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URINARY EXCRETION OF THREE NUCLEIC ACID OXIDATION ADDUCTS AND ISOPROSTANE F2a MEASURED BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY IN SMOKERS, EX-SMOKERS, AND NONSMOKERS
URINARY EXCRETION OF THREE NUCLEIC ACID OXIDATION ADDUCTS AND ISOPROSTANE F2a MEASURED BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY IN SMOKERS, EX-SMOKERS, AND NONSMOKERS
Date: 2002-03-01 00:00:00.0
Contact: david.hanak@kronoslaboratory.com
 
Abstract-Quantification of 8-iso-prostaglandin F2a (8-iso-PGF2a) has been suggested to be a reliable indicator of lipid peroxidation that may be related to in vivo free radical generation, oxidative damage, and antioxidant deficiency. We have developed a LC-MS/MS method to quantify 8-iso- PGF2a and its dinor metabolite, 2,3-dinor-8-iso-prostaglandin F2a (2,3-dinor-8-iso-PGF2a), in human urine samples. After an initial purification step using an automated C18 solid phase extraction procedure, the urine sample was injected directly into a liquid chromatography (LC) system and detected with tandem mass spectrometry. The detection limit of the assay was 9 pg for 8-iso-PGF2a and 3 pg for 2,3-dinor-8-iso-PGF2a with both inter- and intra day variations of less than 12%. The inaccuracies were less than 3% for both analytes at three different levels. The urinary excretion rate of 2,3-dinor-8-iso-PGF2a was higher than that of 8-iso-PGF2, and changed in proportion to the parent compound (R = 0.70, n = 60). Values obtained with this method showed good linear correlation to duplicate 8-iso-PGF2 measurements performed with GCMS (R = 0.97, n = 15). The mean excretion rates of 8-iso-PGF2a and 2,3-dinor-8-iso-PGF2a were significantly higher in smokers than in nonsmokers (0.53 ± 0.37 vs. 0.25 ± 0.15 g/g creatinine, p = 0.002 for 8-iso-PGF2a and 8.9 ± 3.8 vs. 4.6 ± 2.6 ug/g creatinine,p = 0.003 for 2,3-dinor-8-iso-PGF2a, respectively). The excellent accuracy, reproducibility, and high throughput of this method should permit it to be used in large clinical studies and standard clinical laboratories. © 2003 Elsevier Science Inc.

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For further information please contact david.hanak@kronoslaboratory.com or visit www.kronosscience.com


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