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

✍ Scribed by Shuji Kato; Glen C. Post; Veronica M. Bierbaum; Tad H. Koch

• Publisher: Elsevier Science
• Year: 2002
• Tongue: English
• Weight: 89 KB
• Volume: 305
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2002.5682

✦ Synopsis

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 H 3 O ؉ . 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 M, R 2 ‫؍‬ 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 M for sensitive cells and 0.54 M for resistant cells. Treatment with a physiological concentration of doxorubicin (0.5 M) 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.