Liquid chromatography-tandem mass spectrometry analysis of free and esterified fatty acid N-acyl ethanolamines in plasma and blood cells

被引：40
作者：
Balvers, Michiel G. J. [1 ,2 ]
Wortelboer, Heleen M. [1 ]
Witkamp, Renger F. [2 ]
Verhoeckx, Kitty C. M. [1 ]
机构：
[1] TNO, NL-3700 AJ Zeist, Netherlands
[2] Wageningen Univ, Div Human Nutr, NL-6700 EV Wageningen, Netherlands
来源：
ANALYTICAL BIOCHEMISTRY | 2013年 / 434卷 / 02期
关键词：
LC-MS/MS; Endocannabinoids; NAEs; Validation; Plasma; Diet; RAT-BRAIN; ENDOCANNABINOID METABOLOME; ENDOGENOUS CANNABINOIDS; ELECTROSPRAY-IONIZATION; QUANTITATIVE METHOD; ADIPOSE-TISSUE; FISH-OIL; ANANDAMIDE; ACYLETHANOLAMINES; MEDIATORS;
D O I：
10.1016/j.ab.2012.11.008
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
The origin of N-acyl ethanolamides (NAEs) in plasma is not well understood, and it is possible that NAEs are present in plasma in esterified form. To test this hypothesis, a new and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of arachidonoyl ethanolamide, 2-arachidonoyl glycerol, docosahexaenoyl ethanolamide, dihomo-gamma-linolenoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, and stearoyl ethanolamide in 100 mu l of human plasma using a simple acetonitrile extraction step. Using this method, we determined (i) free and esterified NAE levels in human plasma, (ii) free and esterified NAE levels in plasma of mice fed with diets with different amounts of n-3 fatty acids, and (iii) esterified NAE levels in blood cells. Murine and human plasma extracts contained 20- to 60-fold higher levels of esterified NAEs than free NAEs. Moreover, the effect of dietary n-3 fatty acids on murine free plasma NAE profiles was similar for esterified NAEs. Finally, esterified NAEs were also present in murine blood cells, and their pattern followed the same diet effect as observed for free and esterified NAEs in plasma. Together, these data point to the presence of previously ignored pools of esterified NAEs in plasma and blood cells that correlated well with free NAE levels in plasma. (c) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码：275 / 283
页数：9
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