Rapid and sensitive quantification of 8-isoprostaglandin F2alpha in human plasma and urine by liquid chromatography-electrospray ionization mass spectrometry.
The isoprostane, 8-isoprostaglandin F2alpha (8-iso-PGF2alpha), is produced non-enzymatically by direct oxidation of arachidonic acid on the cell surface by oxygen radicals. We developed a new assay method for 8-iso-PGF2alpha using 2H4-8-iso-PGF2alpha as the internal standard (I.S.) by high-performance liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). For this assay, we established a very simple and rapid pretreatment method using a membrane filter-type solid-phase extraction column (Empore disk cartridge) for human urine extracts or intact plasma. LC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 353.24 (8-iso-PGF2alpha) and m/z 357.26 (I.S.) with a resolution of 1,500. The imprecision for this method was below 13.7%. Mean inaccuracy was 8.7% for added levels of 8-iso-PGF2alpha up to 5,000 pg/ml of urine and 500 pg/ml of plasma. Determination of plasma and urinary 8-iso-PGF2alpha concentrations in healthy subjects by the present method revealed that its urinary concentration in smokers tends to be higher than that in nonsmokers.
Ohashi
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N
Discovery Research Laboratory, Tanabe Seiyaku Company Limited, Toda-shi, Saitama, Japan. noriko-o@tanabe.co.jp
Yoshikawa
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M
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015237:Dinoprost / Q000031:analogs & derivatives / Q000032:analysis* / Q000097:blood / Q000652:urine; D028441:F2-Isoprostanes; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization
D028441:F2-Isoprostanes; C075750:8-epi-prostaglandin F2alpha; D015237:Dinoprost