Analysis of doxorubicin in cell culture media and human plasma using solid phase extraction and HPLC

Summary

Doxorubicin is an antineoplastic antibiotic isolated fromStreptomyces peucetius var.cesius clinically used in the treatment of tumors such as lung or breast, Hodgkin's disease and various types of leukemias. The main goal of this study was to develop a simple and sensitive HPLC method with fluorescence detection for the quantitation of doxorubicin in cell culture media collected during an in vitro studies and in human plasma. Solid phase extraction (C₂ silica) was applied. The experiment established five-point standard curve (1 ng mL⁻¹ to 100 ng mL⁻¹). The standard curves prepared in blank cell tissue media were linear over the range of doxorubicin assayed and had a mean correlation coefficient of 0.9973±9.43×10⁻⁴ and slope 0.02545±1.85×10⁻³. The standard curves prepared in human plasma were linear and had mean correlation coefficient of 0.997 and slope 0.01885±5.19×10⁻⁴. The limit of quantitation for doxorubicin in both specimens was arbitrarily established to be 1 ng mL⁻¹. Intra-day variabilities were determined using 3–4 replicates of control solutions of doxorubicin (3 ng mL⁻¹ and 30 ng mL⁻¹) in blank plasma and cell culture media. Inter-day variabilities were determined over a four day period analyzing replicates of controls. All precision and accuracy values fell within the acceptable range.