title: Novel liquid chromatography method based on linear weighted regression for the fast determination of isoprostane isomers in plasma samples using sensitive tandem mass spectrometry detection.
abstract: A simple, fast, sensitive and accurate methodology based on a LLE followed by liquid chromatography-tandem mass spectrometry for simultaneous determination of four regioisomers (8-iso prostaglandin F2α, 8-iso-15(R)-prostaglandin F2α, 11β-prostaglandin F2α, 15(R)-prostaglandin F2α) in routine analysis of human plasma samples was developed. Isoprostanes are stable products of arachidonic acid peroxidation and are regarded as the most reliable markers of oxidative stress in vivo. Validation of method was performed by evaluation of the key analytical parameters such as: matrix effect, analytical curve, trueness, precision, limits of detection and limits of quantification. As a homoscedasticity was not met for analytical data, weighted linear regression was applied in order to improve the accuracy at the lower end points of calibration curve. The detection limits (LODs) ranged from 1.0 to 2.1pg/mL. For plasma samples spiked with the isoprostanes at the level of 50pg/mL, intra-and interday repeatability ranged from 2.1 to 3.5% and 0.1 to 5.1%, respectively. The applicability of the proposed approach has been verified by monitoring of isoprostane isomers level in plasma samples collected from young patients (n=8) subjected to hyperbaric hyperoxia (100% oxygen at 280kPa(a) for 30min) in a multiplace hyperbaric chamber.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, Gdansk University of Technology, 11/12, Narutowicza Street, 80-233 Gdańsk, Poland. Electronic address: aszykjustyna@gmail.com.
authors.forename: Justyna
authors.initials: J
authors.lastname: Aszyk
authors.affiliation: National Centre for Hyperbaric Medicine, Institute of Maritime and Tropical Medicine in Gdynia, Medical University of Gdansk, Powstania Styczniowego 9B, 81-519 Gdynia, Poland.
authors.forename: Jacek
authors.initials: J
authors.lastname: Kot
authors.affiliation: National Centre for Hyperbaric Medicine, Institute of Maritime and Tropical Medicine in Gdynia, Medical University of Gdansk, Powstania Styczniowego 9B, 81-519 Gdynia, Poland.
authors.forename: Yurii
authors.initials: Y
authors.lastname: Tkachenko
authors.affiliation: Chair and Department of Biology and Genetic Medical University of Lublin, W. Chodźki 4A, 20-093 Lublin, Poland.
authors.forename: Michał
authors.initials: M
authors.lastname: Woźniak
authors.affiliation: Chair and Department of Biology and Genetic Medical University of Lublin, W. Chodźki 4A, 20-093 Lublin, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Bogucka-Kocka
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, Gdansk University of Technology, 11/12, Narutowicza Street, 80-233 Gdańsk, Poland.
authors.forename: Agata
authors.initials: A
authors.lastname: Kot-Wasik
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000032:analysis / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D018496:Hyperoxia / Q000097:blood; D007536:Isomerism; D028421:Isoprostanes / Q000032:analysis / Q000097:blood*; D057230:Limit of Detection; D016014:Linear Models; D059625:Liquid-Liquid Extraction / Q000379:methods; D018384:Oxidative Stress; D011453:Prostaglandins / Q000032:analysis / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D015415:Biomarkers; D028421:Isoprostanes; D011453:Prostaglandins
keywords: F2-isoprostanes; LC–MS/MS analysis; Plasma; Prostaglandins; Weighted linear regression