Easy, Fast, and Reproducible Quantification of Cholesterol and Other Lipids in Human Plasma by Combined High Resolution MSX and FTMS Analysis
Easy, Fast, and Reproducible Quantification of Cholesterol and Other Lipids in Human Plasma by Combined High Resolution MSX and FTMS Analysis
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DOI：
10.1007/s13361-017-1829-2
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发表时间：
2018-01-01
影响因子：
3.2
通讯作者：
Ejsing, Christer S.
中科院分区：
文献类型：
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作者：
Gallego, Sandra F.;Hojlund, Kurt;Ejsing, Christer S.
人血浆中非酯化胆固醇的可靠、经济、金标准绝对定量在临床脂质组学和代谢健康监测中至关重要。在这里，我们比较了三种质谱方法的性能，可用于直接检测和定量的人血浆提取物中的胆固醇。这些方法是高分辨率全扫描傅里叶变换质谱（FTMS）分析，平行反应监测（PRM），和新的多重MS/MS（MSX）技术，其中来自选定的前体离子的碎片被同时检测。评价这些方法在动态定量范围、线性和分析精密度方面的性能表明，基于MSX的方法上级基于FTMS和PRM的方法。为了进一步显示该方法的有效性，我们设计了一种简单的常规方法，用于广泛的血浆脂质组表征，仅使用8 μ L血浆，使用具有14种合成脂质标准品的新的市售即加混合物，并执行单次6分钟样品进样，结合MSX分析用于胆固醇定量，FTMS分析用于甾醇酯、甘油脂、甘油磷脂、和鞘脂。使用这种简单的常规方法，可对人血浆样品中包含13种脂质的200种脂质进行可重复的绝对定量。值得注意的是，对于高通量导向的临床脂质组学研究，该程序的分析时间可以缩短，或者用更先进的MSALL技术延长（Almeida R.例如，J. Am.质谱学会26，133-148 [1]）以支持脂质分子的深入结构阐明。
Reliable, cost-effective, and gold-standard absolute quantification of non-esterified cholesterol in human plasma is of paramount importance in clinical lipidomics and for the monitoring of metabolic health. Here, we compared the performance of three mass spectrometric approaches available for direct detection and quantification of cholesterol in extracts of human plasma. These approaches are high resolution full scan Fourier transform mass spectrometry (FTMS) analysis, parallel reaction monitoring (PRM), and novel multiplexed MS/MS (MSX) technology, where fragments from selected precursor ions are detected simultaneously. Evaluating the performance of these approaches in terms of dynamic quantification range, linearity, and analytical precision showed that the MSX-based approach is superior to that of the FTMS and PRM-based approaches. To further show the efficacy of this approach, we devised a simple routine for extensive plasma lipidome characterization using only 8 mu L of plasma, using a new commercially available ready-to-spike-in mixture with 14 synthetic lipid standards, and executing a single 6 min sample injection with combined MSX analysis for cholesterol quantification and FTMS analysis for quantification of sterol esters, glycerolipids, glycerophospholipids, and sphingolipids. Using this simple routine afforded reproducible and absolute quantification of 200 lipid species encompassing 13 lipid classes in human plasma samples. Notably, the analysis time of this procedure can be shortened for high throughput-oriented clinical lipidomics studies or extended with more advanced MSALL technology (Almeida R. et al., J. Am. Soc. Mass Spectrom. 26, 133-148 [1]) to support in-depth structural elucidation of lipid molecules.