Anti-cancer drugs have been developed in response to the global health burden of cancer. Monitoring their concentrations in biological fluids is essential for optimizing therapeutic efficacy, minimizing adverse effects, accurately adjusting dosage, and evaluating clinical outcomes. In parallel, the extensive use of chemotherapy agents has increased concern over the occurrence of anti-cancer drug residues in environmental matrices. Consequently, reliable and sensitive analytical approaches are required for the determination of these compounds in both biological and environmental samples. This review critically summarizes recent advances in the analysis of anti-cancer drugs, covering chromatographic, electrophoretic, electrochemical, and spectroscopic techniques. Particular emphasis is placed on the role of sample preparation, including conventional and modern extraction strategies, in improving analytical reliability and method performance. Comparative evaluation of analytical figures of merit, such as limits of detection and practical applicability, is presented to highlight current capabilities, methodological limitations, and emerging challenges. In addition, recent trends toward greener analytical strategies such as solvent minimization, micro- and miniaturized extraction techniques, and sustainable instrumental approaches are discussed in the context of therapeutic drug monitoring and environmental surveillance. Future perspectives integrating green chemistry principles, nanotechnology, and miniaturized analytical devices are also outlined.

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