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A semi-automated high performance liquid chromatography-tandem mass spectrometric method for determination of LCI699, an 11-hyroxylase inhibitor, in human plasma

Li, Wenkui and Luo, Suyi and Rebello, Sam and Flarakos, Jimmy and Tse, Francis (2014) A semi-automated high performance liquid chromatography-tandem mass spectrometric method for determination of LCI699, an 11-hyroxylase inhibitor, in human plasma. J Chromatogr B Analyt Technol Biomed Life Sci, 960. pp. 182-193.

Abstract

A liquid chromatographic method with tandem mass spectrometric detection (LC-MS/MS) for the determination of LCI699 was developed and validated with concentration ranges of 0.0500-50.0 ng/ml and 1.00-1000 ng/ml using 0.05 ml and 0.1 ml, respectively, of human plasma. LCI699 and the internal standard, [M+6]LCI699, were extracted from fortified human plasma via protein precipitation. After transfer or dilution of the supernatant followed by solvent evaporation and reconstitution, the extract was injected onto the LC-MS/MS system. Optimal chromatographic separation was achieved on an ACE C18 (50 x 4.6 mm, 3 µm) column with 30% aqueous methanol (containing 0.5% acetic acid and 0.05% TFA) as the mobile phase run in isocratic at a flow rate of 1.0 ml/min. The total analysis cycle time is approximately 3.5 min per injection. The addition of an ion-pair reagent, 0.05% TFA, to the mobile phases significantly improved the chromatographic retention and resolution of the analyte on silica based reversed phase column. Although addition of TFA to the mobile phase suppresses the ESI signals of the analyte due to its ion-pairing characteristics in the gas phase of MS source, this negative impact was effectively alleviated by adding 0.5% acetic acid to the mobile phase. The current method was validated for sensitivity, selectivity, linearity, reproducibility, stability and recovery. Additional assessments of incurred sample stability (ISS) and incurred sample reanalysis (ISR) were conducted to demonstrate the ruggedness and robustness of the assay method. Lack of adverse matrix effect and carryover was also demonstrated. The overall precision and accuracy of the quality control samples were  15% (CV) and within 15 % (bias), respectively. The validated method was successfully used in support of rapid turnaround time pharmacokinetic studies in human

Item Type: Article
Keywords: LCI699, 11-hyroxylase inhibitor, LC-MS/MS, Plasma, Quantification
Date Deposited: 13 Oct 2015 13:13
Last Modified: 13 Oct 2015 13:13
URI: https://oak.novartis.com/id/eprint/11120

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