Abstract
Objective:To investigate the in vitro antioxidant activity of different fractions (R1, R2 and R3) obtained from pet ether extract of black pepper fruits (Piper nigrum Linn.)Materials and Methods:The fractions R1, R2 and R3 were eluted from pet ether and ethyl acetate in the ratio of 6:4, 5:5 and 4:6, respectively.1,1-Diphenyl-2-picryl-hydrazyl (DPPH) radical, superoxide anion radical, nitric oxide radical, and hydroxyl radical scavenging assays were carried out to evaluate the antioxidant potential of the extract.Results:The free radical scavenging activity of the different fractions of pet ether extract of P. nigrum (PEPN) increased in a concentration dependent manner. The R3 and R2 fraction of PEPN in 500 µg/ml inhibited the peroxidation of a linoleic acid emulsion by 60.48±3.33% and 58.89±2.51%, respectively. In DPPH free radical scavenging assay, the activity of R3 and R2 were found to be almost similar. The R3 (100µg/ml) fraction of PEPN inhibited 55.68±4.48% nitric oxide radicals generated from sodium nitroprusside, whereas curcumin in the same concentration inhibited 84.27±4.12%. Moreover, PEPN scavenged the superoxide radical generated by the Xanthine/Xanthine oxidase system. The fraction R2 and R3 in the doses of 1000µg/ml inhibited 61.04±5.11% and 63.56±4.17%, respectively. The hydroxyl radical was generated by Fenton's reaction. The amounts of total phenolic compounds were determined and 56.98 µg pyrocatechol phenol equivalents were detected in one mg of R3.Conclusions:P. nigrum could be considered as a potential source of natural antioxidant.
Highlights
The importance of the reactive oxygen species (ROS) has attracted increasing attention over the last decade
The R3 and R2 fraction of pet ether extract of Piper nigrum (PEPN) in 500 μg/ml inhibited the peroxidation of a linoleic acid emulsion by
The R3 (100μg/ml) fraction of PEPN inhibited 55.68±4.48% nitric oxide radicals generated from sodium nitroprusside, whereas curcumin in the same concentration inhibited 84.27±4.12%
Summary
Ten ml of NBT solution (0.025 mM) was added to the reaction mixture From this solution, 995 μl with xanthine was further added to 5μl of each fraction of PEPN and the reference compound, in different concentrations in distilled water. Hydroxyl radical scavenging activity was measured comparing deoxyribose and PEPN for hydroxyl radical generated by the Fe3+-ascorbate-EDTA-H2O2 system (Fenton reaction), using the method of Kunchandy and Rao.[16] The reaction mixture containing (1.0 ml) 100 μl 2-deoxy-2-ribose (28 mM in 20 mM phosphate buffer, pH 7.4), 500 μl of each fraction of PEPN and the reference compound in phosphate buffer (20 mM, pH 7.4), 200 μl 1.04 mM EDTA, and 200 μM FeCl3 (1:1 v/v), 100 μl 1.0.
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