ISOLATION AND MEASUREMENT OF URINARY 8-ISO-PROSTAGLANDIN F2-ALPHA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AND GAS-CHROMATOGRAPHY MASS-SPECTROMETRY

8-iso-Prostaglandin F-2 alpha (8-iso-PGF(2 alpha)) is a product of fre e radical-catalyzed peroxidation of arachidonic acid. Measurement of i ts urinary excretion has been proposed as an index of oxidative status in vivo. A stable isotope dilution method for its quantification by g as chromatography-electron capture chemical ionization mass spectromet ry is described. Sample cleanup required the combined use of high-perf ormance liquid chromatography and thin-layer chromatography. The inter -assay R.S.D. in two separate determinations was 1.6 (n=4) and 2.3% (n =4). The accuracy of the assay was evaluated through recovery experime nts. The equation of the regression plot correlating the amounts added and recovered was y=0.91x-0.31, r=0.9916 (n=12). The pair of fragment ions ([M-181](-)) at m/z 569 and m/z 573 was monitored for quantifica tion. The mean 8-iso-PGF(2 alpha) excretion rate was 528+/-127 (S.D.) ng per day in five male volunteers and 730+/-305 ng per day in six fem ales. Intake of 80 mg of lycopene per day by eleven volunteers for fou r weeks resulted in anon-significant reduction of 8-iso-PGF(2 alpha) e xcretion. (C) 1997 Elsevier Science B.V.