Two-dimensional mass spectroscopy (2D MS) has become a revolutionary tool in the healthcare sector, offering unparalleled insights into the molecular composition of biological samples. This advanced analytical technique allows for the comprehensive analysis of complex mixtures by separating ions based on their mass-to-charge ratio in two dimensions, significantly enhancing resolution and sensitivity compared to traditional mass spectrometry. 2D MS is particularly valuable in healthcare for its ability to identify and quantify disease biomarkers, aiding in early diagnosis and monitoring of conditions like cancer, cardiovascular diseases, and neurological disorders. Furthermore, it plays a critical role in proteomics and metabolomics, enabling researchers to understand the intricate biochemical pathways involved in health and disease. The detailed molecular information provided by 2D MS supports personalized medicine by helping tailor treatments to individual patient profiles, thereby improving therapeutic outcomes and reducing adverse effects. This technology’s precision and depth of analysis make it an indispensable tool in modern medical research and diagnostics.