SBIR Phase I: A Novel High Performance Liquid Chromatography (HPLC) Detector: Generating On-the-Fly Fluorescence Lifetimes Concurrently at Multiple Emission Wavelengths

这个小企业创新研究的第一阶段项目涉及构建和优化一种新型高效液相色谱（HPLC）检测器，该检测器可以解决环境和制药实验室中遇到的化学复杂分析物混合物。这种复杂混合物的标准高效液相色谱分析受到峰重叠的严重阻碍。即使在最佳分离条件下，需要较长的洗脱时间并提高分析成本，感兴趣的物种通常也只能部分解决。达科他科技公司已经获得了一项新检测方案的专利，该方案可以同时记录四个或更多发射波长的荧光衰减曲线，非常适合HPLC荧光检测。该方案使用光纤延迟线来控制不同波长的荧光成分到达单个光电倍增管（PMT）探测器。二维探测器（荧光波长和衰减时间）是简单的，鲁棒的，并且明显便宜的竞争探测器，这仅限于波长或衰减时间域。原理证明已经建立，但需要进一步的技术发展来优化分析多环芳烃（PAH）的方法。在第一阶段的SBIR项目中，DTI将安装一个流动电池，利用全内反射率来传递激发光，从而在PMT处获得更高的光通量。一个新的9位数字荧光粉示波器（DPO）将被纳入市场，以提高数据采集的准确性和速度。将通过实验找到单个多环芳烃的最佳发射波长集，并对化学计量算法进行测试。

This Small Business Innovation Research Phase I project involves construction and optimization of a novel high performance liquid chromatography (HPLC) detector that can resolve the chemically complex analyte mixtures encountered in environmental and pharmaceutical laboratories. Standard HPLC analysis of such complex mixtures is severely hampered by peak overlap. Even under optimal separation conditions, which necessitate long elution times and drive up analysis costs, the species of interest are often only partially resolved. Dakota Technologies, Inc. has patented a novel detection scheme that concurrently records fluorescence decay curves at four or more emission wavelengths and is ideally suited to HPLC fluorescence detection. The scheme uses fiber optic delay lines to control the arrival of different wavelength fluorescence components at the single photomultiplier tube (PMT) detector. The two dimensional detector (fluorescence wavelength and decay time) is simple, robust, and significantly less expensive than competitive detectors, which are limited to just the wavelength or decay time domain. Proof of principle has been established but further technical developments are needed to optimize the approach for analysis of polycyclic aromatic hydrocarbons (PAH). In this Phase I SBIR project DTI will implement a flow cell that that employs total internal reflectance to deliver the excitation light, resulting in higher light fluxes at the PMT. A new-to-the market 9-bit digital phosphor oscilloscope (DPO) will be incorporated to increase the accuracy and speed of data acquisition. The optimal emission wavelength sets for individual PAHs will be found by experiment and chemometric algorithms will be tested.