Quantification of endocannabinoids in rat biological samples by GC/MS: Technical and theoretical considerations
Quantification of endocannabinoids in rat biological samples by GC/MS: Technical and theoretical considerations
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DOI：
10.1016/j.prostaglandins.2006.08.002
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发表时间：
2006-12-01
影响因子：
2.9
通讯作者：
Giuffrida, Andrea
中科院分区：
文献类型：
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作者：
Hardison, Sarah;Weintraub, Susan T.;Giuffrida, Andrea
在过去的几年里，人们对内源性大麻素类和2-花生四烯基甘油的兴趣显著增加，这两种脂类信使激活了大麻素受体。生物样品中这些化合物的量化带来了许多技术挑战。由于其丰度低，内源性大麻素通常通过同位素稀释分析来定量，该分析使用与气相色谱或高效液相色谱相结合的质谱仪。虽然可以通过这些技术直接测定生物体液(如血浆和脑脊液)中的内源性大麻素含量，但脑组织样本复杂的脂谱要求在GC/MS分析之前对脂类提取物进行纯化；当使用高效液相色谱/质谱仪时，这一步骤不是必需的。我们发现，当使用硅胶层析法进行内源性大麻素纯化时，回收率较差以及内标物中氢的损失导致内源性大麻素含量的估计不准确。相比之下，使用顺式固相萃取的纯化策略可以精确和可重复性地定量组织样本中的内源性大麻素。(C)2006 Elsevier Inc.保留所有权利。
In the last several years, interest has increased significantly about the endocannabinoids anandamide and 2-arachidonylglycerol, two lipid messengers that activate cannabinoid receptors. Quantification of these compounds in biological samples presents numerous technical challenges. Because of their low abundance, endocannabinoids are usually quantified by isotope dilution assays using mass spectrometry coupled to either gas chromatography or high-performance liquid chromatography. Although endocannabinoid levels in biological fluids, such as plasma and cerebrospinal fluid, can be directly determined by these techniques, the complex lipid profile of brain tissue samples mandates purification of lipid extracts before GC/MS analysis; this step is not necessary when using HPLC/MS. We have found that when silica gel chromatography is used for endocannabinoid purification, poor recovery and loss of deuterium from the internal standards lead to inaccurate estimation of endocannabinoid levels. By contrast, purification strategies using Cis solid-phase extraction permits precise and reproducible GUMS quantification of endocannabinoids in tissue samples. (c) 2006 Elsevier Inc. All rights reserved.