Citrus peel extract-incorporated gelatin/CMC films as an antiviral active packaging system against hepatitis A virus

Hepatitis A virus (HAV) remains a major foodborne hazard in fresh produce, where conventional decontamination methods often show limited efficacy because of matrix-associated shielding and restricted disinfectant penetration. Although biodegradable active packaging has emerged as a promising strategy, the antiviral application of citrus peel extract (CPE) in biodegradable films against HAV has not been investigated. This study developed CPE-incorporated gelatin/carboxymethyl cellulose (G/CMC) films and evaluated their antiviral efficacy against HAV in suspension, on food-contact surfaces, on bell peppers, and during storage. HPLC analysis identified hesperidin (167.63 mg/g) and gallic acid (136.20 mg/g) as the major bioactive compounds, and CPE was non-cytotoxic to FRhK-4 cells at concentrations of ≤2.5%. In suspension, CPE achieved complete HAV inactivation to the detection limit (1.5 log₁₀ PFU/mL) at ≥2% after 90 min. Maximum reductions of 2.46 and 2.20 log₁₀ PFU/mL were obtained on silicone rubber and stainless steel, respectively, whereas only ∼1.0 log₁₀ reduction was observed on bell peppers, indicating matrix-associated limitations. Incorporation of CPE into G/CMC films significantly enhanced antiviral activity, with the 2% film achieving a 2.51 log₁₀ PFU/mL reduction (p < 0.05). During storage, the films reduced HAV titers to 2.00 log₁₀ PFU/mL after 14 days at 4 °C and 2.15 log₁₀ PFU/mL after 7 days at 25 °C while preserving texture and minimizing color changes (ΔE* ≤ 1.01). These findings demonstrate that CPE-incorporated biodegradable G/CMC films provide a sustainable antiviral active packaging strategy for improving the virological safety and quality preservation of fresh produce.