strategies for developing sensitive and automated lc-msms assays of a pharmaceutical compound and its metabolite from whole blood matrix策略发展敏感和自动化lc-msms制药化合物及其代谢物的检测全血矩阵.pdf

Pharmaceutics 2010, 2, 159–170; doi:10.3390/pharmaceutics2020159 OPEN ACCESS pharmaceutics ISSN 1999-4923 /journal/pharmaceutics Article Strategies for Developing Sensitive and Automated LC-MS/MS Assays of a Pharmaceutical Compound and Its Metabolite from Whole Blood Matrix Raymond N. Xu *, Jill Polzin, Michelle Kranz, Phillip Vaca, Maria Metchkarova, Matthew J. Rieser and Tawakol A. El-Shourbagy Department of Drug Analysis, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064, USA; E-Mails: Jill.Polzin@ (J.P.); michelle.kranz@ (M.K.); phillip.vaca@ (P.V.); maria.metchkarova@ (M.M.); matthew.j.rieser@ (M.J.R.); Tawakol.El-Shourbagy@ (T.A.E.-S.) * Author to whom correspondence should be addressed; E-Mail: raymond.xu@; Tel.: +1-847-938-8158; Fax: +1-847-938-7789. Received: 15 February 2010; in revised form: 9 April 2010 / Accepted: 28 April 2010 / Published: 30 April 2010 Abstract: When compared with biological samples in other matrices (plasma, urine, etc.) that are typically seen in bioanalytical applications, whole blood samples present unique challenges in method development, because of the viscous nature of blood and complexity of its constituents. In this article, we have developed and validated a series of quantitative bioanalytical methods for the determination of a pharmaceutical compound, Compound A, and its phosphate metabolite from whole blood matrices using liquid chromatography tandem mass spectrometry. All methods employed a simple protein precipitation procedure that was automated in 96-well format.