Antibiotic residues in milk represent an important food safety and public health concern, particularly in India where dairy production is largely dominated by smallholder systems and characterized by variable climatic conditions and limited cold-chain infrastructure. Effective residue monitoring therefore requires analytical techniques that are both scientifically reliable and operationally feasible within diverse dairy supply chains. This study presents a comparative evaluation of four major analytical approaches used for detecting antibiotic residues in milk: microbial inhibition assays, immunoassays, biosensor-based platforms, and chromatographic techniques coupled with mass spectrometry. Published literature and validation studies were assessed using comparative criteria including sensitivity, specificity, reproducibility, analytical throughput, cost considerations, infrastructure requirements, and applicability within Indian dairy environments. The analysis indicates that LC–MS/MS provides the highest analytical sensitivity and specificity and is therefore most suitable for confirmatory regulatory testing. Microbial inhibition assays represent the most cost-efficient option for large-scale primary screening, while immunoassays enable rapid intermediate screening at milk collection points. Biosensor technologies demonstrate promising sensitivity and potential for decentralized field detection but remain limited by environmental stability and calibration challenges. The findings support the implementation of a tiered analytical framework integrating screening and confirmatory techniques for effective nationwide antibiotic residue surveillance in Indian dairy systems.