A rapid and accurate method for the determination of tetracycline in human plasma and urine is presented. Determination of tetracycline in plasma is based on precipitation of plasma proteins with trifluoroacetic acid, followed by injection of the centrifuged plasma sample onto a muBondapak C18 column. Acetonitrile in phosphate buffer pH 2.2 is used as mobile phase. Only tetracycline, and no trace of lumecycline can be detected in plasma and urine after administration of lumecycline, indicating that lumecycline is completely degraded to tetracycline, lysine and formaldehyde in the gastrointestinal tract prior to absorption. Determination of tetracycline in urine was performed by injection of urine diluted with phosphoric acid onto a muBondapak Phenyl column. The precision of determination of tetracycline in plasma, expressed as the relative standard deviation, was less than 3% at tetracycline concentrations of 0.05 and 3.7 micrograms/ml. Urine determinations were made with a precision of less than 1.5% at tetracycline concentrations of 0.5 and 6.7 micrograms/ml.