Determination of Protoapigenone in Beagle Dog Plasma by LC–MS/MS: Application to a Pharmacokinetic Study

Protoapigenone, a unique class of natural flavonoid, exerted significant cytotoxic and antitumoral activities. A simple, sensitive, and rapid LC–MS/MS method for the quantification of protoapigenone in plasma was developed and validated to obtain the pharmacokinetic profile of protoapigenone in beagle dogs. Ethyl acetate-10% ammonium hydroxide (2:1, v/v) was employed for liquid–liquid extraction (recovery > 90%). Under optimal conditions, chromatographic resolution was achieved on a C 18 column using acetonitrile and 10 mM ammonium formate in water (9:1, v/v), as mobile phase at a flow rate of 0.5 ml/min. The established method presented good linearity, selectivity, accuracy, and precision in the range of 0.2–100 ng/ml. Finally, this method was successfully applied to a pharmacokinetic study in beagle dogs following oral administration of protoapigenone. Graphical abstract