Beruflich Dokumente
Kultur Dokumente
5(1) 2002 A
Sirwa N. Majid**
Abstract
The isolation and identification of flavonoid compounds, rutin and quercetin, in
the fruit of prunus microcarpa (Rosaceae) are described. The flavonoid compounds have
been extracted and separated by paper chromatography, thin - layer - chromatography,
and flash coloum chromatography. Two compounds, rutin and quercetin, were
identified by 1H-N. M. R, IR, and UV. Rutin was hydrolyzed by 2M HCl /30 min.,
followed by extracting with ethyl acetate. Quercetin was obtained from the organic
layer, and the aqueous layer was examined by paper chromatography and showed the
presence of glucose and rhamnose.
The compound rutin(glycoside flavonoid) was quantified and the concentration
found to be 57.081%.
language
balalook,
arabic
is
OH
Flavonoids
have
a
typical
chemical structure consisting of two
benzene rings enclosing a hetro cyclic six
membered ring containing an oxygen
atom(3)(1).
HO
OC12H21O9
OH
(2)
3
2
8
7
OH
OH
1
2
HO
OH
6
3
6
5
4
OH
O
OH
(1)
Typical chemical structures of the
(3)
Experimental Procedures
The fruit of prunus microcarpa
have been collected in june 2000 in
Sulaimani governerate(Iraqi Kurdistan).
The fruit was allowed to dry in the
laboratory and ground through a wiley
mill and stored tightly at 4Co. 1H-N.M.R.
1
Spectroscopy:
H-N.M.R.
spectra
obtained using a varian 60MHZ
instruments with tetra methyl silane TMS
as internal standard. I.R Spectroscopy:
The infra-red spectra were obtained using
Beijing second optical instruments
factory,
WQF
300
FT
IR
spectrophotometer. UV spectroscopy:
Ultraviolet data were obtained using
PERKIN-ELMER
HITACHI
200
spectrophotometer with recorder. T.L.C :
standard silica gel(T.L.C) plates were
used with 0.25 mm thickness.
Extraction of Flavonoids
In general alcohols are the solvent
of choice for the extraction of flavonoids.
Residue
Ext. with 80%
methanol
Residue
Flash chromatography
Rutin
Quercetin
Flavonoid
10%
HAC in
Flash Chromatography
BAW
BEW
Forestal
CHCl3
Rutin
50
51
50
34
Quercetin
66
84
89
42
Rf 100
Flavonoid
BAW
15% HOAC
Rutin
43
50
Quercetin
64
Chromatography
Rf 100
Glucose
20
13
Brown
Rhamnose
61
62
Pale brown
Solvent key:
Paper Chromatography
The
flavoniod
compounds
(glycoside and a glycon flavonoids) were
separated from prunus microcarpa fruit by
extraction of the defatted fruit powder
with methyl alcohol and then by ethyl
acetate as shown in scheme 1.
Simple two dimensional paper
chromatography and one dimensional T.
L. C examined the complexity of the ethyl
acetate extract, and aqueous methanol
layer respectively in scheme (1).
The paper and thin- layer
chromatography for ethyl acetate extract
showed two spots, first spot which was
corresponding to quercetin was separated
by flash column chromatography and
identified by 1H. N. M. R. , IR. , and UV.
spectroscopy.
Rf ( 100) in
Colour
with
Sugar
Aniline
BAW
BEW
Hydrogen
Phthalate
cm-1
Signal
Assignment
Quercetin
3600-3200
Broad band
1665
Signal band
C=O group
1640
Signal band
1575-1477
Multiplet
1300-1000
Multiplet
C-O group
below 900
Multiplet
3600-3000
Broad band
2900
Signal band
1715
Signal band
C=O group
1638-1484
Multiplet
1433-1368
Multiplet
1300-1000
Multiplet
C-O group
below 900
Multiplet
Rutin
829
Distinct
-D-glycoside linkage
band
signal at (2.5-5.5)for five protons of OH groups and the multiplet signal at (7-
Identification
of
flavoniod
glycoside (Rutin) has been done by paper,
and thin-layer chromatography, and
spectral measurments IR, 1H. N. M. R. ,
UV.
The IR spectrum table (4) of the
aromatic proton.
CH3
glycoside linkage(8).
The 1H. N. M. R. spectrum
table(5) of the compound rutin showed a
multiplet signal at (1.7-2.3) for CH2,
protons.
The
broad
signal
Table (5): 1H. N. M. R. Spectral Data of the Compounds Quercetin and Rutin with
TMS as internal standard , 60 MHZ, D2O .
Signal
Assignment
2.5-5.5
5 H broad
OH groups
7-8
5 H Multiplet
1.7-2.3
Multiplet signal
2.4-5.0
Broad signal
OH groups
7-8
Multiplet signal
compounds
Quercetin
Rutin
Acknowledgements
We
thank
department
of
References
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(Prunus microcarpa)
Flavonoids
P.CPrunus microcarpa
Flash ChromatographyT. L. C.
1H. N. M. R. , IR, UV
Reflex30 min.2MHCl
Prunus microcarpafruit
57.081%
(Prunus microcarpa)
Flavonoids
T. P.Cprunus microcarpa
RutinFlash ChromatographyL. C.
1H. N. M. R. , IR, UVQuercetin
QuercetinEthyl acetate2MHCl
P.C
(Prunus Rutin
57.081%microcarpa