Diamond NMR spectrometer for microfluidic metabolite profiling

项目摘要。核磁共振（NMR）是最强大的分析方法之一。 迄今为止发明的技术，仅方法开发就获得了6项诺贝尔奖。 尽管如此，核磁共振的灵敏度还是很差。最先进的NMR 光谱仪的特点是检测阈值为约1纳摩尔，样品体积为微升（约100 纳克）。这使得NMR灵敏度落后于其他分析方法许多数量级。 化学技术，如质谱、拉曼光谱和荧光 标签。核磁共振的改进通常集中在使用更大的磁铁上，但进展已经 在过去的25年中，基本信号强度仅增加了约2倍。 我们试图通过使用掺杂有金刚石的金刚石膜来从根本上改变NMR硬件。 氮空位中心通过光脉冲非感应地探测核磁化 检测磁共振方法。我们的NMR探测器的外形尺寸易于集成 使用连字符技术，以便样本可以被分成子组件， 分析.最近，我们建立了一个桌面微流控金刚石核磁共振装置与40 pL的检测 体积，并将其用于原理验证分析化学应用，包括第一个2D 由金刚石NMR传感器获得的NMR光谱。在第一阶段，我们将优化传感器光谱， 分辨率和灵敏度，并使用代谢物混合物验证其操作。这项工作将 使我们能够将我们的设备交付给行业（默克）和学术界的最终用户 (UW)并将反馈纳入市场。 如果成功，我们的原型可能会对分析生物化学研究产生深远的影响， 结合质谱级灵敏度和NMR级准确度。我们特别 改进现有的分析方法，提供： 1.更好的性能。我们的灵敏度（pmol而不是nmol）比 当前的NMR光谱仪。该灵敏度接近质谱法的灵敏度，但保留了 NMR的优点，如非破坏性，绝对定量和结构鉴定。 2.与联用分离技术兼容。我们的光谱仪结构紧凑， 易于集成到微流控芯片中，用于在线色谱分析（HPLC）， 有限样本分析（代谢组学、药效学、天然产物）。 3.更低的成本我们的光谱仪中的小样品体积减少了工程设计 成本，导致更大的负担能力相比，目前的NMR光谱仪。

Project Summary. Nuclear magnetic resonance (NMR) is among the most powerful analytical techniques ever invented, as recognized by 6 Nobel Prizes for methods development alone. Nonetheless, NMR is notoriously plagued by poor sensitivity. State-of-the-art NMR spectrometers feature detection thresholds of ~1 nanomole for µL sample volumes (~100 nanograms). This places NMR sensitivity many orders of magnitude behind other analytical chemistry techniques such as mass spectrometry, Raman spectroscopy, and fluorescence labeling. Improvements in NMR often focus on using larger magnets, but progress has plateaued; over the last 25 years, the fundamental signal strength has only increased ~2-fold. We seek to fundamentally change the NMR hardware by using diamond films doped with Nitrogen-Vacancy centers to detect nuclear magnetization non-inductively via pulsed optically detected magnetic resonance methods. The form factor of our NMR detector is easily integrated with hyphenation techniques so that samples can be separated into sub-components before analysis. Recently, we built a tabletop microfluidic diamond NMR apparatus with 40 pL detection volume and used it in proof-of-principle analytical chemistry applications including the first 2D NMR spectra acquired by a diamond NMR sensor. In Phase I, we will optimize sensor spectral resolution and sensitivity and validate its operation using metabolite mixtures. This work will place us in the position to deliver our devices to end-users in industry (Merck) and academia (UW) and incorporate feedback to scale up to market. If successful, our prototype could have a profound impact on analytic biochemistry research, by combining mass-spectrometry-level sensitivity with NMR-level accuracy. Specifically, we improve upon existing analytical methods by offering: 1. Greater performance. We offer 1000-fold better sensitivity (pmol instead of nmol) than current NMR spectrometers. This sensitivity approaches that of mass spectrometry but retains benefits of NMR such as non-destructive, absolute quantitation and structural identification. 2. Compatibility with hyphenated separation techniques. Our spectrometer is compact and easily integrated into microfluidic chips for online chromatography-based assays (HPLC) for sample-limited analyses (metabolomics, pharmacodynamics, natural products). 3. Lower cost. The small sample volume in our spectrometer leads to reduced engineering costs, leading to greater affordability compared to current NMR spectrometers.