Effect of Concentration and Heat Treatment on Anti-inflammatory and Total Antioxidant Capacity of Azadirachta indica, Persea americana, and Mangifera indica Leaves Polyherbal Extract

The traditional methods of preparing polyherbal extracts can induce changes in the phytochemical composition and alter biological activities. This study investigated the effect of heat treatment and concentration on the antioxidant and anti-inflammatory properties of polyherbal extracts derived from Azadirachta indica, Mangifera indica, and Persea americana leaves. The extracts were categorized as heated or fresh and the effect of heat treatment (5, 10, 15, and 20 minutes at 100°C) and concentrations (100–1000 µg/ml) were evaluated in vitro by measuring the inhibition of albumin denaturation, proteinase inhibitory activity, total antioxidant capacity and inhibition of heat-induced haemolysis using standard spectrophotometric procedures. The activities of the extracts were affected by the duration of heat treatment and concentration. The capacity of the extracts to inhibit albumin denaturation and proteinase activity followed a similar trend for both fresh and heated extracts in terms of concentration and duration of heat treatment. Whereas the TAC of the fresh extract increased to 74.06 ± 2.79 mgAAE/g, the heated extract decreased significantly to 21.90 ± 0.70 mgAAE/g. Inhibition of heat-induced haemolysis also reduced significantly from 51.27 ± 2.74% to 39.69 ± 0.87% after 20 minutes. The total antioxidant capacity indicates an optimum range of heat treatment between 5 and 40 minutes at 100°C. These findings underscore the importance of processing conditions and concentration in the optimization of the therapeutic efficacy of this polyherbal formulation..