The isolation of valuable compounds from the byproducts of food processing industries presents a significant contemporary challenge. Thin film nanocomposite (TFN) membranes have been identified as potential media for separating bioactive compounds. In this work, pentahomoserine functionalized graphene oxide (FGO) based TFN membranes were prepared for the effective separation of tea polyphenols epigallocatechin (EGC) and epigallocatechin gallate (EGCG). The nanocomposite selective polyamide (PA) layer was prepared through the dispersion of FGO in an aqueous monomer solution. The prepared membranes possess high average surface roughness with improved hydrophilicity and thermal stability. Additionally, FGO membranes contain more hydroxyl or carboxyl groups with cross-linked PA layer through interfacial polymerization (IP) process. Moreover, FGO membranes featured increased hydroxyl or carboxyl groups, facilitating cross-linking with the PA layer via intermolecular forces. The effects of amine functionality on the membrane characteristics such as surface morphology, roughness and hydrophilicity were also investigated. Remarkably, the membranes demonstrated rejection efficiency of 96.3% towards polyphenols. Furthermore, comprehensive assessments of reusability performance of the membrane highlighted their potential for industrial applications. This study provides valuable insights into the fabrication of highly efficient TFN membranes for biomolecule separation across diverse technological domains.