Comparison of analytical techniques for quantification of 8-iso-PGF2α using HPLC-MS-MS and enzyme immunoassay

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The Ohio State University

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In short, analysis of spiked urine samples by immunoassay and HPLC-MS-MS revealed that there is no significant difference in the recoveries obtained, however, a tendency for higher values obtained through immunoassay analysis was observed. HPLC-MS-MS offers good precision and analysis, given that enough time is spent to optimize the instrument conditions to its full potential and especially when including internal standards. Deuterated standards have shown to be a valuable tool for this purpose. Enzyme immunoassay methods do not require sophisticated instrumentation, but the precision can not be improved beyond its existing parameters. Liquid and solid phase extractions are a necessity for the HPLC-MS-MS analysis, while unpurified urine samples seem to yield good recoveries by immunoassay. Urine, as a matrix, seemingly can influence results in both ways. Urine may create higher recoveries in immunoassay methods due to cross-reactivity, while precipitation and entrapment of the analyte seems a more predominant factor for HPLC-MS-MS assays. Ultimately, the decision between using HPLC-MS-MS and immunoassay would have to be decided by familiarity, cost, and personal preference.



Isoprostane, Comparison, ELISA, immunoassay, HPLC-MS