New Improved UPLC-MS/MS Method for Reliable Determination of Clarithromycin in Human Plasma to Support a Bioequivalence Study
Darshan V. Chaudhary , Daxesh P. Patel, Jaivik V. Shah, Priyanka A. Shah, Mallika Sanyal, Pranav S. Shrivastav
Corresponding Author : Pranav S. Shrivastav,
Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad- 380009, India.
Email ID : email@example.com
Received : 2014-12-28 Accepted : 2015-02-07 Published : 2015-02-07
Abstract : An improved, highly sensitive UPLC-MS/MS method has been developed for the determination of clarithromycin in human plasma. For sample preparation, liquid-liquid extraction with n-hexane: methyl tert-butyl ether (20:80, v/v) mixture was carried out using clarithromycin 13C-d3 as the internal standard (IS). Acquity UPLC BEH C18 (50 mm Ã— 2.1 mm, 1.7 Âµm) analytical column was used for chromatography with methanol-5.0 mM ammonium formate, pH 3.0 (78:22, v/v) as the mobile phase under isocratic conditions. The analysis time was 1.5 min. Quantitation of analyte was done by tandem mass spectrometer using electrospray ionization in the positive mode. The precursor â†’ product ion transitions monitored for clarithromycin and IS were m/z 748.9 â†’ 158.1 and m/z 752.8 â†’ 162.0 respectively. The method was validated over a dynamic concentration range of 0.80-1600 ng/mL with correlation coefficient (r2) â‰¥ 0.9998. The mean extraction recovery of clarithromycin was 96.2 % across six quality control levels. Intra-batch and inter-batch accuracy and precision (% CV) ranged from 96.8 to 103.5 % and 1.28 to 4.85 % respectively. Stability of clarithromycin in plasma was evaluated under different conditions like bench top, auto sampler, dry and wet extract, freeze-thaw and long term. The present method was successfully applied to a bioequivalence study in 20 healthy subjects who received single oral dose 250 mg clarithromycin tablet formulation. The reproducibility of the method was investigated by reanalysis of 100 incurred samples.
Keywords : Clarithromycin, clarithromycin 13C-d3, UPLC-MS/MS, human plasma, sensitive, bioequivalence
Citation : Pranav S. Shrivastav, et al. (2015). New Improved UPLC-MS/MS Method for Reliable Determination of Clarithromycin in Human Plasma to Support a Bioequivalence Study. J. of Advancement in Medical and Life Sciences. V2I3. DOI : 10.5281/zenodo.1000338
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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