Integrating Accelerated Droplet Chemistry with LC-MS for High Throughput Quantitative Analysis

将加速液滴化学与 LC-MS 相结合进行高通量定量分析

• 批准号: 10607125
• 负责人: Abraham Badu-Tawiah
• 金额: $ 30.57万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607125
• 关键词: Acceleration; Acids; Address; Biological Process; Biomedical Research; Blood capillaries; Cell physiology; Cells; Charge; Chemicals; Chemistry; Clinical Research; Complex Mixtures; Coupling; Dedications; Derivation procedure; Development; Devices; Dimensions; Electrospray Ionization; Environment; Exposure to; Gases; Generations; Goals; High Pressure Liquid Chromatography; Human; In Situ; Injections; Ions; Isomerism; Lipids; Liquid Chromatography; Mass Spectrum Analysis; Metabolism; Methods; Modification; Mole the mammal; Molecular Conformation; Monosaccharides; Nature; Noise; Nucleic Acids; Pattern; Performance; Phase; Plasma; Play; Positioning Attribute; Process; Proteins; Reaction; Reactive Oxygen Species; Reagent; Research; Role; Sampling; Signal Transduction; Silicon Dioxide; Solvents; Source; Spectrometry; Spectrometry, Mass, Electrospray Ionization; Stereoisomer; Structure; Techniques; Technology; Time; Triglycerides; adduct; analytical method; anomer; combinatorial; combinatorial chemistry; design; disease diagnosis; experimental study; high throughput analysis; improved; instrument; instrumentation; interest; ion mobility; ion source; ionization; liquid chromatography mass spectrometry; mass spectrometer; method development; milligram; millisecond; monomer; novel; programs; protein structure; sugar; tandem mass spectrometry; voltage

Project Summary/Abstract There is an increasing need to improve the characterization of lipids and saccharides for clinical and biomedical research purposes. NMR is the method of choice for obtaining detailed structural information about saccharides. However, NMR typically requires milligram (micromole) quantities of analyte; this often exceeds biologically relevant levels. For lipids, mass spectrometry (MS) provides an efficient avenue for rapid profiling, but quantitative analysis is challenged by difficulty in isolating species of interest due to wide structural diversity. A new MS approach is proposed that fundamentally addresses challenges in quantitative and qualitative MS by utilizing online accelerated droplet chemistry. Since ion suppression effects in electrospray ionization (ESI) MS occur during droplet formation, our method is designed to tackle this intellectual challenge exactly at the point of droplet formation – not before by adding reagents in solution, and not after by performing gas-phase reactions. This strategy simplifies instrumentation requirements and allows effective coupling to liquid chromatography (LC). Selected droplet-based reactions improve signal-to-noise ratios to enable femtomole sensitivity using <1 µL sample volume. Gas-phase ion intensities generated by our platform reflect the corresponding analyte concentration in solution. Importantly, selected droplet-based reactions allow isomers of lipids and saccharides to be differentiated. We propose to couple online droplet reactions with LC to enable high throughput quantification of lipids and saccharides in complex mixtures. The specific research aims are: Aim 1: To develop a functional contained-electrospray platform for coupling accelerated droplet chemistry on LC-MS for saccharide analysis. A novel contained-ESI source is proposed to couple droplet chemistry with LC-MS. Our method will enable LC mobile phase and ESI spray solvent to be independently optimized. This orthogonal feature is expected to allow effective separation of isomeric saccharides (linkage, anomeric, and position isomers). Selected droplet reactions will improve detectability of saccharides and provide a second layer of identification for isomers that co-elute. The LC-contained-ESI-MS/MS platform will be validated via high throughput combinatorial studies. Aim 2: To develop a plasma-droplet fusing contained-electrospray source for coupling LC-MS for lipid analysis. We propose to include etched silica capillaries on our LC-contained-ESI-MS/MS platform for accurate quantification of all types of lipids, including triglycerides. The device is expected to enable instantaneous determination of degree of unsaturation, C=C bond position, and bond orientation (cis/trans). The tandem development of quantitative analytical methods for lipids and saccharides will result in concomitant creation of versatile platforms for applications in diseases diagnosis and high throughput analysis of rare sugars to effectively guide synthetic method development. The proposed strategy will also be valuable in biomedical research using existing instruments without modification.

项目概要/摘要 人们越来越需要改善脂质和糖的表征，以用于临床和临床研究。 生物医学研究目的。 NMR 是获取详细结构信息的首选方法 糖类。然而，NMR 通常需要毫克（微摩尔）量的分析物；这通常超过 生物学相关水平。对于脂质，质谱 (MS) 提供了快速分析的有效途径， 但由于广泛的结构多样性，定量分析面临着难以分离感兴趣物种的挑战。 提出了一种新的质谱方法，从根本上解决定量和定性质谱方面的挑战 通过利用在线加速液滴化学。由于电喷雾电离 (ESI) 中的离子抑制效应 MS 发生在液滴形成过程中，我们的方法旨在准确地解决这一智力挑战 液滴形成点 – 不是在溶液中添加试剂之前，也不是在执行气相之后 反应。该策略简化了仪器要求并允许与液体有效耦合 色谱法（LC）。选定的基于液滴的反应可提高信噪比，从而实现飞摩尔 使用 <1 µL 样品体积的灵敏度。我们的平台产生的气相离子强度反映了 溶液中相应的分析物浓度。重要的是，选定的基于液滴的反应允许异构体 脂类和糖类有待区分。我们建议将在线液滴反应与 LC 结合起来，以实现高 复杂混合物中脂质和糖类的通量定量。具体研究目标是： 目标 1：开发用于耦合加速液滴的功能性封闭式电喷雾平台 用于糖分析的 LC-MS 化学。提出了一种新颖的包含 ESI 源来耦合液滴 LC-MS 化学分析。我们的方法将使LC流动相和ESI喷雾溶剂能够独立 优化。这种正交特征有望实现异构糖的有效分离（键合、 异头异构体和位置异构体）。选定的液滴反应将提高糖类的可检测性并提供 共洗脱异构体的第二层鉴定。包含 LC 的 ESI-MS/MS 平台将得到验证 通过高通量组合研究。 目标 2：开发一种用于耦合 LC-MS 的等离子体液滴熔融包含电喷雾源 脂质分析。我们建议在我们的包含 LC 的 ESI-MS/MS 平台上加入蚀刻二氧化硅毛细管，用于 准确定量所有类型的脂质，包括甘油三酯。该设备预计能够启用 瞬时测定不饱和度、C=C 键位置和键方向（顺式/反式）。 脂质和糖类定量分析方法的串联发展将导致 同时创建用于疾病诊断和高通量分析应用的多功能平台 稀有糖的研究有效指导合成方法的开发。拟议的战略也将很有价值 使用现有仪器进行生物医学研究，无需修改。