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J Chromatogr A. 2014 Sep 12;1359:60-9. doi: 10.1016/j.chroma.2014.07.006. Epub 2014 Jul 12.

Comprehensive ultra-performance liquid chromatographic separation and mass spectrometric analysis of eicosanoid metabolites in human samples.

Author information

1
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093-0601, USA.
2
Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093-0601, USA.
3
Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093-0601, USA; Department of Medicine, University of California at San Diego, La Jolla, CA 92093-0601, USA.
4
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: chaoyan@sjtu.edu.cn.
5
Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093-0601, USA; Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, CA 92093-0601, USA. Electronic address: edennis@ucsd.edu.

Abstract

Over the past decade, the number of known eicosanoids has expanded immensely and we have now developed an ultra-performance liquid chromatography-electrospray ionization triple quadrupole mass spectrometric (UPLC-QTRAP/MS/MS) method to monitor and quantify numerous eicosanoids. The UPLC-QTRAP/MS/MS approach utilizes scheduled multiple reaction monitoring (MRM) to optimize sensitivity, number of metabolites that can be analyzed and the time requirement of the analysis. A total of 184 eicosanoids including 26 deuterated internal standards can be separated and monitored in a single 5min UPLC run. To demonstrate a practical application, human plasma samples were analyzed following solid-phase extraction (SPE) and the recovery rate and matrix effects were determined for the 26 deuterated internal standards added to the plasma. The method was validated and shown to be sensitive with the limit of quantitation at pg levels for most compounds, accurate with recovery rates of 70-120%, and precise with a CV<30 for all compounds. Also, the method showed a linear response over a range spanning several orders of magnitude. In a QC human plasma sample, we identified and rigorously quantified over 120 eicosanoids.

KEYWORDS:

Eicosanoids; Lipidomics; Plasma; Tandem mass spectrometry; Ultra-performance liquid chromatography

PMID:
25074422
PMCID:
PMC4592635
DOI:
10.1016/j.chroma.2014.07.006
[Indexed for MEDLINE]
Free PMC Article
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