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Pharmaceutics 2010, 2(2), 159-170; doi:10.3390/pharmaceutics2020159

Strategies for Developing Sensitive and Automated LC-MS/MS Assays of a Pharmaceutical Compound and Its Metabolite from Whole Blood Matrix

Department of Drug Analysis, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064, USA
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Received: 15 February 2010 / Revised: 9 April 2010 / Accepted: 28 April 2010 / Published: 30 April 2010
(This article belongs to the Special Issue What's on Board in Pharmaceutics)
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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. The methods were subjected to vigorous tests in precision, accuracy, matrix effect, reproducibility, and robustness. Monolithic chromatography was used to improve sample throughput in one of the methods. The results also demonstrated that proper sample preparation procedures, such as sample transfer and lysing of blood cells prior to the extraction, are key to reproducible results for pharmacokinetic parameter determination. View Full-Text
Keywords: high throughput analysis; whole blood matrix; liquid chromatography; tandem mass spectrometry; automation; prodrug; incurred sample reproducibility; metabolite; extraction high throughput analysis; whole blood matrix; liquid chromatography; tandem mass spectrometry; automation; prodrug; incurred sample reproducibility; metabolite; extraction
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Xu, R.N.; Polzin, J.; Kranz, M.; Vaca, P.; Metchkarova, M.; Rieser, M.J.; El-Shourbagy, T.A. Strategies for Developing Sensitive and Automated LC-MS/MS Assays of a Pharmaceutical Compound and Its Metabolite from Whole Blood Matrix. Pharmaceutics 2010, 2, 159-170.

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