Quantification of 8-iso-prostaglandin-F and 2,3-dinor-8-iso-prostaglandin-F in human urine using liquid chromatography-tandem mass spectrometry

Yuanling Liang, Ping Wei, Russell W. Duke, Peter D. Reaven, S. Mitchell Harman, Richard G. Cutler, Christopher B. Heward

Research output: Contribution to journalArticle

87 Scopus citations

Abstract

Quantification of 8-iso-prostaglandin F (8-iso-PGF) 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- PGF and its dinor metabolite, 2,3-dinor-8-iso-prostaglandin F (2,3-dinor-8-iso-PGF), 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-PGF and 3 pg for 2,3-dinor-8-iso-PGF with both inter- and intraday 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-PGF was higher than that of 8-iso-PGF, 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-PGF measurements performed with GCMS (R = 0.97, n = 15). The mean excretion rates of 8-iso-PGF and 2,3-dinor-8-iso-PGF were significantly higher in smokers than in nonsmokers (0.53 ± 0.37 vs. 0.25 ± 0.15 μg/g creatinine, p = 0.002 for 8-iso-PGF and 8.9 ± 3.8 vs. 4.6 ± 2.6 μg/g creatinine, p = 0.003 for 2,3-dinor-8-iso-PGF, 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.

Original languageEnglish (US)
Pages (from-to)409-418
Number of pages10
JournalFree Radical Biology and Medicine
Volume34
Issue number4
DOIs
StatePublished - Feb 15 2003
Externally publishedYes

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Keywords

  • 2,3-dinor-8-iso-prostaglandin F
  • 8-Iso-prostaglandin F
  • Free radicals
  • Human urine
  • LC-MS/MS
  • Oxidative stress
  • Quantification

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

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