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

这个小型企业创新研究第一阶段项目涉及建造和优化一种新型的高效液相色谱(HPLC)检测器，该检测器可以解析环境和制药实验室中遇到的化学复杂的分析物混合物。这种复杂混合物的标准高效液相色谱分析因峰重叠而严重受阻。即使在需要较长洗脱时间和增加分析成本的最佳分离条件下，感兴趣的物种通常也只能部分分离。达科塔技术公司已经为一种新的检测方案申请了专利，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，非常适合于高效液相色谱荧光检测。该方案使用光纤延迟线来控制不同波长的荧光分量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)简单、坚固，而且比竞争探测器便宜得多，后者仅限于波长或衰减时间域。已经确立了原则证明，但还需要进一步的技术发展，以优化多环芳烃(PAH)的分析方法。在第一阶段SBIR项目中，DTI将实施一个流动池，该池使用全内部反射来传递激发光，从而在光电倍增管上产生更高的光通量。新上市的9位数字磷光示波器(DPO)将被纳入，以提高数据采集的精度和速度。将通过实验找到单个多环芳烃的最佳发射波长集，并测试化学计量学算法。

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.