<p>This work presents zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) methods employed in conjunction with ultraviolet-visible (UV-Vis) techniques to analyze and quantify three anticancer medications, namely 6-mercaptopurine (6-MP), pemetrexed (PMX), and capecitabine (CAP), in both their bulk forms and pharmaceutical preparations. Two ZIC-HILIC stationary phases were created by incorporating sulfobetaine monomers into polystyrene-divinylbenzene particles. The techniques were formulated by analyzing the impact of HILIC mode variables such as pH, acetonitrile concentration, and acetate buffer concentration in the eluent. To examine the retention characteristics of anticancer medications, two ZIC-HILIC columns are employed. The current methodology has demonstrated an outstanding linear range (0.12–20, 0.12–18, 0.12–8 μg mL^-1) for detecting target anticancer medications (6-MP, PMX, and CAP). The proposed techniques exhibited excellent precision (relative standard deviation &lt; 1.01), strong linearity (R² ≥ 0.9985), and high accuracy (ranging from 98.43% to 101.25%) for the detection and quantification of all target pharmaceuticals. The limits of detection by using ZIC1 were 0.0092, 0.0722, and 0.0534 μg mL^-1, whereas by using ZIC5 they were 0.0063, 0.0503, and 0.0365 μg mL^-1 for 6-MP, PMX, and CAP respectively. Statistical analysis was employed to examine the methodologies utilized in the British Pharmacopoeia protocol, and no significant disparity in precision was observed.</p>