Clinical analysis for the anti-neoplastic agent 1,4-dihydroxy-5,8-bis((2-[(2-hydroxyethyl)amino]ethyl)-amino)9,10-anthracenedio ne dihydrochloride (NSC 301739) in plasma. Application of temperature control to provide selectivity in paired-ion high-performance liquid chromatography.

An analytical method is described which permits monitoring of plasma level of the anti-tumor agent 1,4-dihydroxy-5,8-bis((2-[(2-hydroxyethyl)amino]ethyl)amino)9,10-antracenedione dihydrochloride (DHAD) following its intravenous administration to cancer patients. The drug cannot be efficiently extracted from plasma into water-immiscible solvents, but is effectively separated from the biological matrix by retention on hydrophobic XAD-2 beads packed in a disposable glass cartridge. DHAD is subsequently selectively eluted from this column and then analyzed by reversed-phase partition chromatography with spectrophotometric detection of the analyte. Resolution of overlapping bands during high-performance liquid chromatographic separation was achieved by systematic optimization of mobile phase, ion-pairing agent and temperature. A possible explanation for the observed selectively provided by temperature adjustment is offered. Plasma levels in the range of 75--3000 ng of DHAD per ml (7.5--300 ng applied to the column) can be analyzed with a precision of less than +/- 10%. Total recovery of drug from plasma is ca. 95%.