Automated 96-Well SPE and LC-MS-MS for Determination of Protease Inhibitors in Plasma and Cartilage Tissues

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• 作者：Sean X. Peng ; Salane L. King ; David M. Bornes ; David J. Foltz ; Timothy R. Baker ; and Michael G. Natchus
• 刊名：Analytical Chemistry
• 出版年：2000
• 出版时间：April 15, 2000
• 年：2000
• 卷：72
• 期：8
• 页码：1913 - 1917
• 全文大小：70K
• 年卷期：v.72,no.8(April 15, 2000)
• ISSN：1520-6882

文摘

Bioanalytical methods based on automated solid-phaseextraction (SPE) and high-performance liquid chromatography with electrospray tandem mass spectrometry(LC-MS-MS) have been developed and utilized for thedetermination of MMP inhibitors in plasma and cartilagetissues. The SPE methods were automated using a 96-well extraction plate and a 96-channel programmableliquid-handling workstation. The LC-MS-MS methodswere developed using a rapid gradient LC separation,followed by sample introduction through an ionsprayinterface in the positive ion mode and tandem massspectrometric detection with selected reaction monitoring.In the optimized SPE methods, crude plasma or groundcartilage supernatant samples were loaded onto an SPEplate to remove proteins and other interfering componentsin the matrixes to render relatively clean extracts for LC-MS-MS analysis. Compared to the simple plasma proteinprecipitation method, the automated SPE method afforded significant time-saving in sample preparation andimproved sensitivity in MS detection. The methods werevalidated and successfully applied to the analysis ofprotease inhibitors in plasma and cartilage tissues.