A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosate

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• 作者：Hongfei Yue ; Susan A. Jansen ; Kenneth I. Strauss ; Michael R. Borenstein ; Mary F. Barbe ; Luella J. Rossi ; Elise Murphy
• 关键词：LC/MS ; Brain tissue ; Arachidonic acid ; Prostaglandins (PGs) ; Hydroxyeicosatetraenoic acids (HETEs) ; Dihydroxyeicosatrienoic acids (DiHETrEs) ; Epoxyeicosatrienoic acids (EETs)
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：2007
• 期刊代码：65_07317085
• 类别：ch
• 出版时间：19 February 2007
• 卷：43
• 期：3
• 页码：1122-1134
• 文件大小：910 K

摘要

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_2α, PGE₂, PGD₂, PGJ₂, 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_2α-d₄, PGD₂-d₄, 15(S)-HETE-d₈, 14,15-EET-d₈, 11,12-EET-d₈, 8,9-EET-d₈, and AA-d₈ 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 PGE₂ and PGD₂, and 20–2000 ng for AA, respectively.