Validation of an isocratic HPLC method to detect 2-fluoro-beta-alanine for the analysis of dihydropyrimidine dehydrogenase activity.
Kinta M Serve, Jennifer L Darnell, Jody K Takemoto, Neal M Davies, Margaret E Black
Index: J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci. 878(21) , 1889-92, (2010)
Full Text: HTML
Abstract
The efficacy of the chemotherapeutic drug 5'-fluorouracil is reduced by catabolism to 2'-fluoro-beta-alanine (FBAL), a three-step reaction in which dihydropyrimidine dehydrogenase (DPD) catalyzes the rate-limiting step. To study in vitro DPD activity, we developed and validated an isocratic, reverse-phase HPLC method to detect and quantify FBAL without using multiple columns or radiolabeled substrates. Pre-column derivatization of FBAL was performed using o-phthalaldehyde in the presence of two sulfur donors, ethanthiol or beta-mercaptoethanol, and the resulting products assayed. Calibration curves were linear over a range of 10-200 microg/ml and the method was successfully applied to the examination of DPD activity in cultured cells.Copyright 2010 Elsevier B.V. All rights reserved.
Related Compounds
Related Articles:
1999-01-01
[Am. J. Physiol. 276(1 Pt 1) , G206-10, (1999)]
Proton-decoupled 19F spectroscopy of 5-FU catabolites in human liver.
1997-03-01
[Magn. Reson. Med. 37(3) , 321-6, (1997)]
2011-04-15
[J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci. 879(13-14) , 915-20, (2011)]
1999-11-01
[Magn. Reson. Med. 42(5) , 936-43, (1999)]
2007-10-01
[Cancer Sci. 98(10) , 1604-8, (2007)]