Setting up a 2D-LC/MS/MS method for the rapid quantitation of the prostanoid metabolites 6-oxo-PGF1α and TXB2 as markers for hemostasis assessment

✍ Scribed by Bruno Casetta; Gennaro Vecchione; Michela Tomaiuolo; Maurizio Margaglione; Elvira Grandone

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 449 KB
• Volume: 44
• Category: Article
• ISSN: 1076-5174
• DOI: 10.1002/jms.1511

✦ Synopsis

Abstract

6‐oxo‐PGF~1α~ and TXB~2~ are the metabolites of the prostanglandin PGI~2~ and of the thromboxane TXA~2~, respectively. PGI~2~ and TXA~2~ are arachidonic acid‐derived compounds which regulate the blood hemostasis. Their quick metabolism leads to the 6‐oxo‐PGF~1α~ and TXB~2~ metabolites in plasma. In order to study on a large base the external factors influencing the hemostatic conditions, there is a need for a fast and reliable assay for quantitating these metabolites. Some methods have been published for the analysis of the arachidonic acid‐derived compounds and some are dealing with mass spectrometry but nonspecifically centered on these specific compounds with a fast and cheap protocol, amenable for large‐scale studies. Here we describe an analytical strategy that incorporates a two‐dimensional chromatography running coupled to tandem mass spectrometry that minimizes the sample preparation and addresses the presence of the TXB~2~ anomers for a robust quantitation measurement. After a protein precipitation, 100 µl of the supernatant (corresponding to 50 µl of the original plasma) was injected in a two‐dimensional chromatographic system which operates an on‐line clean‐up and a subsequent chromatographic separation of the targeted analytes with a limit of quantitation (LOQ) of 22 pg/ml for 6‐oxo‐PGF~1α~, and and a LOQ of 25 pg/ml for TXB~2~. Copyright © 2008 John Wiley & Sons, Ltd.