The Large‐Scale Measurement of F2‐Isoprostanes

The measurement of in vivo oxidative damage is an important step toward understanding the etiology and prevention of chronic disease. While several indicators of oxidative damage have been proposed, F 2 ‐isoprostanes have been recognized as the most sensitive, specific and reliable indicators of in vivo oxidative damage. We report the evaluation of analytical procedures for the measurement of F 2 ‐isoprostanes with the objective of further defining analytical procedures that are essential for large‐scale F 2 ‐isoprostanes analysis. Successful analysis depends on careful attention to sample collection and handling procedures, reagent and material quality and extraction procedures. Some critical aspects of analysis are the following. Sample concentrations remain stable for at least 4 hours when maintained at 4 degrees C. The most reliable measurements of F 2 ‐isoprostanes have utilized an extensive extraction procedures and analysis with gas chromatography‐mass spectrometry, equipped with negative chemical ionization. Consistently good recoveries of the F 2 ‐isoprostanes depend on careful performance of solid phase extractions and thin layer chromatorgraphy. Quantitation should be approached by one of several isotope dilution methods. Analytical coefficients of variation are less than 10%. Additional detailed evaluations of critical procedures of analysis will be presented.