A Comprehensive Review of Analysis Strategies for 25-Hydroxyvitamin D₃: Mechanisms, Platforms, and Future Perspectives.

Vitamin D₃ is an essential fat-soluble vitamin for the human body. Its metabolite, 25-hydroxyvitamin D₃ (25(OH)D₃), serves as the primary biomarker to assess vitamin D levels. The monitoring of 25(OH)D₃ concentration is crucial for human health assessment. While traditional detection methods offer high sensitivity and accuracy, they are operationally complex and costly. This review systematically summarizes the most recent progress in 25(OH)D₃ detection technologies. Special attention is given to the recognition modes of 25(OH)D₃ by antibodies, nucleic acids, and molecularly imprinted recognition elements. Subsequently, the design strategies of diverse types of biosensors, including fluorescent, colorimetric, and electrochemical biosensors, are analyzed. Moreover, the development of portable devices, smartphone software, and flexible wearable devices for detection applications is also examined. Biosensing detection platforms are compared from the perspectives of target recognition, signal conversion, signal output, and application scenarios. Additionally, the potential of biosensor detection platforms in clinical diagnosis, health management, and community health surveillance is further investigated. Finally, the future trends of intelligent, portable, accurate, and home-use 25(OH)D₃ detection systems are delineated. This review offers a comprehensive reference for researchers developing next-generation 25(OH)D₃ diagnostic sensors and provides insights for the early prevention and treatment of vitamin D deficiency-related diseases.