Determination of donepezil in human plasma using ultra performance liquid chromatography-tandem mass spectrometry

Abstract

An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the quantification of donepezil in human plasma. Donepezil and donepezil-D4 were extracted from human plasma by liquid-liquid extraction using a mixture of hexane and ethyl acetate (70:30 v/v). The extracted samples were analyzed using a Thermo Hypersil Gold C18 column with 5% acetic acid in 20 mM ammonium acetate buffer (pH 3.3) and 100% acetonitrile as a mobile phase with the 60:40 (v:v) isocratic method, at a flow rate of 0.3 mL/min. The injection volume was 3 μL, and the total run time was 3 min. Inter-and intra-batch accuracies ranged from 98.0% to 110.0%, and the precision was below 8%. The developed method was successfully applied to the quantification of donepezil in human plasma. The mean (standard deviation) maximum concentration and the median (range) time to maximum concentration were 8.6 (2.0) ng/mL and 2.0 h (1.0~5.0 h), respectively, in healthy Koreans after oral administration of 5 mg donepezil.

Ultrafast liquid chromatography-tandem mass spectrometry determination of donepezil in human plasma: application to a bioequivalence study

“…The objective of this study was to develop a high-efficiency liquid chromatography with tandem mass spectrometry (LC-MS/MS) method to support a comparative pharmacokinetics of the test and reference formulations of donepezil, an acetylcholinesterase (AChE) inhibitor as a selective treatment for Alzheimer’s disease [ 1 ], in healthy volunteers. In previous literature, high-performance liquid chromatography (HPLC) hyphenated with tandem mass spectrometry (MS/MS) methods have become the method of choice for donepezil quantitation in various biological fluids due to its inherent selectivity and high sensitivity [ 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 ]. However, for the determination of donepezil in human plasma, these methods conventionally demanded lengthy sample preparation procedures such as solid-phase extraction (SPE) [ 8 14 ] or liquid extraction [ 6 7 10 12 17 ] followed by drying the final extract at an elevated temperature and reconstitution steps before injection into LC-MS/MS systems to avoid ionization suppression, the requirement for longer chromatographic run times (≥ 4 minutes run time) to resolve interference by endogenous constituents and/or for a longer lifespan of chromatographic columns.…”