2.7. COX Inhibition Assay

This test was performed as per the Viji and Helen assay. According to the given protocol, a mixture was prepared using tris-HCl buffer enzyme, haemoglobin, glutathione, arachidonic acid and Trichloroacetic acid (TCA; 10% in 1N HCl, 0.2 mL). This mixture was then incubated for approximately 20 min at 37 °C. Thereafter, 0.2 mL of thiobarbituric acid (TBA reagent) was added to the mixture, which was subsequently kept on boiling water for 20 min. Afterward, it was cooled and centrifuged at 1000 rpm for 3 min. The final supernatant was used for the measurement of COX activity at 560 nm [27].

T is the absorbance of the inhibitor well (at 560 nm) and C denotes the initial absorbance activity without the inhibitor well (at 560 nm).