Advion Triversa Nanomate/ABSciex Nanospray III Ion Source for Targeted Lipidomics

描述（由申请人提供）：脂质是能量代谢的中间体，是生物膜的结构成分，也是细胞内和细胞外的信号。异常脂质代谢在人类病理生理中起着核心作用，包括心血管疾病、肥胖、糖尿病和癌症。系统水平的脂质分析落后于基因组学和蛋白质组学的快速进展。使用串联液相色谱法和多维质谱法的新开发的分析方法催生了“脂质组学”领域-代谢组学的一个重点领域，涉及脂质和脂质依赖过程的系统级分析。作为我们肥胖和心血管疾病研究中心核心实验室的组成部分，肯塔基大学操作ABSciex 4000 Q-Trap混合型三重四极线性离子阱质谱仪，主要用于分析脂类和脂类相关分子，包括甘油和鞘磷脂、脂肪酸、神经酰胺、鞘磷脂、甾醇和胆固醇酯、二甘油和三甘油、酰基肉碱、氧化脂类及其反应产物。肽激素、花生四烯酸衍生的信号分子及其代谢物采用HPLC、电喷雾电离（ESI）选择性反应监测模式串联质谱分析。除了能够作为三重四极杆质谱仪的组成部分外，该仪器的Q3质量分析仪还可以作为线性离子阱，赋予仪器显著的附加功能，使其成为代谢物鉴定的极其强大的工具。虽然使用高流量Turbo V ESI离子源进行样品分离和引入的高效液相色谱配置对于通过选择性反应监测进行分析物定量是理想的，但这种配置与新开发的强大的脂质分析策略不兼容。这是因为通常很难（或不可能）通过柱层析法在给定类别中分离有时超过30-50种不同的脂质分子种类，因此窄峰宽限制了仪器进行扫描以识别紧密洗脱的脂质种类成分的时间。解决这一限制的方法要求直接输注样品，允许仪器通过一系列脂类特定前体离子或中性损失扫描来识别脂类。由于灵敏度、样品可用性和分析物丰度的问题，在这些实验中，通过小体积浓缩样品的低流量（10-200 nl/min）输注，可以大大增强对脂质的检测。本申请的目的是申请资金升级仪器，配备一个advisory Nanomate Triversa自动芯片输注系统和ABI Nanospray III离子源。这次升级将使我们能够使用我们的仪器进行脂质分析研究，通过自动化纳米喷雾ESI质谱，使用这些新开发的仪器操作和数据分析平台。

DESCRIPTION (provided by applicant): Lipids are intermediates in energy metabolism, structural components of biological membranes and as intra- and extra- cellular signals. Aberrant lipid metabolism plays a central role human pathophysiologies, including cardiovascular disease, obesity, diabetes, and cancer Systems-level analysis of lipids has lagged behind rapid progress in genomics and proteomics. Newly developed analytical approaches using tandem liquid chromatography and multi- dimensional mass spectrometry have spawned the field of "lipidomics"- a focused area of metabolomics that concerns the systems-level analysis of lipids and lipid-dependent processes. As a component of a core laboratory of our Center for Research on Obesity and Cardiovascular Disease, the University of Kentucky operates an ABSciex 4000 Q-Trap hybrid triple quadrupole linear ion trap mass spectrometer which is used primarily for analysis of lipids and lipid-related molecules including glycero and sphingo phospholipids, fatty acids, ceramides, sphingomyelins, sterols and cholesterol esters, di and tri glycerides, acyl carnitines, oxidized lipids and their reaction products, peptide hormones, arachidonic acid derived signaling molecules and their metabolites by HPLC, electospray ionization (ESI) selective reaction monitoring mode tandem mass spectrometry. In addition to its ability to operate as a component of a triple quadrupole mass spectrometer, the Q3 mass analyzer of this instrument can function as a linear ion trap conferring significant additional capabilities to the instrument that make it an extremely powerful tool for metabolite identification. While the HPLC configuration for sample separation and introduction using a high flow Turbo V ESI ion source is ideal for analyte quantitation by selective reaction monitoring, this arrangement is not compatible with newly developed powerful strategies for lipid profiling. This is because it is often difficult (or not possible) to separate the sometimes greater than 30-50 distinct lipid molecular species within a given class by column chromatography so narrow peak widths limit the time the instrument has to conduct scans to identify components of closely eluting lipid species. Methods that address this limitation mandate direct sample infusion allowing the instrument to identify lipid species conducting a series of lipid class specific precursor ion or neutral loss scans. Because of issues of sensitivity, sample availability and analyte abundance, detection of lipids in these experiments is greatly enhanced by low flow (10-200 nl/min) infusion of small volumes of concentrated samples. The purpose of the present application is to request funds to upgrade the instrument with an Advion Nanomate Triversa automated chip infusion system and an ABI Nanospray III ion source. This upgrade will allow us to use our instrument for lipid profiling studies by automated nanospray ESI mass spectrometry using these newly developed platforms for instrument operation and data analysis.