Development And Validation of Novel LC-MS Method For Quantification Of Donepezil From Human plasma

Donepezil is used worldwide for the treatment of Alzheimer’s disease, where it is used to increase cortical acetylcholine. We present a simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method developed and validated for the quantification of Donepezil in Human Plasma. Quetiapine was used as the internal standard. The simple and cost effective Liquid-Liquid Extraction method is applied for the extraction of Donepezil and Internal standard from Plasma. A rapid isocratic separation of Donepezil is achieved by a short C18 column using mobile phase of Acetonitrile and 1mM Ammonium acetate buffer (92:8 v/v, (%) ) at flow rate of 0.5 ml/min. The run time is 2.5 minutes suggests high throughput of the proposed method. The collision-induced transition m/z 380à 91 was used to analyze donepezil in selected reaction monitoring mode. The signal intensity of this transition was found to relate linearly with donepezil concentrations in plasma from 0.142 ng/ml. The lower limit of quantification of the LC-MS/MS method was 0.1ng/ml. The intra-and inter-day precisions were below 10.0% and the accuracy was between + 5.0% and 5.0%. The validated LC/MS/MS method was applied to pharmacokinetic study on healthy male volunteer who received single oral dose of 10 mg Donepezil hydrochloride. The maximum plasma concentration observed was 17.05ng/ml at 2.5 hours post dosing. This validated LCMS/MS is rapid, sensitive, specific and cost-effective method for determining donepezil in human plasma samples.