This paper provides an extensive overview of chromatography techniques utilized in the analysis of Alcaftadine, a pharmaceutical compound used in ophthalmic formulations. Various chromatographic methods including UV Spectrophotometric, HPLC, RP-HPLC, HPTLC, and UPLC have been developed and validated for the accurate quantification of Alcaftadine in bulk and dosage forms. The methods described offer specific advantages such as ease of use, precision, and sensitivity, ensuring reliable results in pharmaceutical analysis. Key parameters of method validation, including accuracy, precision, linearity, limit of detection, limit of quantitation, specificity, range, and robustness, are comprehensively discussed, highlighting the stringent criteria required for analytical method validation in accordance with regulatory guidelines. The importance of stability-indicating methods for assessing the stability of Alcaftadine formulations is emphasized, underscoring the need for robust analytical techniques to ensure product quality and efficacy. The review underscores the crucial role of chromatography in pharmaceutical research, providing valuable insights for researchers and practitioners involved in drug development and quality control.