This paper describes a systematic approach to the development of a solid-phase extraction method for simultaneous extraction of 10 antibiotic residues in bovine milk, belonging to groups of sulfonamides, tetracyclines, and pyrimethamine. The sample preparation steps include acidic deproteinization of milk proteins followed by sample enrichment and cleanup using a polymer-based Oasis HLB solid-phase extraction cartridge. The analyses were carried out by using a method based on liquid chromatography-electrospray ionisation-mass spectrometry with positive ion mode. The parameters affecting the extraction efficiency such as sample loading pH, SPE wash solvent composition, and eluting solution pH were carefully investigated and optimized. The developed solid-phase extraction procedure coupled to multiresidue liquid chromatography-electrospray ionization-mass spectrometry method was applied for the analysis of 10 antibiotic residues in milk samples, and it proved to be simple, sensitive, and selective providing a recovery ranging from 70 to 106%.