In the past decade high-performance liquid chromatography (HPLC) has developed into the most powerful tool available for the analysis of anthracycline antitumour drugs in all kinds of biological matrices. The results of drug level monitoring studies have contributed significantly to a better understanding of the pharmacokinetics and metabolic disposition of these drugs. Some aspects of anthracycline HPLC analysis in plasma samples are discussed, including the chromatographic separation and detection, sampling procedures, storage of plasma samples and sample pre-treatment.