Continuous purification of reaction products by micro free-flow electrophoresis enabled by large area deep-UV fluorescence imaging
Microreactors have gained increasing attention in their application toward continuous micro flow synthesis. An unsolved problem of continuous flow synthesis is the lack of techniques for continuous product purification. Herein, we present a micro free-flow electrophoresis device and accompanying setup that enables the continuous separation and purification of unlabeled organic synthesis products. The system is applied to the separation and purification of triarylmethanes. For imaging of the unlabeled analytes on-chip a novel setup for large area (3.6 cm2) deep ultra violet excitation fluorescence detection was developed. Suitable separation conditions based on low conductivity electrophoresis buffers were devised to purify the product. With the optimized conditions, starting materials and product of the synthesis were well separated (R > 1.2). The separation was found to be very stable with relative standard deviations of the peak positions smaller than 3.5% over 15 min. The stable conditions enabled collection of the separated compounds, and purity of the product fraction was confirmed using capillary electrophoresis and mass spectrometry. This result demonstrates the great potential of free-flow electrophoresis as a technique for product purification or continuous clean-up in flow synthesis.
KeywordsContinuous flow Free-flow separation Flow microreactor synthesis Ultraviolet fluorescence
The authors gratefully acknowledge funding by the Deutsche Forschungsgemeinschaft (DFG) through grant FOR 2177.
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
- 4.Plutschack MB, Pieber B, Gilmore K, Seeberger PH. The Hitchhiker’s Guide to Flow Chemistry. Chem Rev. 2017; https://doi.org/10.1021/acs.chemrev.7b00183.
- 22.Herzog C, Poehler E, Peretzki AJ, Borisov SM, Aigner D, Mayr T, Nagl S. Continuous on-chip fluorescence labelling, free-flow isoelectric focusing and marker-free isoelectric point determination of proteins and peptides. Lab Chip. 2016;16:1565–72.Google Scholar
- 24.Novo P, Janasek D. Current advances and challenges in microfluidic free-flow electrophoresis – a critical review. Anal Chim Acta. 2017; https://doi.org/10.1016/j.aca.2017.08.017.
- 30.Poehler E, Herzog C, Lotter C, Pfeiffer SA, Aigner D, Mayr T, Nagl S. Label-free microfluidic free-flow isoelectric focusing, pH gradient sensing and near real-time isoelectric point determination of biomolecules and blood plasma fractions. Analyst. 2015;140:7496–502.Google Scholar