Visualization of Streams of Small Organic Molecules in Continuous-Flow Electrophoresis

Continuous-flow electrophoresis (CFE) separates a stream of a multicomponent mixture into multiple streams of individual components inside a thin rectangular chamber. CFE will be able to benefit flow chemistry when it is both compatible with nonaqueous solvents utilized in organic synthesis and capable of generically detecting streams of small organic molecules. While stable nonaqueous CFE has been demonstrated, generically detecting molecular streams has not been achieved yet. Here we propose a general approach for molecular stream visualization in CFE via analyte-caused obstruction of excitation of a fluorescent layer underneath the separation chamber-fluorescent sublayer-based visualization (FSV). The concept of FSC-based visualization has been adapted from visualization of small organic molecules on fluorescent plates in thin-layer chromatography. We designed and fabricated a CFE device with one side made of quartz and another side made of UV-absorbing visibly fluorescent, chemically inert, machinable plastic. This device was demonstrated to support nonaqueous CFE of small organic molecules and quantitative detection of their streams in real-time with a limit of detection below 100 μM. Thus, CFE may satisfy conditions required for its seamless integration with continuous flow organic synthesis in flow chemistry.