JOHN. C. ANIONYE

The widespread use of herbal supplements in Sub-Saharan Africa has created an urgent need for safety evaluation of commercially available polyherbal formulations. Miracle Seed Ultima® (MSU) is a popular liquid extract marketed for immune enhancement, fertility improvement, and metabolic regulation, yet it lacks comprehensive toxicological data to support its safety claims. This study investigated the acute and subacute toxicity profile of MSU in male albino Wistar rats to establish its safety. The acute toxicity assessment was conducted using the Lorke method, which involved administering MSU at doses ranging from 10 mg/kg to 5000 mg/kg. Animals were observed for 14 days for mortality, behavioral changes, and clinical signs of toxicity. The subacute toxicity study followed OECD guideline 407, where rats received daily oral doses of 100 mg/kg, 300 mg/kg, and 1000 mg/kg for 28 days. Body weight, food consumption, and clinical signs were monitored throughout the study period. Blood samples were collected for hematological analysis and biochemical assessment of liver function (ALT, AST, ALP, total protein, albumin, bilirubin), kidney function (urea, creatinine, electrolytes), lipid profile (total cholesterol, triglycerides, LDL, HDL, VLDL), and fasting blood glucose. Phytochemical analysis identified high concentrations of cardiac glycosides (310.59 ± 2.36 mg/g), steroids (174.22 ± 5.01 mg/g), flavonoids (151.82 ± 3.58 mg/g), coumarins (113.27 ± 4.81 mg/g), and alkaloids (93.81 ± 7.86 mg/g). Elemental analysis indicated high levels of calcium (6,304.00 ± 2.08 mg/kg), potassium (5,252.33 ± 1.20 mg/kg), and magnesium (3,704.67 ± 2.40 mg/kg), with no detectable levels of lead or cadmium, suggesting low contamination risk. In vitro antioxidant tests showed moderate DPPH radical scavenging at 55.77 ± 3.67% and FRAP reducing activity at 51.95 ± 1.31%, with notably strong nitric oxide scavenging at 88.15 ± 2.21%, comparable to ascorbic acid (92.38 ± 0.41%). Acute toxicity tests at doses up to 5,000 mg/kg showed no mortality or clinical signs over 14 days, classifying MSU as practically non-toxic (LD50 > 5,000 mg/kg). However, kidney cysts were observed in some animals at higher doses. Subacute exposure (100, 300, 1,000 mg/kg/day for 28 days) revealed dose-dependent hepatocellular stress, with elevated serum ALT at 300 mg/kg (135.00 ± 8.70 U/L) and 1,000 mg/kg (200.56 ± 22.20 U/L). Conversely, kidney function improved, with decreased plasma urea at 1,000 mg/kg (68.62 ± 2.90 mg/dL) versus control (102.72 ± 5.70 mg/dL), suggesting nephroprotective effects. All hematological parameters and metabolic markers remained normal, indicating a No Observed Adverse Effect Level (NOAEL) of 100 mg/kg/day. This study provides the first comprehensive toxicological evaluation of Miracle Seed Ultima®, demonstrating that the formulation has acceptable safety at moderate doses but may cause mild liver stress at higher doses. The findings suggest that MSU can be safely used at doses up to 100 mg/kg, but long-term users should undergo periodic liver function monitoring, particularly those with pre-existing liver conditions. These results provide essential baseline data to inform regulatory decisions, guide clinical use, and protect consumer safety while supporting the continued use of this traditional herbal formulation.