Determination of doxorubicin and doxorubicinol in plasma of cancer patients by high-performance liquid chromatography.
A high-performance liquid chromatographic assay with fluorescence detection has been developed for the determination of doxorubicin and its metabolite doxorubicinol in plasma of cancer patients. Quantitative extraction was achieved by a single protein-precipitation step of 1-ml samples with 500 microl of acetone in the presence of 100 microl of zinc sulfate [70% (w/v) in water]. Doxorubicin and doxorubicinol were separated isocratically on a column packed with Inertsil ODS-80A material and a mobile phase composed of water:acetonitrile:tetrahydrofuran (76:24:0.5, v/v/v). The related compound daunorubicin was used as internal standard. The column effluent was monitored fluorimetrically at an excitation wavelength of 480 nm and an emission wavelength of 560 nm, with a band width of 40 nm. The calibration graphs of doxorubicin and doxorubicinol were linear over a range of 1.0 to 100 and 0.50 to 50. 0 ng/mL, respectively, with lower limits of quantitation of 1.0 and 0.50 ng/ml. Results obtained from a 4-day validation study demonstrated excellent accuracy (91.0-106%) and precision (0.90-10. 2%) across the calibration ranges for both compounds. The developed method has been applied extensively to a clinical study to examine the pharmacokinetics and metabolism of doxorubicin in patients cotreated with a potent inhibitor of MDR1 P-glycoprotein activity, GF120918.
de Bruijn
P
P
Department of Medical Oncology, Rotterdam Cancer Institute (Daniel den Hoed Kliniek), Rotterdam, 3008 AE, The Netherlands.
Verweij
J
J
Loos
W J
WJ
Kolker
H J
HJ
Planting
A S
AS
Nooter
K
K
Stoter
G
G
Sparreboom
A
A
D000970:Antineoplastic Agents / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000097:blood* / Q000378:metabolism / Q000493:pharmacokinetics; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000097:blood* / Q000378:metabolism; D012680:Sensitivity and Specificity; D013997:Time Factors
D000970:Antineoplastic Agents; D004317:Doxorubicin; C010013:adriamycinol