Four antibiotics, oxytetracycline, tetracycline, doxycycline, and chlortetracycline were separated and quantified in medicated feed. Tetracyclines from feed samples were extracted with 0.01 M citric buffer and acetonitrile (pH 3.0) and further purified with 0.45 µm syringe filters. The purified extract was separated on commercial RP-C18 column and analyzing using liquid chromatograph (LC) with two different detectors: diode array detector (DAD) and mass spectrometry (MS). These methods provided average recoveries from 72.2 to 101.8% for high pressure liquid chromatography (HPLC) with diode array detection and from 45.6 to 87.0% for liquid chromatography with mass spectrometry. The limit of detection in medicated feed ranged from 4.2 to 10.7 mg kg−1 for HPLC–DAD and 5.6 to 10.8 mg kg−1 for LC–MS. Our experiment showed that using the same extraction mixture we do not obtain the same recovery values for the analyzed compounds using two different detection techniques such as DAD and MS.