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Chemical Ionization Mass Spectrometric Determination of Acrolein in Human Breast Cancer Cells

Authors
Journal
Analytical Biochemistry
0003-2697
Publisher
Elsevier
Publication Date
Volume
305
Issue
2
Identifiers
DOI: 10.1006/abio.2002.5682
Keywords
  • Chemical Ionization Mass Spectrometry
  • Acrolein
  • Breast Cancer Cells
  • Doxorubicin
  • Drug Resistance
Disciplines
  • Biology
  • Chemistry

Abstract

Abstract A selected ion flow tube–chemical ionization mass spectrometric method is presented for the first determination of acrolein metabolically produced in biological tissues. Acrolein in aqueous samples (2.5 ml) is preconcentrated by distillation and directly analyzed using gas-phase proton transfer from H3O+. This method provides sensitive detection of acrolein with the method detection limit of 15 nM at the 99% confidence level. Detection is linear up to the highest concentration studied (13.5 μM, R2 = 0.998). Acrolein levels are determined in doxorubicin-sensitive (MCF-7) and doxorubicin-resistant (MCF-7/Adr) human breast cancer cells in vitro. The intracellular acrolein concentrations differ insignificantly: 0.61 μM for sensitive cells and 0.54 μM for resistant cells. Treatment with a physiological concentration of doxorubicin (0.5 μM) for 24 h at 37°C increased acrolein levels by factors of 2.6 and 1.9 for MCF-7 and MCF-7/Adr cells, respectively. The differential enhancement observed is consistent with the lower levels of enzymes that neutralize oxidative stress in sensitive MCF-7 cells and overexpression of an active drug efflux pump P-170 glycoprotein in resistant MCF-7/Adr cells.

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