In vivo and in vitro evaluation of antioxidant, antihemolytic, acute toxicity effects and high-performance liquid chromatography analysis of an aqueous extract of Moroccan Punica granatum L.

Abstract

Introduction: The use of pomegranates in Moroccan pharmacopeia is due to their healing and nutritional properties because of their richness in secondary molecules. The following study analyses the composition of the aqueous extract of Punica granatum peel and evaluates in vivo and in vitro antioxidant effects, hemolytic protection, and acute toxicity. Methods: Quantification of the plant extract was realized by high-performance liquid chromatography (HPLC). The hemolytic assay was used for erythrocyte protection, while the in vitro antioxidant effect was evaluated by 2, 20-azinobis-(3-ethylbenzothiazoneline-6-sulphonic acid) (ABTS) and reducing ferric power (RFP) assays. The in-vivo antioxidant activity was tested by measuring levels of lipid peroxidation (LPO) in serum. The toxicological study was tested by oral administration of the extract to four groups of mice for 21 days, followed by a histopathological examination of the spleen. Results: HPLC analysis showed the presence of some phenolic compounds such as coumarin, caffeic, gallic and syringic acids. The IC50 of the antioxidant assays were 254.49 ± 62.17 μg/mL and 40.265 ± 2.9 μg/mL for ABTS and reducing power, respectively. Furthermore, the thiobarbituric acid reactive substances (TBARS) assay showed the lowest levels at 150 mg/mL concentration. All of the concentrations used for hemolytic protection did not exceed 15% of hemolysis. Moreover, the toxicity test showed no sign of mortality, signs of weakness, or weight loss; also the histopathological examination of the spleen tissues showed the absence of any damage. Conclusion: The peel extract of P. granatum showed good potential and could be exploited as a natural antioxidant and antihemolytic remedy, leading to the development of new drugs.