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Unripe apples are discarded due to thinning out or falling in orchards, however they are abundant in functional ingredients. In this study, polyphenols were efficiently enzymatically extracted and isolated from unripe apples. Enzymatic extraction of polyphenols over 12 hours followed using Viscozyme L with an enzyme concentration of 2%(v/w) based on the total phenolic content. In order to isolate the polyphenols, extracts of unripe apples were eluted with water, 25, 50, 75 and 100% ethanol using Diaion HP-20 gel column chromatography. Polyphenol compounds of each fraction were analyzed by high performance liquid chromatography (HPLC), with the 50% ethanol fraction containing the most polyphenols (387.88 μg/mL of caffeic acid, 226.34 μg/mL of p-coumaric acid, and 187.33 μg/mL of phloridzin). The 50% ethanol fraction also contained the most antioxidants (total penolic and flavonoid content; p<0.05). Antioxidant assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity and ferric ion reducing antioxidant power (FRAP) activity exhibited the highest activities in the 50% ethanol fraction (p<0.05). Antimicrobial activities were also assessed as being highest in 50% ethanol fraction (p<0.05). The 50% ethanol fraction of unripe apple enzyme extracts can be used as a functional polyphenol material to improve the utilization of unripe apple.

Antioxidant and antimicrobial activities of polyphenols isolated from unripe apples (Malus pumila cv. Hongro)