Assay for determination of daunorubicin in cancer cells with multidrug resistance phenotype
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
21641880
DOI
10.1016/j.jchromb.2011.05.008
PII: S1570-0232(11)00313-8
Knihovny.cz E-resources
- MeSH
- K562 Cells MeSH
- Drug Resistance, Neoplasm MeSH
- Leukemia, Myelogenous, Chronic, BCR-ABL Positive drug therapy metabolism MeSH
- Daunorubicin analysis pharmacokinetics MeSH
- DNA, Neoplasm analysis MeSH
- Fluorescence MeSH
- Intracellular Space chemistry metabolism MeSH
- Humans MeSH
- Linear Models MeSH
- Drug Resistance, Multiple MeSH
- Antibiotics, Antineoplastic analysis pharmacokinetics MeSH
- Flow Cytometry MeSH
- Sensitivity and Specificity MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Daunorubicin MeSH
- DNA, Neoplasm MeSH
- Antibiotics, Antineoplastic MeSH
A sensitive assay for direct determination of intracellular level of daunorubicin (DRN) in resistant leukemia cells with overexpressed P-glycoprotein has been developed. This assay is based on a rapid separation of cells from media and fast cut-off of DRN transportation by centrifugation of cells through a layer of silicone oil. Cell pellets were extracted using 1% (v/v) formic acid in 50% (v/v) ethanol in water. The cell extracts were subsequently analysed by liquid chromatography (HPLC) coupled a low-energy collision tandem mass spectrometer equipped with an electrospray ionization source (ESI-CID-MS/MS) operated in the multiple-reaction monitoring (MRM) mode. Calibration curve was linear from 0.4 to 250nM with correlation coefficient (r²) better than 0.998. The limit of quantitation (LOQ) was 0.4 nM. The assay has been successfully applied to a determination of intracellular content of daunorubicin in sensitive K562 and resistant K562/Dox and K562/HHT300 cells.
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