HPLC analysis of mitoxantrone in mouse plasma and tissues: Application in a pharmacokinetic study

A simple, fast and economical HPLC assay for the determination of mitoxantrone in mouse plasma and tissue homogenates is described. Protein precipitation with sequential addition of sulfosalicylic acid and acetonitrile was used for sample preparation. The resolution of mitoxantrone and the I.S. were achieved by using acetonitrile and 10 mM sodium phosphate buffer with 0.1% TEA. The separation was performed on a Nucleosil C18, 250 mm × 4 mm I.D. column with UV detection at 610 nm. The inter-day and intra-day precision and accuracy of quality control (QC) samples, evaluated both in plasma and tissue homogenates, were all within 15%. The lower limit of quantification (LLOQ) was 5 ng/ml in plasma, 25 ng/ml in liver homogenate and 12.5 ng/ml in other tissue homogenates. This assay was successfully applied in a pharmacokinetic and tissue distribution study of mitoxantrone in mice.