Pc. Schmid et al., A sensitive endocannabinoid assay. The simultaneous analysis of N-acylethanolamines and 2-monoacylglycerols, CHEM PHYS L, 104(2), 2000, pp. 185-191
Mammalian cells produce both N-arachidonoylethanolamine (20:4n-6 NAE, anand
amide) and 2-arachidonoylglycerol (2-AG), lipid signaling molecules that ac
tivate cannabinoid receptors. Because both agonists occur in the presence o
f 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 e
xtraction and converted to tert-butyldimethylsilyl (tBDMS) derivatives in t
he presence of deuterated analogs. The tBDMS derivatives are analyzed by ga
s chromatography/mass spectrometry using selected ion monitoring programs s
pecific 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 o
rgans and cultured JB6 cells are reported. (C) 2000 Elsevier Science Irelan
d Ltd. All rights reserved.