High-performance liquid chromatography of anthracycline antibiotics with electrochemical detection. Application to the clinical pharmacokinetics of 4'-deoxydoxorubicin.
The high-performance liquid chromatography with electrochemical detection (HPLC-EC) of six anthracycline antibiotics is described. All the compounds of interest may be analysed using a reversed-phase column (ODS Hypersil or Zorbax ODS) and a mobile phase consisting of acetonitrile-isopropanol-0.1 M phosphate buffer (pH 4.5). The use of short columns (7.5 cm) allowed short analysis times and improved sensitivity, such that detection limits of between 1 and 2 ng ml(-1) could be achieved. Sample preparation involved an alkaline extraction into chloroform, followed by a back extraction into acid solution. Final clean-up of the samples is achieved by shaking the final acidic extract with an organic solvent. The precision of the assay for the compounds studied ranges between 1.22 and 6.46% and the accuracy ranges between 94.5 and 106.0%. The analytical methodology developed is applied to the clinical pharmacokinetics of 4'-deoxydoxorubicin. The correlation between plasma levels found by HPLC-EC and those found by HPLC with fluorescence detection is excellent (r(2) = 0.990).
Riley
C M
CM
Department of Pharmaceutics, College of Pharmacy, J. Hillis Miller Health Center (Box J-4), University of Florida, Gainesville, FL 32610, USA.
Runyan
A K
AK