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Abstract
A sensitive, rapid, and specific liquid chromatography/tandem mass spectrometry assay has been established and validated for the quantitation of evodiamine and evodine in Beagle dog plasma. Plasma samples of 0.2 ml were processed by liquid–liquid extraction with n-hexane/ethyl acetate (2:1, v/v). Chromatographic separations were done on a Symmetry C18 column (100 mm × 4.6 mm, ID, 5 μm) at 35°C with a linear gradient of methanol and 20 mM ammonium formate containing 0.2% formic acid. Evodiamine, evodine, and glibenclamide [internal standard (IS)] were ionized with an electrospray ionization source operated in positive ion mode. The MS/MS transitions were m/z 304.1 → 161.1 for evodiamine, m/z 471.2 → 425.1 for evodine, and m/z 494.1 → 369.1 for IS. Calibration curves were linear over the concentration range of 0.1–100 ng/ml for evodiamine and 0.5–500 ng/ml for evodine. The mean extraction recoveries were 88.10 ± 3.21% for evodiamine and 81.24 ± 4.07% for evodine. The intra- and inter-day precisions were less than 11.10% and 12.81%, and the accuracy was within ± 11.76% for both analytes. Evodiamine and evodine were stable during storage and analytical periods. The validated method has been successfully applied to a pharmacokinetic study of evodiamine and evodine in beagle dogs after oral administration.
Acknowledgements
This research was financially supported by National Basic Research Programs of China (Nos 2010CB933904, 2010CB735602, and 2012CB724002).