A sensitive endocannabinoid assay. The simultaneous analysis of N-acylethanolamines and 2-monoacylglycerols.
Mammalian cells produce both N-arachidonoylethanolamine (20:4n-6 NAE, anandamide) and 2-arachidonoylglycerol (2-AG), lipid signaling molecules that activate cannabinoid receptors. Because both agonists occur in the presence of receptor-inactive congeners, we have developed a sensitive method for the simultaneous assay of N-acylethanolamines (NAEs) and 2-monoacylglycerols (2-MAG). These lipid classes are isolated from total lipids by solid phase extraction and converted to tert-butyldimethylsilyl (tBDMS) derivatives in the presence of deuterated analogs. The tBDMS derivatives are analyzed by gas chromatography/mass spectrometry using selected ion monitoring programs specific for NAE and 2-MAG. Individual NAEs and 2-MAGs can be quantified in the nanogram and subnanogram range. The NAE and 2-MAG compositions of rat organs and cultured JB6 cells are reported.
Schmid
P C
PC
The Hormel Institute, University of Minnesota, Austin 55912, USA.
Schwartz
K D
KD
Smith
C N
CN
Krebsbach
R J
RJ
Berdyshev
E V
EV
Schmid
H H
HH
D000818:Animals; D001095:Arachidonic Acids / Q000032:analysis* / Q000302:isolation & purification; D063385:Cannabinoid Receptor Modulators; D002186:Cannabinoids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D063388:Endocannabinoids; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D005989:Glycerides / Q000032:analysis* / Q000302:isolation & purification; D007202:Indicators and Reagents; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008297:Male; D009206:Myocardium / Q000737:chemistry; D018377:Neurotransmitter Agents / Q000032:analysis*; D053284:Polyunsaturated Alkamides; D051381:Rats; D011924:Rats, Zucker; D012680:Sensitivity and Specificity; D013154:Spleen / Q000737:chemistry; D013737:Testis / Q000737:chemistry
D001095:Arachidonic Acids; D063385:Cannabinoid Receptor Modulators; D002186:Cannabinoids; D063388:Endocannabinoids; D005989:Glycerides; D007202:Indicators and Reagents; D018377:Neurotransmitter Agents; D053284:Polyunsaturated Alkamides; C094503:glyceryl 2-arachidonate; C078814:anandamide