Quantitative subcellular study of doxorubicin in MCF-7/Adr cells using liquid chromatography-tandem mass spectrometry.
J Chromatogr B Analyt Technol Biomed Life Sci, 2015/12/15;1007:18-22.
Ma W[1], Wang J[2], Guo Q[1], Tu P[3]
Affiliations
PMID: 26562803
Impact factor: 3.318
Abstract
A rapid, sensitive and selective high-performance liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed and validated for the determination of doxorubicin in intracellular compartments using glibenclamide as internal standard (IS). MCF-7/Adr cancer cells (1×10(6)) were incubated with doxorubicin (8μg/mL) for 0.5, 1, 2 and 4h and then subjected to sequential extraction of cytosolic, membrane/organelle, nuclear and cytoskeleton soluble protein. Samples were extracted using protein precipitation with methanol. Chromatographic separation was carried out on a C18 column with acetonitrile and 0.1% formic acid water as mobile phase and with gradient elution at a flow rate of 0.2mL/min. The method was linear over the range of 1-300ng/mL with a lower limit of quantification (LLOQ) of 1ng/mL. The distribution of doxorubicin in subcellular components of MCF-7/Adr cancer cells was mainly in nucleic protein fraction.
Keywords: Doxorubicin; Intracellular distribution; LC–MS/MS; MCF-7/Adr cells
MeSH terms
Antibiotics, Antineoplastic; Chromatography, Liquid; Doxorubicin; Humans; Limit of Detection; MCF-7 Cells; Reference Standards; Subcellular Fractions; Tandem Mass Spectrometry
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