A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, and epoxyeicosatrienoic acids in rat brain tissue
| Autor: | Kenneth I. Strauss, Susan A. Jansen, Elise Murphy, Luella J. Rossi, Michael R. Borenstein, Hongfei Yue, Mary F. Barbe |
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| Rok vydání: | 2007 |
| Předmět: |
Male
Quality Control Metabolite Electrospray ionization Lipoxygenase Clinical Biochemistry Pharmaceutical Science Arachidonic Acids Mass Spectrometry Article Analytical Chemistry Rats Sprague-Dawley chemistry.chemical_compound Cytochrome P-450 Enzyme System Liquid chromatography–mass spectrometry Hydroxyeicosatetraenoic Acids Drug Discovery Animals Solid phase extraction Spectroscopy Unsaturated fatty acid Brain Chemistry Cerebral Cortex Arachidonic Acid Chromatography Chemistry Reproducibility of Results Hydroxyeicosatetraenoic acid Reference Standards Rats Biochemistry Eicosanoid Prostaglandin-Endoperoxide Synthases Calibration Prostaglandins Eicosanoids Arachidonic acid Chromatography Liquid |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis. 43:1122-1134 |
| ISSN: | 0731-7085 |
| DOI: | 10.1016/j.jpba.2006.10.009 |
| Popis: | A sensitive, specific, and robust liquid chromatography/mass spectrometric (LC/MS) method was developed and validated that allows simultaneous analysis of arachidonic acid (AA) and its cyclooxygenase, cytochrome P450, and lipoxygenase pathway metabolites prostaglandins (PGs), dihydroxyeicosatrienoic acids (DiHETrEs), hydroxyeicosatetraenoic acids (HETEs) and epoxyeicosatrienoic acids (EETs), including PGF(2alpha), PGE(2), PGD(2), PGJ(2), 14,15-DiHETrE, 11,12-DiHETrE, 8,9-DiHETrE, 5,6-DiHETrE, 20-HETE, 15-HETE, 12-HETE, 9-HETE, 8-HETE, 5-HETE, 14,15-EET, 11,12-EET, 8,9-EET, and 5,6-EET in rat brain tissues. Deuterium labeled PGF(2alpha)-d(4), PGD(2)-d(4), 15(S)-HETE-d(8), 14,15-EET-d(8), 11,12-EET-d(8), 8,9-EET-d(8), and AA-d(8) were used as internal standards. Solid phase extraction was used for sample preparation. A gradient LC/MS method using a C18 column and electrospray ionization source under negative ion mode was optimized for the best sensitivity and separation within 35 min. The method validation, including LC/MS instrument qualification, specificity, calibration model, accuracy, precision (without brain matrix and with brain matrix), and extraction efficiency were performed. The linear ranges of the calibration curves were 2-1000 pg for PGs, DiHETrEs, HETEs, and EETs, 10-2400 pg for PGE(2) and PGD(2), and 20-2000 ng for AA, respectively. |
| Databáze: | OpenAIRE |
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