In vitro and in vivo inhibition of Hass avocado polyphenol oxidase enzymatic browning by paeonol, beta-cyclodextrin, and paeonol:beta-cyclodextrin inclusion complex

Abstract

Polyphenol oxidase (PPO) was extracted from Hass avocados and its physicochemical properties were analyzed. The optimum pH and temperature of the enzyme were pH 7.5 and 20°C. This PPO showed a high thermal stability, since 26% of the initial activity was retained by the enzyme after heating at 60°C for 40 min. Inhibition studies were performed using different chemical reagents, and the order in the inhibition efficiency was paeonol > 4-hydroxybenzaldehyde > β-cyclodextrin (β-CD). The first two inhibitors presented a non-competitive mechanism while the inhibition by β-CD results from a mixed type mechanism. Since the aqueous solubility of paeonol (a natural compound) is very low, the inclusion complex between this drug and β-CD was obtained in solution and solid state. The stoichiometry of the paeonol:β-CD complex was 1:1 and its ΔG° of formation was −26 kJ/mol. The complexation of paeonol by β-CD not only enhances the aqueous solubility and thermal stability of the drug, but also improves the in vitro inhibition efficiency against PPO. Colorimetric analysis on avocados pulp (in vivo) showed that the inclusion complex does not increase the inhibitory effect of paeonol, remaining practically unchanged. However, the formulation of paeonol:β-CD inclusion complex allows employing this compound as PPO inhibitor in aqueous solutions.