Development and validation of a high-performance liquid chromatography–electrospray mass spectrometry method for the simultaneous determination of 23 eicosanoids
| Autor: | Anthony J. Blewett, Susan A. Jansen, Joseph R. Libonati, Tiffany Gilles, Deepti Varma |
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| Rok vydání: | 2008 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Formic acid Electrospray ionization Eicosatetraenoic acid Clinical Biochemistry Pharmaceutical Science Kidney Mass spectrometry Sensitivity and Specificity Analytical Chemistry chemistry.chemical_compound Liquid chromatography–mass spectrometry Rats Inbred SHR Drug Discovery Animals Chromatography High Pressure Liquid Spectroscopy Unsaturated fatty acid Arachidonic Acid Chromatography Rats chemistry Eicosanoid Hypertension Eicosanoids lipids (amino acids peptides and proteins) Arachidonic acid |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis. 46:653-662 |
| ISSN: | 0731-7085 |
| DOI: | 10.1016/j.jpba.2007.11.047 |
| Popis: | Inflammation is implicated in the pathogenesis of a number of diseases, including cardiovascular disease. Current research is focused on developing assays to search for biomarkers for inflammation. Eicosanoids are the oxidative metabolites of arachidonic acid (eicosatetraenoic acid, AA), a long chain polyunsaturated fatty acid common in Western diets. AA can be oxidized by one of three pathways to form prostaglandins (PGs), leukotrienes (LTs), or a number of hydroxyl and epoxy compounds. These eicosanoids have a variety of physiological functions, including regulating inflammation. We have developed a method utilizing LC-MS to separate and quantitate 23 different eicosanoids from all the three oxidative pathways. The eicosanoids were separated using a gradient elution of acetonitrile with 0.1% formic acid (v/v) and water with 0.1% formic acid (v/v) at a flow rate of 1 mL/min with a Symmetry C18 column (250 mm x 4.6 mm). Deuterated eicosanoids were used as internal standards for quantitation. Mass spectrometric detection was carried out using an Agilent 1100-series LC-MSD with an electrospray ionization interface. Electrospray ionisation (ESI) mass spectra were acquired using negative ionization and selective ion monitoring. The method was validated and shown to be sensitive (LOQ at pg levels for most compounds), accurate (recovery values 75-120%) and precise (R.S.D.20 for all compounds) with a linear range over several orders of magnitude. The method was applied to rat kidney tissue and shown to be indicative of the eicosanoid levels within a specific organ. The analysis of eicosanoids can provide insight into the inflammatory mechanisms associated with cardiovascular disease. |
| Databáze: | OpenAIRE |
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