DSpace at VNU: Two new flavonol glycosides from the leaves of Cleome viscosa L.

Phytochemistry Letters 18 (2016) 10–13

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Phytochemistry Letters
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Short communication

Two new flavonol glycosides from the leaves of Cleome viscosa L.
Nhat Minh Phana,* , Tan Phat Nguyena , Tien Dung Lea , Thanh Chi Maia ,
Mai Thanh Phongb , Dinh Tri Maia
a
b

Institute of Chemical Technology, Vietnam Academy of Science and Technology, Ho Chi Minh city, Viet Nam
University of Technology, Vietnam National University, Ho Chi Minh city, Viet Nam

A R T I C L E I N F O

Article history:
Received 25 May 2016
Received in revised form 18 August 2016
Accepted 26 August 2016
Available online xxx
Keywords:
Cleome viscosa
Capparaceae
Visconoside A
Visconoside B

A B S T R A C T

From the leaves of Cleome viscosa L., two new flavonol glycosides, named visconoside A (1) and
visconoside B (2), together with six known flavonol glycosides, vincetoxicoside A (3), vincetoxicoside B
(4), kaempferitrin (5), kaempferide 3-O-b-D-glucopyranoside 7-O-a-L-rhamnopyranoside (6), kaempferol 3-O-b-D-glucopyranoside 7-O-a-L-rhamnopyranoside (7), and isorhamnetin 3-O-b-D-glucopyranoside (8) were isolated by various chromatography methods. Its chemical structure was elucidated by IR,
UV, HR-ESI-MS, NMR 1D and 2D experiments and compared with literatures.
ã 2016 Phytochemical Society of Europe. Published by Elsevier Ltd. All rights reserved.

1. Introduction

2. Results and discussion

Cleome viscosa L. commonly known as “wild or dog mustard”, is
an annual, sticky herb belonging to family Capparaceae found as a
common weed throughout the tropics of the world. The whole
plant is used as drugs by the traditional medical practitioners with
beneficial action for the treatment of diarrhoea, fever, inflammation, liver diseases, bronchitis, skin diseases, and malarial fever
(Mali, 2010). This plant has been screened scientifically for various
pharmacological activities such as anthelmintic, antimicrobial,
anti-inflammatory, hepatoprotective (Mali, 2010), antifibrotic
(Kumar et al., 2009), anticonvulsant (Mishra et al., 2010),
antioxidant (Gupta et al., 2011), antinociceptive, antibacterial
(Bose et al., 2011), antitumor (Gopal et al., 2012) and antibacterial
activities (Donkor et al., 2014). Previous phytochemical investigations of C. viscosa have yielded a number of flavanones
(Srivastava et al., 1979), flavonol (Chauhan et al., 1979; Senthamilselvi et al., 2012), sterol (Srivastava, 1980), coumarinolignoid
(Ray et al., 1985; Kumar et al., 1988), diterpene (Jente et al., 1990),
lactam derivative (Jana and Biswas, 2011), and dipyridodiazepinone
derivative (Chatterjee et al., 2013). In this paper, we describe the
isolation and structure elucidation of two new flavonol glycosides
(1, 2) and six known flavonol glycosides (3–8) from Cleome viscosa

L. collected in Ben Cat, Binh Duong province, Viet Nam.

The MeOH extract from the dried leaves of Cleome viscosa L. was
subjected to column chromatography over silica gel normal-phase
and reversed-phase RP-18 to give two new flavonol glycosides,
named visconosides A (1), visconosides B (2), and six known
flavonol glycosides, vincetoxicoside A (3) (Gaind et al., 1981),
vincetoxicoside B (4) (Ishiguro et al., 1991), kaempferitrin (5) (Rao
et al., 2009), kaempferide 3-O-b-D-glucopyranoside 7-O-a-Lrhamnopyranoside (6) (Imperato, 1984), kaempferol 3-O-b-Dglucopyranoside 7-O-a-L-rhamnopyranoside (7) (Song et al.,
2007), and isorhamnetin 3-O-b-D-glucopyranoside (8) (Wang
et al., 2012).
Compound (1) was isolated as a yellow amorphous powder, and
the HR-ESI–MS showed the quasimolecular ion at m/z 821.2116
[M + Na]+ consistent with the molecular formula C35H42O21. The
aglycone of 1 was identified as quercetin, according to observation
of fifteen carbon signals in 13C-NMR and DEPT spectrum (Table 1)
including one carbonyl carbon at dC 178.0 (C-4), seven oxygenated
aromatic carbons, two quaternary aromatic carbons and five nonoxygenated aromatic carbons together with two AX-type aromatic
protons at dH 6.45 (d, 2.0, H-6) and 6.76 ; three ABX-type aromatic
protons at dH 6.93 (d, 8.5, H-50 ), 7.30 (d, 2.0, H-60 ) and 7.31 (dd, 2.0,
8.5, H-20 ) in 1H-NMR data. Moreover, three anomeric carbons at dH
5.36 (br s, H-1”), 4.27 (d, 8.0, H-000 ), 5.54 (d, 1.0, H-10000 ) corresponded
to three anomeric carbons at dC 101.1 (C-100 ), 104.8 (C-1000 ), 96.8
(C-10000 ) were assigned to a-L-rhamnose (Rha I), b-D-glucose (Glc)
and a-L-rhamnose (Rha II) units, respectively. The COSY and HSQC
spectrum allowed analysis of their spin systems and assignment of

* Corresponding author.
E-mail address: (N.M. Phan).

/>1874-3900/ã 2016 Phytochemical Society of Europe. Published by Elsevier Ltd. All rights reserved.


N.M. Phan et al. / Phytochemistry Letters 18 (2016) 10–13
Table 1
1
H (500 Hz) and
Aglycone

13

C (125 Hz) NMR spectral data for compounds 1 and 2 in DMSO-d6.

1

2

Sugar

dH

dC

dH

dC

2
3
4

5
6
7
8

–
–
–
–
6.45 d (2.0)
–
6.76 d (2.0)

158.2
133.9
178.0
161.1
99.8
161.9
94.8

–
–
–
–
6.27 d (1.5)
–
6.50 br s

156.8

133.2
177.4
160.5
99.5
161.7
93.6

Glc
1000
2000
3000
4000
5000
6000

9
10
1'
2'
3'
4'
5'
6'

–
–
–
7.31 dd (2.0, 8.5)
–
–

6.93 d (8.5)
7.30 d (2.0)

156.3
105.9
120.4
115.8
145.5
149.1
115.8
121.6

–

7.26 d (1.5)
–
–
6.93 d (7.5)
7.33 dd (2.0, 8.0)

155.6
105.3
120.5
115.6
145.4
148.8
115.2
121.5

Rha II

1””
2””
3””
4””
5””
6””
Sinapinoyl

5.36 br s
4.20 s
3.87 d (2.5)
4.82 t (10.0)
3.37 dd (6.5, 10.0)
0.74 d (6.5)
–
1.96 s

101.2
69.7
76.9
71.5
68.5
17.3
170.2
21.0

5.54 br s
4.08 br s
3.86 d (2.5)
4.79 t (10.0)

3.07 m
0.66 d (6.5)
–
1.94 s

99.9
69.7
76.4
71.3
68.4
17.1
169.8
20.8

Rha I
1”
2”
3”
4”
5”
6”
4”-OAc

11

their proton resonances to determine clearly every sugar unit.
Beside, the sugar moiety was identified as L-rhamnose and Dglucose by the acidic hydrolysis and using TLC to compare the
hydrolysate with the authentic sugars (see Experimental). Further,
1 revealed an acetyl group [dC 170.2–21.0 with methyl proton at dH
1.96 (s, CH3CO)]. The HMBC spectrum (Fig. 1), showed correlations

between anomeric proton at dH 5.36 (br s, H-100 ) of Rha I and
carbons at dC 133.9 (C-3) of aglycone; between anomeric proton at
dH 4.27 (d, 8.0, H-1000 ) of Glc and carbons at dC 76.9 (C-300 ) of Rha I;
between oxygenated methine proton at dH 3.87 (d, 2.5, H-300 ) and

1

2

dH

dC

dH

dC

4.27 d (8.0)
2.99 t (8.5)
3.17 dd (8.5, 9.0)
3.10 t (9.0)
3.22 m
3.72 dd (1.5, 11.5)
3.54 dd (9.0, 11.5)

104.8
73.4
77.0
70.1
76.6

61.2

4.32 d (7.5)
3.07 m
3.24 t (9.0)
3.07 m
3.63 m
4.52 dd (8.5, 11.0)
4.25 dd (2.0, 11.0)

105.0
73.0
76.7
70.6
74.1
63.6

5.54 d (1.0)
3.84 br s
3.63 dd (3.5, 9.5)
3.31 t (9.5)
3.44 dd (3.5, 6.0)
1.11 d(6.0)

98.6
70.0
70.4
71.8
70.3
18.1

5.54 br s
3.89 d (2.5)
3.63 m
3.31 t (9.0)
3.40 dd (6.0, 9.0)
1.15 d (6.0)

98.6
69.8
70.3
71.7
69.9
18.0

6.30 d (15.5)
7.41 d (15.5)
–
6.50 s
–
–
–
6.50 s
–
3.54 d (6.5)

114.3
145.1
123.8
104.9

147.3
137.8
147.3
104.9
166.4
55.3

a
b

1000 00
2000 00
3000 00
4000 00
5000 00
6000 00
ÀÀCOO
ÀÀOCH3

carbon acetal at dC 104.8 (C-1000 ). On the other hands, anomeric
proton at dH 5.54 (d, 1.0, H-10000 ) correlated with oxygenated
aromatic carbon at dC 161.9 (C-7). Based on data of HR-ESI-MS, 1D,
2D-NMR and compared with previous published data (Minh et al.,
2015), the structure of 1 was determined as quercetin 3-O-[b-Dglucopyranosyl-(1 ! 3)]-a-L-(4-O-acetyl)-rhamnopyranoside 7-Oa-L-rhamnopyranoside, and named visconoside A.
Compound (2) was obtained as a yellow amorphous powder.
The molecular formula was established as C46H52O25 by HR-ESI-MS
data ([M + Na]+ m/z 1027.2680). The 1H and 13C-NMR data (Table 1)

Fig. 1. Chemical structures and selected HMBC and COSY correlations of compounds 1 and 2.

12

N.M. Phan et al. / Phytochemistry Letters 18 (2016) 10–13

demonstrated that 2 has the same aglycone and sugar chains as 1,
except for presence of an sinapoyl unit, including two oxymethyl at
dC 55.3 (–OCH3), one carbonyl carbon at dC 166.0 (–COO), three
oxygenated aromatic carbons at dC 147.3 (C-300000 , C-50000 '), 137.8 (C400000 ), one quaternary aromatic carbon at dC 123.8 (C-100000 ), four
olefinic carbons at dC 104.9 (C-200000 , C-600000 ), 114.3 (C-a), 145.1 (C-b)
correlated with two aromatic protons at dH 6.50 (s, H-200000 , H-600000 ),
two trans protons at dH 6.30 (d, 15.5, H-a) and 7.41 (d, 15.5, H-b),
respectively. Moreover, the sinapoyl moiety was also confirmed by
correlations observed in the HMBC spectrum (Fig. 1), between
protons at dH 6.30 (H-a), 7.41 (H-b) and carbons at dC 166.0 (-COO),
123.8 (C-100000 ), between protons at dH 6.50 (H-200000 , H-600000 ) and
carbons at dC 145.1 (C-b), 123.8 (C-100000 ), 137.8 (C-400000 ). On the other
hands, two methylene protons at dH 4.52 (dd, 8.5, 11.0, H-6a000 ), 4.25
(dd, 2.0, 11.0, H-6b000 ) correlated with carbonyl at dC 166.0. So, the
sinapoyl moiety attached to C-6 of the glucose unit. Based on data
of HR-ESI-MS, 1D, 2D-NMR and compared with previous published
data (Minh et al., 2015; Han et al., 2015); the structure of 2 was
identified as quercetin 3-O-[sinapoyl-(1 ! 6)-b-D-glucopyranosyl(1 ! 3)]-a-L-(4-O-acetyl)-rhamnopyranoside 7-O-a-L-rhamnopyranoside, and named visconoside B.

(25 mg). Fraction M3 (5 g) was separated by silica gel chromatographic column using CHCl3-MeOH (5:1), and further
separated by RP-18 using gradient mixtures of MeOH–H2O (5:1,
v/v) to affrord compound 7 (268 mg). The same manner was
applied to fraction M4 (75 g), eluted CHCl3-MeOH (6:1 ! 3:1) to
give four subfractions (M4.1–M4.4). Subfraction M4.1 (12 g) was
further purifed by RP-18 with MeOH–H2O (4:1, v/v) to give

compounds 5 (25 mg) and 6 (56 mg). Subfraction M4.2 (18 g) was
done as the same manner, further separated by RP-18 with MeOH–
H2O (4:1, v/v) to affrord 1 (250 mg) and 3 (7 g). Fraction M5 (5 g)
was applied on a silica gel chromatographic column and eluted
with CHCl3-MeOH (2:1) in silica gel column chromatography to
yield compound 2 (40 mg).

3. Experimental

3.3.2. Visconoside B (2)

3.1. General experimental procedures
The optical rotations were measured on a ADP220 polarimeter
(Bellingham + Stanley Ltd., RG224BA, UK). The IR data were
recorded on a Bruker Tensor 27 FT-IR spectrometer (Bremen,
Germany). The UV spectra were performed with a Jasco V-630
spectrophotometer (Tokyo, Japan). The high resolution electrospray ionisation mass spectroscopy (HR-ESI-MS) was recorded on a
Bruker MicroTOF-QII spectrometer (Bruker Daltonik GmbH, Bremen, Germany). The 1H-NMR (500 MHz), 13C-NMR (125 MHz),
DEPT, COSY, HSQC and HMBC spectra were recorded on a Bruker
AM500 FT-NMR spectrometer using tetramethylsilane (TMS) as
internal standard. Column chromatography was carried out using
silica gel normal-phase (230–400 mesh) and reversed-phase
(Merck KGaA, 64271 Darmstadt, Germany). Analytical TLC was
carried out in silica gel plates (Kieselgel 60 F254, Merck).
Compounds were visualized by spraying with aqueous 10%
H2SO4 and heating for 3–5 min.
3.2. Plant material
The leaves of Cleome viscosa L. were collected in Ben Cat, Binh
Duong province, Viet Nam, in May 2015; and identified by Prof. Vo
Van Chi. A voucher specimen (No. VH/MINH-0515) was deposited

in the Institute of Chemical Technology, Vietnam Academy of
Science and Technology.
3.3. Extraction and isolation
Powdered leaves of Cleome viscosa L. (8 kg) were extracted with
95% EtOH for three times (3 Â 30 L, total amount 90 L) at room
temperature, filtered residue, removed solvents under low
pressure, obtained crude extract (980 g). Then, crude extract was
applied to solid-phase extraction procedures and successively
partitioned into n-hexane (70 g), CHCl3 (150 g), EtOAc (260 g) and
MeOH (450 g). The MeOH extract was subjected to silica gel
column chromatography and eluted with gradient solvent system
of chloroform – methanol (95:5–5:95) to afford seven fractions:
M1 (25 g), M2 (30 g), M3 (86 g), M4 (75 g), M5 (60 g), and M6 (72 g).
Fraction M2 (5 g) was chromatographed on silica gel and eluted
with CHCl3-MeOH (10:1) to obtain compounds 4 (72 mg) and 8

3.3.1. Visconoside A (1)
Yellow amorphous powder; ½aŠ25
D À 0.94 (c 0.01, MeOH); IR nmax
(MeOH): 3317, 2943, 2831, 1449, 1417, 1114, 1022 cmÀ1; UV (MeOH)
lmax: 257 and 348 nm; HR-ESI–MS: m/z 821.2116 [M+Na]+ (calcd
for C35H42O21Na, 821.2116); 1H and 13C NMR data (DMSO-d6), see
Table 1.

Yellow amorphous powder; ½aŠ25
D À 1.23 (c 0.01, MeOH); IR nmax
(MeOH): 3317, 2943, 2831, 1449, 1416, 1115, 1022 cmÀ1; UV (MeOH)
lmax: 249 and 336 nm; HR-ESI–MS: m/z 1027.2680 [M+Na]+ (calcd
for C46H52O25Na, 1027.2695); 1H and 13C NMR data (DMSO-d6), see
Table 1.

3.4. Acid hydrolysis
Each new compounds (2 mg) was refluxed with 2N aq.
CH3COOH (5 mL) for 2 h at 100  C. After extraction with CH3Cl
(3 Â 5 mL), the aqueous layer was repeatedly evaporated to dryness
with MeOH until neutral, and then analyzed by TLC over silica gel
(MeCOEt–isoPrOH–Me2CO–H2O 20:10:7:6) by comparison with
authentic samples (L-rhamnose Rf 0.65; D-glucose Rf 0.40) (Nguyen
et al., 2015).

Acknowledgments
This research work has been financially supported by the
Vietnam Academy of Science and Technology, Project No. VAST.
DLT.09/16-17.
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