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A Very Sensitive Bioanalytical Method for the Estimation of Escitalopram in Rat Plasma Using Liquid Chromatography with Tandem Mass Spectrometry-Application to Animal Pharmacokinetic Study

Author(s): Amita Rai, Raghavendra Shetty*, Harish Prabhu, Aruna Theerthahalli, N. Kumar, N. Krishnadas and A. H. Khan
Ecron Acunova Limited, 1Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal 576104, Karnataka, 2National Institute of Pharmaceutical Education and Research, Export Promotion Industrial Park (EPIP), Hajipur, Bihar 844102, India

Correspondence Address:
Raghavendra Shetty, Ecron Acunova Limited, Export Promotion Industrial Park (EPIP), Hajipur, Bihar 844102, India, E-mail:

Escitalopram is a widely used antidepressant and antianxiety agent in clinical setup. This study was aimed at developing and validating a very sensitive and specific bioanalytical method for the estimation of escitalopram in rat plasma using liquid chromatography coupled with tandem mass spectrometry as per the current United States Food and Drug Administration (May 2018) guideline. The mass spectrometer was operated in heated electrospray ionization mode with positive ion detection. The compounds were tuned and monitored by selected reaction monitoring scan mode. Imipramine hydrochloride was used as an internal standard. The extraction technique used to extract the analyte from the biological matrix was a liquidliquid extraction technique. The chromatographic separation was done by using methanol:ammonium formate (2 mM, pH 3.00) (90:10 %v/v) as mobile phase with the flow rate of 0.500 ml/min with a run time of 2.10 min. Detection and quantification were performed under selective reaction monitoring using heated electron spray ionization mode. The mass transition monitored for escitalopram was 325.200 m/z (fragments 108.98 and 262.00 m/z) and for internal standard imipramine 281.200 m/z (fragments 58.20 and 86.19 m/z). This method demonstrated acceptable performance and was successfully applied for the estimation of escitalopram in rat plasma at 0.050 to 39.708 ng/ml range.

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