Objectives: A convenient method based on anion-exchange HPLC was recently developed to determine cholesterol levels of lipoproteins (HDL, LDL, IDL, VLDL, and chylomicron). The present study was performed to compare this HPLC method to homogenous assay in

Objectives: Increased lipid peroxidation (i.e. “oxidative stress”) has been identified as a central mechanism in the development of atherosclerosis and inflammatory vascular damage. Measurement of 8-iso-PGF2α has demonstrated to be a reliable indicator of in vivo oxidative stress levels. The purpose of this study was to develop a rapid, sensitive, and specific LC-MS/MS method for detection of urinary 8-iso-PGF2α, establish reference intervals, and correlate isoprostane levels with cardiac troponin I. Design and methods: Urinary 8-iso-PGF2α was detected after direct injection onto a C18 silica column and monitored in the MRM mode using m/z transitions of 353.2 > 193.25 (8-iso-PGF2α) and 357.2 > 197.25 (8-iso-PGF2α-d4). The LC-MS/MS method was also compared to an ELISA kit. Reference interval studies were evaluated against a separate population of patients presenting with chest pain that had positive cTnI values. Results: Elution of 8-iso-PGF2α was achieved after 7 min, with a total run time of 10 min. Inter-assay CVs were 13.8–20.0% and intra-assay CVs were 10.9–17.0%. Linearity ranged from 100 pg/mL to 100 ng/mL. Deming regression of ELISA and LC-MS/MS methods for 8-iso-PGF2α levels yielded poor correlation, with a slope of 0.0265, y-intercept of 0.255 ng/mL, and R2 value of 0.0434. Urine 8-iso-PGF2α concentrations in samples obtained from healthy individuals (n = 34) ranged from 57 to 390 ng/g creatinine with a mean of 221 ng/g creatinine. 8-iso-PGF2α levels were statistically significant in troponin-positive (n = 35) versus troponin-negative (n = 36) patients (p < 0.0049). Conclusions: This LC-MS/MS method provides a rapid, accurate, sensitive, and cost-effective alternative to other methods for detection of 8-iso-PGF2α in urine. 8-iso-PGF2α has potential to be a great prognostic risk indicator in individuals with a high probability for future coronary events.