The determination of antibiotics in biological samples is compromised by their interaction with proteins. It is showed here that the water resistant zeolite imidazolate framework 67 (ZIF-67) represents a useful sorbent for solid-phase extraction (SPE) of fluoroquinolones (FQs) from biological samples. ZIF-67 is unique in possessing permanent nanoscale porosity and a high surface area. As a result, it efficiently excludes proteins from the inner network but can well adsorb small organic molecules. Its extraction capacity for FQs exceeds that of isostructural ZIF-8, probably due to high affinity for Co(II) ions in ZIF-67. The findings were exploited to design a method for the analysis of FQs, which were extracted from bio-samples by ZIF-67 via SPE. Following elution of the absorbed FQs with NaOH/methanol, they were quantified by HPLC with diode array detection. The assay has a wide linear range that extends from 3.9 to 4000 μg⋅kg‾ 1 , with detection limits between 1.2 and 2.9 μg⋅kg‾ 1 . The method was successfully applied to the determination of FQs in manure.