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Vol3-No5-1 (Flavonoid NMR) PDF
Vol3-No5-1 (Flavonoid NMR) PDF
J OURNAL OF FOREST PRODUCTS & INDUSTRIES, 2014, 3(5), 211-215 ISSN:2325–4513( PRINT) ISSN 2325 - 453X (ONLINE )
Research Article
organisms. the cup-plate agar was adopted with some minor III. RESULTS AND DISCUSSION
modification. (2 ml) Of the standard bacteria stock suspension Spectral data of compound A1
were mixed with 200 ml of sterile molten nutrient agar which The UV λmax (MeOH nm) 260, 295sh, 360 nm
was maintained at 45 C. (MeOH+NaOMe) 270, 326 and 410 nm (MeOH+NaOAc) 271,
(20 ml) Aliquots of incubated agar were distributed into sterile 323sh, 386 nm (MeOH+NaOAc+H3BO3) 263, 295sh, 379 nm
Petri dishes. The agar was left to settle and each plate was cut (MeOH+AlCl3) 275, 325sh, 425 nm (MeOH+AlCl3+HCl) 269,
and agar discs were removed. Alternated cups were filled with 371, 400 nm.
0.1 ml sample of each of the extract (pure component) and IR νmax (cm-1, KBr disc): 632- 759 (C-H, Ar bending), 1253
allowed to diffuse at room temperature for two hours. The Petri (C-O, ether), 1620 (C= C, Ar), 1705 (C=O), 2927 (C-H,
dishes were then incubated in the upright position at 37 C for alkanes) and 3301- 3452 cm-1 (O-H).
1
18 hours. After incubation, the diameter of the resultant growth H-NMR (500 MHz, DMSO-d6): δ (ppm): showed (figure: 1)
inhibition zones were measured, and averaged [12]. 3.0- 3.5 (m, 6H) assigned for sugar protons; 6.15 (s, 1H) and
6.35 (s, 1H) for C6- and C8- H's respectively. The resonances at
6.79 and 7.53 were assigned for the aromatic H's of B-ring.
Table 1. The antibacterial activity of crude and pure compounds extracted from Acacia nilotica.
Inhibition zone diameter (mm / mg sample)
Sample
Bacillus Escherichia Neisseria Pseudomonas Staphylococcus
subtilis coli gonorrhoeae (G-) aeruginosa aureus
(G+) (G-) (G-) (G+)
Tetracycline
Standard
27 31 33 29 26
Antibacterial agent
Plant crude 14 15 14 15 15
AL3 10 10 10 10 12
AL4 12 12 12 10 12
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J OURNAL OF FOREST PRODUCTS & INDUSTRIES, 2014, 3(5), 211-215 ISSN:2325–4513( PRINT) ISSN 2325 - 453X (ONLINE )
The UV spectrum of compound A1 gave λ max (MeOH) 260, aromatic H's of B-ring. 13C NMR 400 MHz, DMSO-d6) δ C:
360 nm due to the benzoyl and cinnamoyl chromophores of 133.8 – 163.9 (C7, C5, C3, C2, C9, C4` and C3`), 177.9 (C4),
the flavonol moiety [13]. The sodium methoxide spectrum 156.89 (C2), 156.66 (C9). The non-oxygenated carbon atoms
exhibited a bathochromic shift of 50 nm without decrease in (C1`, C2`, C5`, C6`, C6, C8 and C10) appear in the range of
intensity indicating a 4`-hydroxyl group. The addition of 121.6- 93.6. Other carbon atoms bearing one oxygen
sodium acetate gave a bathochromic shift of about +11 nm function in sugar moiety appear at 76.8 – 67.1 ppm. The mass
indicating the presence of 7-hydroxyl group. 19 spectrum of compound (A1) exhibited a molecular ion peak
bathochromic shift was observed in the boric acid spectrum at m/z 465 corresponding to the molecular formula C15H5 O7.
of A1 indicating a catechols moiety. The fragments at m/z 152, 126 and m/z 150 indicated that the
The 1H-NMR spectrum of compound A1 showed: δ 3.0- 3.5 A ring contained two hydroxyl groups and that the B-ring
(m, 6H) assigned for sugar protons; δ 6.15 (s, 1H) and δ 6.35 also has two hydroxyl groups according to following retro
(s, 1H) accounting for C6- and C8- H's respectively. The Diels- Alder process:
resonances at δ 6.79 and 7.53 ppm were assigned for the
OH
OH
HO O
OH
OH O
The complete acid hydrolysis of compound A1 and H3BO3. Thus catechol systems are absent. The AlCl3 spectrum
subsequent UV studies of aglycone using the shift reagent did not affored any detectable bathochromic shift suggesting
AlCl3 indicated a 3 -OH group in the resultant aglycone and absence of C5-, C3-hydroxylation. The 1H-NMR spectrum of
this is precisely the site of glycosylation. compound A2 (figure 3) showed: δ 6.68 (d, 2H) assigned for
The following cumulative data suggest that compound A1 C2`- and C6`-H's; δ 6.5 (d, 1H) assigned for C8- H. The signals at
(may be because the positions of hydroxyls are not 5.84 and 5.65 ppm are characteristic of C3` and C5`-H's, while
confirmed) is a 5, 7, 3`, 4`-tetrahydroxyflavone the resonances at δ 3.3, 3.54 and 3.78 ppm (m, 15H) accounts
-3-O-galactosyl. for a methoxyl function.
OH The mass spectrum of compound A2 exhibited a molecular ion
peak at m/z 372 corresponding to a penta-methoxyflavone. The
HO O retro Diels- Alder fission supported the substitution pattern
OH proposed for assigned for A2, where the ions m/z 162 and m/z
210 were recorded in the electron beam
O
galactosyl
OH O
The UV spectrum of compound A2 gave λ max (MeOH) 279,
355 nm due to the benzoyl and cinnamoyl chromophores of the
flavonol moiety [13]. Band I in A2 absorbs at 355 nm
indicating a flavone or chalcone. But, chalcones are
distinguished (in their1H NMR spectrum) by a double doublet
(J = 17 Hz) in the range delta 6.7 – 7.4 and 7.3 – 7.7 ppm for α-
and β- H's respectively [14]. The 1H NMR spectrum of A2
(figure 3) did not reveal any signals for α- and β- protons. Thus
A2 is probably a flavone.
Only a slight bathochromic shift was observed in the sodium
methoxide spectrum indicating absence of a 3- and 4`-OH. The
sodium acetate spectrum was devoid of bathochromic shifts
indicating the absence of a 7-hydroxyl group. Also no
bathochromic shift was observed on addition of NaOAc/
215
J OURNAL OF FOREST PRODUCTS & INDUSTRIES, 2014, 3(5), 211-215 ISSN:2325–4513( PRINT) ISSN 2325 - 453X (ONLINE )
nilotica in table (1) showed moderate inhibition against all comprehensive review on ethnopharmacological claims.
five organisms, also compounds A1 and A2 showed moderate International Journal of Pharmacy and Life Sciences, 2(6),
inhibition against all five organisms. 0976-7126.
[4] Saini, M. L. (2008): Comparative Pharmacognostical
IV. CONCLUSION and antimicrobial studies of Acacia species (Mimosaceae).
In the present study, two flavonoids: querstin 3-galactosyl Journal of Medicinal Plants Research, 2(12), 378-386.
and flavone: were isolated from ethyl acetate extract of [5] Duke, J. A. (1983): Medicinal plants of the Bible.
acacia nioltica (Leguminosae) leaves, and purified using Trado-Medic Books, Owerri.
different chromatographic techniques. These compounds [6] Bohm, B. A. (1998): "Introduction to flavonoids"
(A1 and A2) were identified via spectroscopic tools: IR, UV, Harwood Academic Publishers, Amsterdam.
1
H NMR, 13C NMR and MS spectroscopy. These [7] Rasmussem, S. E. and Breinholt, V. M. (2003): Int. J.
compounds showed potent antibacterial activity. This would Vitamin Nutr. Res, 73(2), 111.
be helpful to create awareness among people for taking [8] Cushnie, T. P. and Andrew, J. and Lamb A. J. (2005):
control measures based on, herbal plants against infectious Antibacterial activity of flavonoids. International Journal of
diseases. Antimicrobial Agents, 26, 343–356.
[9] Tsuchiya, H. (2010): Food Chemistry, 120, 1089-1096.
ACKNOWLEDGMENTS [10] Pal, R. S., Ariharasivakumar, G., Girhepunjhe, K. and
We are grateful to Professor Dr. Salwa Abd El Mgsoud, for Upadhay, A. (2009): International Journal of Pharmacy
her interest in this study and for her valuable help and and Pharmaceutical Sciences, 1, 136-140.
support; our thanks also extend to the Cairo University for [11] Marchand, L. L. (2002): Cancer preventive affects of
technical assistance during this work. flavonoids- a review. Biomedicine and Pharmacotherapy,
56, 296-301.
REFERENCES [12] Kavanagh, F. (1972): "Analytical Microbiology" Vol.
[1] Maslin, B. R., Miller, J. T. and Seigler, D. S. (2003): II, Academic Press (Pub) New York and London, P. 11.
Australian Systematic Botany. 16(1), 1-18. [13] Harbone, J. B. (1973): "Phytochemical Methods",
[2] Orchard, A. E. and Maslin, B. R. (2003): Taxon, 52(2), Chapman and Hall, London.
362-363.
[14] Harbone, J. B., Mabry, T. J. and Mabry, H. "The
[3] Sapna, M., Swati, R., Anil, K. and Meena, V. (2011):
flavonoids", part 1, Chapman and Hall, London (1975).
Medicinal attributes of Acacia nilotica Linn. A