Rapid simultaneous analysis of prostaglandin E2, 12-hydroxyeicosatetraenoic acid and arachidonic acid using high performance liquid chromatography/electrospray ionization mass spectrometry.
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.
Newby
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CS
St. John's Instutute of Dermatology, UMDS, University of London, St. Thomas' Hospital, UK.
Mallet
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AI
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000032:analysis* / Q000378:metabolism; D016718:Arachidonic Acid / Q000032:analysis* / Q000378:metabolism; D002138:Calibration; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D015232:Dinoprostone / Q000032:analysis* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry
D003470:Culture Media; D016718:Arachidonic Acid; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D015232:Dinoprostone