Tetracycline and analogues are among the most used antibiotics in the dairy industry. Besides the therapeutic uses, tetracyclines are often incorporated into livestock feed as growth promoters. A considerable amount of antibiotics is released unaltered through milk from dairy animals. The presence of antibiotic residues in milk and their subsequent consumption can lead to potential health impacts, including cancer, hypersensitivity reactions, and the development of antibiotic resistance. Thus, it is important to monitor residual levels of tetracyclines in milk. The purpose of this study is to develop a quick and simple method for simultaneously extracting five tetracycline analogues from bovine milk. Specifically, five tetracycline analogues: Chlortetracycline (CTC), demeclocycline (DEM), doxycycline (DC), minocycline (MC), and tetracycline (TC) were simultaneously extracted from milk using trifluoroacetic acid. Subsequently, the extracted analogues were separated by reverse-phase high-performance liquid chromatography (RP-HPLC) and detected at 355 nm using UV/Vis. Calibration curves for all five tetracycline analogues show excellent linearity (r2 value > 0.99). Percent recovery for MC, TC, DEM, CTC, and DC were: 31.88%, 96.91%, 151.29, 99.20%, and 85.58% respectively. The developed extraction method has good precision (RSD < 9.9% for 4 of the 5 analogues). The developed method with minimal sample preparation and pretreatment has the potential to serve as an initial screening test.
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