Diamond NMR spectrometer for microfluidic metabolite profiling

项目摘要。核磁共振是最强大的分析技术之一。 曾经发明过的技术，仅在方法开发方面就获得了6项诺贝尔奖的认可。 尽管如此，核磁共振还是出了名的受到低灵敏度的困扰。最先进的核磁共振 光谱仪对L样品体积(~100)的检测阈值为~1纳摩尔 纳克)。这使得核磁共振的灵敏度比其他分析方法低了许多个数量级 化学技术，如质谱学、拉曼光谱和荧光 贴标签。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年里，基本信号强度只增加了~2倍。 我们寻求从根本上改变核磁共振硬件，通过使用掺杂了 通过光脉冲无感探测核磁化的氮空位中心 检测到了磁共振方法。我们的核磁共振探测器的外形很容易集成 使用连字技术，以便样品可以在 分析。最近，我们建立了一台台式微流控金刚石核磁共振装置，可以检测到40Pl 卷，并将其用于原理证明分析化学应用，包括第一个2D 由钻石核磁共振传感器获得的核磁共振光谱。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工业(默克)和学术界的最终用户 (UW)，并纳入反馈，以扩大到市场。 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影响，通过 结合了质谱级的灵敏度和核磁共振级的准确度。具体来说，我们 通过提供以下功能改进现有的分析方法： 1.更好的性能。我们提供1000倍以上的灵敏度(pmoL而不是nmoL)。 目前的核磁共振波谱仪。这种灵敏度接近于质谱学，但保留了 核磁共振的优点，如无损、绝对定量和结构鉴定。 2.与联用分离技术的兼容性。我们的光谱仪结构紧凑， 轻松集成到微流控芯片中，用于基于在线色谱的分析(HPLC) 样本限量分析(代谢组学、药效学、天然产物)。 3.成本较低。我们的光谱仪样品体积小，导致工程量减少。 成本，与目前的核磁共振光谱仪相比，价格更实惠。

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.