This Issue is Dedicated to the Memory of Professor Ivano Morelli

Flavonoid glycosides from Chrozophora senegalensis Natural Product Communications Vol. 1 (12) 2006 1093
chelating activity capturing ferrous ions before
ferrozine, with an IC 50 value (concentration that
inhibited the ferrozine-Fe 2+ by 50%) of 13.65, 6.19
and 18.31 μg/mL, respectively (Table 4).
These data also suggest that the biological effect of
C. senegalensis observed from ethnopharmacological
studies is due in part to the anti-oxidant action of its
active components.
Experimental
General: Optical rotations were measured on a
Perkin-Elmer 241 polarimeter equipped with a
sodium lamp (589 nm) and a 10 cm microcell.
Elemental analysis was obtained from a Carlo Erba
1106 elemental analyzer. UV spectra were recorded
on a Perkin-Elmer-Lambda 12 spectrophotometer. A
Bruker DRX-600 NMR spectrometer using the
UXNMR software package was used for NMR
experiments. ESIMS (negative mode) were obtained
using a Finnigan LC-Q Advantage Termoquest
spectrometer, equipped with Xcalibur software. TLC
was performed on precoated Kieselgel 60 F 254 plates
(Merck, Darmstadt, Germany); compounds were
detected by spraying with Ce(SO 4 ) 2 /H 2 SO 4 (Sigma-
Aldrich, St. Louis, Mo, USA) and NTS (Naturstoffe
reagent)-PEG (Polyethylene glycol 4000) solutions.
Column chromatography was performed over
Sephadex LH-20 (Pharmacia); reversed-phase (RP)
HPLC separations were conducted on a Waters 515
pumping system equipped with a Waters R401
refractive index detector and Waters U6K injector,
using a C 18 μ-Bondapak column (30 cm x 7.8 mm)
and a mobile phase consisting of MeOH-H 2 O
mixtures at a flow rate of 2 mL/min. GC analyses
were performed using a Dani GC 1000 instrument. A
Hitachi U-2000 spectrophotometer (Hitachi, Tokyo,
Japan) was used for all antioxidant assays.
Plant material and chemicals: The leaves of
Chrozophora senegalensis were collected in
Bandiagara, Mali, in 1999 and identified by Prof.
N’Golo Diarra of the Departement Medicine
Traditionelle (DMT), Bamako, Mali where a voucher
specimen (DMT n. 0074 ) is deposited. pBR322
plasmid DNA, 1,1-diphenyl-2-picryl-hydrazyl radical
(DPPH), diethylenetriaminepentaacetic acid (DTPA)
and 3-(2-pyridyl)-5,6-bis (4-phenyl-sulfonic acid)-
1,2,4-triazine (ferrozine) were obtained from Sigma
Aldrich Co (St. Louis, USA); β-nicotinamide-adenine
dinucleotide (NADH) was obtained from Boehringer
Mannheim GmbH (Germany). All other chemicals
were purchased from GIBCO BRL Life Technologies
(Grand Island, NY, USA).
Extraction and isolation: The air-dried powdered
leaves of C. senegalensis (600 g) were defatted with
n-hexane and extracted successively by exhaustive
maceration (3 x 1 L, for 48 h) with CHCl 3 , CHCl 3 -
MeOH 9:1, and MeOH. The extracts were
concentrated under reduced pressure to afford 13.4,
14.0, 13.8, and 62.4 g of dried residues, respectively.
A portion of the MeOH extract (27.0 g) was
partitioned between n-BuOH and H 2 O to give a
n-BuOH soluble portion (9.0 g); 5.0 g of this residue
were chromatographed over a Sephadex LH-20
column (100 cm x 5 cm) with MeOH as the eluent. A
total of 115 fractions were collected (10 mL each).
These were combined according to TLC analysis
[silica 60 F 254 gel-coated glass sheets with n-BuOH-
AcOH-H 2 O (60:15:25) and CHCl 3 -MeOH-H 2 O
(40:9:1)] to give nine pooled fractions (A-I).
Fractions G, H, and I yielded compounds 3 (19.2
mg), 4 (40 mg), and 9 (30 mg), respectively. Fraction
A (90 mg) was purified by RP-HPLC using MeOH-
H 2 O (45:55) to give compounds 10 (6 mg, t R = 10
min) and 12 (5 mg, t R = 20 min). Fraction B (36 mg)
was purified by RP-HPLC using MeOH-H 2 O (1:1) to
give compounds 2 (8 mg, t R = 10 min) and 11 (12 mg,
t R = 20 min). Fraction C (50.5 mg) was purified by
RP-HPLC using MeOH-H 2 O (45:55) to give
compounds 5 (28 mg, t R = 10 min) and 7 (10.8 mg,
t R = 20 min). Fraction D (100 mg) was purified by
RP-HPLC using MeOH-H 2 O (45:55) to give
compounds 1 (14.5 mg, t R = 10 min) and 6 (6.5 mg,
t R = 20 min), while fraction E (70 mg) was purified by
RP-HPLC using MeOH-H 2 O (55:45) to yield
compound 3 (11 mg, t R = 10 min). Finally, fraction F
(85 mg) was chromatographed on a RP-HPLC using
MeOH-H 2 O (1:1) as the eluent to afford compounds
8 (5 mg, t R = 28 min) and 4 (6.3 mg, t R = 46 min).
Quercetin 3-O-(6''-caffeoyl)-β-D-glucopyranoside-
3'-O-β-D-glucopyranoside (1)
Yellow amorphous powder.
[α] D : -27° (c 0.1, MeOH).
UV/Vis λ max (MeOH) nm (log ε): 267 (3.99), 344
(4.32)
1 H NMR (600 MHz, CD 3 OD): Table 1.
13 C NMR (600 MHz, CD 3 OD): Table 1.
ESIMS: m/z 787 [M - H] - .
Anal. Calcd for C 36 H 36 O 20 : C, 54.83; H, 4.60. Found
C, 54.79; H 4.62.