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Analysis of doxorubicin distribution in MCF-7 cells treated with drug-loaded nanoparticles by combination of two fluorescence-based techniques, confocal spectral imaging and capillary electrophoresis

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Abstract

The intracellular distribution of the antiancer drug doxorubicin (DOX) was followed qualitatively by fluorescence confocal spectral imaging (FCSI) and quantitatively by capillary electrophoresis (CE). FCSI permits the localization of the major fluorescent species in cell compartments, with spectral shifts indicating the polarity of the respective environment. However, distinction between drug and metabolites by FCSI is difficult due to their similar fluorochromes, and direct quantification of their fluorescence is complicated by quantum yield variation between different subcellular environments. On the other hand, capillary electrophoresis with fluorescence detection (CE-LIF) is a quantitative method capable of separating doxorubicin and its metabolites. In this paper, we propose a method for determining drug and metabolite concentration in enriched nuclear and cytosolic fractions of cancer cells by CE-LIF, and we compare these data with those of FCSI. Significant differences in the subcellular distribution of DOX are observed between the drug administered as a molecular solution or as a suspension of drug-loaded iron oxide nanoparticles coated with polyethylene glycol. Comparative analysis of the CE-LIF vs FCSI data may lead to a tentative calibration of this latter method in terms of DOX fluorescence quantum yields in the nucleus and more or less polar regions of the cytosol.

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Abbreviations

7-DeoxyDOXone:

7-Deoxydoxorubicinone

CE:

Capillary electrophoresis

CE-LIF:

Capillary electrophoresis with laser-induced fluorescence detection

DLPS:

Doxorubicin-loaded PEGylated SPION

DOX:

Doxorubicin

DOXol:

Doxorubicinol

DOXone:

Doxorubicinone

EOF:

Electroosmotic flow

FCSI:

Fluorescence confocal spectral imaging

MEKC:

Micellar electrokinetic chromatography

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Acknowledgments

This study was supported by grants from ARC (Association pour la Recherche contre le Cancer).

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Authors and Affiliations

  1. Universite Francois-Rabelais de Tours, EA6295 “Nanomedicaments et Nanosondes”, 31 Avenue Monge, 37200, Tours, France

    Juliette Gautier, Emilie Munnier, Martin Soucé, Igor Chourpa & Laurence Douziech Eyrolles

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  1. Juliette Gautier
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Correspondence to Laurence Douziech Eyrolles.

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Gautier, J., Munnier, E., Soucé, M. et al. Analysis of doxorubicin distribution in MCF-7 cells treated with drug-loaded nanoparticles by combination of two fluorescence-based techniques, confocal spectral imaging and capillary electrophoresis. Anal Bioanal Chem 407, 3425–3435 (2015). https://doi.org/10.1007/s00216-015-8566-9

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