Speciation of arsenic contaminants plays an important role in environmental fate, risk assessment, and engineering management. It is essential to identify the arsenic speciation accurately. Synchrotron-based X-ray absorption spectroscopy (XAS), including X-ray Absorption Near Edge Structure (XANES) and Extended X-ray Absorption Fine Structure (EXAFS), provides detailed insights into arsenic oxidation states and local coordination environments. This review summarizes recent advances in the application of XAS to characterize both inorganic and organic arsenic species in environmental and biological systems. XANES was commonly used to distinguish arsenic oxidation states and identify complex arsenic-bearing minerals and organoarsenic compounds such as arsenopyrite (FeAsS) and dimethylarsinic acid (DMA). EXAFS provides quantitative structural parameters, bond lengths, coordination numbers, and adsorption configurations. The geometric structure includes binuclear bidentate, corner-sharing (2C), mononuclear monodentate, corner-sharing (1 V), and binuclear bidentate, edge-sharing (2E) configurations. Applications of XAS to metal oxides, natural minerals, and metal–organic frameworks (MOFs) have revealed how surface chemistry and functional group modifications influence arsenic binding mechanisms. In biological contexts, XAS also identifies arsenic species in tissues and biomolecules. XANES and EXAFS enable comprehensive arsenic speciation analysis, supporting mechanistic understanding of arsenic transformation, mobility, and stability. This review highlights the critical role of XAS in arsenic speciation analysis and its potential in advancing environmental monitoring and remediation strategies.