Schmid P C, Kuwae T, Krebsbach R J, Schmid H H
Hormel Institute, University of Minnesota, Austin 55912, USA.
Chem Phys Lipids. 1997 Jul 14;87(2):103-10. doi: 10.1016/s0009-3084(97)00032-7.
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.
在由[1,1,2,2 - 2H4]乙醇胺合成的内标存在的情况下，通过叔丁基二甲基甲硅烷基衍生物的气相色谱 - 质谱联用（GC - MS）对小鼠腹腔巨噬细胞中的N - 酰基磷脂酰乙醇胺（N - acyl PE）和游离N - 酰基乙醇胺（NAE）进行鉴定和定量。N - acyl PE的含量为123 - 187 pmol/μmol脂质P（521 - 768 pmol/10⁸细胞），花生四烯酸约占N - 酰基部分的3 - 4%。另一方面，NAE的含量仅为17 - 30 pmol/μmol脂质P（70 - 121 pmol/10⁸细胞），N - 花生四烯酰乙醇胺（花生四烯酸乙醇胺）占总NAE的比例不到1%。使用氘标记的内标和优化GC - MS条件能够在脂质提取物中检测到低至0.1 ng的饱和NAE和1 ng（3 pmol）的多不饱和NAE。本方法可用于确定激动剂诱导的N - acyl PE和NAE水平及组成的变化。
