Quantitative Determination of ABT-925 in Human Plasma by On-Line SPE and LC-MSMS Validation and Sample Analysis in Phase II Studies 英文参考文献.docVIP

Pharmaceutics 2010, 2, 171–181; doi:10.3390/pharmaceutics2020171 OPEN ACCESS pharmaceutics ISSN 1999-4923 /journal/pharmaceutics Article Quantitative Determination of ABT-925 in Human Plasma by On-Line SPE and LC-MS/MS: Validation and Sample Analysis in Phase II Studies Katty Wan *, Matthew Rieser and Tawakol El-Shourbagy Abbott/100 Abbott Park Road, Abbott Park, IL 60064, USA; E-Mails: matthew.j.rieser@ (M.R.); tawakol.a.elshourbagy@ (T.E.) ? Author to whom correspondence should be addressed; E-Mail: xia.wan@; Tel.: +1-847-938-0495; Fax: +1-847-938-7789. Received: 6 April 2010; in revised form: 29 April 2010 / Accepted: 30 April 2010 / Published: 4 May 2010 Abstract: A fully automated 96-well On-Line Solid Phase Extraction (SPE) followed by High Performance Liquid Chromatography (HPLC)-Tandem Mass Spectrometric (MS/MS) method for the determination of ABT-925 (2-{3-[4-(2-tert-Butyl-6- trifluoromethyl-pyrimidin-4-yl)-piperazin-1-yl)-propyl-sulfanyl}-3H-pyrimidin-4-one fumarate) in human plasma was developed, validated and utilized in Phase II clinical studies. 50 μL of plasma sample was fortified with internal standard (IS, d8-ABT-925) and extracted on-line with Cohesive Turbo Flow Cyclone P HTLC column. The chromatographic separation was performed on Aquasil C18 (3 μm 50 × 3 mm) HPLC column with a mobile phase consisting of 50/50/0.1 (v/v/v) ACN/H2O/formic acid. The mass spectrometric measurement was conducted under positive ion mode using multiple reaction monitoring (MRM) of m/z 457.4 → 329.4 for analyte and m/z 465.5 → 337.5 for IS. The peak area ratio (analyte/IS) was used to quantitate ABT-925. A dynamic range of 0.0102 μg/mL to 5.24 μg/mL was established after the validation. The validated method was then used for two Phase II studies. To demonstrate the method reproducibility, approximately 10% of the incurred samples from one