Fitoterapia 71 Ž2000.

46]49

Quercetin triglycoside from Capparis

spinosa
M. Sharaf U , M.A. El-Ansari, N.A.M. Saleh
Phytochemistry and Plant Systematics Department, National Research Centre, Dokki-12311,
Cairo, Egypt

Received 29 March 1999; accepted in revised form 16 July 1999

Abstract

In addition to rutin, quercetin 3-O-glucoside and quercetin 3-O-glucoside-7-O-rhamno-

side, the methanolic extract of the aerial parts of Capparis spinosa yielded the new flavonoid
quercetin 3-O-w690-a-L-rhamnosyl-60-b-D-glucosyl x-b-D-glucoside (1). Q 2000 Elsevier Sci-
ence B.V. All rights reserved.

Keywords: Capparis spinosa; Flavonoids

1. Introduction

In a previous chemosystematic study, we have reported the isolation and identifi-

cation of 13 flavonoid compounds from three Capparis species w1x. In continuation
of this study, the present communication describes the isolation and structure
elucidation of a new flavonol glycoside, the structure of which was identified as
quercetin 3-O-w690-a-L-rhamnosyl-60-b-D-glucosyl x-b-D-glucoside Ž1., from the
aerial parts of Capparis spinosa L. ŽCapparidaceae..

2. Results and discussion

The methanolic extract of C. spinosa was fractionated on a polyamide column.

U
Corresponding author. Fax: q202-3370931.
E-mail address: sharafali@hotmail.com ŽM. Sharaf.

0367-326Xr00r$ - see front matter Q 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 3 6 7 - 3 2 6 X Ž 9 9 . 0 0 1 1 6 - 1
 M. Sharaf et al. r Fitoterapia 71 (2000) 46]49 47

Purification, achieved by a combination of PPC, silica gel TLC and Sephadex

LH-20, afforded compound 1 which was identified as follows.
Acid hydrolysis of 1 afforded quercetin, glucose and rhamnose. Traces of
gentiobiose were also observed. Mild acid hydrolysis for 15 days w2,3x gave quercetin
3-O-glucoside and quercetin 3-O-gentiobioside.
The 1 H-NMR spectrum suggested that 1 is a triglycoside of quercetin on the
basis of the signal of Hs-1 glucose and rhamnose Ž d 5.30, 5.10 and 4.40.. The
signals at d 5.30 and 5.10 were assigned to the anomeric proton ŽH-10 and H-109 .
of two glucose moieties, with a coupling constant Ž J 7.5 Hz. indicating a b-config-
uration w4x. The doublet at d 4.40 was assigned to the anomeric proton of rhamnose
ŽH-100 .. The diequatorial coupling Ž J 2 Hz. between the H-1 and H-2 rhamnose
indicated a-configuration w5x. The three protons doublet Ž J 6 Hz. for the rhamnose
methyl was located at d 0.98. This chemical shift confirmed the presence of
rutinosyl moiety wrhamnosyl-Ž1 ª 6.-glucosylx in 1 w4x.
The 13 C-NMR spectrum of 1 confirmed that the compound is a triglycoside of
quercetin on the basis of two substituted glucose Cs-6 w68.50 and 66.98 Žunsub-
stituted C-6 of glucose resonates at 60]62 ppm.x w6x, and confirming no terminal
glucose. The 13 C-NMR of the aglycone part corresponds well to that of quercetin
w7,8x, the only differences being an upfield shift of the C-3 Žapprox. 3 ppm. and a
downfield shift of the ortho related C-2 Žapprox. 9 ppm.. These shifts are analogous
to those reported when the C-3 hydroxyl group is substituted in flavonoids w7,8x.
Also, the 13 C-NMR spectrum showed the presence of signals at d 100.60 and
101.00 for C-10 and C-190 of two glucose moieties, and signal for one rhamnose
unit Žindicated by the presence of one signal at d 17.70, methyl-rha... These
observations support the 1 H-NMR data. The assignments of the aglycone and the
sugar carbons in 1 are based on those given in the literature w8]11x. Furthermore,
the signal at d 68.50 was assigned to C-60. This is in agreement with the
observation that b-glucosidation of the glucose C-6 in gentiobioside provides a
downfield shift of approximately 6 ppm w12,13x. Rhamnosylation of the glucose C-6
Že.g. rutinoside. infers a 4-ppm downfield shift of the glucose C-6 w2,7x. Thus, the
signal at 66.98 was assigned to C-60. Compound 1 was therefore identified as
quercetin 3-O-w690-a-L-rhamnosyl-60-O-b-D-glucosyl x-b-D-glucoside.
48 M. Sharaf et al. r Fitoterapia 71 (2000) 46]49

3. Experimental

3.1. Plant material

C. spinosa aerial parts were collected in May 1998 from Wadi Firan, Southern
Sinai and authenticated by Prof. L. Bolous. A voucher specimen was deposited in
the Herbarium of NRC, Cairo.

3.2. Extraction and isolation

The dried plant Ž500 g. was extracted with 80% MeOH. The concentrated
extract Ž20 g. was subjected to polyamide column eluted with H 2 O]EtOH mixtures
of decreasing polarities. PPC using H 2 O, AcOH]H 2 O Ž3:17., BAW Ž n-
BuOH]AcOH]H 2 O 4:1:5, upper phase. afforded pure samples of quercetin 3-O-
rutinoside Ž22 mg., quercetin 3-O-glucoside Ž30 mg. and quercetin 3-O-glucoside-7-
O-rhamnoside Ž28 mg.. A combination of TLC Želuted with CHCl 3 ]MeOH]H 2 O
60:60:12. and Sephadex LH-20 afforded 92 mg of purified compound 1.

Quercetin 3-O-[690 -a -L-rhamnosyl-60 -b -D -glucosyl]-b -D -glucoside Ž1.. UVm ax

ŽMeOH.: 257, 295sh, 360; qNaOMe 270, 327, 405; qAlCl 3 272, 300sh, 420;
qAlCl 3rHCl 267, 300sh, 357, 395; qNaOAc 265, 385; qNaOAcrH 3 BO 3 262,
295sh, 375 nm; 1 H-NMR Ž270 MHz, DMSO-d6 .: d 7.50 Ž2H, m, H-29,69., 6.80 Ž1H,
d, J 9 Hz, H-59., 6.40 Ž1H, d, J 2 Hz, H-8., 6.20 Ž1H, d, J 2 Hz, H-6., 5.30 Ž1H, d, J
7 Hz, H-10 ., 5.10 Ž1H, d, J 7 Hz, H-190 ., 4.40 Ž1H, d, J 2 Hz, H-100 ., 3.10]3.50 Ž m,
sugar proton, hidden by hydroxyl signals., 0.98 Ž3H, d, J 6 Hz, CH 3-rha.; 13 C-NMR
Ž270 MHz, DMSO-d6 .: 156.50 ŽC-2., 133.50 ŽC-3., 177.50 ŽC-4., 161.00 ŽC-5., 98.60
ŽC-6., 164.00 ŽC-7., 93.50 ŽC-8., 156.50 ŽC-9., 104.50 ŽC-10., 121.50 ŽC-19., 114.50
ŽC-29., 144.50 ŽC-39., 149.00 ŽC-49., 116.50 ŽC-59., 121.50 ŽC-69., 100.60 ŽC-10 ., 74.20
ŽC-20 ., 76.50 ŽC-30 ., 70.40 ŽC-40 ., 76.80 ŽC-50 ., 68.50 ŽC-60 ., 101.00 ŽC-190 ., 74.20
ŽC-290 ., 76.00 ŽC-390 ., 70.50 ŽC-490 ., 76.50 ŽC-590 ., 66.98 ŽC-690 ., 101.40 ŽC-1909 .,
70.55 ŽC-2909 ., 70.40 ŽC-4909 ., 71.60 ŽC-4909 ., 69.90 ŽC-5909 ., 17.70 ŽC-6909 ..

3.3. Hydrolyses

Acid hydrolysis: 2 N HCl, 1008C, 2 h. Mild acid hydrolysis: 0.2 N HCl, room temp,
15 days. Enzymatic hydrolysis: 0.5 ml of b-glucosidase ŽBDH, Poole, England. in
0.05 M acetate buffer, pH 5.1, room temp, 4 h.

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