Rapid simultaneous analysis of prostaglandin E2, 12-hydroxyeicosatetraenoic acid and arachidonic acid using high performance liquid chromatography/electrospray ionization mass spectrometry

✍ Scribed by Craig S. Newby; Anthony L. Mallet

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 100 KB
• Volume: 11
• Category: Article
• ISSN: 0951-4198
• DOI: 10.1002/(sici)1097-0231(19971015)11:15<1723::aid-rcm71>3.0.co;2-k

✦ Synopsis

Arachidonic acid (AA) can be metabolized to a variety of lipid mediators including prostaglandins (PGE), and hydroxyeicosatetraenoic acids (HETE) by cyclooxygenase, lipoxygenase and cytochrome P450-dependent monooxygenase enzymatic pathways. Traditional experimental procedures to quantify these lipid mediators require purification, often by high performance liquid chromatography (HPLC), prior to derivatization for gas chromatography/mass spectrometry (GC/MS) analysis. This paper describes a rapid and simple technique for the simultaneous quantitative analysis of PGE2, 12-HETE, and AA by HPLC/electrospray ionization mass spectrometry on cultured human dermal fibroblast supernatants. Extension of the method to analyse 5-HETE and 15-HETE was investigated. The advantages of this method include minimal sample preparation and elimination of the problem associated with thermal stability for GC/MS analysis. A detection limit of 20pg on column for PGE2 and 5pg on column for 12-HETE and AA was determined.