Poly(ethylene terephthalate)‐based molecularly imprinted polymers (MIPs) were synthesized, and their recognition capability was evaluated. Adsorption isotherm was described by the Langmuir model and the maximum adsorption capacity of MIPTy reached 172.4 mg g−1 in water at pH 6.2. A recognition coefficient of 1.17 was obtained. A solid‐phase extraction cartridge was manufactured and its behavior was evaluated for tylosin extraction from aqueous and milk samples. An off‐line SPE‐UV method was applied. An acceptable linearity was obtained in the range of 1–20 μg ml−1 and the average recovery at three spike levels in milk samples was higher than 92%. The limit of quantification was 2.6 × 10−2 μg ml−1. The manufactured SPE cartridge has a great potential for clean‐up processes in complex media. The cartridge offers a fast and sensitive option to the existing sorbents for extracting this drug from milk samples.
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