Structure Elucidation and Evaluation of Antioxidant and Tyrosinase Inhibitory Effect and Mechanism of Proanthocyanidins from Leaf and Fruit of Leucaena Leucocephala

Abstract

This study aimed to investigate the structural features, antioxidant activity, tyrosinase inhibitory effect, and mechanism of proanthocyanidins from leaf and fruit of Leucaena leucocephala. MALDI-TOF-MS, thiolysis coupled with HPLC-ESI-MS, and 13C-NMR confirmed that these proanthocyanidins were complex mixtures of propelargonidins, procyanidins, and prodelphinidins. (Epi)catechin, (epi)gallocatechin, (epi)catechin gallate, and (epi)gallocatechin gallate were the main constitutive units. The findings obtained from enzyme analysis revealed that the proanthocyanidins had inhibitory effects on both monophenolase and diphenolase of tyrosinase. The IC50 values of leaf and fruit proanthocyanidins were 73.5 ± 2.7 and 52.3 ± 3.5 µg/mL for monophenolase activity, and 27.2 ± 1.4 and 16.1 ± 1.1 µg/mL for diphenolase activity, respectively. The inhibition of diphenolase by proanthocyanidins were proved to be reversible and mixed type. In addition, it was also found from various antioxidant methods that the proanthocyanidins in leaf and fruit possessed potent DPPH radical scavenging activity with IC50 values of 103.4 ± 0.8 and 87.8 ± 1.1 µg/mL, ABTS radical scavenging activity with IC50 values of 72.9 ± 0.4 and 65.7 ± 0.9 µg/mL, and ferric reducing antioxidant power with FRAP values of 3.74 ± 0.03 and 4.02 ± 0.15 mmol AAE/g, respectively. The results obtained suggested that the proanthocyanidins from leaf and fruit of L. leucocephala might have the potential to be exploited as natural tyrosinase inhibitors and antioxidants.