Automated Chip-Based Metabolomic Analysis

描述(由申请人提供)：此R21应用程序的目的是开发一种自动化的、基于芯片的代谢分析，用于实践点应用程序。当前形式的细胞内代谢物测量是繁琐、不准确和昂贵的。然而，随着基因组学、转录组学和蛋白质组学的各种高通量技术的发展，它们的重要性已经显著增加。在整合各种“组学”数据以在系统层面上解释复杂的生理学之前，代谢物图谱的测量成为下一个挑战。通常，细胞内代谢物的测量需要大量的操作来抑制新陈代谢并从细胞中提取代谢物。由于代谢物的周转较快，猝灭和提取效率成为其测量的主要障碍。此外，代谢物的测量通常需要液相色谱/质谱仪(LC/MS)或气相色谱/质谱仪(GC/MS)，这可能不是每个实验室都能随时获得的，而且仪器的成本往往令人望而却步。本申请提出通过开发一种低成本、自动化的、基于芯片的样品制备和高效液相系统来解决这些问题，该系统可以在靠近实践点(无论是实验还是临床)的普通实验室中使用。芯片上的分馏器与其他实验室的MS/MS分离，但随时可以与MS/MS对接。在该方案中，由于自动化，样品制备更加准确和方便；代谢物分离更加高效，因为串联高效液相色谱可以建立在芯片上；由于芯片上高效液相色谱的低成本和将MS/MS操作外包给MS专家，大大降低了测量成本。

DESCRIPTION (provided by applicant): The purpose of this R21 application is to develop an automated, chip-based metabolomic analysis for point of-practice applications. Intracellular metabolite measurements in current forms are tedious, inaccurate, and expensive. However, their importance has grown significantly as various high-throughput technologies for genomics, transcriptomics, and proteomics have been developed. The measurement of metabolite profiles becomes the next challenge before integrating various "omic" data to explain complex physiology at the systems level. Typically, intracellular metabolite measurements require extensive operations for quenching the metabolism and extracting metabolites from the cell. Because of the relatively fast turnover of metabolites, the quenching and extraction efficiency become major hurdles in their measurements. In addition, metabolite measurements typically require liquid chromatography/mass spectrometry (LC/MS) or gas chromatography (GC)/MS, which may not be readily available in each laboratory, and the cost of instrumentation, is often prohibitive. This application proposes to solve these problems by developing a low cost, automated, chip-based sample preparation and HPLC system, which can be used in common laboratories next to the point-of-practice (either experimental or clinical). The HPLC output is fractionated by an on-chip fractionator, which is physically separated from, but ready to interface with, MS/MS in other laboratories. In this scheme, sample preparation is much more accurate and convenient because of automation; metabolite separation is more efficient because tandem HPLC can be build on chip; cost of measurement is significantly reduced because of the low cost of on-chip HPLC and outsourcing of the MS/MS operation to MS specialists.