An herbal formulation composed of Allium cepa (Onion), Zingiber officinale (Ginger), Allium sativum (Garlic), Aloe vera, Andrographis paniculata (King of bitters), Allium ascalonicum (Shallot), reportedly used as anti-inflammatory agent was investigated. This study evaluated the physicochemical, phytochemical properties and in vivo anti-inflammatory effect of the herbal formulation. The herbal formulation (Hp-01) was subjected to physicochemical analysis; visual and organoleptic test, thin layer chromatography (TLC), pH, trace elements, phytochemical screening, antioxidant activity using DPPH inhibition and total phenolic acid (TPA) content determination. The biological effect at three doses (half, normal and double) on acute inflammation using carrageenan-induced paw oedema, weight, haematological parameters and liver functions were investigated in Wistar rats. The herbal formulation (Hp-01) was an acidic (pH 3.18), bitter preparation with pungent odour and sediments. The TLC analysis showed a maximum of three spots. Trace elements content of Pb, Cd, Fe, and Zn were within acceptable limits at 0.053, 0.003, 1.190 and 4.190 mg/l respectively. Phytochemical screening showed the presence of anthraquinones, tannins, terpenoids, saponins (2.12%w/w), alkaloids (2.45%w/w) and flavonoids (49.45%w/w). The IC 50 for DPPH radical scavenging activity was 2646.82±58.24 µg/ml indicating low activity, while TPA content was 0.0184±0.0005 mgGAE/g. The antiinflammatory activity of the herbal formulation was comparable acetylsalicylic acid (reference) at the three doses tested (p>0.05). Also, there was no significant effect on the weight, liver function, some haematological parameters; PCV, RBC, Hb. The herbal formulation has comparable anti-inflammatory activity with acetylsalicylic acid and may be explored for new anti-inflammatory agents.