Duca Zachary A, Speller Nicholas C, Cantrell Thomas, Stockton Amanda M
Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
Rev Sci Instrum. 2020 Oct 1;91(10):104101. doi: 10.1063/5.0008734.
Microcapillary electrophoresis (μCE) enables high-resolution separations in miniaturized, automated microfluidic devices. Pairing this powerful separation technique with laser-induced fluorescence (LIF) enables a highly sensitive, quantitative, and compositional analysis of organic molecule monomers and short polymers, which are essential, ubiquitous components of life on Earth. Improving methods for their detection has applications to multiple scientific fields, particularly those related to medicine, industry, and space science. Here, a modular benchtop system using μCE with LIF detection was constructed and tested by analyzing standard amino acid samples of valine, serine, alanine, glycine, glutamic acid, and aspartic acid in multiple borate buffered solutions of increasing concentrations from 10 mM to 50 mM, all pH 9.5. The 35 mM borate buffer solution generated the highest resolution before Joule heating dominated. The limits of detection of alanine and glycine using 35 mM borate buffer were found to be 2.12 nM and 2.91 nM, respectively, comparable to other state-of-the-art μCE-LIF instruments. This benchtop system is amenable to a variety of detectors, including a photomultiplier tube, a silicon photomultiplier, or a spectrometer, and currently employs a spectrometer for facile multi-wavelength detection. Furthermore, the microdevice is easily exchanged to fit the desired application of the system, and optical components within the central filter cube can be easily replaced to target alternative fluorescent dyes. This work represents a significant step forward for the analysis of small organic molecules and biopolymers using μCE-LIF systems.
微毛细管电泳(μCE)能够在小型化的自动化微流控设备中实现高分辨率分离。将这种强大的分离技术与激光诱导荧光(LIF)相结合,能够对有机分子单体和短聚合物进行高灵敏度、定量和成分分析,而这些是地球上生命中必不可少且普遍存在的成分。改进它们的检测方法在多个科学领域都有应用,特别是与医学、工业和空间科学相关的领域。在此,构建了一个使用μCE与LIF检测的模块化台式系统,并通过在浓度从10 mM到50 mM(均为pH 9.5)递增的多种硼酸盐缓冲溶液中分析缬氨酸、丝氨酸、丙氨酸、甘氨酸、谷氨酸和天冬氨酸的标准氨基酸样品进行了测试。在焦耳热占主导之前,35 mM硼酸盐缓冲溶液产生了最高分辨率。使用35 mM硼酸盐缓冲液时,丙氨酸和甘氨酸的检测限分别为2.12 nM和2.91 nM,与其他最先进的μCE-LIF仪器相当。这个台式系统适用于多种探测器,包括光电倍增管、硅光电倍增管或光谱仪,目前采用光谱仪进行便捷的多波长检测。此外,微器件可轻松更换以适应系统的预期应用,并且中央滤光片盒内的光学组件可轻松更换以针对其他荧光染料。这项工作代表了使用μCE-LIF系统分析小分子有机化合物和生物聚合物方面向前迈出的重要一步。