Arachidonic acid (AA) is metabolized in human platelets by two main pathways: via cyclooxygenase (COX-1) to prostaglandins and thromboxane (TX)A2 and via 12-lipoxygenase (12-LOX) to 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE). While COX products are known to regulate platelet reactivity, the role of 12-LOX metabolites is still controversial. To better understand the platelet enzymatic pathways, we developed a simple and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the simultaneous measurement of both platelet metabolites in human serum. After the addition of deuterated d4-TXB2 and d8-12(S)-HETE as internal standards and the solid-phase extraction of serum samples, analytes were resolved using reversed-phase C18 column and quantified using negative ion electrospray ionization-tandem mass spectrometry. Intra and interassay imprecisions were less than 10% for both analytes. The lower limits of quantification were 0.244ng/ml and 0.976ng/ml for TXB2 and 12(S)-HETE, respectively. This method was applied to measure platelet metabolites in healthy subjects (n=35). LC-MS/MS allows rapid, simultaneous, sensitive and accurate quantification of both platelet AA products in human serum with a small sample volume required and a minimal sample preparation. © 2014 Elsevier B.V.
Liquid chromatography-tandem mass spectrometry for simultaneous measurement of thromboxane B2 and 12(S)-hydroxyeicosatetraenoic acid in serum / I. Squellerio, B. Porro, P. Songia, F. Veglia, D. Caruso, E. Tremoli, V. Cavalca. - In: JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS. - ISSN 0731-7085. - 96(2014 Aug 05), pp. 256-262.
|Titolo:||Liquid chromatography-tandem mass spectrometry for simultaneous measurement of thromboxane B2 and 12(S)-hydroxyeicosatetraenoic acid in serum|
SQUELLERIO, ISABELLA (Primo)
PORRO, BENEDETTA (Secondo)
TREMOLI, ELENA (Penultimo)
CAVALCA, VIVIANA MARIA (Ultimo)
|Parole Chiave:||12(S)-hydroxyeicosatetraenoic acid ; Mass spectrometry ; Platelet ; Serum ; Analytical Chemistry ; Drug Discovery3003 Pharmaceutical Science ; 3003 ; Spectroscopy ; Clinical Biochemistry|
|Settore Scientifico Disciplinare:||Settore BIO/14 - Farmacologia|
Settore BIO/12 - Biochimica Clinica e Biologia Molecolare Clinica
|Data di pubblicazione:||5-ago-2014|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.jpba.2014.04.004|
|Appare nelle tipologie:||01 - Articolo su periodico|