3.5. Antiglycation Non-Enzymatic Assay (BSA-Methylglyoxal Model)
REGRESSION
For this purpose, 21.2 μM bovine serum albumin was incubated with 0.5 mM methylglyoxal and 1–1.5 mM of the test compound or peppermint leaf dry extract (1–3 mg/mL) in 100 mM PBS solution, pH 7.4, with 0.02% sodium azide. The reaction mixture was shaken (50 rpm) at 37 °C, for 7 days, in closed vials without light. The fluorescent intensity of MGO-mediated AGEs formed during incubation was analyzed using a Synergy HTX Multi-Mode Microplate Reader (BioTek Instruments Inc., Winooski, VT, USA) at a wavelength of 360 nm for excitation (λThe percentage contribution of each component to the activity of the peppermint leaf dry extract was calculated from regression equations for the relationship between the concentration [μM] and % inhibition. These were then related to the overall effect exerted by the extract.