A Comparative Study of Phenolics in Green Husks of Selected Hungarian Walnut Cultivars

Green husks, which are the fleshy pericarp of Juglans regia L. fruit, are an abundant yet under-utilized source of bioactive compounds. They are useful for plant defense and have potential for valorization to multiple commercial products. This study characterized total phenolic content (TPC) and individual phenolics in green husks of four Hungarian-bred cultivars (Milotai 10, Milotai intenzív, Milotai kései, Esterhazy kései) and one U.S. cultivar (Chandler). Phenolic compounds were extracted with aqueous organic solvents, quantified by HPLC-DAD and qualitatively identified by HPLC-MS. Linear mixed-effects models were used to assess the effects of cultivar, year, sampling time, and cumulative growing degree days (GDDs) on TPC and compound profiles. Mean TPC ranged from 34.9 to 57.2 mg GAE g^-1 DW, with significantly higher values in the warmest year, 2024, and in cultivar Esterhazy kései compared with Chandler. Across cultivars and years, phenolic levels were generally elevated at early lignification (S1, BBCH 73-75) and at full maturity (S5-S6, BBCH 87-88), with depressed concentrations during mid-fruit development (S2-S4, BBCH 77-86). Several hydroxycinnamic acids, flavonoids, and naphthoquinones showed cultivar-specific and year-dependent patterns. Thermal conditions (cumulative GDDs) explained a substantial proportion of residual variation in TPC. These results highlight the combined roles of genotype, seasonal climate, and developmental stage dependencies in biosynthetic processes of phenolics in walnut green husks despite the diversity in factor effects.