SBIR Phase II: Novel Multi-Wavelength Time-Resolved Laser Induced Fluorescence Detector

这个小型企业创新研究（SBIR）二期项目将在使用激光诱导荧光（LIF）进行色谱检测方面取得突破。商用独立LIF探测器基于连续波激光器，以固定波长收集数据。因此，除了色谱分离本身固有的能力之外，它们对复杂混合物的溶解能力增加得很少。在一个单一的发射波长上的荧光寿命测量已被提出作为一种更好的方法来解决共洗脱物种的信号。我们的方法要强大得多，因为它提供了在飞行中同时在几个波长下的寿命。第一阶段介绍了一种新型的棱镜流池光纤耦合发射光谱仪，此外，开发并演示了两种不同的算法策略来分析多维荧光数据。在第二阶段，二极管泵浦激光器将取代闪光灯泵浦激发激光器，从而提供100倍高的脉冲重复频率，10倍短的脉冲持续时间，10倍更好的单次稳定性。新的数字化技术将被纳入以适应激光的高重复频率。重要的第二阶段活动包括开发荧光方法，以扩大药物和药物代谢物的应用范围，并详细说明化学计量算法。通过II期研究实现的仪器方法将对药物纯度、生物等效性和药代动力学研究的QA/QC评估以及人类和动物的研究调查产生深远影响。预计每年向制药商、合同研究机构和大学销售数百台。这项技术随后将用于更快、更准确的DNA测序。

This Small Business Innovation Research (SBIR) Phase II project will lead to a breakthrough in the use of laser-induced fluorescence (LIF) for chromatographic detection. Commercial standalone LIF detectors are based on CW lasers and collect data at a fixed wavelength. Consequently, they add minimal capability for resolving complex mixtures beyond that inherent in the chromatographic separation itself. On-the-fly fluorescence lifetime measurements at a single emission wavelength have been proposed as a better way to resolve the signals of co-eluting species. Our approach is far more powerful because it provides lifetimes on-the-fly and at several wavelengths simultaneously. A new prism flow cell fiber optically coupled to the emission spectrograph was introduced in Phase I. In addition, two different algorithms strategies for analyzing the multi-dimensional fluorescence data were developed and demonstrated. In Phase II a diode-pumped laser will replace the flashlamp pumped excitation laser, thereby providing 100 times higher pulse repetition frequency, 10 times shorter pulse duration, and 10 times better shot-to-shot stability. New digitizer technology will be incorporated to accommodate the laser's high repetition frequency. Important Phase II activities include fluorescence methods development to extend the range of applications to drugs and drug metabolites and elaboration of the chemometric algorithms.The instrumental approach to be realized through the Phase II research will have a profound impact on QA/QC assessments of drug purity, bioequivalence and pharmacokinetic studies, and research investigations in humans and animals. Sales of several hundred units per year to pharmaceutical manufacturers, contract research organizations, and universities are anticipated. The technology will later be adapted for faster and more accurate DNA sequencing.