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

M.G.J. Balvers, H.M. Wortelboer, R.F. Witkamp, K.C.M. Verhoeckx

Research output: Contribution to journalArticleAcademicpeer-review

23 Citations (Scopus)

Abstract

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-¿-linolenoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, and stearoyl ethanolamide in 100 µl 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.
Original languageEnglish
Pages (from-to)275-283
JournalAnalytical Biochemistry
Volume434
Issue number2
DOIs
Publication statusPublished - 2013

Fingerprint

Ethanolamines
Plasma Cells
Nonesterified Fatty Acids
Mass spectrometry
Blood Cells
Mass Spectrometry
Blood
Fatty Acids
Cells
Plasmas
Plasma (human)
Liquids
Omega-3 Fatty Acids
Nutrition
Diet

Keywords

  • rat-brain
  • endocannabinoid metabolome
  • endogenous cannabinoids
  • electrospray-ionization
  • quantitative method
  • adipose-tissue
  • fish-oil
  • anandamide
  • acylethanolamines
  • mediators

Cite this

@article{190a670ad458486f85250e29e8de6387,
title = "Liquid chromatography–tandem mass spectrometry analysis of free and esterified fatty acid N-acyl ethanolamines in plasma and blood cells",
abstract = "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-¿-linolenoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, and stearoyl ethanolamide in 100 µl 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.",
keywords = "rat-brain, endocannabinoid metabolome, endogenous cannabinoids, electrospray-ionization, quantitative method, adipose-tissue, fish-oil, anandamide, acylethanolamines, mediators",
author = "M.G.J. Balvers and H.M. Wortelboer and R.F. Witkamp and K.C.M. Verhoeckx",
year = "2013",
doi = "10.1016/j.ab.2012.11.008",
language = "English",
volume = "434",
pages = "275--283",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Elsevier",
number = "2",

}

Liquid chromatography–tandem mass spectrometry analysis of free and esterified fatty acid N-acyl ethanolamines in plasma and blood cells. / Balvers, M.G.J.; Wortelboer, H.M.; Witkamp, R.F.; Verhoeckx, K.C.M.

In: Analytical Biochemistry, Vol. 434, No. 2, 2013, p. 275-283.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Liquid chromatography–tandem mass spectrometry analysis of free and esterified fatty acid N-acyl ethanolamines in plasma and blood cells

AU - Balvers, M.G.J.

AU - Wortelboer, H.M.

AU - Witkamp, R.F.

AU - Verhoeckx, K.C.M.

PY - 2013

Y1 - 2013

N2 - 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-¿-linolenoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, and stearoyl ethanolamide in 100 µl 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.

AB - 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-¿-linolenoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, and stearoyl ethanolamide in 100 µl 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.

KW - rat-brain

KW - endocannabinoid metabolome

KW - endogenous cannabinoids

KW - electrospray-ionization

KW - quantitative method

KW - adipose-tissue

KW - fish-oil

KW - anandamide

KW - acylethanolamines

KW - mediators

U2 - 10.1016/j.ab.2012.11.008

DO - 10.1016/j.ab.2012.11.008

M3 - Article

VL - 434

SP - 275

EP - 283

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 2

ER -