Journal
ANALYTICAL BIOCHEMISTRY
Volume 404, Issue 2, Pages 211-216Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2010.05.023
Keywords
Mass spectrometry; Enzyme immunoassay; Quantitative analysis; Biomarker; 8-iso-Prostaglandin F-2 alpha
Funding
- National Cancer Institute [5R01CA101052]
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A rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was developed for the measurement of urinary 8-iso-prostaglandin F-2 alpha (8-iso-PGF(2 alpha)), a biomarker of lipid peroxidation. Because urine contains numerous F-2 prostaglandin isomers, each with identical mass and similar mass spectrometric fragmentation patterns, chromatographic separation of 8-iso-PGF(2 alpha) from its isomers is necessary for its quantitative analysis using MS/MS. We were able to achieve this separation using an isocratic LC method with a run time of less than 9 min, which is at least threefold faster than previous methods, while maintaining sensitivity, accuracy, precision, and reliability. The limits of detection and quantitation were 53 and 178 pg/ml urine, respectively. We compared our method with a commercially available affinity purification and enzyme immunoassay kit and found both assays to be in agreement. Despite the high sensitivity of the enzyme immunoassay method, it is more expensive and has a narrower dynamic range than LC-MS/MS. Our method was optimized for rapid measurement of 8-iso-PGF(2 alpha) in urine, and it is ideally suited for clinical sample analysis. (C) 2010 Elsevier Inc. All rights reserved.
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