Document Detail


Rapid, simultaneous quantitation of mono and dioxygenated metabolites of arachidonic acid in human CSF and rat brain.
MedLine Citation:
PMID:  19892608     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Currently, there are few biomarkers to predict the risk of symptomatic cerebral vasospasm (SV) in subarachnoid hemorrhage (SAH) patients. Mono and dioxygenated arachidonic acid metabolites, involved in the pathogenesis of ischemic injury, may serve as indicators of SV. This study developed a quantitative UPLC-MS/MS method to simultaneously measure hydroxyeicosatetraenoic acid (HETE), dihydroxyeicosatrienoic acid (DiHETrE), and epoxyeicosatrienoic acid (EET) metabolites of arachidonic acid in cerebrospinal fluid (CSF) samples of SAH patients. Additionally, we determined the recovery of these metabolites from polyvinylchloride (PVC) bags used for CSF collection. Linear calibration curves ranging from 0.208 to 33.3 ng/ml were validated. The inter-day and intra-day variance was less than 15% at most concentrations with extraction efficiency greater than 73%. The matrix did not affect the reproducibility and reliability of the assay. In CSF samples, peak concentrations of 8,9-DiHETrE, 20-HETE, 15-HETE, and 12-HETE ranged from 0.293 to 24.9 ng/ml. In rat brain cortical tissue samples, concentrations of 20-, 15-, 12-HETE, 8,9-EET, and 14,15-, 11,12-DiHETrE ranged from 0.57 to 23.99 pmol/g wet tissue. In rat cortical microsomal incubates, all 10 metabolites were measured with formation rates ranging from 0.03 to 7.77 pmol/mg/min. Furthermore, 12-HETE and EET metabolites were significantly altered by contact with PVC bags at all time points evaluated. These data demonstrate that the simultaneous measurement of these compounds in human CSF and rat brain can be achieved with a UPLC-MS/MS system and that this method is necessary for evaluation of these metabolites as potential quantitative biomarkers in future clinical trials.
Authors:
Tricia M Miller; Mark K Donnelly; Elizabeth A Crago; Dana M Roman; Paula R Sherwood; Michael B Horowitz; Samuel M Poloyac
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Validation Studies     Date:  2009-10-14
Journal Detail:
Title:  Journal of chromatography. B, Analytical technologies in the biomedical and life sciences     Volume:  877     ISSN:  1873-376X     ISO Abbreviation:  J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.     Publication Date:  2009 Dec 
Date Detail:
Created Date:  2009-11-16     Completed Date:  2010-03-02     Revised Date:  2014-09-22    
Medline Journal Info:
Nlm Unique ID:  101139554     Medline TA:  J Chromatogr B Analyt Technol Biomed Life Sci     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  3991-4000     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
8,11,14-Eicosatrienoic Acid / analogs & derivatives,  cerebrospinal fluid,  metabolism
Animals
Arachidonic Acid / cerebrospinal fluid,  metabolism*
Brain / metabolism*
Chromatography, Liquid / methods*
Humans
Hydroxyeicosatetraenoic Acids / cerebrospinal fluid,  metabolism
Male
Rats
Rats, Sprague-Dawley
Reference Standards
Subarachnoid Hemorrhage / cerebrospinal fluid,  metabolism
Tandem Mass Spectrometry / methods*
Vasospasm, Intracranial / cerebrospinal fluid
Grant Support
ID/Acronym/Agency:
R01 GM073031/GM/NIGMS NIH HHS; R01 GM073031-04/GM/NIGMS NIH HHS; R01 NR004339/NR/NINR NIH HHS; R01 NR004339-12/NR/NINR NIH HHS; R01 NS052315/NS/NINDS NIH HHS; R01 NS052315-05/NS/NINDS NIH HHS; R01NR004339/NR/NINR NIH HHS; R01NS052315/NS/NINDS NIH HHS; S10 RR023461/RR/NCRR NIH HHS; S10 RR023461-01/RR/NCRR NIH HHS; S10RR023461/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Hydroxyeicosatetraenoic Acids; 27YG812J1I/Arachidonic Acid; 7324-41-6/8,11,14-Eicosatrienoic Acid; 79551-86-3/20-hydroxy-5,8,11,14-eicosatetraenoic acid; 81246-85-7/8,9-epoxyeicosatrienoic acid
Comments/Corrections

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