
3.66 (m) and an acetyl methyl singlet at d 2.18 were observed, in
addition to the signals characteristic to a fatty acyl group (methyl
triplet at d 0.88, methylene protons adjacent to a carboxyl group at
d 2.37 and longer-chain methylene protons centered at d 1.26). The
13C NMR spectroscopic data (Table 2) confirmed the presence of a
di-O-acylated glucopyranosyl moiety (ester carbonyl resonances at
d 174.35 and 172.57). It also further established that the corre-
sponding fatty acyl group was linear (terminal methyl carbon at
d 14.13) and mono-oxygenated along the methylene chain (pres-
ence of an oxymethine carbon at d 80.05 associated with a proton
at d 3.66). The two-ester functionalities and the glucopyranosyl
moiety accounted for three of the four unsaturations of 1,
indicating the presence of an additional ring structure. The HMBC
spectrum provided evidence that the acetyl group was attached to
30-O of the glucosyl moiety (the acetyl methyl singlet at d 2.18 and
30-H at d 4.92 were correlated with the ester carbonyl carbon at d
172.57) and the oxymethine carbon of the fatty acyl group was
linked to the sugar through a glycosidic bond (10-H at d 4.52 were
correlated with the oxymethine carbon at d 80.05). The 60-O-fatty
acyl linkage was deduced from the chemical shifts of 60-H2 of the
glucosyl moiety. The carbon number of the oxygenated fatty acyl
moiety was calculated as being C22 based on the molecular for-
mula. All data indicated that compound 1 was a 1,60-cyclic ester
of (3-O-acetyl-b-D-glucopyranosyloxy)docosanoic acid.
However, the exact position and the absolute configuration of
the oxymethine center in the docosanoyl moiety remained un-
known. These issues were solved via a series of chemical transfor-
mations of 1. Alkaline hydrolysis of compound 1, followed by
treatment with acidic conditions, gave a hydroxy-docosanoic acid
that was then converted to the methyl ester 15 (EIMS m/z: 352
[MꢀH2O]+) (Fig. 2) by treatment with trimethylsilyldiazomethane.
The EIMS spectrum of the trimethyl (TMS) ether derivative of 15
Table 1
1H NMR spectroscopic data (500 MHz, CDCl3) for compounds 1–8 and 9m–14m.a
Position
1
2
3
4
5
6
7
8
2
3
2.37 (m)
1.65 (m)
2.37 (m)
1.66 (m)
2.39 (m)
1.58 (m)
2.38 (m)
1.66 (m)
2.38 (m)
1.67 (m)
2.38 (brt)
1.63 (m)
2.38 (m)
1.66 (m)
2.38 (m)
1.58 (m)
1.72 (m)
1.39–1.20 (m)
1.52 (m)
1.72 (m)
1.72 (m)
1.42–1.20 (m)
1.48 (m)
1.71 (m)
1.42–1.20 (m)
1.51 (m)
1.73 (m)
4–21b
1.38–1.20 (m)
1.59-1.41 (m)
3.66 (m)
1.40–1.20 (m)
1.60–1.40 (m)
3.58 (m)
1.39–1.20 (m)
1.59-1.41 (m)
3.59 (m)
1.74–1.20 (m)
1.58-1.41 (m)
3.67 (m)
1.41–1.20 (m)
1.60–1.40 (m)
3.57 (m)
c
9/10/11 3.66 (m)
3.57 (m)
3.57 (m)
22
0.88 (t, 7.0)
0.88 (t, 6.9)
4.42 (d, 7.7)
3.49 (dd, 9.1,
7.7)
0.88 (t, 6.9)
4.37 (d, 7.8)
3.49 (ddd, 9.5, 7.8,
2.6)
0.88 (t, 7.0)
4.47 (d, 7.8)
4.72 (dd, 9.6,
7.8)
0.88 (t, 7.0)
4.50 (d, 7.8)
4.71 (dd, 9.4,
7.8)
0.88 (t, 7.0)
4.33 (d, 7.9)
3.35 (dd, 8.8,
7.9)
0.88 (t, 7.0)
4.35 (d, 7.7)
3.36 (dd, 8.8,
7.7)
0.88 (t, 7.0)
4.31 (d, 7.7)
3.35 (dd, 9.0,
7.7)
3.56 (dd, 9.6,
9.0)
10
4.38 (d, 7.7)
3.48 (ddd, 9.1, 7.7,
2.5)
20
30
40
4.92 (t, 9.1)
4.92 (t, 9.1)
4.90 (t, 9.5)
3.59 (t, 9.6)
3.58 (t, 9.4)
3.57 (brt, 8.8)
3.57 (brt, 8.8)
3.57 (td, 9.1, 4.8)
3.49 (td, 9.1,
4.8)
3.53 (m)
3.49 (td, 9.5, 5.1)
3.50 (brt, 9.6)
3.53 (t, 9.4)
3.48 (m)
3.49 (m)
3.42 (brt, 9.6)
50
60
3.52 (m)
4.37 (dd,12.1, 1.8)
3.64 (m)
4.42 (dd, 11.9, 1.9)
3.46 (m)
4.35 (dd, 12.1,
1.9)
3.48 (m)
4.43 (dd, 12.0,
2.0)
3.48 (m)
4.35 (dd, 12.0,
1.4)
3.49 (m)
4.43 (brd, 12.0)
3.61 (m)
4.41 (dd, 12.0,
2.0)
4.45 (brd, 11.9)
4.30 (dd,12.1, 4.7)
4.19 (dd, 11.9,
5.3)
2.18 (s)
4.23 (dd, 11.9, 7.8)
4.32 (dd, 12.1,
4.4)
2.12 (s)
4.20 (dd, 12.0,
5.3)
2.12 (s)
4.33 (dd, 12.0,
4.21 (dd, 12.0,
4.23 (dd, 12.0,
5.4)
4.9)
7.8)
–
–
–
CH3CO
20/30-
OH
40-OH
2.18 (s)
2.18 (s)
2.31 (d, 2.5)
2.36 (brs)
2.30 (d, 2.6)
2.99 (brs)
2.72 (brs)
2.45 (brs)
2.99/2.62 (brs)
3.14 (brs)
2.98/2.50 (brs)
2.87 (d, 4.8)
2.91 (d, 4.8)
2.76 (d, 5.1)
2.90 (brs)
2.77 (brs)
2.88 (brs)
2.77 (brs)
Position
9m
10m
11m
12m
13m
14m
2
3
2.38 (m)
1.66 (m)
1.73 (m)
1.41–1.20 (m)
1.47 (m)
2.38 (t, 6.4)
1.67 (m)
2.37 (m)
1.67 (m)
1.72 (m)
1.41–1.20 (m)
1.46 (m)
2.38 (t, 6.5)
1.67 (m)
2.37 (m)
1.66 (m)
1.72 (m)
1.41–1.20 (m)
1.47 (m)
3.57 (m)
2.38 (t, 6.5)
1.67 (m)
4–21b
1.38–1.20 (m)
1.58-1.39 (m)
3.57 (m)
1.38–1.20 (m)
1.58-1.39 (m)
3.58 (m)
1.39–1.20 (m)
1.58-1.39 (m)
3.57 (m)
c
9/10/11
3.57 (m)
3.57 (m)
22
10
20
30
40
50
60
0.88 (t, 7.0)
4.52 (d, 7.7)
4.97 (dd, 9.8, 7.7)
5.02 (t, 9.8)
3.64 (td, 9.8, 5.0)
3.51 (m)
4.36 (dd, 12.1, 2.2)
4.32 (dd, 12.1, 4.1)
2.14 (s)
0.88 (t, 6.9)
4.55 (d, 7.4)
4.98 (dd, 9.7, 7.4)
5.01 (t, 9.7)
3.64 (td, 9.7, 5.0)
3.52 (m)
4.45 (dd, 12.0, 2.0)
4.21 (dd, 12.0, 5.0)
2.11 (s)
0.88 (t, 6.9)
4.53 (d, 7.8)
4.92 (dd, 9.8, 7.8)
5.09 (t, 9.8)
3.70 (td, 9.8, 3.2)
3.57 (m)
4.39 (dd, 12.1, 1.7)
4.26 (dd, 12.1, 5.0)
2.05 (s)
0.88 (t, 7.0)
4.56 (d, 7.9)
4.93 (dd, 9.8, 7.9)
5.09 (t, 9.8)
3.71 (td, 9.8, 3.4)
3.58 (m)
4.40 (dd, 12.1, 1.7)
4.28 (dd, 12.1, 5.0)
2.05 (s)
3.50 (d, 16.3)
3.39 (d, 16.3)
0.88 (t, 7.0)
0.88 (t, 7.0)
4.52 (d, 7.9)
4.82 (dd, 9.3, 7.9)
3.65 (td, 9.3, 3.0)
3.59 (td, 9.3, 2.2)
3.51 (m)
4.49 (d, 7.9)
4.81 (dd, 9.5, 7.9)
3.65 (td, 9.5, 3.1)
3.57 (t, 9.5, 2.0)
3.50 (m)
4.38 (dd, 12.1, 1.8)
4.46 (dd, 12.0, 2.0)
4.29 (dd, 12.1, 5.0)
4.19 (dd, 12.0, 5.5)
–
–
CH3CO
200
3.35 (s)
3.35 (s)
3.36 (s)
3.36 (s)
3.49 (d, 16.3)
3.40 (d, 16.3)
3.47 (d, 16.1)
3.42 (d, 16.1)
3.78 (s)
3.41 (d, 3.1)
2.74 (d, 2.0)
3.47 (d, 16.2)
3.42 (d, 16.2)
3.78 (s)
3.44 (d, 3.0)
2.65 (d, 2.2)
CO2Me
20/30-OH
40-OH
3.73 (s)
3.73 (s)
3.77 (s)
3.77 (s)
–
–
–
–
2.81 (d, 5.0)
2.74 (d, 5.0)
3.47 (d, 3.2)
3.39 (d, 3.4)
a
b
c
Multiplicity and coupling constants (J in Hz) are in parentheses.
Methylene protons excluding those coupling to the oxymethine proton.
Methylene protons (4H) coupling to the oxymethine proton.

T. Asai et al. / Phytochemistry 82 (2012) 149–157
151
Table 2
13C NMR spectroscopic data (125 MHz, in CDCl3) for 1–8 and 9m–15m.a
No.
1
2
3
4
5
6
7
8
9m
10m
11m
12m
13m
14m
1
174.35
80.05
34.53
34.30
34.04
29.84
29.65
29.64
29.62
29.62
29.59
29.35
27.99
27.49
27.35
26.81
25.37
24.47
24.27
31.92
22.69
14.13
101.84
72.20
77.85
69.17
74.03
63.29
21.07
172.57
174.14
79.12
34.37
33.99
33.91
29.85
29.67
29.65
29.65
29.63
29.61
29.36
27.12
26.98
26.48
26.38
25.32
24.43
23.86
31.93
22.70
14.13
101.53
72.27
78.06
69.40
73.75
63.07
21.07
172.66
173.97
82.02
35.26
34.41
34.27
29.78
29.70
29.67
29.65
29.65
29.61
29.59
29.36
27.66
27.27
27.02
25.65
24.47
23.23
31.93
22.70
14.13
102.61
72.21
78.16
70.11
73.87
64.02
21.05
172.78
174.56
80.54
34.57
34.41
34.08
29.96
29.71
29.67
29.67
29.67
29.62
29.34
29.08
27.49
27.28
26.83
25.14
24.40
24.21
31.91
22.68
14.10
100.24
76.75
74.78
70.68
73.24
63.35
20.95
171.22
174.21
79.95
34.40
34.13
33.86
29.91
29.72
29.67
29.67
29.67
29.64
29.34
27.12
26.02
26.70
26.23
25.05
24.36
23.80
31.91
22.67
14.10
100.07
76.01
75.01
70.93
72.87
63.08
20.94
171.38
174.57
79.72
34.62
34.30
34.06
29.84
29.65
29.63
29.63
29.61
29.60
29.34
28.04
27.47
27.30
26.85
25.41
24.42
24.27
31.91
22.68
14.11
101.64
73.62
76.08
69.81
74.08
63.34
174.27
78.88
34.43
33.99
33.96
29.84
29.68
29.66
29.65
29.63
29.61
29.35
27.13
26.98
26.52
26.41
25.36
24.41
23.94
31.92
22.68
14.11
101.33
73.31
76.28
70.00
74.15
63.21
174.06
81.70
35.34
34.44
34.30
29.78
29.69
29.67
29.65
26.65
29.62
29.60
29.36
27.70
27.31
27.07
25.67
24.46
23.31
31.92
22.69
14.11
102.44
73.31
76.30
70.61
74.12
64.15
174.41
80.47
34.45
34.28
34.05
29.90
29.68
29.66
29.66
29.62
29.34
27.99
27.55
27.53
27.27
26.82
25.20
24.41
24.13
31.90
22.68
14.11
99.90
72.32
75.71
69.03
73.81
62.99
20.85
172.04
164.92
41.08
166.42
52.56
174.13
79.70
34.28
34.01
33.92
29.87
29.69
29.67
29.67
29.67
29.65
29.35
27.16
27.02
26.60
26.27
25.12
24.38
23.77
31.91
22.68
14.11
99.67
72.34
76.02
69.30
73.53
62.78
20.85
172.20
164.92
41.08
166.44
52.56
174.25
80.76
34.51
34.41
34.08
29.95
29.68
29.66
29.66
29.62
29.34
28.05
27.54
27.52
27.34
26.82
25.10
24.46
24.23
31.91
22.68
14.10
100.41
71.27
77.84
68.85
73.22
63.23
20.77
169.59
166.34
41.19
168.05
52.96
174.05
80.17
34.40
34.11
33.85
29.92
29.70
29.67
29.67
29.67
29.64
29.34
27.17
27.07
26.70
26.21
25.03
24.20
23.76
31.91
22.68
14.10
100.31
71.32
77.87
69.08
72.91
62.99
20.77
169.60
166.36
41.18
168.17
52.98
174.37
80.38
34.45
34.38
34.07
29.89
29.73
29.73
29.68
29.67
29.64
29.35
28.07
27.51
27.33
26.82
25.18
24.43
24.21
31.91
22.68
14.10
99.77
75.77
75.49
69.86
73.43
63.3
174.13
79.78
34.33
34.10
33.89
29.84
29.73
29.69
29.69
29.68
29.65
29.35
27.12
27.03
26.65
26.25
25.14
24.38
23.78
31.91
22.68
14.10
99.62
75.86
75.66
70.03
73.12
63.12
9/10/11
2–19b
20
21
22
10
20
30
40
50
60
Ac (CH3)
Ac (CO)
100
200
300
165.53
41.18
168.16
52.91
165.56
41.18
168.26
52.92
MeO
a
Signal assignments for 1–4, 7, 9m–11m and 13m were based on 2D experiments including HSQC and HMBC.
Except for the oxymethine carbon.
b
showed intense fragment ions at m/z 257 ([TMSOCH(CH2)10CH3]+
due to C-10/C-11 cleavage) and 287 ([CH3OOC(CH2)10CH(OTMS)]+
due to C-11/C-12 cleavage) in addition to a [M–Me]+ ion peak at
m/z 427. The diagnostic fragment ions unequivocally established
that compound 15 was methyl 11-hydroxydocosanoate. GLC anal-
ysis of the TMS ether derivative of 15 showed a single peak, indi-
cating the absence of homologous compounds. The absolute
configuration at C-11 of 15 was determined to be R by application
of the Ohrui–Akasaka method (Ohtaki et al., 2005) (vide infra).
Hence, the structure of glomeraside A (1) was determined to be
fatty acid methyl esters 16 and 17, respectively, as described for
1. The methyl esters 16 (EIMS m/z: 352 [M–H2O]+) and 17 (EIMS
m/z: 352 [M–H2O]+) (Fig. 2) were determined to be methyl 10-
hydroxydocosaonoate and methyl 9-hydroxydocosanoate, respec-
tively, on the basis of the EIMS spectra of the corresponding TMS
ether derivatives (16-TMS m/z 427 [M–Me]+, 271 ([TMS-
OCH(CH2)11Me]+ due to C-9/C-10 cleavage), 273 ([MeO-
OC(CH2)9CH(OTMS)]+ due to C-10/C-11 cleavage; 17-TMS m/z
427 [M–Me]+, 285 ([TMSOCH(CH2)12Me]+ due to C-8/C-9 cleavage),
259 ([CH3OOC(CH2)7CH(OTMS)]+ due to C-9/C-10 cleavage). The
absolute configurations at C-10 of 16 and C-9 of 17 were deter-
mined as being R (vide infra). It was therefore concluded that the
structures of glomerasides B (2) and C (3) were the 1,60-cyclic ester
the 1,60-cyclic ester of 11(R)-(3-O-acetyl-b-
D-glucopyranosyl-
oxy)docosanoic acid, as depicted in Fig. 1.
Compounds 2 and 3, named glomerasides B and C, respectively,
had the same molecular formula as 1. The 1H NMR spectra of 2 and
3 (Table 1) resembled that of 1, thus indicating the presence of a
glucopyranosyl moiety that was esterified at the 30-O and 60-O
positions and glycosylated at the 10-O position (dH 3.66, dC 79.12
for 2 and dH 3.58, dC 82.02 for 3) with an oxymethine carbon of a
fatty acyl group, as in 1. The HMBC spectra of 2 and 3 confirmed
their 30-O-acetylation, 60-O-fatty acylation, and glycosidic linkage.
The 13C NMR spectra (Table 1) of 2 and 3 were also similar to that
of 1, but small differences were observed for most signals, com-
pared to those of compound 1. The resonance of the oxymethine
carbon in the fatty acyl moiety displayed the largest difference
(an upfield shift by 0.93 ppm for 2 and a downfield shift by
1.97 ppm for 3, compared to that of 1). These deviations observed
in the 13C spectroscopic data suggested that compounds 2 and 3
could be either a positional isomer or a stereoisomer of 1 with re-
spect to the oxygenated position in the fatty acyl moiety. To solve
this problem, compounds 2 and 3 were converted to the hydroxy
of 10(R)-(3-O-acetyl-b-D-glucopyranosyloxy)docosanoic acid and
the 1,60-cyclic ester of 9(R)-(3-O-acetyl-b-
D-glucopyranosyl-
oxy)docosanoic acid, as depicted in Fig. 1.
Compounds 1, 2 and 3 were treated with Ac2O and pyridine to
afford the corresponding peracetates 1a, 2a and 3a, respectively,
which were helpful for the structure elucidation of compounds
4–8. The three peracetates could not be separated by HPLC, how-
ever, with standard reversed-phase columns (ODS). However, they
were effectively separated with an Inertsil ODS-P column (see
Experimental).
The configuration at the oxymethine center in 15–17 was deter-
mined using the Ohrui–Akasaka method (Ohtaki et al., 2005). Com-
pound 15 was thus converted into the ester derivative 15a of
(1R,2R)-2-(2,3-anthracenedicarboximide)cyclohexanecarboxylic
acid ((R)-ACC acid) and into the (S)-ACC ester 15b (Fig. 2). Com-
pounds 16 and 17 were similarly converted into the (R)-ACC 16a
and (S)-ACC ester 16b and (R)-ACC ester 17a and (S)-ACC ester

152
T. Asai et al. / Phytochemistry 82 (2012) 149–157
1
O
O
O
(CH2)6
O
(CH2)5
(CH2)4
O
11R
10R
9R
O
O
O
22
6′
H
H
H
R
(CH2)10CH3
(CH2)11CH3
(CH2)12CH3
O
R3O
R2O
R3O
R2O
R3O
R2O
1′
O
O
O
OR1
OR1
OR1
1: R2=Ac, R1=R3=H
1a: R1=R2=R3=Ac
4: R1=Ac, R2=R3=H
6: R1=R2=R3=H
2: R2=Ac, R1=R3=H
2a: R1=R2=R3=Ac
5: R1=Ac, R2=R3=H
7: R1=R2=R3=H
3: R2=Ac, R1=R3=H
3a: R1=R2=R3=Ac
8: R1=R2=R3=H
O
O
(CH2)6
(CH2)5
O
11R
10R
O
O
H
H
(CH2)10CH3
(CH2)11CH3
O
R3O
R2O
R3O
R2O
O
O
OCOCH2COOR1
OCOCH2COOR1
1′′ 2′′
3′′
9: R1=H, R2=Ac, R3=H
9m: R1=Me, R2=Ac, R3=H
9a: R1=Me, R2=R3=Ac
13: R1=H, R2=R3=H
10: R1=H, R2=Ac, R3=H
10m: R1=Me, R2=Ac, R3=H
10a: R1=Me, R2=R3=Ac
14: R1=H, R2=R3=H
13m: R1=Me, R2=R3=H
14m: R1=Me, R2=R3=H
O
O
(CH2)6
O
(CH2)5
11R
10R
O
O
H
H
(CH2)10CH3
(CH2)11CH3
O
HO
HO
R1OOCCH2COO
O
O
R1OOCCH2COO
3′′ 2′′
1′′
OR2
OR2
11: R1=H, R2=Ac
11m: R1=Me, R2=Ac
12: R1=H, R2=Ac
12m: R1=Me, R2=Ac
Fig. 1. Compounds 1–14 and their derivatives.
ing to H2-2 were consistently observed at higher field in the spec-
22
11
tra of the (R)-ACC esters than in those of the corresponding (S)-ACC
esters. The d(R)–(S) values for H2-2 of 15ab–17ab were negative and
the magnitudes of the values (ꢀ0.074, ꢀ0.058, ꢀ0.030 ppm for
17ab, 16ab and 15ab, respectively) were in good agreement with
those described in the paper (Ohtaki et al., 2005). The results per-
mitted the assignments of 11R, 10R and 9R configurations for com-
pounds 15, 16 and 17, respectively. Ohrui reported that anisotropic
effect of anthracene derivative reaches protons attached to the car-
bon 10 bond apart from the oxymethine carbon (Ohrui, 2008). The
d(R)–(S) value for the COOMe signal (10 bond apart from the oxyme-
thine carbon) of 17ab also showed an expected negative small d(R)–(S)
value (ꢀ0.006 ppm). Careful interpretation of d(R)–(S) values in the
paper of the Ohrui–Akasaka method (Ohtaki et al., 2005) indicated
that the anthracene ring causes a small but unique deshielding ef-
fect on the protons attached to the carbon 12–14 bond apart from
the oxymethine carbon. Thus, the signs of the d(R)–(S) values for the
COOMe signal of 15a/b (12 bond) and for the terminal methyl trip-
let signals of 17a/b (13 bond) and 16a/b (12 bond) were rational-
ized by this anomalous anisotropic effect.
MeOOC
R
1
H
RO
: R=H
a: R=(R)-ACC
b: R=(S)-ACC
MeOOC
10
R
H
RO
: R=H
a: R=(R)-ACC
b: R=(S)-ACC
9
MeOOC
R
H
RO
: R=H
a: R=(R)-ACC
b: R=(S)-ACC
The assignment was verified in the case of methyl 10-hydroxy-
docosanoate. Authentic samples of 16a and 16b with known
absolute configuration at C-10 were prepared from methyl
(R)-10-hydroxydocosanoate. This compound was synthesized by
reacting undecanylmagnesium bromide with methyl (S)-10,11-
epoxyundecanoate in the presence of CuI. The chiral epoxide was
obtained by kinetic resolution of the racemic epoxide (Schaus
et al., 2002). The chemical shifts of H2-2, COOMe and H3-22 of
the derivatives were in complete agreement with those observed
for 16a and 16b, thus confirming the 10(R) configuration of 16.
Compound 4, named glomeraside D, had the molecular formula
(R)-ACC =
(S)-ACC =
O
O
R
S
N
N
C
O
C
O
R
S
O
O
Fig. 2. Structures of the hydroxy fatty acid methyl esters 15–17 and their
derivatives.
17b (Fig. 2). The chemical shifts of the key proton signals of these
derivatives are listed in Table 3. The triplet resonances correspond-
C
30H54O8, which was identical to that of 1. The 1H NMR spectrum

T. Asai et al. / Phytochemistry 82 (2012) 149–157
153
Table 3
Pertinent 1H NMR (500 MHz, CDCl3) spectroscopic data for the ACC esters 15ab, 16ab and 17ab.
Compound
CO2Me
d(R)–(S)
+0.005
+0.001
–0.006
H2-2
d(R)–(S)
–0.030
–0.058
–0.074
H3-22
d(R)–(S)
15
15a
15b
3.667 (s)
3.666 (s)
3.661 (s)
2.300 (t, J = 7.5 Hz)
2.198 (t, J = 7.5 Hz)
2.218 (t, J = 7.5 Hz)
0.887 (t, J = 7.2 Hz)
0.867 (t, J = 7.2 Hz)
0.866 (t, J = 7.2 Hz)
+0.001
–0.005
–0.005
16
3.667 (s)
3.654 (s)
3.653 (s)
2.300 (t, J = 7.5 Hz)
2.122 (t, J = 7.5 Hz)
2.180 (t, J = 7.5 Hz)
0.887 (t, J = 7.2 Hz)
0.877 (t, J = 7.2 Hz)
0.882 (t, J = 7.2 Hz)
16aa
16ba
17
17a
17b
3.667 (s)
3.635 (s)
3.641 (s)
2.300 (t, J = 7.5 Hz)
2.073 (t, J = 7.5 Hz)
2.147 (t, J = 7.5 Hz)
0.887 (t, J = 7.2 Hz)
0.882 (t, J = 7.2 Hz)
0.887 (t, J = 7.2 Hz)
a
The 1H NMR data was identical with that of synthetic authentic sample.
of 4 resembled that of 1 (Table 1). However, analysis of the H–H
COSY spectrum indicated that the signals at d 4.72 and 3.59 corre-
sponded to H-20 and H-30 of the glucopyranosyl moiety, respec-
tively. The downfield shift of H-20 indicated a 20-O-acetylation of
the glucose ring in place of a 30-O-acylation. This was confirmed
by HMBC correlations between the acetyl methyl (d 2.12) and H-
20 (d 4.72) resonances and the acetyl carbonyl carbon (d 171.22).
The acetyl derivative of 4 was identified as the peracetate 1a.
Glomeraside D (4) was thus found to be the 1,60-cyclic ester of
(d 3.57) and an acetyl methyl singlet (d 2.14). In addition, reso-
nances corresponding to a malonate methyl ester were observed
at d 3.35 (s, 2H) and d 3.73 (s, 3H) (Table 1). The 13C NMR spectrum
exhibited 34 signals among which four resonances were assigned
to the malonate methyl ester (d 166.42, 164.92, 52.56 and 41.08)
(Table 2). The HMBC spectrum gave evidence for the 20-O-malony-
lation (H-20 at d 4.97 and H2-200 at d 3.35 were correlated with the
malonate C-100 at d 164.92) and the 30-O-acetylation (the acetyl
methyl singlet at d 2.14 and 30-H at d 5.02 were correlated with
the ester carbonyl carbon at d 172.04). The remaining ester linkage
was therefore located at 60-O-position. A glycosidic bond with an
oxygenated carbon in the fatty acyl moiety was also determined
by the HMBC correlation between H0-1 (d 4.52) and the oxyme-
thine carbon (d 80.47). The hydroxy fatty acid methyl ester that
was obtained from 9m was identified as methyl 11(R)-hydroxy-
docosanoate (15). The 1H NMR spectrum of the (R)-ACC ester of
the methyl ester was superimposable to that of 15. Thus, com-
pound 9 was determined to be the 1,60-cyclic ester of 11(R)-(3-O-
11(R)-(2-O-acetyl-b-D-glucopyranosyloxy)docosanoic acid, as de-
picted in Fig. 1.
Compounds 5, named glomeraside E, had the same molecular
formula as 1. Analysis of the 1H NMR spectrum of 5, together with
that of the H–H COSY spectrum, indicated the same acylation pat-
tern on the b-glucopyranosyl moiety (H-20 at d 4.71, H-30 at d 4.71)
as that found for compound 4 (Table 1). The 13C NMR spectroscopic
data of 5 (Table 2) was only slightly different from that of 4 and
evidenced from the presence of the glucopyranosyl, oxygenated
docosanoyl, and acetyl moieties. The acetate derivative of 5 was
identified as the peracetate 2a. Thus, glomeraside E (5) was deter-
acetyl-2-O-malonyl-b-D-glucopyranosyloxy)docosanoic acid, as
depicted in Fig. 1. It was named glomeraside I.
mined to be the 1,60-cyclic ester of 10(R)-(2-O-acetyl-b-
D-glucopyr-
Compound 10m had the same molecular formula as 9m. The 1H
and 13C NMR spectroscopic data indicated that the b-glucopyrano-
syl moiety was acylated at the 20-O, 30-O and 60-O positions, simi-
larly to 9m (Tables 1 and 2). HMBC correlations indicated that
the acetyl, malonyl and fatty acyl groups were located at the same
position as in compound 10m. The hydroxy fatty acid methyl ester
obtained from 10m was identified as methyl 10(R)-hydroxydoco-
sanoate (16). The 1H NMR spectrum of the (R)-ACC ester of the
methyl ester was superimposable to that of 16. Compound 10
was thus determined to be the 1,60-cyclic ester of 10(R)-(3-O-acet-
anosyloxy)docosanoic acid, as depicted in Fig. 1.
Compound 6, named glomeraside F, showed a pseudo-molecu-
lar ion at m/z 501.3804 [M+H]+ in the positive HRFABMS that cor-
responded to the molecular formula C28H52O7. The 1H and 13C NMR
spectra of 6 indicated that there is no acetyl group in the molecule
and that the glucopyranosyl moiety was only acylated at the 60-O
position (Tables 1 and 2). The oxymethine proton and carbon sig-
nals (dH 3.65 and dC d 79.72) indicated the presence of a glycosidic
linkage to an oxymethine carbon of the docosanoyl moiety. The
acetate derivative of 6 was identified as the peracetate 1a. Thus,
the structure of glomeraside F (6) was determined to be the 1,60-
yl-2-O-malonyl-b-D-glucopyranosyloxy)docosanoic acid, as de-
picted in Fig. 1. It was named glomeraside J. Compounds 9m and
10m were converted into the peracetates 9a and 10a, respectively,
to help elucidate the structures of 13 and 14 (vide infra).
cyclic ester of 11(R)-(b-D-glucopyranosyloxy)docosanoic acid, as
depicted in Fig. 1.
Compounds 7 and 8, named glomerasides G and H, respectively,
had the same molecular formula as 6. The 1H and 13C NMR spectra
of 7 and 8 resembled those of 6, indicating a 1,60-cyclic ester struc-
ture as in 6 (Tables 1 and 2). The acetate derivatives obtained from
7 and 8 were identified as the peracetates 2a and 3a, respectively.
Thus, the structures of glomerasides G (7) and H (8) were deter-
Compound 11m had the same molecular formula as 9m. The 1H
NMR spectroscopic data resembled that of 9m, thus indicating the
presence of a b-glucopyranosyl moiety that was acylated at the 20-
O, 30-O and 60-O positions (H-20 at d 4.92, H-30 at d 5.09 and H-60 at d
4.40/4.28) with acetyl, malonyl and fatty acyl groups and glycosyl-
ated at the 10-O position with an oxymethine carbon of a fatty acyl
group. The 13C NMR spectroscopic data was similar to that of com-
pound 9m. However, the signals of the ester carbonyl groups
showed small but distinct differences when compared to those of
9m and 10m (Table 2). The differences were found to be due to
the interchange of the acyl groups at the 20-O and 30-O positions
in compound 11m, compared to compounds 9m and 10m. The
HMBC spectrum of 11m established the 20-O-acetyation (the acetyl
methyl singlet at d 2.05 and H-20 at d 4.92 were correlated with the
acetyl carbonyl carbon at d 169.59) and the 30-O-malonylation (H-
30 at d 5.09 and H2-300 at 3.49/3.40 were correlated with the malon-
ate C-100 at d 166.34). The hydroxy fatty acid methyl ester obtained
mined to be the 1,60-cyclic esters of 10(R)-(b-
oxy)docosanoic acid and 9(R)-(b- -glucopyranosyloxy)docosanoic
D-glucopyranosyl-
D
acid, respectively, as depicted in Fig. 1.
Compound 9m showed a pseudo-molecular ion at m/z 643.4080
[M+H]+ in the positive HRFABMS that corresponded to the molec-
ular formula C34H58O11. The 1H NMR spectrum of 9m presented se-
ven proton signals that correspond to
a 2,3,6-tri-O-acyl-b-
glucopyranosyl moiety [d 4.52 (d, J = 7.7 Hz, H-10), 4.97 (dd,
J = 9.8, 7.7 Hz, H-20), 5.02 (dd, J = 9.8 Hz, H-30), 3.64 (td, J = 9.8,
5.0 Hz, H-40), 3.51 (m, H-50), 4.36 (dd, J = 12.1, 2.2 Hz, Ha-60) and
4.32 (dd, J = 12.1, 4.1 Hz, Hb-60)], an oxygenated fatty acid moiety

154
T. Asai et al. / Phytochemistry 82 (2012) 149–157
Table 4
from 11m was identified as methyl 11(R)-hydroxydocosanoate
(15). Hence, compound 11 was determined to be the 1,60-cyclic es-
ter of 11(R)-(2-O-acetyl-3-O-malonyl-b-D-glucopyranosyloxy)doc-
osanoic acid, as depicted in Fig. 1. It was named glomeraside K.
Compound 12m had the same molecular formula as 9m. The 1H
and 13C NMR spectroscopic data indicated that 12m had the same
acylation pattern on the glucose ring as in compound 11m (Tables
1 and 2). The hydroxy fatty acid methyl ester obtained from 12m
was identified as methyl 10(R)-hydroxydocosanoate (16). Thus,
compound 12 was determined to be the 1,60-cyclic ester of
Relative abundance of glomerasides in the glandular trichome
exudate.
Compound
Relative abundance (%)a
Glomeraside A (1)
Glomeraside B (2)
Glomeraside C (3)
Glomeraside D (4)
Glomeraside E (5)
Glomeraside F (6)
Glomeraside G (7)
Glomeraside H (8)
Glomeraside I (9)
Glomeraside J (10)
Glomeraside K (11)
Glomeraside L (12)
Glomeraside M (13)
Glomeraside N (14)
3.7
5.9
3.7
24.8
5.4
9.6
16.1
11.7
3.7
1.0
8.9
10(R)-(2-O-acetyl-3-O-malonyl-b-
noic acid, as depicted in Fig. 1. It was named glomeraside L.
Compound 13m showed pseudo-molecular ion at m/z
D-glucopyranosyloxy)docosa-
a
601.3933 [M+H]+ in the positive HRFABMS that corresponded to
the molecular formula C32H56O10. The 1H NMR spectrum had seven
signals that corresponded to a 2,6-di-O-acyl-b-glucopyranosyl
moiety (d 4.81 for H-20 and d 4.38/4.29 for H2-60), in addition to sig-
nals for a malonate methyl ester [d 3.78 (s, 3H) and 3.42 (d,
J = 16.1 Hz)/3.47 (d, J = 16.1 Hz)] and an oxygenated fatty acyl
group (d 3.57) (Table 1). The 20-O-malonylation was confirmed by
HMBC correlations between H-20 and H2-200 and the malonate C-
100 at d 165.53. These NMR spectroscopic data, together with the
fact that acetylation of 13m furnished the peracetate 9a allowed
us to assign the structure of compound 13 as the 1,60-cyclic ester
1.3
3.1
1.1
a
The remainder (19.2% of the whole extract) of the exu-
date constituents was exclusively a mixture of wax (hydro-
carbons) that was not included in the % calculation.
llaceous plants can further convert glycosyloxy-fatty acids to mac-
rocyclic compounds via an intramolecular lactonization. The initial
hydroxylation step is reminiscent of the action of a cytochrome
P450 enzyme that is associated with the formation of a hydroxy
fatty acid as one of the components of cutin polyester lipids (Li-
Beisson et al., 2009). Resin glycosides, most of which have a mac-
rocyclic ring across disaccharides, are reported from the Convolvul-
aceae family (Pereda-Miranda et al., 2010). Investigation of
secondary metabolites of glandular trichome exudates may help
understand the relationships between chemical structures of gly-
colipids and the genera of Caryophyllaceae. Such studies are cur-
rently in progress in our laboratory.
of 11(R)-(2-O-malonyl-b-D-glucopyranosyloxy)docosanoic acid, as
depicted in Fig. 1. It was named glomeraside M.
Compound 14m had the same molecular formula as 13m. The
13
1H and C NMR spectra of 13m indicated 20-O-malonylation (H-
20 at d 4.82) and the 60-O-fatty acylation (H2-60 at d 4.46/4.19) on
a b-glucopyranosyl moiety. These spectroscopic data and the fact
that acetylation of 14m furnished 10a established that compound
14m was the 1,60-cyclic ester of 10(R)-(2-O-malonyl-b-
D-glucopyr-
anosyloxy)docosanoic acid, as depicted in Fig. 1. Compound 14 was
named glomeraside N.
4. Experimental
4.1. General experimental procedures
3. Concluding remarks
The apparatus and conditions used for the experiments are de-
scribed in our previous paper (Asai and Fujimoto, 2010), unless
otherwise cited. HPLC was performed using an ODS column (Shi-
madzu Shim-Pack CLC-ODS, 15 cm ꢁ 4.6 mm i.d.) under isocratic
solvent conditions, unless otherwise stated.
A glandular trichome exudate of C. glomeratum specifically con-
tained the 14 new macrocyclic glycolipids (compounds 1–14). The
relative abundance of these constituents is summarized in Table 4.
Glomeraside D (4) was the most abundant constituent (25%), fol-
lowed by glomeraside G (7) (16%). The glomerasides were found
to be unique 1,60-cyclic esters with 19-, 18- or 17-membered ring.
The oxygenation position in the docosanoyl moiety varied from C-9
to C-11, whereas the absolute configurations at the oxymethine
centers were consistently R.
This is the first report that presents evidence of the presence of
1,60-cyclic esters of glycosyloxy-fatty acids in the plants. The
structures of glomerasides were in sharp contrast with those of
gallicasides isolated from S. gallica, which were 1,20-cyclic esters
4.2. Plant material
C. glomeratum (Caryophyllaceae) were collected in May 2010 in
Kawasaki city in the Kanagawa prefecture. The plant was identified
by Prof. S. Kohshima (Department of Biological Sciences, Graduate
School of Bioscience and Biotechnology, Tokyo Institute of Technol-
ogy). A voucher specimen (CMS22–02) was deposited in the
Department of Chemistry and Materials Science of the Tokyo Insti-
tute of Technology.
of (b-
D
-glucopyranosyloxy)fatty acids (Asai and Fujimoto, 2010).
-xylopyranosyloxy)fatty acids were,
1,20-Cyclic esters of 5-(b-
D
however, reported from the aerial part of Stellaria dichotoma
(Caryophyllaceae) (Ganenko et al., 2001). The 1,20-cyclic esters is
also likely to be of glandular trichome origin, since S. dichotoma
is known to be rich in glandular trichomes. These findings thus
suggest that cyclic glycolipids are specific in glandular trichome
exudates of the Caryophyllaceae family and are interesting from
a chemotaxonomic point of view. Our present and previous find-
ings also suggest that glandular trichomes have a unique capacity
to introduce a hydroxy group at various positions in fatty acids and
to form a glycosidic bond with the introduced hydroxy group.
Examples of glycosyloxy-fatty acids have been reported in glandu-
lar trichome exudates of I. lutea and P. louisiana (Martyniaceae)
(Asai et al., 2010). It appears that glandular trichomes of Caryophy-
4.3. Extraction and isolation
Fresh upper aerial parts of C. glomeratum (fresh wt. 400 g) were
briefly (ca. 10 s) rinsed in a beaker containing Et2O (1.5 L), and the
Et2O solution was filtered and concentrated to dryness under re-
duced pressure. The residue (788 mg) was subjected to silica gel
(80 g) column chromatography (CC). The column was eluted with
a discontinuous gradient of hexane–EtOAc (10:1 ? 0:1, total vol-
ume 1.4 L) and EtOAc–MeOH (50:1 ? 3:1, total volume 450 mL).
The fractions (50 mL each, a total of 36 fractions) were combined
to give a non-polar fraction (152 mg, frs. 1–7, eluted with hex-
ane–EtOAc, 10:1 ? 1:1, the major substance was identified as a

T. Asai et al. / Phytochemistry 82 (2012) 149–157
155
mixture of hydrocarbons) and six pools (I–VI) according to the TLC
profile: Pool I (70 mg, frs. 18–20, a mixture of 1, 2 and 3, eluted
with hexane–EtOAc, 1:1), Pool II (159 mg, a mixture of 4 and 5,
frs. 21–22, eluted with hexane–EtOAc, 1:2), Pool III/IV (222 mg,
frs. 23–27, eluted with hexane–EtOAc, 1:2), Pool V (54 mg, a mix-
ture of 11 and 12, frs. 28–33, eluted with EtOAc-MeOH, 5:1) and
Pool VI (22 mg, a mixture of 13 and 14, frs. 34–36, eluted with
EtOAc–MeOH, 5:1 ? 3:1). The Pool III/IV was further separated
by silica gel CC with increasing polarity of hexane-EtOAc to give
Pool III (195 mg, a mixture of 6, 7 and 8, eluted with hexane–EtOAc,
1:2) and Pool IV (23 mg, a mixture of 9 and 10, eluted with EtOAc).
A portion of Pool I (26 mg) was further separated by HPLC (MeOH–
H2O, 20:1; flow rate, 1.0 mL/min) to afford 1 (6.8 mg), 2 (11.1 mg)
and 3 (6.8 mg) at 12.4, 12.8 and 13.3 min, respectively. A portion of
Pool II (10 mg) was further separated by HPLC (MeOH–H2O, 20:1;
flow rate, 1.0 mL/min) to afford 4 (7.8 mg) and 5 (1.7 mg) at 13.9
and 14.7 min, respectively. A portion of Pool III (18 mg) was further
separated by HPLC (MeOH–H2O, 20:1; flow rate, 1.0 mL/min, UV
detection at 215 nm) to afford 6 (3.6 mg), 7 (6.0 mg) and 8
(4.4 mg) at 12.4, 12.8and 13.4 min, respectively.
PTLC (hexane–EtOAc, 3:1) gave the peracetyl derivative 1a
(2.5 mg): Colorless oil; ½a D25
ꢂ
– 1.0 (c = 0.22, CHCl3); IR (CHCl3) mmax
2930, 2860, 1720 cm–1; (+)-HRFABMS m/z: 649.3950 [M+Na]+
(calcd for C34H58O10Na, 649.3928); 1H NMR d: 0.88 (t, J = 6.9 Hz,
H3-22), 1.52-1.20 (m), 1.63 (m, Ha-3), 1.75 (m, Hb-3), 2.00 (s, Ac),
2.02 (s, Ac), 2.03 (s, Ac), 2.32 (ddd, J = 11.7, 9.4, 4.5 Hz, Ha-2),
2.43 (ddd, J = 11.7, 8.0, 4.5 Hz, Hb-2), 3.56 (m, H-11), 3.69 (m, H-
50), 4.09 (dd, J = 12.3, 4.6 Hz, Ha-60), 4.15 (dd, J = 12.3, 1.9 Hz, Hb-
60), 4.53 (d, J = 8.0 Hz, H-10), 4.98 (dd, J = 9.7, 8.0 Hz, H-20), 5.10 (t,
J = 9.7 Hz, H-40), 5.20 (t, J = 9.7 Hz, H-30), 13C NMR d: 14.11, 20.63,
20.65, 20.68, 22.68, 24.33, 24.35, 25.09, 26.88, 27.29, 27.52,
27.64, 28.14, 29.34, 29.61, 29.65 (x2), 29.68, 29.93, 31.90, 34.12,
34.44, 34.60, 62.22, 68.28, 71.46, 71.77, 73.14, 80.85, 100.34,
169.28, 169.31, 170.43, 173.94.
4.5. Glomeraside B (2)
Colorless oil; (+)-HRFABMS m/z: 543.3919 [M+H]+ (calcd for
C30H55O8, 543.3897); ½a D25
ꢂ
– 6.3 (c = 1.1, CHCl3); IR (CHCl3) mmax
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
A portion of Pool IV (12 mg) in Et2O (100
l
L) and MeOH (40
lL)
was treated with TMSCH2N2 in hexane (0.6 M solution, 40
l
L) at
Methyl 10(R)-hydroxydocosanoate 16 (0.8 mg) was obtained
from 2 (2.0 mg) in the same manner as described for 1. EIMS m/z:
352 ([M–H2O]+, 1), 339 ([M–MeO]+, 2), 320 ([M–H2O–MeO]+, 10),
201 (C-10/C-11 cleavage, 53), 172 (46), 169 (201–MeOH) (100),
143 (14), 87 (42), 55 (24); 1H NMR (CDCl3) d: 3.67 (s, OMe), 3.59
(m, H-10), 2.30 (t, J = 7.6 Hz, H2-2), 1.65-1.25 (m), 0.88 (t,
J = 6.7 Hz, H3-22). A portion of 16 (0.1 mg) was converted into
the corresponding TMS ether in the same manner as that described
for 15 and was analyzed by GLC and EIMS. EIMS of the TMS deriv-
ative of 16: m/z 427 ([M–Me]+, 9), 411 ([M–MeO]+, 10), 395 (28),
287 (C-11/C-12 cleavage, 100), 257 (C-10/C-11 cleavage, 100),
129 (31), 73 (65), 57 (36).
room temperature for 30 min. Removal of the solvent by flushing
with N2, followed by purification by silica gel CC (eluted with hex-
ane–EtOAc 2:1), yielded a mixture of methyl esters 9m and 10m
(11 mg). A portion of Pool V (10 mg) was similarly converted to a
mixture of methyl esters 11m and 12m (9.0 mg). A portion of Pool
VI (14 mg) was dissolved in EtOAc (200 lL) and ethereal CH2N2
was added until a yellow color persisted. Removal of the solvent
by flushing with N2 followed by purification by silica gel CC (eluted
with hexane–EtOAc 1:1) yielded a mixture of methyl esters 13m
and 14m (11 mg). An aliquot (10 mg) of these was separated by
HPLC (MeOH–H2O, 20:1; flow rate, 1.0 mL/min) to afford 9m
(7.4 mg) and 10m (1.8 mg) at 13.4 and 14.0 min, respectively.
The mixture of the methyl esters 11m and 12m (17 mg) was sep-
arated by HPLC (MeOH–H2O, 20:1; flow rate, 1.0 mL/min) to afford
11m (12 mg) and 12m (1.8 mg) at 15.3 and 15.9 min, respectively.
The mixture of the methyl esters 13m and 14m (10 mg) was sep-
arated by HPLC (MeOH–H2O, 20:1; flow rate, 1.0 mL/min) to afford
13m (5.7 mg) and 14m (2.0 mg) at 11.8 and 12.3 min, respectively.
Compound 2 (2.2 mg) was converted to the corresponding ace-
tate 2a (2.4 mg) following the procedure described for compound
1: Colorless oil; ½a D25
ꢂ
– 5.0 (c = 0.24, CHCl3); IR (CHCl3) mmax 2930,
2860, 1720 cm–1; (+)-HRFABMS m/z: 649.3906 [M+Na]+ (calcd for
C
34H58O10Na, 649.3928); 1H NMR (CDCl3) d: 0.88 (t, J = 6.9 Hz,
H3-22), 1.49-1.20 (m), 1.65 (m, Ha-3), 1.71 (m, Hb-3), 2.01 (s, Ac),
2.03 (s, Ac (x2)), 2.37 (m), 3.58 (m, H-10), 3.69 (m, H-50), 3.97 (dd,
J = 12.1, 5.5 Hz, Ha-60), 4.23 (dd, J = 12.1, 1.9 Hz, Hb-60), 4.57 (d,
J = 8.0 Hz, H-10), 4.98 (dd, J = 9.7, 8.0 Hz, H-20), 5.07 (t, J = 9.7 Hz,
H-40), 5.21 (t, J = 9.7 Hz, H-30); 13C NMR d:14.13, 20.66 (x2),
20.70, 22.69, 23.66, 24.37, 25.05, 26.21, 26.59, 27.00, 27.09,
29.36, 29.65, 29.67 (x3), 29.71, 29.90, 31.92, 33.82, 34.01, 34.36,
62.28, 68.59, 71.26, 71.83, 73.18, 80.20, 100.24, 169.30, 169.37,
170.45, 173.91.
4.4. Glomeraside A (1)
Colorless oil; (+)-HRFABMS m/z: 543.3894 [M+H]+ (calcd for
C
30H55O8, 543.3897); ½a D25
ꢀ 9.2 (c = 0.60, CHCl3); IR (CHCl3) mmax
ꢂ
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
Compound 1 (3.0 mg) was hydrolyzed as described in our pre-
vious paper (Asai and Fujimoto, 2010) to give glucose and a hydro-
xy-fatty acid. Glucose was identified by TLC and GC–MS as
described in our previous paper (Asai and Fujimoto, 2010; Asai
et al., 2009). Treatment of the hydroxy-fatty acid with ethereal
CH2N2 gave, after PTLC purification (hexane–EtOAc, 6:1), methyl
11(R)-hydroxydocosanoate 15 (1.3 mg). Amorphous; EIMS m/z:
352 ([M–H2O]+, 1), 339 ([M–MeO]+, 2), 320 ([M–H2O–MeO]+, 10),
215 (C-11/C-12 cleavage, 42), 186 (45), 183 (215–MeOH, 42), 143
(23), 87 (42); 1H NMR (CDCl3) d: 3.67 (s, OMe), 3.59 (m, H-11),
2.30 (t, J = 7.6 Hz, H2-2), 1.65-1.25 (m), 0.88 (t, J = 7.8 Hz, H3-22).
A portion of 15 was converted to the corresponding TMS ether
as described previously (Asai and Fujimoto, 2010) and analyzed
by GLC and EIMS. EIMS of TMS derivative of 15: m/z 427 ([M–
Me]+, 9), 411 ([M–MeO]+, 10), 395 (28), 287 (C-11/C-12 cleavage,
100), 257 (C-10/C-11 cleavage, 100), 129 (31), 73 (65), 57 (36).
4.6. Glomeraside C (3)
Colorless oil; (+)-HRFABMS m/z: 543.3807 [M+H]+ (calcd for
C
30H55O8, 543.3897); ½a D25
ꢂ
0 (c = 0.68, CHCl3); IR (CHCl3) mmax
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
Methyl 9(R)-hydroxydocosanoate 17 (0.8 mg) was obtained
from 3 (2.0 mg) in the same manner as that described for 1. 17:
EIMS m/z: 352 ([M–H2O]+, 1), 339 ([M–MeO]+, 2), 320 ([M–H2O–
MeO]+, 10), 187 (C-9/C-10 cleavage, 45), 158 (45), 155 (188–MeOH,
100), 87 (30), 55 (20); 1H NMR (CDCl3) d: 3.67 (s, OMe), 3.59 (m, H-
9), 2.30 (t, J = 7.6 Hz, H2-2), 1.65-1.25 (m), 0.88 (t, J = 7.8 Hz, H3-22).
A portion of the methyl ester 17 was converted to the correspond-
ing TMS ether in the same manner as described for 15 and ana-
lyzed by GLC and EIMS. EIMS of the TMS derivative of 17: m/z
427 ([M–Me]+, 9), 411 ([M–MeO]+, 12), 395 (12), 285 (C-8/C-9
cleavage, 100), 259 (C-9/C-10 cleavage, 100), 129 (28), 73 (55),
57 (53).
Compound 1 (2.2 mg) was treated with Ac2O (20
dine (40 L) at room temperature overnight. Removal of the sol-
vent by flushing with N2 and purification of the crude product by
lL) and pyri-
l

156
T. Asai et al. / Phytochemistry 82 (2012) 149–157
Compound 3 (3.0 mg) was converted into the corresponding
4.11. Glomeraside H (8)
acetate 3a (2.1 mg) following the procedure described for com-
pound 1. Colorless oil; ½a D25
ꢂ
– 18.1 (c = 0.21, CHCl3); IR (CHCl3) mmax
Colorless oil; (+)-HRFABMS m/z: 501.3782 [M+H]+ (calcd for
2930, 2860, 1720 cm–1; (+)-HRFABMS m/z: 649.3953 [M+Na]+
(calcd for C34H58O10Na, 649.3928); 1H NMR d: 0.88 (t, J = 6.9 Hz,
H3-22), 1.50-1.20 (m), 1.60 (m, Ha-3), 1.70 (m, Hb-3), 2.00 (s, Ac),
2.03 (s, Ac), 2.04 (s, Ac), 2.39 (t, J = 6.0 Hz, H2-2), 3.50 (m, H-9),
3.79 (m, H-50), 4.08 (dd, J = 12.1, 7.2 Hz, Ha-60), 4.12 (dd, J = 12.1,
2.6 Hz, Hb-60), 4.54 (d, J = 8.0 Hz, H-10), 4.95 (t, J = 9.8 Hz, H-40),
4.96 (dd, J = 9.8, 8.0 Hz, H-20), 5.21 (t, J = 9.8 Hz, H-30); 13C NMR d:
14.11, 20.63 (x2), 20.68, 22.68, 23.25, 24.42, 25.38, 26.90, 27.17,
27.54, 29.35, 29.65 (x4), 29.68 (x2), 29.87, 31.91, 34.25 (x2),
35.24, 63.21, 69.01, 71.54, 71.85, 72.99, 82.57, 100.86, 169.19,
169.46, 170.37, 173.77.
C
28H53O7, 501.3791); ½a D25
ꢂ
– 16.0 (c = 0.44, CHCl3); IR (CHCl3) mmax
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
Compound 8 (2.2 mg) was converted to the corresponding per-
acetate (2.2 mg) in the same manner as described for 1. The perac-
etate was identified as 3a by 1H and 13C NMR, FABMS and HPLC.
4.12. Glomeraside I methyl ester (9m)
Colorless oil; (+)-HRFABMS m/z: 643.4080 [M+H]+ (calcd for
C
34H59O11, 643.4057); ½a D25
ꢂ
– 17.2 (c = 0.29, CHCl3); IR (CHCl3) mmax
3500, 2930, 2860, 1760, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
4.7. Glomeraside D (4)
Compound 9m (3.0 mg) was converted into a hydroxy fatty acid
methyl ester (1.1 mg) that was identified as methyl 11(R)-
hydroxydocosanoate 15 by TLC, EIMS of the TMS ether derivative
and 1H NMR of the (R)-ACC ester.
Colorless oil; (+)-HRFABMS m/z: 543.3881 [M+H]+ (calcd for
C
30H55O8, 543.3897); ½a D25
ꢂ
– 26.1 (c = 1.1, CHCl3); IR (CHCl3) mmax
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
Compound 9m (2.5 mg) was converted into the corresponding
peracetate 9a (2.6 mg) in the same manner as described for 1.
9a: Colorless oil; (+)-HRFABMS m/z: 685.4143 [M+H]+ (calcd for
Compound
4 (2.0 mg) was converted into the peracetate
(2.1 mg) in the same manner as described for 1. The peracetate
was identified as 2a by 1H and 13C NMR, FABMS and HPLC. HPLC
identification of the peracetate with 1a was carried out as follows.
The peracetates 1a, 2a and 3a could not be separated using routine
ODS columns (with either MeOH–H2O or CH3CN–H2O as the elu-
tion solvents), but were separated with an Inertsil ODS-P column
(GL Science Inc., 4.6 ꢁ 15 cm, solvent CH3CN, flow rate 0.8 L/min,
UV detection at 215 nm). The retention times for 1a, 2a and 3a
were 9.43, 10.05 and 10.80 min, respectively.
C
36H61O12, 685.4163); ½a D25
ꢂ
– 5.9 (c = 0.34, CHCl3); IR (CHCl3) mmax
2930, 2860, 1750 cm–1
;
1H NMR d: 0.88 (t, J = 7.0 Hz, H3-22),
1.50-1.20 (m), 1.63 (m, Ha-3), 1.76 (m, Hb-3), 2.02 (s, Ac), 2.04 (s,
Ac), 2.32 (ddd, J = 14.5, 9.0, 4.0 Hz, Ha-2), 2.43 (ddd, J = 14.5, 8.0,
4.5 Hz, Hb-2), 3.34 (s, H-200), 3.56 (m, H-11), 3.69 (m, H-50), 3.72
(s, OMe), 4.09 (dd, J = 12.0, 4.5 Hz, Ha-60), 4.16 (dd, J = 12.0,
2.0 Hz, Hb-60), 4.54 (d, J = 8.0 Hz, H-10), 5.03 (dd, J = 9.5, 8.0 Hz, H-
20), 5.10 (t, J = 9.5 Hz, H-40), 5.23 (t, J = 9.5 Hz, H-30); 13C NMR d:
14.10, 20.60, 20.62, 22.68, 24.34 (x2), 25.20, 26.89, 27.32, 27.53,
27.66, 28.14, 29.35, 29.62, 29.67 (x3), 29.89, 31.90, 34.09, 34.38,
34.53, 41.08, 52.55, 62.16, 68.37, 71.56, 72.43, 72.61, 80.76,
99.99, 164.73, 166.29, 169.31, 170.44, 173.90.
4.8. Glomeraside E (5)
Colorless oil; (+)-HRFABMS m/z: 543.3901 [M+H]+ (calcd for
C
30H55O8, 543.3897); ½a D25
ꢂ
– 21.3 (c = 0.15, CHCl3); IR (CHCl3) mmax
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
4.13. Glomeraside J methyl ester (10m)
Colorless oil; (+)-HRFABMS m/z: 643.4030 [M+H]+ (calcd for
Compound 5 (1.7 mg) was converted into the corresponding
peracetate (1.8 mg) in the same manner as described for 1. The
peracetate was identified as 2a by 1H and 13C NMR, FABMS and
HPLC.
C
34H59O11, 643.4057); ½a D25
ꢂ
– 13.3 (c = 0.07, CHCl3); IR (CHCl3) mmax
3500, 2930, 2860, 1760, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
Compound 10m (1.7 mg) was converted into the corresponding
peracetate 10a (1.7 mg) in the same manner as described for 1.
10a: Colorless oil; (+)-HRFABMS m/z: 685.4176 [M+H]+ (calcd for
4.9. Glomeraside F (6)
C
36H61O12, 685.4163); ½a D25
ꢂ
0 (c = 0.15, CHCl3); IR (CHCl3) mmax
Colorless oil; (+)-HRFABMS m/z: 501.3804 [M+H]+ (calcd for
2930, 2860, 1750 cm–1 1H NMR d: 0.88 (t, J = 7.0 Hz, H3-22),
;
C
28H53O7, 501.3791); ½a D25
ꢂ
– 27.8 (c = 0.36, CHCl3); IR (CHCl3) mmax
1.49-1.20 (m), 1.65 (m, Ha-3), 1.70 (m, Hb-3), 2.03 (s, Ac), 2.04 (s,
Ac), 2.36 (m, H2-2), 3.34 (s, H-200), 3.59 (m, H-10), 3.69 (m, H-50),
3.72 (s, OMe), 3.97 (dd, J = 12.0, 5.5 Hz, Ha-60), 4.24 (dd, J = 12.0,
2.0 Hz, Hb-60), 4.57 (d, J = 8.0 Hz, H-10), 5.03 (dd, J = 9.5, 8.0 Hz, H-
20), 5.07 (t, J = 9.5 Hz, H-40), 5.23 (t, J = 9.5 Hz, H-30); 13C NMR d:
14.11, 20.62 (x2), 22.68, 23.67, 24.37, 25.14, 26.27, 26.58, 27.00,
27.11, 29.35, 29.65, 29.67, 29.68 (x3), 29.86, 31.92, 33.85, 33.96,
34.29, 41.08, 52.56, 62.19, 68.66, 71.33, 72.47, 72.65, 80.00,
99.82, 164.74, 166.31, 169.36, 170.46, 173.87.
Compound 10a (1.6 mg) was converted into a hydroxy fatty
acid methyl ester (0.4 mg) that was identified as methyl 10(R)-
hydroxydocosanoate 16 by TLC, EIMS of the TMS ether derivative
and 1H NMR of the (R)-ACC ester.
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
Compound 6 (1.8 mg) was converted into the corresponding
peracetate (2.0 mg) in the same manner as described for 1. The
peracetate was identified as 1a by 1H and 13C NMR, FABMS and
HPLC.
4.10. Glomeraside G (7)
Colorless oil; (+)-HRFABMS m/z: 501.3825 [M+H]+ (calcd for
C28H53O7, 501.3791); ½a D25
ꢂ
– 25.5 (c = 0.60, CHCl3); IR (CHCl3) mmax
3590, 3440, 2930, 2860, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2, respectively.
Compound 7 (2.1 mg) was converted into the corresponding
peracetate (2.2 mg) in the same manner as described for 1. The
peracetate was identified as 2a by 1H and 13C NMR, FABMS and
HPLC.
4.14. Glomeraside K methyl ester (11m)
Colorless oil; (+)-HRFABMS m/z: 643.4069 [M+H]+ (calcd for
C
34H59O11, 643.4057); ½a D25
ꢂ
– 24.7 (c = 0.60, CHCl3); IR (CHCl3) mmax

T. Asai et al. / Phytochemistry 82 (2012) 149–157
157
3500, 2930, 2860, 1760, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2.
Compound 11m (3.0 mg) was converted into a hydroxy-fatty
acid methyl ester (1.1 mg) that was identified as 11(R)-hydroxy-
docosanoate 15 by TLC, EIMS of TMS ether derivative and 1H
NMR of the (R)-ACC ester.
8.08 ((2H, m), 7.61 (2H, m), 4.73 (1H, m), 4.48 (1H, td, 12.0,
3.5 Hz), 3.67 (3H, s, OCH3), 3.56 (1H, td, 12.0, 3.5 Hz), 2.20 (2H, t,
J = 7.5 Hz, H2-2), 2.18 (1H, m), 1.87 (2H, m), 1.50-0.84 (5H, m),
0.867 (3H, t, J = 7.2 Hz). The chemical shifts of the key 1H signals
of these derivatives are summarized in Table 3.
Stereochemically defined methyl 10(R)-hydroxydocosanoate
16, ½a 2D5
ꢂ
– 0.2 (c = 2.8, CHCl3), was synthesized by reacting (S)-
10,11-epoxyundecanoic acid methyl ester with undecanylmagne-
sium bromide in the presence of CuI according to a published pro-
tocol (Ohtaki et al., 2005). The (S)-epoxide was obtained by kinetic
resolution of the racemic epoxide according to a published method
(Schaus et al., 2002), which was prepared by m-CPBA treatment of
methyl undec-10-enoate.
4.15. Glomeraside L methyl ester (12m)
Colorless oil; (+)-HRFABMS m/z: 643.4086 [M+H]+ (calcd for
C
34H59O11, 643.4057); ½a D25
ꢂ
– 36.7 (c = 0.03, CHCl3); IR (CHCl3) mmax
3500, 2930, 2860, 1760, 1720 cm–1; for 1H and 13C NMR spectro-
scopic data, see Tables 1 and 2. Compound 12m (1.7 mg) was con-
verted into a hydroxy fatty acid methyl ester (0.5 mg) that was
identified as methyl 10(R)-hydroxydocosanoate 16 by TLC, EIMS
of the TMS ether derivative and 1H NMR of the (R)-ACC ester.
Acknowledgments
The authors thank Prof. Shiro Kohshima (Department of
Biological Sciences, Graduate School of Bioscience and Biotechnol-
ogy, Tokyo Institute of Technology) for the botanical identification.
4.16. Glomeraside M methyl ester (13m)
Colorless oil; (+)-HRFABMS m/z: 601.3933 [M+H]+ (calcd for
C32H57O10, 601.3952); ½a D25
ꢂ
– 16.9 (c = 0.38, CHCl3); IR (CHCl3) mmax
References
3580, 3500, 2930, 2860, 1760, 1720 cm–1; for 1H and 13C NMR
spectroscopic data, see Tables 1 and 2, respectively.
Asai, T., Hara, N., Kobayashi, S., Kohshima, S., Fujimoto, Y., 2009. Acylglycerols in the
glandular trichomes on the leaves of Paulownia tomentosa. Helv. Chim. Acta 92,
1473–1497.
Asai, T., Hara, N., Fujimoto, Y., 2010. Fatty acid derivatives and dammarane
triterpenes from the glandular trichome exudates of Ibicella lutea and
Proboscidea louisiana. Phytochemistry 71, 877–894.
Asai, T., Fujimoto, Y., 2010. Cyclic fatty acyl glycosides in the glandular trichome
exudate of Silene gallica. Phytochemistry 71, 1410–1417.
Asai, T., Fujimoto, Y., 2011. 2-Acety-1-(3-glycosyloxyoctadecanoyl)glycerol and
dammarane triterpenes in the exudates from glandular trichome-like secretory
organs on the stipules and leaves of Cerasus yedoensis. Phytochemistry Lett. 4,
38–42.
Compound 13m (2.0 mg) was converted into the corresponding
peracetate (1.7 mg) in the same manner as described for 1. The
peracetate was identified as 9a by 1H and 13C NMR, FABMS and
HPLC. HPLC identification of the peracetate with 9a was carried
out as follows. The peracetates 9a and 10a were separated using
an Inertsil ODS-P column (conditions: CH3CN–H2O 40:1, at a flow
rate of 0.8 L/min, UV detection at 215 nm). The retention times
for 9a and 10a were 9.95 and 10.52 min, respectively.
Duke, S.O., Canel, C., Rimando, A.M., Tellez, M.R., Duke, M.V., Paul, R.N., 2000.
Current and potential exploitation of plant glandular trichome productivity.
Adv. Bota. Res. 31, 121–151.
4.17. Glomeraside N methyl ester (14m)
Ganenko, T.V., Semoenov, A.A., Vereschagin, A.L., Ushakov, I.A., Chuvashov, Y.A.,
´
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C
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ꢂ
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Compound 14m (2.0 mg) was converted to the corresponding
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oily (R)-ACC derivative 15b in good yield. Reaction of 15 with the
(S)-ACC acid instead of (R)-ACC acid gave the (S)-ACC derivative
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