Bioanalytical Method Development and Validation for the Estimation of Lumefantrine and Desbutyl Lumefantrine in Human Plasma by LC-MS/MS

Research Article

Bioanalytical Method Development and Validation for the Estimation of Lumefantrine and Desbutyl Lumefantrine in Human Plasma by LC-MS/MS

Shanti Swaroop Yadav, Jaivik V. Shah, Priyanka A. Shah, Pavan Patel, Mallika Sanyal, Mukesh C. Patel, Pranav S. Shrivastav

Corresponding Author : Pranav S. Shrivastav,

Department of Chemistry, Pacific University, Debari, Udaipur-313024, Rajasthan, India.

Email ID : pranav_shrivastav@yahoo.com

Received : 2015-10-29 Accepted : 2015-12-15 Published : 2015-12-15

Abstract : A liquid chromatography–electrospray ionization–tandem mass spectrometry (LC–ESI–MS/MS) method for the simultaneous quantitation of lumefantrine (LF) and its active metabolite, desbutyl lumefantrine (DBL) metabolite in human plasma is developed and validated. The chromatographic separation was carried out on Hypersil Gold C18 (50 × 4.6 mm, 5 µm) using 5.0 mM ammonium formate (pH 3.0 adjusted with formic acid): acetonitrile (10:90, v/v) as the mobile phase. For sample preparation, solid phase extraction was carried out using artemisinin as the internal standard (IS) from 100 µL human plasma. Quantitation of analytes was done by tandem mass spectrometer using electrospray ionization in the positive mode. The precursor to product ion transitions monitored were m/z 530.5→ 512.1, m/z 472.7→512.1 and m/z 300.2 →219.5 for LF, DBL and IS respectively. The calibration curve was linear over the concentration range of 5.0–5000 ng/mL for both the analytes with a correlation coefficient (r2) ≥ 0.9992. The intra-day and inter-day assay precision ranged from 1.77 to 7.22 % and 0.96 to 3.90 % for LF and 2.36 to 8.12 % and 0.57 to 4.64 % for DBL respectively. Similarly, the intra-day and inter-day assay accuracy was between 98.3–101.2 % and 98.8–102.1 % for LF and 97.5–101.6 % and 98.2–101.7 % for DBL respectively. The mean extraction recovery of LF and DBL was 98.5 % and 98.4 % across four quality control levels. Stability study in plasma was evaluated under different conditions like bench top, auto sampler, freeze-thaw and long term. The application of this assay was demonstrated through a bioequivalence with 12 healthy subjects using 20/120 mg artemether/lumefantrine tablet formulation.

Keywords : Lumefantrine, desbutyl lumefantrine, LC-MS/MS, solid phase extraction, bioequivalence

Citation : Pranav S. Shrivastav et al. (2015). Bioanalytical Method Development and Validation for the Estimation of Lumefantrine and Desbutyl Lumefantrine in Human Plasma by LC-MS/MS J. of Advancement in Medical and Life Sciences. V3I4.02 DOI : 10.5281/zenodo.890255

Copyright : © 2015 Pranav S. Shrivastav. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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