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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 12, pp 3425–3435 | Cite as

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

  • Juliette Gautier
  • Emilie Munnier
  • Martin Soucé
  • Igor Chourpa
  • Laurence Douziech EyrollesEmail author
Research Paper

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.

Keywords

Doxorubicin Nanovectors Capillary electrophoresis Cancer 

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

Notes

Acknowledgments

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

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Juliette Gautier
    • 1
  • Emilie Munnier
    • 1
  • Martin Soucé
    • 1
  • Igor Chourpa
    • 1
  • Laurence Douziech Eyrolles
    • 1
    Email author
  1. 1.Universite Francois-Rabelais de Tours, EA6295 “Nanomedicaments et Nanosondes”ToursFrance

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