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

Bruno Casetta, Gennaro Vecchione, Michela Tomaiuolo, Maurizio Margaglione, Elvira Grandone

Research output: Contribution to journalArticle


6-oxo-PGF and TXB2 are the metabolites of the prostanglandin PGI2 and of the thromboxane TXA2, respectively. PGI2 and TXA2 are arachidonic acid-derived compounds which regulate the blood hemostasis. Their quick metabolism leads to the 6-oxo-PGF and TXB2 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 TXB2 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,and anda LOQof25 pg/ml forTXB 2.

Original languageEnglish
Pages (from-to)346-352
Number of pages7
JournalJournal of Mass Spectrometry
Issue number3
Publication statusPublished - Mar 2009



  • Electrospray ionization
  • Hemostasis
  • On-line SPE
  • Prostaglandin
  • Prostanoids
  • Tandem mass spectrometry
  • Thromboxane

ASJC Scopus subject areas

  • Spectroscopy

Cite this