Analysis of intracellular doxorubicin and its metabolites by ultra-high-performance liquid chromatography.
Doxorubicin, a highly effective anticancer drug, produces severe side effect such as cardiotoxicity, which is mainly caused by its metabolite, doxorubicinol. While in vitro studies by measuring cellular concentration of doxorubicin have been reported, there have been no reports on measuring cellular concentration of the metabolites. In this report, we developed a sensitive and high-throughput method for measuring cellular concentrations of doxorubicin and its metabolites by ultra-high-performance liquid chromatography. The method achieved more than 96% recovery of doxorubicin and its metabolites from cell homogenates. Using simple separation conditions, doxorubicin and its three main metabolites, and the internal standard, were separated within 3 min. The method has a limit of quantification of 17.4 pg (32.0 fmol) injected doxorubicin. This high sensitivity enables the detection and intracellular quantification of doxorubicin and its metabolite, doxorubicinol, in cell homogenates, and its use will facilitate studies of the relationship between doxorubicin pharmacokinetics and therapeutic outcome.
Sakai-Kato
Kumiko
K
Division of Drugs, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. kumikato@nihs.go.jp
Saito
Eiko
E
Ishikura
Keiko
K
Kawanishi
Toru
T
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004317:Doxorubicin / Q000031:analogs & derivatives / Q000032:analysis / Q000737:chemistry; D019073:HT29 Cells; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space; D016014:Linear Models; D012680:Sensitivity and Specificity
D004317:Doxorubicin