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Analysis of perillic acid in plasma by reversed-phase high-performance liquid chromatography with ultraviolet
E Ikechukwu Ezennia1, L R Phillips, T L Wolfe
1Drug Formulation Laboratory, National Cancer Institute, Frederick Cancer Research and Development Center, MD 21702-1201, USA.
Abstract:
A simple and sensitive isocratic high-performance liquid chromatographic (HPLC) method with UV detection for the quantitation of perillic acid, a major circulating metabolite of perillyl alcohol and d-limonene, in plasma is described. Sample preparation involved protein precipitation and subsequent transfer and dilution with 10 mM NaHCO3. The mobile phase consisted of acetonitrile (36%) and 0.05 M ammonium acetate buffer pH 5.0 (64%). Separations were achieved on a C18 column and the effluent monitored for UV absorption at the analytes' respective UVmax. Separation was excellent with no interference from endogenous plasma constituents. This method was found suitable for quantifying drug concentrations in the range of 0.25 to 200.0 micrograms/ml using a 0.05-ml plasma sample, and was used to study the plasma pharmacokinetics of perillic acid in mice.