Combined Effects of Spinach Leaf Powder and Pomegranate Peel Powder on Hematological Recovery and Iron Status in Phenylhydrazine-Induced Hemolytic Anemia in Rats

Abstract

Iron deficiency anemia (IDA) is a critical public health problem in the world and effective, affordable and sustainable nutritional interventions are needed. This study investigated the hematological efficacy and safety of spinach leaf powder (SLP), pomegranate peel powder (PPP) and their combination in anemic Sprague-Dawley rats induced with phenylhydrazine. Forty-eight male rats (n= 12/ group) with confirmed anemia (baseline Hb < 10.5 g/dL, representing a >30% reduction from pre-induction levels) were divided into four groups: Anemic control, SLP (130 mg/kg/day), PPP (55 mg/kg/day), and SLP+PPP (65 + 28 mg/kg/day) for 48 days. Proximate analysis showed that SLP is very rich in iron (109.2 mg/100 g) and protein (24.5%) while PPP is rich in dietary fiber (32.4%) and bioactive polyphenols. Hematological parameters including hemoglobin, RBC, PCV, serum iron and TIBC showed significant improvements in all treatment groups compared with the anemic control (p = 0.001 for Hb, RBC, and PCV; p < 0.05 for serum iron and TIBC). The SLP+PPP treatment has the best hematological recovery of Hb (15.3 ± 1.0 g/dL), RBC (8.5 ± 0.5 × 10⁶/µL), PCV (45.5 ± 2.3%), and serum Fe (0.88 ± 0.03 μg/dL), despite each component being administered at only half the individual doses. No noticeable toxicity was observed, with no significant changes in WBC (p = 0.450), platelet count (p = 0.620) and relative liver and kidney weights (p > 0.05). These findings provide evidence for synergistic interaction between spinach and pomegranate peel, suggesting that this sustainable plant-based combination is a promising functional food strategy to combat iron deficiency anemia.