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Published on:March 2021
Indian Journal of Pharmaceutical Education and Research, 2021; 55(1s):s303-s311
Original Article | doi:10.5530/ijper.55.1s.63

Quantification of Fluoxetine in Human Plasma by the Development of Liquid Chromatography-tandem Mass Spectrometry Method

Authors and affiliation (s):

Geeta Negi1, Jaseela Majeed2,*, Saurabh Arora3, Sachin Rana3, Manoj Singh3, Jaideep Tomar3

1Department of Pharmaceutical Chemistry, School of Pharmaceutical Sciences, Delhi Pharmaceutical Sciences and Research University, New Delhi, INDIA.

2School of Allied Health Sciences, Delhi Pharmaceutical Sciences and Research University, New Delhi, INDIA.

3Arbro Analytical Division, Arbro Pharmaceuticals Private Limited, Kirti Nagar Industrial Area, Kirti Nagar, New Delhi, INDIA.


Aim: The aim of this research paper is the development and validation of an easy, selective and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of fluoxetine in human K3EDTA plasma and application of this method on bioequivalence studies of fluoxetine. Methods: As Amitriptyline belongs to same category drug so, it was selected as an internal standard for the quantification of fluoxetine. The protein precipitation (PPT) method was used to extract analyte from 250 μl aliquot of human plasma. Chromatographic separation was achieved on BDS Hypersil C18 (50 x 4.6 mm, 5 μm) column at in 4.0 min run time using isocratic mobile phase consisting of acetonitrile and mixture of ammonium acetate containing 0.15% formic acid (55:45 % v/v) at a flow rate of 0.5 ml/min. The ionization was carried out through Electron spray ionization (ESI) operating in positive ion mode and detection was via multiple reaction monitoring (MRM) acquisition mode using the respective m/z 310.1→44.2 for fluoxetine and 278.1→233.1 for IS. The method was validated to be linear over the concentration range 0.25 to 40.00 ng/ml. Results: This LC-MS/MS method was found to be accurate and precise with intra-batch and inter-batch accuracy (% Nominal) of ±15 % and precision of <15 % and the method was successfully applied in analyzing human plasma samples of fluoxetine.

Key words: Fluoxetine, LC-MS/MS, Amitriptyline, Human plasma, Protein Precipitation, Pharmacokinetic.



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Indian Journal of Pharmaceutical Education and Research (IJPER) [ISSN-0019-5464] is the official journal of Association of Pharmaceutical Teachers of India (APTI) and is being published since 1967.


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