alexa Effect of disease state on ionization during bioanalysis of MK-7009, a selective HCV NS3 NS4 protease inhibitor, in human plasma and human Tween-treated urine by high-performance liquid chromatography with tandem mass spectrometric detection.


Journal of Antivirals & Antiretrovirals

Author(s): Anderson MD, Breidinger SA, Woolf EJ

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Abstract HPLC-MS/MS methods for the determination of a Hepatitis C NS3/NS4 protease inhibitor (MK-7009) in human plasma and Tween-treated urine were developed and validated over the concentration range 1-1000 ng/mL and 0.2-100 microg/mL respectively. A stable isotope labeled internal standard (ISTD), D(4)-MK-7009, was employed. Analytes were chromatographed by reversed phase HPLC and quantified by an MS/MS system. Electrospray ionization in the positive mode was employed. Multiple reaction monitoring of the precursor to product ion pairs m/z 758.6-->637.4 MK-7009 and m/z 762.5-->637.4 ISTD was used for quantitation. Analyte and internal standard were extracted from 250 microL of plasma using an automated 96-well liquid-liquid extraction. Plasma pH adjustment prior to extraction minimized ionization suppression in plasma samples from patients with Hepatitis C. The urine method involved direct dilution in the 96-well format of 0.020 mL Tween-treated urine. These methods have supported several clinical studies. Incurred plasma sample reanalysis demonstrated adequate assay reproducibility and ruggedness. This article was published in J Chromatogr B Analyt Technol Biomed Life Sci and referenced in Journal of Antivirals & Antiretrovirals

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