title: A simple and robust UPLC-SRM/MS method to quantify urinary eicosanoids.
abstract: Eicosanoids are key mediators and regulators of inflammation and oxidative stress often used as biomarkers for diseases and pathological conditions such as cardiovascular and pulmonary diseases and cancer. Analytically, comprehensive and robust quantification of different eicosanoid species in a multi-method approach is problematic because most of these compounds are relatively unstable and may differ in their chemical properties. Here we describe a novel ultra-performance liquid chromatography-selected reaction monitoring mass spectroscopy (UPLC-SRM/MS) method for simultaneous quantification of key urinary eicosanoids, including the prostaglandins (PG) tetranor PGE-M, 8-iso-, and 2,3-dinor-8-iso-PGF(2α); the thromboxanes (TXs) 11-dehydro- and 2,3-dinor-TXB₂; leukotriene E₄; and 12-hydroxyeicosatetraenoic acid. In contrast to previous methods, which used time-consuming and complex solid phase extraction, we prepared samples with a simple liquid/liquid extraction procedure. Because collision-induced dissociation produced characteristic product ions for all analytes, no derivatization step for SRM/MS analysis was necessary. Analytes were separated with a short UPLC reversed-phase column (1.7 µm particles), allowing shorter run times than conventional HPLC columns. The method was validated and applied to human urine samples showing excellent precision, accuracy, detection limits, and robustness. In summary, the developed method allows robust and sensitive profiling of urinary eicosanoid species, making it a useful and valuable tool for biomarker profiling in clinical/toxicological studies.
authors.affiliation: ABF Analytisch-Biologisches Forschungslabor GmbH, 80336 Munich, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Sterz
authors.affiliation: ABF Analytisch-Biologisches Forschungslabor GmbH, 80336 Munich, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Scherer
authors.affiliation: ABF Analytisch-Biologisches Forschungslabor GmbH, 80336 Munich, Germany. Electronic address: josef.ecker@abf-lab.com.
authors.forename: Josef
authors.initials: J
authors.lastname: Ecker
mesh_terms: D053000:Analytic Sample Preparation Methods; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015777:Eicosanoids / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine*; D006801:Humans; D018836:Inflammation Mediators / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine; D015203:Reproducibility of Results; D012907:Smoking / Q000652:urine; D053719:Tandem Mass Spectrometry / Q000379:methods*; D016482:Urinalysis / Q000379:methods*
chemical_list: D015777:Eicosanoids; D018836:Inflammation Mediators