info:eu-repo/semantics/article
Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
Fecha
2018-12Registro en:
Micheloni, Oscar Bernardo; Farroni, Abel Eduardo; García, Paula; Furlan, Ricardo Luis Eugenio; Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization; Elsevier; Food Chemistry; 269; 12-2018; 638-643
0308-8146
CONICET Digital
CONICET
Autor
Micheloni, Oscar Bernardo
Farroni, Abel Eduardo
García, Paula
Furlan, Ricardo Luis Eugenio
Resumen
This work describes a TLC-coupled bioautographic assay suitable for the separation and detection of apple polyphenol oxidase (PPO) inhibitors from natural extracts. PPO was immobilised in agar containing L-DOPA as substrate and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) to enhance colour development. The inhibition was detected as white spots on reddish background. Minimum amount of PPO inhibitors detected was 0.0125 µg of 4-hexylresorcinol, 0.025 µg of ascorbic acid, 0.5 µg of cysteine and 1 µg of kojic acid. The assay was compatible with normal and reverse phase TLC systems and allows detecting compounds that directly had action on the enzyme as well as agents that could convert quinones back to their reduced form. The chromatographic run evidenced the different nature of enzymatic browning inhibitory compounds from garlic and onion extracts. Using natural enzymes will provide a fast and cheap alternative for target specific exploration of natural enzymatic inhibitors.