Anandamide and other N-acylethanolamines in mouse peritoneal macrophages.
N-acyl phosphatidylethanolamine (N-acyl PE) and free N-acylethanolamine (NAE) in mouse peritoneal macrophages were identified and quantified by gas chromatography-mass spectrometry (GC-MS) of tertbutyldimethylsilyl derivatives in the presence of internal standards synthesized from [1,1,2,2-2H4]ethanolamine. N-acyl PE was present at a level of 123-187 pmol/mumol lipid P (521-768 pmol/10(8) cells), with arachidonic acid making up about 3-4% of the N-acyl moieties. NAE, on the other hand, was present at a level of only 17-30 pmol/mumol lipid P (70-121 pmol/10(8) cells), with N-arachidonoylethanolamine (anandamide) making up less than 1% of total NAE. Use of deuterium labeled internal standards and optimization of GC-MS conditions makes it possible to detect as little as 0.1 ng of saturated and 1 ng (3 pmol) of polyunsaturated NAEs in a lipid extract. The present method can be used to determine agonist-induced changes in the levels and compositions of N-acyl PE and NAE.
Schmid
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Hormel Institute, University of Minnesota, Austin 55912, USA.
Kuwae
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Krebsbach
R J
RJ
Schmid
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HH
D000217:Acyltransferases / Q000378:metabolism; D000818:Animals; D001095:Arachidonic Acids / Q000032:analysis*; D003903:Deuterium; D063388:Endocannabinoids; D004983:Ethanolamines / Q000032:analysis* / Q000302:isolation & purification; D005227:Fatty Acids / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D017737:Macrophages, Peritoneal / Q000737:chemistry*; D051379:Mice; D017646:Organosilicon Compounds / Q000138:chemical synthesis; D010714:Phosphatidylethanolamines / Q000032:analysis / Q000302:isolation & purification; D053284:Polyunsaturated Alkamides
D001095:Arachidonic Acids; D063388:Endocannabinoids; D004983:Ethanolamines; D005227:Fatty Acids; C022203:N-acylethanolamines; C109143:N-arachidonylphosphatidylethanolamine; D017646:Organosilicon Compounds; D010714:Phosphatidylethanolamines; D053284:Polyunsaturated Alkamides; C009660:t-butyldimethylsilyl compounds; D003903:Deuterium; D000217:Acyltransferases; C042139:phosphatidylethanolamine N-acyltransferase; C078814:anandamide