Table of Contents
ISRN Spectroscopy
Volume 2012, Article ID 659257, 9 pages
Research Article

Method Development and Validation of Darifenacin in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry: Application to a Pharmacokinetic Study

1Pacific University of Higher Education and Research, Udaipur, Rajasthan 313001, India
2Department of Pharmaceutical Chemistry, Nirmala College of Pharmacy, Madras Road, Kadapa 516002, Andhra Pradesh, India

Received 12 August 2012; Accepted 4 October 2012

Academic Editors: F.-F. Hsu, E. Saint-Aman, and E. Stathatos

Copyright © 2012 Thejomoorthy Karavadi and B. R. Challa. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


A selective, sensitive, and high-throughput liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS) method has been developed and validated for the quantitation of darifenacin in rat plasma. Sample clean up involved liquid-liquid extraction (LLE) and used 100 μL of rat plasma. Zorbax, SB C18,  mm, 3.5 μm particle size analytical column using 10 mM ammonium acetate buffer (pH 5) and methanol (10 : 90, v/v) as the mobile phase was used. The parent → product ion transitions for the drug (m/z 427.3 → 147.3) and IS (m/z 431.4 → 151.2) were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) and positive ion mode. The method was validated over the concentration range of 10.00–20000.00 pg/mL for darifenacin. The method was successfully applied into a pharmacokinetic study in rat plasma under fasting conditions.