Abstract
A reverse-phase, high-performance liquid chromatography method is described for the quantification of 1′-hydroxybufuralol formed enzymatically by the incubation of bufuralol with cDNA-expressed CYP2D6 or human liver microsomes. Analytical separation is achieved using a C18 column and a mobile phase consisting of 30% acetonitrile and 2 mM perchloric acid, with detection by fluorescence using an excitation wavelength of 252 nm and an emission wavelength of 302 nm. This method is applicable to enzymatic studies for determination of CYP2D6-catalyzed bufuralol 1′-hydroxylation activity.
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Acknowledgments
This work was supported in part by the Canadian Institutes of Health Research (grant MOP-42385) and by the National Institutes of Health (grant 5 P42 ES07381).
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Crespi, C.L., Chang, T.K.H., Waxman, D.J. (2006). CYP2D6-Dependent Bufuralol 1′-Hydroxylation Assayed by Reverse-Phase Ion-Pair High-Performance Liquid Chromatography With Fluorescence Detection. In: Phillips, I.R., Shephard, E.A. (eds) Cytochrome P450 Protocols. Methods in Molecular Biology, vol 320. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-998-2:121
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DOI: https://doi.org/10.1385/1-59259-998-2:121
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