2.3.3. Inhibition of Albumin Denaturation (IAD)

The anti-inflammatory efficacy was evaluated using an inhibition of albumin denaturation (IAD) in vitro study. Sakat method [14] with a modest modification [15] was used to finish the analysis.

Human albumin was used in the experiment. In distilled water, an albumin (1%) solution was prepared (pH 7.4). The tested compounds/standard were first dissolved in 1.2 mL DMF and PBS up to 25 mL, resulting in a stock solution with a final concentration of 1000 μg/mL. Then, in PBS, a series of working solutions with varying concentrations (20–500 μg/mL) were prepared. The reaction mixture contained 2 mL of different concentrations of test sample/standard and 1 mL of albumin (1%). The mixture was incubated at 37 ◦C for 15 min before being heated in a water bath at 70 ◦C for 15 min. After cooling, the turbidity was measured with a spectrophotometer at 660 nm (Camspec M508, Leeds, UK). The experiment was carried out three times. The inhibition of albumin denaturation (IAD) was calculated as a percentage of the control. The control sample was albumin dissolved in distilled water at the same concentration.
