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

• Published in: Biomedical chromatography Vol. 26; no. 5; pp. 636 - 649
• Main Authors: Khuroo, Arshad H., Gurule, Sanjay J., Monif, Tausif, Goswami, Dipanjan, Saha, Arabinda, Singh, Santosh K.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-05-2012
• Subjects: Area Under Curve; Chromatography, Liquid; clinical impact; donepezil and its metabolites; Drug Stability; Humans; Indans - blood; Indans - chemistry; Indans - pharmacokinetics; LC-MS/MS simultaneous method development; Least-Squares Analysis; Male; Piperidines - blood; Piperidines - chemistry; Piperidines - pharmacokinetics; plasma stability; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry - methods; validation
• ISSN: 0269-3879; 1099-0801
• DOI: 10.1002/bmc.1709

ABSTRACT
A bioanalytical method was developed and validated to estimate donepezil, 6‐desmethyl donepezil and 5‐desmethyl donepezil simultaneously in human plasma using galantamine as an internal standard (IS). The chromatographic separation was achieved on a reverse‐phase XTerra RP (150 × 4.6 mm, 5 µm) column without affecting recovery (mean recovery > 60% with CV < 10%) for all analytes. ESI‐MS/MS multiple reaction monitoring in positive polarity was used to detect mass pairs for donepezil (m/z 380.3 → 91.3), 6‐desmethyl donepezil (m/z 366.4 → 91.3), 5‐desmethyl donepezil (m/z 366.4 → 91.3) and galantamine m/z (288.1 → 213.0). The linearity was established over a dynamic range of 0.339–51.870, 0.100–15.380 and 0.103–15.763 ng/mL for donepezil, 6‐desmethyl donepezil and 5‐desmethyl donepezil, respectively. The current method shows that minimal conversion of labile metabolites to parent donepezil in plasma as stability was successfully achieved for 211 days at −15 °C storage temperature. The method was successfully applied to a clinical study after administration of 10 mg donepezil tablets to healthy male Indian volunteers. Copyright © 2011 John Wiley & Sons, Ltd.