Study on the browning mechanism of betel nut (Betel catechu L.) kernel

Taking betel nut (Betel catechu L.) kernel as raw materials, to analyze the browning mechanism of betel nut kernel. First, we extract and separate the phenolic substances and polyphenol oxidase (PPO) from the betel nut kernel and then find out how O₂ penetrates into the kernel, study the above 3 key factors of enzymatic browning so as to prove the possibility of enzymatic browning, and further study the browning products to clarify the mechanism of browning of betel nut kernel during storage process. The results showed that 11 kinds of phenolic compounds were isolated and identified from the betel nut kernel by liquid chromatography-mass spectrometry, among which chlorogenic acid was the highest, followed by dopamine and L-epicatechin, and other contents were relatively low. Meanwhile, PPO was also separated from the betel nut kernel by DEAE-Sepharose Fast Flow and Phenyl-Sepharose 6 Fast Flow column chromatography. On this basis, scanning electron microscopy showed that the damage of the betel nut tissue was aggravated with the prolonged storage time, the wax was gradually decomposed, and the stratum corneum of the peel is destroyed in a honeycomb-shaped; the lignification of the flesh was aggravated, and the interstitial space was increased; and the crack of the kernel membrane was also enlarged. These changes of structure contribute to increase gas exchange within and outside the organization, including the entry of O₂. Finally, the oxidation products generated from simulated reaction of chlorogenic acid, dopamine, and epicatechin with purified PPO under aerobic conditions in vitro were compared with the products extracted from naturally brown betel nut, and the same absorption spectra were found. Therefore, it indicates that the browning of betel nut kernel is an enzymatic browning caused by the reaction of phenolic substrates were oxidized by PPO.