Flavonoids from the aerial part of Vicia subvillosa

Article abstract of DOI:10.1007/s10600-007-0058-5

The new flavone glucoside viscioside, luteolin-4-O-β-D- galactopyranoside, in addition to the known flavonoids apigenin, luteolin, quercetin, cinaroside, luteolin-4-O-β-D-glucoside, and isoquercitrin were isolated from the aerial part of Vicia subvillosa. Springer Science+Business Media, Inc. 2007.

Full text of DOI:10.1007/s10600-007-0058-5

Chemistry of Natural Compounds, Vol. 43, No. 1, 2007
FLAVONOIDS FROM THE AERIAL PART OF
Vicia subvillosa
M. P. Yuldashev,1 B. A. Muminova,1
A. A. Drenin,2 and E. Kh. Botirov2
UDC 547.972
′ O β
The new flavone glucoside viscioside, luteolin-4 - - -D-galactopyranoside, in addition to the known
′
O β
flavonoids apigenin, luteolin, quercetin, cinaroside, luteolin-4 - - -D-glucoside, and isoquercitrin were
Vicia subvillosa.
isolated from the aerial part of
Key words:
, apigenin, luteolin, quercetin, cinaroside, luteolin-4′- -β-D-glucoside, isoquercitrin,
Vicia subvillosa
O
luteolin-4′-O-β-D-galactopyranoside.
Plants of the genus Vicia (Fabaceae) contain flavonoids, anthocyans, carotinoids, and vitamins. Some ofthem are used
in folk medicine for ascites, paralyses, epilepsy, flu, and cholic [1, 2].
Vicia subvillosa (Ledeb.) Trautv. is a perennial indigenous to steppes and sometimes alkaline areas and foothills of
Central Asia [3].
Herein we report results of a study of flavonoids from the aerial part of V. subvillosa collected during flowering (April
1997) near the Alimtaumountains(ChimkentskOblast, Republic ofKazakhstan). Column chromatographyofvarious fractions
of the ethanol extract isolated a new flavone glycoside (1) of composition C H O , which we called viscioside, in addition
21 20 11
to the known flavonoids apigenin [4, 5], luteolin [4, 6], quercetin [7, 8], cinaroside [4, 7], luteolin-4′-O-β-D-glucoside [9], and
isoquercitrin [7, 8].
The UV spectrum of 1 was characteristic of flavone derivatives [5, 10].
The IR specrum of 1 had absorption bands for hydroxyls, γ-pyrone carbonyls, aromatic C=C bonds, and glycoside C–O
vibrations.
The PMR spectrum of 1 in DMSO-d exhibited signals for five aromatic protons, H-3, an anomeric proton (H-1′′), and
6
phenolic hydroxyls on C-5 and C-7.
OR
7
RO
O
O
4'
O
5
CH OH
2
OR
RO
O
OR
OR
1, 2
1: R = H; 2: R = COCH3
The chromatic mobility and IR and PMR spectra indicated that 1 was a glycoside. Acid hydrolysis of 1 to produce
luteolin (5,7,3′,4′-tetrahydroxyflavone) [6, 7] and D-galactose confirmed this.
Acetylation of 1 by acetic anhydride in pyridine produced the heptaacetate derivative 2 of formula C H O
([M] 742).
35 34 18
+
1) S. Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan,
Tashkent, fax (99871) 120 64 75; 2) Surgut State University, Russia, 628412, Surgut, ul. Energetikov, 14, e-mail: botirov-
nepi@mail.ru. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 30-31, January-February, 2007. Original article
submitted December 1, 2006.
0009-3130/07/4301-0034 ©2007 Springer Science+Business Media, Inc.
34

A bathochromic shift of UV band I of 1 upon addition of sodium acetate and a proton signal at 10.85 ppm in the PMR
spectrum indicated that 1 had a free hydroxyl in the 7-position [5, 10].
The site of attachment of the carbohydrate to the aglycon was found bycomparing the UV spectra of 1 and luteolin that
were recorded with ionizing and complexing additives [2, 5, 10].
The bathochromic shift of band I in the spectrum of 1 upon addition of AlCl and AlCl /HCl (+46 nm) indicated that
3
3
ring B did not have an ortho dihydroxy group. Band I in the UV spectrum of 1 underwent a bathochromic shift by 55 nm with
a decreased intensity for the absorption maximum if CH ONa was added. This indicated that the luteolin 4′-OH group was
3
glycosylated in 1 [5, 10].
The signal for the anomeric proton of D-galactose in the PMR spectrum of 1 appeared at 4.90 ppm as a doublet with
SSCC 7.0 Hz. This indicated that the glycosidic D-galactose had the -configuration [5]. Therefore, 1 had the structure
β
luteolin-4 -O- -D-galactopyranoside.
′
β
Although luteolin 4 -O-arabinoside, 4 -O-glucoside, and 4 -O-glucuronide have been reported, the 4 -O- -D-
′
′
′
′
β
galactopyranoside has not [11].
Viscioside (1) is a new natural compound. Apigenin, luteolin, quercetin, cinaroside, luteolin-4 -O- -D-glucoside, and
′
β
isoquercitrin were isolated for the first time from V. subvillosa.
EXPERIMENTAL
The solvent systems CHCl :CH OH(19:1, 1; 9:1, 2; 85:15, 3) and n-butanol:pyridine:water (6:4:3, 4) were used. TLC
3
3
used Silufol UV-254 plates. Column chromatography was performed over KSK silica gel of particle size 100/160 m. Paper
µ
chromatography (PC) used Filtrak No. 12 chromatography paper.
Spots of flavonoids on TLC were developed using ammonia vapors and vanillin (1%) in conc. H SO .
2
4
Monosaccharides were developed by spraying chromatograms with anilinium acid phthalate with subsequent heating at 90-
100°C.
PMR spectra were recorded on a Tesla BS-567A instrument at working frequency 100 MHz ( , ppm, 0 = HMDS).
δ
IR spectra were recorded on a Perkin—Elmer System 2000 FT—IR Fourier spectrophotometer in KBr disks; UV spectra, on
a Perkin—Elmer Lambda 16 spectrophotometer. Melting points were determined on a Boetius stage with a PHMK 0.5 visual
device.
Extraction and Isolation of Flavonoids. Air-dried ground aerial part of V. subvillosa (1.0 kg) was extracted eight
times at room temperature with ethanol. The combined alcohol extract was condensed in vacuo to 0.7 L and diluted with water
to 1.4 L. The aqueous alcohol extract was successively shaken with CHCl (5 × 0.5 L), ethylacetate (8 × 0.5 L), and n-butanol
3
(10 × 0.5 L). Solvents were distilled to produce chloroform (20.5 g), ethylacetate (14.0), and butanol (25.0) fractions.
The ethylacetate fraction (14.0 g) was chromatographed over a silica-gel (350 g) column (2.8 × 160 cm) using systems
1-3. Fractions of 400 mL were collected. Elution of the column by system 1 isolated apigenin (0.15 g), luteolin (0.24), and
quercetin (0.22). Elution by system 2 afforded viscioside (0.10 g), cinaroside (0.19), and luteolin-4′-O- -D-glucoside (0.16);
β
by system 3, isoquercitrin (0.12).
Apigenin (5,7,4 -trihydroxyflavone), C H O , mp 347-348°C,
(ethanol): 270, 311 nm [4].
′
λ
max
15 10
5
Luteolin (5,7,3 ,4 -tetrahydroxyflavone), C H O , mp 328-330°C (dec.),
: 260, 270, 356 nm [4, 6].
′ ′
λ
15 10
6
max
Quercetin (3,5,7,3 ,4 -pentahydroxyflavone), C H O , mp 313-314°C,
′ ′
: 257, 268, 371 nm [7, 8].
λ
15 10
7
max
Cinaroside (luteolin-7-O- -D-glucopyranoside), C H O , mp 240-242°C (dec.),
: 256, 268, 350 nm [4, 6].
β
λ
21 20 11
max
Luteolin-4 -O- -D-glucopyranoside, C H O , mp 187-190°C,
λ
max
: 271, 290, 342 nm [9].
′
β
21 20 11
Isoquercitrin (quercetin-3-O- -D-glucopyranoside), C H O , mp 238-239°C,
: 255, 266, 362 nm;
λ
max
β
21 20 12
+CH COONa: 274, 323, 374 nm [7, 8].
3
Viscioside (1) (luteolin-4 -O- -D-galactopyranoside), C H O , mp178-180°C,
λ
max
: 272, 292, 340nm; +CH ONa:
′
β
21 20 11
3
-1
273, 395; +CH COONa: 271, 370; +AlCl : 279, 351, 386; +AlCl /HCl: 280, 352, 385. IR spectrum (
, cm ): 3490-3310
ν
3
3
3
max
(OH), 1660 ( -pyrone C=O), 1575, 1520 (aromatic C=C), 1090, 1025 (glycoside C–O). PMR spectrum (DMSO-d , , ppm,
γ
δ
6
J/Hz): 3.40-3.83 (sugar protons), 4.90 (1H, d, J = 7.0, H-1 ), 6.21 (1H, d, J = 2.0, H-6), 6.52 (1H, d, J = 2.0, H-8), 6.85 (1H,
′′
s, H-3), 7.25 (1H, d, J = 8.5, H-5 ), 7.50 (1H, d, J = 1.5, H-2′), 7.54 (1H, dd, J = 1.5, 8.5, H-6′), 10.85 (1H, s, 7-OH), 12.85 (1H,
′
br.s, 5-OH).
35

Acid Hydrolysis of 1. A solution of 1 (25 mg) in HCl (5%, 20 mL) was boiled for 4 h. The resulting precipitate of
the aglycon was filtered off and recrystallized from ethanol to afford luteolin (8 mg), mp 327-330°C. PC (system 4) of the
hydrolysate identified D-galactose.
Heptaacetate (2). A solution of 1 (20 mg) in pyridine (1.5 mL) and acetic anhydride (4 mL) was held at room
temperature for 4 h. The usual workup afforded the heptaacetate (24 mg), mp 118-120°C.
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