A high performance LC-MS/MS method for the estimation of Erlotinib in human plasma
Abstract
A High Performance Liquid Chromatography Mass Spectrometric Method for the estimation of Erlotinib in human plasma has been developed and validated using Erlotinib D6 as an internal standard (IS). The extraction of analyte and IS was accomplished by Liquid-Liquid Extraction technique. The method has been validated over a concentration range of 1.00ng/mL to 2502.02 ng/mL. Chromatographic separations was achieved using Waters X Bridge C18, 75mm x 4.6mm, 3.5μ, column eluted at flow rate of 0.8 mL/minute with 1:1 splitted post column with mobile phase Acetonitrile: Ammonium acetate buffer pH 9.2 (70:30 v/v).The overall run time of method was about 3.0 min. with elution times of Erlotinib and its internal standard Erlotinib D6 at around 1.3 min. The multiple reaction monitoring transitions were set at 394.2 > 278.1 (m/z) and 400.4 > 278.1 (m/z) for Erlotinib and Erlotinib D6 respectively. The calibration curves were linear (r2 ≥ 0.99) over the range of 1.0-2502.02 ng/mL with lower limit of quantitation validated at 1.0 ng/mL. Extraction recoveries were > 80 % for both Erlotinib and its stable labeled IS Erlotinib D6. The within run and between run precisions were within 0.62%-7.07%, while accuracy ranged from 94.4 to 103.3%.
Keywords:
High performance liquid chromatography, Mass spectrometry, Tyrosine kinase inhibitor, Anticancer, ErlotinibDownloads
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Copyright (c) 2016 Mirza Layeeq Ahmed Baig, Syed Ayaz Ali
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