Mizahn Ethnomedicinal plant resources At sites of inflammation, the oxidation of Cl — ions by the neutrophil enzyme myeloperoxidase results in the production of another harmful ROS, hypochlorous acid [ 41 ]. Scavenging of reactive oxygen species and iron chelating activity IC 50 radcial of Spondias pinnata and reference compounds. As shown in figure 3, the IC50 values performed six times. Amelioration of chronic ileitis by nitric oxide synthase inhibition. Antioxidant and free radical scavenging activity of Spondias pinnata The results suggest that the plant extract is a more potent scavenger of superoxide radical than the standard quercetin.
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Bibhabasu Hazra: moc. This article has been cited by other articles in PMC. Abstract Background Many diseases are associated with oxidative stress caused by free radicals.
Current research is directed towards finding naturally-occurring antioxidants of plant origin. The aim of the present study was to evaluate the in vitro antioxidant activities of Spondias pinnata stem bark extract. Results The extract showed total antioxidant activity with a trolox equivalent antioxidant concentration TEAC value of 0.
The IC50 values for scavenging of free radicals were The IC50 for hydrogen peroxide scavenging was For the peroxynitrite, singlet oxygen and hypochlorous acid scavenging activities the IC50 values were The reducing power was increased with increasing amounts of extract.
The plant extract mg yielded Oxidative stress is initiated by free radicals, which seek stability through electron pairing with biological macromolecules such as proteins, lipids and DNA in healthy human cells and cause protein and DNA damage along with lipid peroxidation.
These changes contribute to cancer, atherosclerosis, cardiovascular diseases, ageing and inflammatory diseases [ 1 , 2 ]. All human cells protect themselves against free radical damage by enzymes such as superoxide dismutase SOD and catalase, or compounds such as ascorbic acid, tocopherol and glutathione [ 3 ]. Sometimes these protective mechanisms are disrupted by various pathological processes, and antioxidant supplements are vital to combat oxidative damage.
Recently, much attention has been directed towards the development of ethnomedicines with strong antioxidant properties but low cytotoxicities. Spondias pinnata Linn. In India it is commonly seen in the deciduous to semi-evergreen forests of the Western Ghats. The genus Spondias includes 17 described species, 7 of which are native to the neotropics and about 10 are native to tropical Asia. The phytochemistry of this plant has been studied [ 4 ].
The gum exudate of the species has been found to contain acidic polysaccharides [ 5 ]. A crude extract of S. In ethnomedicine, equal quantities of bark juice of S. An aqueous extract of this plant inhibits the citrus canker of lime [ 8 ].
However, there has been no report on the antioxidant properties of this species. The extract was examined for different reactive oxygen species ROS scavenging activities including hydroxyl, superoxide, nitric oxide, hydrogen peroxide, peroxynitrite, singlet oxygen and hypochlorous acid, and for phenol and flavonoid contents, iron chelating capacity and total antioxidant activity.
Ltd, Mumbai, India. Gallic acid and curcumin were obtained from MP Biomedicals, France. Ferrous sulfate and catalase were obtained from HiMedia Laboratories Pvt.
Plant material The bark of the S. Extraction The stem bark of S. The pellet was mixed again with ml methanol-water and the entire extraction process was repeated. The supernatants collected from the two phases were mixed in a round bottom flask and concentrated under reduced pressure in a rotary evaporator. The concentrated extract was then lyophilized. The ABTS.
The absorbance of the ABTS. All experiments were repeated six times. The percentage inhibition of absorbance was calculated and plotted as a function of the concentration of standard and sample to determine the trolox equivalent antioxidant concentration TEAC. To calculate the TEAC, the gradient of the plot for the sample was divided by the gradient of the plot for trolox.
Hydroxyl radical scavenging This was assayed as described by Elizabeth and Rao [ 10 ] with a slight modification. The assay is based on quantification of the degradation product of 2-deoxyribose by condensation with TBA. The reaction mixture contained, in a final volume of 1 ml, 2-deoxyribose 2. After cooling, the absorbance was measured at nm against an appropriate blank solution. All tests were performed six times.
Mannitol, a classical OH. Percentage inhibition was evaluated by comparing the test and blank solutions. Superoxide radical scavenging This activity was measured by the reduction of NBT according to a previously reported method [ 11 ]. The 1 ml reaction mixture contained phosphate buffer 20 mM, pH 7. After incubation for 5 min at ambient temperature, the absorbance at nm was measured against an appropriate blank to determine the quantity of formazan generated. Quercetin was used as positive control.
Nitric oxide radical scavenging At physiological pH, nitric oxide generated from aqueous sodium nitroprusside SNP solution interacts with oxygen to produce nitrite ions, which may be quantified by the Griess Illosvoy reaction [ 12 ]. Then 1 ml of napthylethylenediamine dihydrochloride NED 0.
The pink chromophore generated during diazotization of nitrite ions with sulphanilamide and subsequent coupling with NED was measured spectrophotometrically at nm against a blank sample. Curcumin was used as a standard. Hydrogen peroxide scavenging This activity was determined according to a previously described method [ 13 ] with minor changes.
The reaction mixture was then vortexed and incubated at room temperature for 30 min. The absorbance of the ferric-xylenol orange complex was measured at nm. All tests were carried out six times and sodium pyruvate was used as the reference compound [ 14 ]. An acidic solution 0. Excess H2O2 was removed by treatment with granular MnO2 prewashed with 1. An Evans Blue bleaching assay was used to measure peroxynitrite scavenging activity. The assay was performed by a standard method [ 16 ] with a slight modification.
The reaction mixture contained 50 mM phosphate buffer pH 7. The percentage scavenging of ONOO- was calculated by comparing the results of the test and blank samples. Gallic acid was used as the reference compound. Singlet oxygen scavenging The production of singlet oxygen 1O2 was determined by monitoring N, N-dimethylnitrosoaniline RNO bleaching, using a previously reported spectrophotometric method [ 17 , 18 ].
The reaction mixture contained 45 mM phosphate buffer pH 7. The scavenging activity of sample was compared with that of lipoic acid, used as a reference compound.
The assay was carried out as described by Aruoma and Halliwell [ 19 ] with minor changes. The scavenging activity was evaluated by measuring the decrease in absorbance of catalase at nm. The reaction mixture contained, in a final volume of 1 ml, 50 mM phosphate buffer pH 6.
Ascorbic acid, a potent HOCl scavenger, was used as a reference [ 20 ]. The mixture was shaken vigorously and incubated for 20 min at room temperature, then the absorbance was measured at nm. EDTA was used as a positive control. Different concentrations 0.
After incubation, 0. The upper portion of the solution 1 ml was mixed with 1 ml distilled water, and 0. The reaction mixture was left for 10 min at room temperature and the absorbance was measured at nm against an appropriate blank solution. A higher absorbance of the reaction mixture indicated greater reducing power. Butylated hydroxytoluene BHT was used as a positive control. Determination of total phenolic content Total phenolic content was determined using Folin-Ciocalteu FC reagent according to the method of Singleton and Rossi [ 23 ] with a slight modification.
Briefly, the plant extract 0. The phenolic content was evaluated from a gallic acid standard curve. Determination of total flavonoid content The total flavonoid content was determined with aluminium chloride AlCl3 according to a known method [ 24 ] using quercetin as a standard.
The plant extract 0. After a further 5 min, the reaction mixture was treated with 0. Finally, the reaction mixture was diluted to 1 ml with water and the absorbance was measured at nm.
The flavonoid content was calculated from a quercetin standard curve. Statistical analysis was performed using KyPlot version 2. The IC50 values were compared by paired t tests. Results Total antioxidant activity The total antioxidant activity of the extract was calculated from the decolorization of ABTS. Interaction with the extract or standard trolox suppressed the absorbance of the ABTS. The TEAC value of the extract was 0. Figure 1 Total antioxidant activity.
Total antioxidant activity of plant extract and trolox. Effect of a Spondias pinnata extract and b reference compound trolox on decolorization of ABTS radical cation. The percentage inhibition was plotted against the concentration of sample. The IC50 value of the extract was less than that of the standard. Figure 2 Hydroxyl radical scavenging assay.
ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF SPONDIAS PINNATA PDF
Abstract Background Many diseases are associated with oxidative stress caused by free radicals. Current research is directed towards finding naturally-occurring antioxidants of plant origin. The aim of the present study was to evaluate the in vitro antioxidant activities of Spondias pinnata stem bark extract. Results The extract showed total antioxidant activity with a trolox equivalent antioxidant concentration TEAC value of 0. The IC50 values for scavenging of free radicals were
Antioxidant and free radical scavenging activity of Spondias pinnata.
Bibhabasu Hazra: moc. This article has been cited by other articles in PMC. Abstract Background Many diseases are associated with oxidative stress caused by free radicals. Current research is directed towards finding naturally-occurring antioxidants of plant origin.