綜上所述，本論文展示了奈米酵素光催化抗氧化劑檢測平台由概念驗證逐步發展至兼具廣泛分析功能與生物應用價值之完整研究歷程。所建立之奈米酵素催化平台具有優異之穩定性、高靈敏度及廣泛適用性，不僅可作為抗氧化劑分析之有效工具，亦可為生物體氧化還原調控機制及相關疾病研究提供重要之分析策略與應用價值。;Reactive oxygen species (ROS)-mediated oxidative imbalance is closely associated with cellular damage and the progression of various diseases， making the sensitive and selective detection of antioxidants essential for biomedical research and diagnostic applications. However， accurate quantification of antioxidant levels， which is critical for monitoring oxidative stress， remains challenging due to limitations of existing analytical methods， including poor responsiveness toward thiol-containing antioxidants and restricted applicability in complex biological environments. In this context， nanozyme-based catalytic systems have emerged as promising alternatives to natural enzymes owing to their superior stability， tunable catalytic activity， and functional versatility. This thesis focuses on development and systematic evaluation of advanced nanozyme-mediated catalytic platforms for efficient antioxidant detection through photo-responsive catalytic oxidation of thiamine， with emphasis on improving analytical performance， expanding detection scope， and enabling biological and real-sample validation. In the first study， a light-activated DTT-assisted BSA-stabilized gold nanocluster (DTT@BSA-AuNCs) system was developed for antioxidant detection under neutral conditions. The platform enabled efficient detection of both thiol and non-thiol antioxidants and was further validated for intracellular antioxidant detection in cancer cells. Mechanistic investigations revealed a Type II photosensitization pathway， where singlet oxygen (¹O₂) served as the dominant reactive species responsible for catalytic oxidation. In the second study， the analytical scope and functional applicability of the system were further expanded using bimetallic gold–silver nanoclusters (AuAgNCs)， where Ag incorporation modulated the catalytic properties and enhanced catalytic efficiency. This platform enabled the detection of a wide range of physiologically relevant antioxidants and total antioxidant capacity (TAC) assessment in complex matrices， including simulated saliva， human saliva， and commercial antioxidant-containing products such as vitamin C tablets and fruit juices. Analytical reliability was confirmed through spike-and-recovery experiments in human saliva with near-quantitative recoveries， indicating minimal matrix effects， while strong correlation with the Ferric Reducing Antioxidant Power (FRAP) assay (R² = 0.99)， a conventional standard antioxidant assay， validated its quantitative robustness. In addition， intracellular applications were extended to evaluate oxidative stress alterations， antioxidant intervention， recovery processes， and steatosis-associated pathological conditions relevant to non-alcoholic fatty liver disease (NAFLD). Mechanistic analysis indicated a Type I photosensitization pathway， with peroxyl radicals (ROO•) identified as the dominant reactive species. Overall， this thesis demonstrates a progressive evolution from feasibility demonstration to broadened analytical functionality and biological relevance. The developed nanozyme-based platforms provide robust， sensitive， and versatile tools for antioxidant detection and offer valuable insights into redox regulation in biological and disease-relevant systems.
