Simultaneous solid phase extraction, derivatization, and gas chromatographic mass spectrometric quantification of thromboxane and prostacyclin metabolites, prostaglandins, and isoprostanes in urine.
The current analytical methods for the various prostanoids require a separate and extended sample workup, derivatization, and gas chromatographic/mass spectrometric detection of each compound. Therefore, we developed and validated a rapid method for the common purification, derivatization, and GC/MS determination of 11-dehydrothromboxane B2, 2,3-dinor-6-keto-PGF1a, PGF2A, PGE2, PGD2, and isoprostanes in urine. A single reversed-phase solid-phase extraction step and modified reaction conditions yielded excellent sample purification at high recoveries and efficient derivatization for all compounds in one vial. The method allows, for the first time, the simultaneous quantification of these index metabolites of systemic thromboxane and prostacyclin synthesis, renal prostaglandin formation, and nonenzymatic in vivo lipid peroxidation in a single GC/MS run with high sensitivity and precision.
Wübert
J
J
Institute for Prophylaxis and Epidemiology of Cardiovascular Disease, University of Munich, Germany.
Reder
E
E
Kaser
A
A
Weber
P C
PC
Lorenz
R L
RL
D015121:6-Ketoprostaglandin F1 alpha / Q000031:analogs & derivatives* / Q000652:urine; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000652:urine*; D015232:Dinoprostone / Q000652:urine*; D028441:F2-Isoprostanes; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D013929:Thromboxane B2 / Q000031:analogs & derivatives* / Q000652:urine
D028441:F2-Isoprostanes; C075750:8-epi-prostaglandin F2alpha; D013929:Thromboxane B2; D015121:6-Ketoprostaglandin F1 alpha; C028290:2,3-dinor-6-ketoprostaglandin F1alpha; C049235:11-dehydro-thromboxane B2; D015237:Dinoprost; D015232:Dinoprostone