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

这个小型企业创新研究(SBIR)第二阶段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。商用的独立LIF探测器基于连续波激光，并在固定波长收集数据。因此，除了色谱分离本身所固有的能力之外，它们增加了最低限度的解析复杂混合物的能力。在单一发射波长下的实时荧光寿命测量已经被认为是解决共洗脱物种信号的一种更好的方法。我们的方法要强大得多，因为它同时提供了几个波长的动态寿命。在第一阶段引入了一种新的棱镜流动单元光纤耦合到发射光谱仪。此外，还提出了分析多维荧光数据的两种不同的算法策略。在第二阶段，二极管泵浦激光器将取代闪光灯泵浦激励激光器，从而提供100倍高的脉冲重复频率、10倍短的脉冲持续时间和10倍更好的快照对快照稳定性。将采用新的数字化技术来适应激光的高重复频率。第二阶段的重要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，以及开发化学计量学算法。通过第二阶段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。这项技术稍后将被应用于更快、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.