Antibiotics in biological materials, including tetracyclines, are determined usually be means of microbiological methods. Because of the low specificity and the time consuming procedure of the microbiological assays, we developed a fluorimetric method, which enables us to determine specifically the tetracyclines and to reduce the time for the assay down to 4 hours. Our method is based on the determination of tetracyclines published by Kohn (1961), which we modified for the determination of chlortetracycline in meat and bone. Tetracyclines are fluorescent compounds, whose quantum yields are very much enhanced in most cases by forming a complex with Ca2+ ions and by extracting it into an organic solvent. This transfer into acetic acid ethylester separates the chlortetracycline from interfering substances. The yield of chlortetracycline with the described method was about 80%, the sensitivity of the detection was about 0.5 ppm.