<p>The emergence of non-invasive biomarker discovery represents a transformative paradigm in contemporary medical diagnostics. Traditional invasive sampling methods pose significant patient discomfort and procedural risks, necessitating innovative approaches that enable continuous, pain-free health monitoring. Skin, as the largest human organ, harbors complex biofluids including sweat, sebum, and interstitial fluid, which offer unprecedented opportunities for comprehensive molecular profiling with applications extending to systemic health monitoring. This comprehensive review aims to elucidate the current landscape of skin-derived biofluid analysis, exploring advanced collection methodologies, sophisticated analytical techniques, and potential clinical applications across diverse health conditions with emphasis on both dermatological and systemic health monitoring. Recent technological advancements have revolutionized skin biofluid analysis through multifaceted innovations. Cutting-edge wearable sensors, microneedle-based extraction techniques, and microfluidic devices now enable real-time, continuous biomarker monitoring. Sophisticated analytical platforms, including metabolomics, proteomics, transcriptomics, and lipidomics, facilitate unprecedented molecular characterization. Emerging applications span multiple medical domains, including metabolic disorders, cardiovascular diseases, neurological conditions, cancer detection, infectious diseases, and stress monitoring. Artificial intelligence and machine learning algorithms have dramatically enhanced data interpretation, enabling complex pattern recognition and predictive modeling. Multiplex biomarker panels combining metabolites, proteins, and nucleic acids promise enhanced diagnostic accuracy and personalized therapeutic strategies. Skin-derived biofluid analysis represents a rapidly evolving frontier with transformative potential in healthcare delivery. By offering minimally invasive, continuous health monitoring and enabling early disease detection, this approach promises to revolutionize personalized medicine, providing comprehensive molecular insights into both skin health and systemic physiological status with minimal patient intervention.</p>