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、生物膜的结构组分以及细胞内和细胞外信号。脂质代谢异常在包括心血管疾病、肥胖症、糖尿病和癌症在内的人类病理生理学中起着核心作用。脂质的系统水平分析落后于基因组学和蛋白质组学的快速进展。使用串联液相色谱和多维质谱的新开发的分析方法已经催生了“脂质组学”领域-代谢组学的关注领域，其涉及脂质和脂质依赖性过程的系统水平分析。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，肯塔基州大学使用ABSciex 4000 Q-Trap混合三重四极线性离子阱质谱仪，主要用于分析脂质和脂质相关分子，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、甘油二酯和甘油三酯，酰基肉毒碱、氧化脂质及其反应产物、肽类激素、花生四烯酸衍生的信号分子及其代谢产物。除了能够作为三重四极杆质谱仪的组件运行外，该仪器的Q3质量分析仪还可以作为线性离子阱，为仪器提供显著的附加功能，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HPLC配置是通过选择性反应监测进行分析物定量的理想选择，但这种安排与新开发的强大的脂质分析策略不兼容。这是因为通常难以（或不可能）通过柱色谱法分离给定类别内的有时大于30-50种不同的脂质分子种类，因此窄峰宽度限制了仪器必须进行扫描以鉴定紧密洗脱的脂质种类的组分的时间。解决这一限制的方法要求直接样品输注，允许仪器进行一系列脂质类特异性前体离子或中性损失扫描来鉴定脂质种类。由于灵敏度、样品可用性和分析物丰度的问题，这些实验中的脂质检测通过低流量（10-200 nl/min）输注小体积的浓缩样品而大大增强。本申请的目的是申请资金以升级具有Advion Nanomate Triversa自动芯片注入系统和ABI Nanospray III离子源的仪器。此次升级将使我们能够使用我们的仪器通过自动化纳米喷雾ESI质谱法进行脂质分析研究，使用这些新开发的平台进行仪器操作和数据分析。

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.