High Resolution Metabolomics

高分辨率代谢组学

• 批准号: 9065541
• 负责人: ROBERT T KENNEDY
• 金额: $ 22.29万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9065541
• 关键词: Anions; Back; Basic Science; Biological; Biological Assay; Biological Markers; Body Fluids; Caliber; Cell Extracts; Cell physiology; Cellular Metabolic Process; Chromatography; Clinical Research; Complex; Complex Mixtures; Complications of Diabetes Mellitus; Coupled; Coupling; Data Analyses; Detection; Development; Diabetic Nephropathy; Dimensions; Disease; Effectiveness; Ensure; Goals; Health; High Pressure Liquid Chromatography; Length; Liquid Chromatography; Mass Spectrum Analysis; Measurement; Measures; Metabolic Pathway; Metabolism; Methods; Michigan; Modeling; Mus; Obesity; Outcome; Particle Size; Performance; Pharmaceutical Preparations; Phase; Plasma; Procedures; Property; Pump; Reproducibility; Research; Resolution; Route; Sampling; Signal Transduction; Silicon Dioxide; Speed; System; Techniques; Technology; Testing; Time; Work; analytical method; base; biological systems; exercise capacity; falls; improved; insight; instrumentation; ionization; lipid biosynthesis; metabolic phenotype; metabolome; metabolomics; new technology; novel; novel strategies; particle; pressure; success; tool; ultra high pressure

DESCRIPTION (provided by applicant): Metabolomics is emerging as an important approach to study biological systems. Measurement of many metabolites at one time has led to important discoveries of disease biomarkers, drug effects, and mechanism of cellular function. Despite the success of metabolomics, current analytical methods are limited in their ability to measure a large fraction of the metabolome. As a result, many changes in the metabolome are not detected by current techniques. High pressure liquid chromatography coupled to mass spectrometry (HPLC-MS) is a prominent method for metabolomics. In this work, we seek to develop new approaches to HPLC that will vastly improve the resolving power for complex mixtures and apply these methods to metabolomics. The resolving power of HPLC increases with column length and with decreasing diameter of stationary phase particles. Changing these dimensions of the HPLC column also increases the pressure required to pump mobile phase through the column. HPLC has traditionally been limited to about 8,000 psi (lbs/in2), but advanced commercial systems can generate up to 19,000 psi. Application of these "ultra" high pressure LC (UHPLC) systems has yielded significant improvement in the ability to detect metabolites in complex mixtures; however, these systems still fall far short of the goal of resolving all metabolites in common biological samples. We will develop UHPLC systems with capability of 100,000 psi and compatible LC columns. We will investigate use of both reversed phase and hydrophilic interaction liquid chromatography at these extreme pressures to measure metabolites across a broad polarity range. We hypothesize that the new technology will allow measurement of thousands of more metabolites in a given sample than currently possible. The new method should also enhance sensitivity and reproducibility of metabolite detection in complex samples. The new methods will be used to identify metabolic pathways involved in models of adipogenesis, adaptation to high exercise capacity, and diabetic complications. Further, newly developed instrumentation and methods will be incorporated into a national center for metabolomics to ensure widespread impact of the new technology.

描述（由申请人提供）：代谢组学已成为研究生物系统的重要方法。一次测量许多代谢物已导致疾病生物标志物，药物作用和细胞功能机制的重要发现。尽管代谢组学取得了成功，但当前的分析方法的测量能力很大。结果，当前技术未检测到代谢组的许多变化。高压液相色谱与质谱法（HPLC-MS）耦合是代谢组学的重要方法。在这项工作中，我们试图开发新的HPLC方法，这些方法将大大提高复杂混合物的解决能力，并将这些方法应用于代谢组学。 HPLC的分辨能力随列长度和固定相颗粒直径降低而增加。更改HPLC列的这些维度也增加了通过列泵流相需的压力。传统上，HPLC仅限于约8,000 psi（LBS/IN2），但是先进的商业系统最多可以生成19,000 psi。这些“ Ultra”高压LC（UHPLC）系统的应用已在复杂混合物中检测代谢物的能力显着提高。但是，这些系统仍然远远远远没有解决常见生物样品中的所有代谢产物的目标。我们将开发具有100,000 psi和兼容LC列的能力的UHPLC系统。我们将在这些极端压力下研究相反的相和亲水相互作用液相色谱，以测量跨极性范围内的代谢产物。我们假设新技术将允许在给定样本中的数千种代谢产物的测量。新方法还应提高复杂样品中代谢物检测的灵敏度和可重复性。新方法将用于识别参与脂肪生成模型，适应高运动能力和糖尿病并发症的代谢途径。此外，新开发的仪器和方法将被纳入国家代谢组学中心，以确保新技术的广泛影响。