A new multi-residue method for the analysis of the tetracycline antibiotics oxytetracycline, tetracycline, chlortetracycline, and doxycycline was developed and validated for animal feed. After extraction with 0.1M Na2EDTA-McIlvaine buffer (pH 4), the samples were centrifuged, purified by SPE (Strata-X-CW cartridges) and analysed by HPLC-MS. Validation of the method was performed according to the guidelines indicated in European Commission Decision 2002/657/EC. The procedure was validated by spiking feed samples at three different levels (300, 1000, and 5000 µg/kg). Average recoveries for tetracyclines were in the range 79.70-98.8%, with RSD for repeatability and reproducibility in the ranges of 5.0-9.5% and 6.5-11.0%, respectively. The method was successfully validated and proved to be efficient, precise, and useful for quantification of tetracyclines in animal feed. The validated method was successfully applied to 65 suspect feed samples collected from different regions of Poland in 2015-16 and obtained from farms and feed manufactures. Of these 65 purportedly non-medicated feeds, eight (12.3%) were positive for the presence of doxycycline or chlortetracycline.