Flavonoids and anthocyans from Alhagi pseudoalhagi

Full text of DOI:10.1007/s10600-009-9287-0

Chemistry of Natural Compounds, Vol. 45, No. 2, 2009
FLAVONOIDS AND ANTHOCYANS FROM Alhagi pseudoalhagi
E. N. Novruzov, G. A. Abdullaeva,
UDC 547.581.972
L. A. Shamsizade, and N. Sh. Mustafaev*
The plant Alhagi pseudoalhagi is relatively little studied. Acidic sulfatidylated polygalactones, tanning agents [1],
catechins, and proanthocyanidines [2] have been observed in its aerial part. It was found that tincture prepared from runners
exhibits diuretic, litholytic [3], hypoazotemic, antioxidant [4], and cholegogic activity [5].
Herein we present data on the flavonoid composition of the aerial part and the qualitative composition and quantitative
content of anthocyans from flowers of A. pseudoalhagi (Fabaceae Lindl.) growing in the Republic of Azerbaidzhan [6].
Material was collected near Baku (Ramana) during budding (June 2007). Spectrophotometry [7] showed that the aerial part of
the plant contained 2.95% flavonoids (per dry weight) [8].
Pure flavonoids were isolated from ground air-dried aerial part by exhaustive extraction with ethanol on a water bath
at 70°C. The extract was filtered and condensed in vacuo to a small volume. The evaporated extract was diluted with water
and treated successively with CHCl , ether, EtOAc, and n-BuOH. The ether extract contained two compounds; EtOAc, 3;
3
BuOH, 2.
Preparative paper chromatography (Whatman-3) of the ether extract isolated two compounds from MeOH with
mp 312°C, which were identified as quercetin (1) and 2 with mp 300-303°C. UV spectrum (MeOH, λmax, nm): 256 and
2
76 nm. Alkaline destruction of 2 produced phloroglucinol and vanillinic acid. Analyses suggested that 2 was identical to
isorhamnetin.
The EtOAc extract was separated using column chromatography over polyamide (Woelm) with elution by
CH OH:CHCl and CH OH:H O. Compounds 3 and 4 were isolated from the CHCl :CH OH (90:10) and CH OH:H O
3
3
3
2
3
3
3
2
(
95:5) fractions.
Compound 3 was yellow needles (from aqueous CH OH), mp 179-180°C. UV spectrum (MeOH, λmax, nm): 258 and
3
3
60 nm. The algycon was obtained in 42% yield by hydrolysis with H SO (5%) in ethanol (50%). The aglycon was identified
2
4
as isorhamnetin. Paper chromatography of the hydrolysate detected D-glucose and L-rhamnose. The sugar in 3 was bonded
to the C position according to brown fluorescence of a spot in UV light and a positive zirconium-citrate sample. The
3
bathochromic shift of the long-wavelength absorption maximum in CH COONa by 12 nm compared with that in EtOH
3
indicated that C was unsubstituted [9]. Thus, 3 was identified as isorhamnetin 3-rutinoside.
7
Compound 4, mp 184-185°C. UV spectrum (MeOH, λmax, nm): 258 and 364 nm. Addition of AlCl and CH COOH
3
3
and CH COONa + H BO gave a bathochromic shift, indicating that free hydroxyls were located in the 5, 7, 3′, and 4′
3
3
3
positions. Hydrolysis of 4 by H SO (2%) produced quercetin in 47% yield. The hydrolysate of 4 contained D-glucose and
2
4
L-rhamnose.
Based on these data, 4 was characterized as 5,7,3′,4′-tetrahydroxyflavon-3-β-D-rutinoside (rutin).
Fresh petals were separated from the bases, homogenized with MeOH (1:1 ratio), and left to stand for 2 h in order to
analyze the anthocyans. The homogenate was passed through a laboratory screw press. The solids were treated three times
with MeOH containing HCl (1%) for 2 h. The extracts were combined, filtered, and condensed in vacuo to an aqueous residue
that was treated successively with ether and EtOAc. Two-dimensional paper chromatography (FN 16) using CH CO H:HCl:H O
3
2
2
(
15:3:82, system I) and n-BuOH:CH CO H:H O (4:1:2, system II) detected three anthocyans in the purified extract.
3 2 2
Total purified anthocyans were separated by distribution chromatography over a column packed with cellulose activiated
by HCl (1%) using system I in descending mode. The column showed three clearly resolved bands. The adsorbent was
Institute of Botany, National Academy of Sciences of Azerbaidzhan, Baku, AZ1073, Patamdarskoe Shosse, 40,
e-mail: mustafayevn02@yahoo.co.uk. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 213-214, March-April,
2
009. Original article submitted November 27, 2008.
0
009-3130/09/4502-0249 ©2009 Springer Science+Business Media, Inc.
249

removed from the column. The layers were separated. Anthocyans were eluted by MeOH containing HCl (1%) and precipitated
by Et O. The precipitates of pure anthocyans were rechromatographed on paper using system II to afford three pure anthocyans
2
that were arbitrarily designated anthocyan 5, 6, and 7.
Anthocyan 5 was fine dark-violet crystals, R 0.20 and 0.42 (systems I and II, respectively). It was purple in daylight
f
and dull purple in UV light. UV spectrum [MeOH containing HCl (0.01%), λmax, nm): 535 nm; +AlCl (5%), 546 nm;
3
E440/Emax = 18%. Acid hydrolysis formed the aglycon with R 0.32 (system III, CH CO H:HCl:H O, 30:3:10). UV spectrum
f
3
2
2
[
MeOH containing HCl (0.01%), λmax, nm): 546 nm; +AlCl , 568 nm; E440/Emax = 18%. The aglycon was identified as
3
delphinidin using chromatography and UV spectra. The sugar part of the hydrolysate contained D-glucose. The aglycon:sugar
ratio was 1:1. The bonding site of the carbohydrate was found from the hypochromic shift of λmax of the glycoside compared
with the maximum of the aglycon (535 and 546 nm). This effect was consistent with a carbohydrate in the C position [10].
3
Anthocyan 6 was fine dark-red crystals with R 0.17 and 0.28 (systems I and II, respectively). UV spectrum [MeOH
f
containing HCl (0.01%), λmax, nm): 525 nm; +AlCl , 540 nm; E440/Emax = 13%. Acid hydrolysis produced the aglycon with
3
Rf 0.48 (system III). UV spectrum [MeOH containing HCl (0.01%), λmax, nm): 535 nm; +AlCl , 543 nm; E440/Emax = 18%.
3
The aglycon was cyanidin according to these data. The sugar part of the hydrolysate contained D-glucose. The aglycon:sugar
ratio was 1:2. Stepwise hydrolysis formed first an anthocyan with R 0.38 and 0.43 (systems I and II). Further hydrolysis
f
cleaved the aglycon cyanidin and D-glucose.
Anthocyan 7 was microcrystalline dark-violet powder with a metallic sparkle and R 0.32 and 0.15 (systems I and II,
f
respectively). It was purple in daylight and dark-purple in UV light. UV spectrum [MeOH containing HCl (0.01%),
λmax, nm): 534 nm; +AlCl , 545; E440/Emax = 11%. Total acid hydrolysis gave the aglycon, which was identified as delphinidin,
3
and D-glucose. The aglycon:sugar ratio was 1:2. Stepwise acid hydrolysis formed after 30 min an anthocyan corresponding
to anthocyan A and D-glucose; after 60 min, the aglycon corresponding to delphinidin and D-glucose.
Thus, anthocyan 5 was identified as delphinidin-3-monoglucoside; 6, cyanidin-3,5-diglucoside; 7, delphinidin-3,5-
diglucoside based on the results and a comparison with authentic samples and the literature [10, 11].
In both instances the qualitative composition of the anthocyans from flowers was almost the same with a difference in
the ratio of the individual components. The content of 6 was elevated at a contaminated site. Photoelectrocalorimetry [12]
established that flowers collected at a radioactive contaminated site had elevated amounts of 6 compared with samples collected
at the Botanical Garden. The anthocyan content from Ramana was 1.58%; from the Botanical Garden, 1.23%.
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