Determination of epoxyeicosatrienoic acids in human red blood cells and plasma by GC/MS in the NICI mode.
Epoxyeicosatrienoic acids (EETs) are cytochrome P450 metabolites of arachidonic acid involved in the regulation of vascular tone. Despite the importance of EETs in a variety of physiological effects, few methods have been developed to quantify them in human blood. This led us to develop a method by GC/MS with negative ion chemical ionization. As EETs are primarily located in phospholipids, red blood cells (RBCs) and plasma phospholipids were hydrolyzed with phospholipase A(2) after a solid phase extraction. Then, EETs were derivatized as pentafluorobenzyl esters, and [(2)H(8)]-arachidonic acid was used as internal standard for quantification. EETs were found to be at concentrations of 106+/-37ng mL(-1) in plasma and 33.4+/-8.5 ng/10(9) RBCs (mean+/-S.D.) in 10 healthy volunteers. Their amount in RBCs was 3-fold that in plasma; both parameters proved to be well correlated.
Goulitquer
Sophie
S
Université de Brest, INSERM U613, ECLA, Brest, France.
Dréano
Yvonne
Y
Berthou
François
F
Corcos
Laurent
L
Lucas
Danièle
D
D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives / Q000097:blood; D015777:Eicosanoids / Q000097:blood*; D004912:Erythrocytes / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D010949:Plasma / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D035501:Uncertainty
D015777:Eicosanoids; D015126:8,11,14-Eicosatrienoic Acid