Determination of amsacrine in human nucleated hematopoietic cells

A new method has been developed for the determination of amsacrine (AMSA) in human nucleated hematopoietic cells. In order to prevent efflux during the cell separation procedure, white blood cells (WBCs) were separated from red blood cells by dextran sedimentation, leaving the WBCs in their natural environment. After cell counting, pelletting the cell suspension and correcting for the admixture of supernatant, AMSA was extracted from the WBCs and determined by high-performance liquid chromatography. Linearity of extraction was observed up to 40.10(6) cells. The inter-assay variation was 4.7%. Plasma and cellular concentrations were measured in five patients at the end of a 3-h infusion of 100 mg/m2 AMSA. A pharmacokinetic study of plasma and cellular AMSA concentrations up to 19 h after infusion was carried out. AMSA concentrations in WBCs correlated well with the plasma levels (n = 20, r = 0.967) with an accumulation factor compared to the plasma concentration of 2.6-9.8 in the patients studied. The method described is useful for studying cellular pharmacokinetics of AMSA in man.