Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 43, Issue 3, Pages 1122-1134Publisher
ELSEVIER
DOI: 10.1016/j.jpba.2006.10.009
Keywords
LC/MS; brain tissue; arachidonic acid; prostaglandins (PGs); hydroxyeicosatetraenoic acids (HETEs); dihydroxyeicosatrienoic acids (DiHETrEs); epoxyeicosatrienoic acids (EETs)
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Funding
- NINDS NIH HHS [R01 NS038654, R01 NS038654-04] Funding Source: Medline
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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(2 alpha), 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(2 alpha)-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 intemal standards. Solid phase extraction was used for sample preparation. A gradient LUMS 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 LUMS 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. (c) 2006 Published by Elsevier B.V.
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