title: An efficient sample preparation method for high-throughput analysis of 15(S)-8-iso-PGF2α in plasma and urine by enzyme immunoassay.
abstract: Although several methods have been reported on the analysis of the oxidative stress marker 15(S)-8-iso-prostaglandin-F2alpha (8-iso-PGF2α) in biological fluids, they either involve extensive sample preparation and costly technology or require high sample volume. This study presents a sample preparation method that utilizes low sample volume for 8-iso-PGF2α analysis in plasma and urine by an enzyme immunoassay (EIA). In brief, 8-iso-PGF2α in deproteinized plasma or native urine sample is complexed with an antibody and then captured by molecular weight cut-off filtration. This method was compared with two other sample preparation methods that are typically used in the analysis of 8-iso-PGF2α by EIA: Cayman's affinity column purification method and solid-phase extraction on C-18. The immunoaffinity purification method described here was superior to the other two sample preparation methods and yielded recovery values of 99.8 and 54.1% for 8-iso-PGF2α in plasma and urine, respectively. Analytical precision (relative standard deviation) was ±5% for plasma and ±15% for urine. The analysis of healthy human plasma and urine resulted in basal 8-iso-PGF2α levels of 31.8 ± 5.5 pg/mL and 2.9 ± 2.0 ng/mg creatinine, respectively. The robustness and analytical performance of this method makes it a promising tool for high-throughput screening of biological samples for 8-iso-PGF2α.
authors.affiliation: Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, ON K1A 0K9, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Bielecki
authors.forename: G
authors.initials: G
authors.lastname: Saravanabhavan
authors.forename: E
authors.initials: E
authors.lastname: Blais
authors.forename: R
authors.initials: R
authors.lastname: Vincent
authors.forename: P
authors.initials: P
authors.lastname: Kumarathasan
mesh_terms: D015415:Biomarkers / Q000097:blood / Q000652:urine; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000097:blood / Q000652:urine; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D018384:Oxidative Stress; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; C075750:8-epi-prostaglandin F2alpha; D015237:Dinoprost