title: Quantitative high performance liquid chromatography/tandem mass spectrometric analysis of the four classes of F(2)-isoprostanes in human urine.
abstract: Isoprostanes (iPs) are free radical catalyzed prostaglandin isomers. Analysis of individual isomers of PGF(2alpha)-F(2)-iPs-in urine has reflected lipid peroxidation in humans. However, up to 64 F(2)-iPs may be formed, and it is unknown whether coordinate generation, disposition, and excretion of F(2)-iPs occurs in humans. To address this issue, we developed methods to measure individual members of the four structural classes of F(2)-iPs, using liquid chromatography/tandem mass spectrometry (LC/MS/MS), in which sample preparation is minimized. Authentic standards of F(2)-iPs of classes III, IV, V, and VI were used to identify class-specific ions for multiple reaction monitoring. Using iPF(2alpha)-VI as a model compound, we demonstrated the reproducibility of the assay in human urine. Urinary levels of all F(2)-iPs measured were elevated in patients with familial hypercholesterolemia. However, only three of eight F(2)-iPs were elevated in patients with congestive heart failure, compared with controls. Paired analyses by GC/MS and LC/MS/MS of iPF(2alpha)-VI in hypercholesterolemia and of 8, 12-iso-iPF(2alpha)-VI in congestive heart failure were highly correlated. This approach will permit high throughput analysis of multiple iPs in human disease.
authors.affiliation: Center for Experimental Therapeutics, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: H
authors.initials: H
authors.lastname: Li
authors.forename: J A
authors.initials: JA
authors.lastname: Lawson
authors.forename: M
authors.initials: M
authors.lastname: Reilly
authors.forename: M
authors.initials: M
authors.lastname: Adiyaman
authors.forename: S W
authors.initials: SW
authors.lastname: Hwang
authors.forename: J
authors.initials: J
authors.lastname: Rokach
authors.forename: G A
authors.initials: GA
authors.lastname: FitzGerald
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015237:Dinoprost / Q000652:urine*; D005260:Female; D006801:Humans; D015227:Lipid Peroxidation; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008875:Middle Aged; D015203:Reproducibility of Results
chemical_list: D015237:Dinoprost