HPLC profiling and studies on Copaifera salikounda methanol leaf extract on phenylhydrazine-induced hematotoxicity and oxidative stress in rats

Anemia is a clinical disorder orchestrated by factors such as drugs, including phenylhydrazine (PHZ). This study explored the protective roles of Copaifera salikounda methanol leaf extract (CSMLE) against PHZ-induced hematotoxicity and oxidative stress in rats. In vitro antioxidants studies of CSMLE using 1,1-diphenyl-2-picryl hydrazyl (DPPH) and ferric reducing antioxidant potential (FRAP) and quantification of CSMLE bioactive compounds using High-Performance-Liquid-Chromatography (HPLC) were done. In vivo, thirty rats in five groups (n = 6) were used. Anemia was induced intraperitoneally in groups 2 to 5 with 40 mg/kg body-weight/day PHZ for two days. Group 1 received normal saline, group 2 (untreated: negative-control), group 3 received 1 ml/kg-body-weight Vit B₁₂ syrup while groups 4 and 5 received 200 mg/kg and 400 mg/kg-body-weight CSMLE, respectively, daily for fourteen days. Result of CSMLE profiling showed luteolin (76.49 ng/Ml) and squalene (38.31 ng/Ml) as the most abundant flavonoid and terpenoid, respectively. In vitro CSMLE demonstrated dose-dependent antioxidant effects on DPPH and FRAP with IC₅₀ of 0.1085 mg/ml and 0.07344 mg/ml (EC₅₀), respectively. In vivo, CSMLE improved the body weight, red blood cell, hematocrit and hemoglobin concentrations, significantly (P < 0.05), reduced serum malondialdehyde and nitic oxide but increased reduced glutathione levels, superoxide dismutase and catalase activities and attenuated the levels of Interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in dose-dependent manner in treated groups relative to the negative control. The spleen tissues histology results corroborated biochemical results. The results indicated that CSMLE possesses anti-anemic potentials and attenuate spleenotoxic effects of PHZ.