Quantification of diacylglycerol molecular species in biological samples by electrospray ionization mass spectrometry after one-step derivatization
Quantification of diacylglycerol molecular species in biological samples by electrospray ionization mass spectrometry after one-step derivatization
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DOI：
10.1021/ac0615910
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发表时间：
2007-02-15
影响因子：
7.4
通讯作者：
Gross, Michael L.
中科院分区：
文献类型：
--
作者：
Li, Ying L.;Su, Xiong;Gross, Michael L.
二酰基甘油 (DAG) 是细胞运输和信号传导中重要的脂质中间体。它们的浓度在糖尿病、癌症和其他疾病状态下会发生变化。生物样品中 DAG 的定量可以为揭示导致各种细胞功能紊乱的分子机制提供关键信息。使用质谱法进行脂质组学的最新进展极大地加速了全球脂质分析和定量。然而，通过电喷雾质谱 (ESI/MS) 定量 DAG 面临着以下挑战：分子上不存在永久电荷、其质子亲和力和酸性低、以及在正常生物条件下丰度低。我们在这里描述了使用 N-氯甜菜酰氯将季铵阳离子引入 DAG 分子，以提供衍生的 DAG，其信号强度比未衍生的钠加合物高 2 个数量级。通过使用 ESI/MS 以二月桂酰甘油作为内标，可以绘制出峰强度比与摩尔比的线性校准曲线。对这种分析物采用这种新方法，我们发现肥胖 db/db 小鼠肝脏中的总 DAG 比杂合瘦对照小鼠增加了 9 倍。这一经过验证的策略可用于在一步衍生化后使用 ESI/MS 检测和量化生物样品中的 DAG 分子种类。
Diacylglycerols (DAGs) are important lipid intermediates in cellular trafficking and signaling. Their concentrations are altered in diabetes, cancer, and other disease states. Quantification of DAGs in biological samples may provide critical information to uncover molecular mechanisms leading to various cellular functional disorders. Recent advances in lipidomics using mass spectrometry have greatly accelerated global lipid analysis and quantification. Quantification of DAGs by electrospray mass spectrometry (ESI/MS), however, is challenged by the absence of a permanent charge on the molecule, its low proton affinity and acidity, and its low abundance under normal biological conditions. We describe here the introduction of a quaternary ammonium cation to DAG molecules, using N-chlorobetainyl chloride, to afford a derivatized DAG that gives 2 orders of magnitude higher signal intensities than their underivatized sodium adducts. A linear calibration curve in which peak intensity ratios are plotted versus molar ratios can be achieved by using ESI/MS with dilauroyl glycerol as the internal standard. Employing this new approach to this analyte, we found a 9-fold increase of total DAGs in the livers of obese db/db mice as compared to their heterozygous lean controls. This proven strategy can be used to detect and quantify DAG molecular species from biological samples using ESI/MS after one-step derivatization.