A rapid and sensitive LC‐MS/MS method for quantification of donepezil and its active metabolite, 6‐o‐desmethyl donepezil in human plasma and its pharmacokinetic application

Abstract

A rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay method has been developed and fully validated for simultaneous quantification of donepezil and its active metabolite, 6-o-desmethyl donepezil in human plasma. Analytes and the internal standard were extracted from human plasma by liquid-liquid extraction technique using a 30:70 v/v mixture of ethyl acetate and n-hexane. The reconstituted samples were chromatographed on a C(18) column by using a 70:30 v/v mixture of acetonitrile and ammonium formate (5 mm, pH 5.0) as the mobile phase at a flow rate of 0.6 mL/min. The calibration curve obtained was linear (r ≥ 0.99) over the concentration range of 0.09-24.2 ng/mL for donepezil and 0.03-8.13 ng/mL for 6-o-desmethyl donepezil. The results of the intra-day and inter-day precision and accuracy studies were well within the acceptable limits. The proposed method was successfully applied for the estimation of the drug in real time plasma samples for pharmacokinetic studies.

ESI‐MS/MS stability‐indicating bioanalytical method development and validation for simultaneous estimation of donepezil, 5‐desmethyl donepezil and 6‐desmethyl donepezil in human plasma

“…This investigation for the first time presented the stability effect when all drug and dealkylated metabolites were spiked together. Moreover the lower C max value for donepezil reported only in Indian volunteers (Patel et al ., ; Pilli et al ., ) could be due to loss or degradation of donepezil during blood sample collection as room temperature might be conducive to activating plasma esterase enzyme, causing degradation.…”

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

“…The current method showed reliable results with a liquidliquid extraction (LLE) preparation method and a C18 column with a relatively short run time of 3 min as well as a small injection volume of 3 μL compared to previous studies (Table 5). [13][14][15][16] First, as a comparison of the sample preparation method, Shah, H. J. et al used solid-phase extraction (SPE) for sample preparation. Although SPE in known to be highly selective compared to liquid-liquid extraction (LLE) or protein precipitation (PP), it is considered to be difficult for handling, and it has a higher associated cost.…”

Review of the validated HPLC and LC‐MS/MS methods for determination of drugs used in clinical practice for Alzheimer's disease

“…have been used by several researchers for chromatographic separation of donepezil. In quantitation of donepezil most of the researchers have used C 18 as a stationary phase both on HPLC (Yasui‐Furukori et al ., ; Abonassif et al ., ) and LC‐MS/MS (Lu et al ., ; Thevis et al ., ; Xie et al ., ; Apostolou et al ., ; Patel et al ., ; Kim et al ., ; Pilli et al ., ; Khuroo et al ., ; Noetzli et al ., ). Other columns reported in the literature are phenyl (Koeber et al ., ), RP‐5 (Nakashima et al ., ), C 8 (Shah et al ., ) and HILIC (Park et al ., ; Meier‐Davis et al ., ).…”

“…Several approaches have been published for direct liquid–liquid extraction of donepezil from plasma samples using ACN (Radwan et al ., ), EtOAc– n ‐hexane (Pilli et al ., ), isopropyl alcohol (IPA)– n ‐hexane (Matsui et al ., ) or extracting from plasma after bringing pH of the plasma to basic pH using IPA– n ‐hexane (Yasui‐Furukori et al ., ; Lu et al ., ; Nakashima et al ., ), TBME (Kim et al ., ; Iordachescu et al ., ), ACN (Noetzli et al ., ), hexane (Apostolou et al ., ) and EtoAc (Xie et al ., ). Park et al .…”

“…Labeled donepezil was used as IS by several researchers to avoid complexities in quantification approach on LC‐MS/MS (Matsui et al ., , ; Iordachescu et al ., ; Meier‐Davis et al ., ; Noetzli et al ., ). On the other hand, few researchers have selected diverse series of compounds (galantamine – Patel et al ., ; Khuroo et al ., ; pioglitazone – Kim et al ., ; dipyridamole – Pilli et al ., ; escitalopram – Shah et al ., ; cisapride – Park et al ., ; clenbuterol – Thevis et al ., ; diphenhydramine – Xie et al ., and loratadine – Lu et al ., ; Apostolou et al ., ) and do not show relevance in selection strategy in comparison with others.…”