In-situ nanoelectrospray for high-throughput screening of enzymes and real-time monitoring of reactions

• In-situ nanoelectrospray was designed for direct sampling and ionization for MS.
• It was fabricated by only inserting capillary into high-voltage applied liquid phase.
• Analytes in liquid reaction phase were directly extracted and ionized for MS.
• It achieved high-throughput enzyme evaluation and fast optimizations.
• It has been used for real-time monitoring of enzyme catalyzed reactions.

The in-situ and high-throughput evaluation of enzymes and real-time monitoring of enzyme catalyzed reactions in liquid phase is quite significant in the catalysis industry. In-situ nanoelectrospray, the direct sampling and ionization method for mass spectrometry, has been applied for high-throughput evaluation of enzymes, as well as the on-line monitoring of reactions. Simply inserting a capillary into a liquid system with high-voltage applied, analytes in liquid reaction system can be directly ionized at the capillary tip with small volume consumption. With no sample pre-treatment or injection procedure, different analytes such as saccharides, amino acids, alkaloids, peptides and proteins can be rapidly and directly extracted from liquid phase and ionized at the capillary tip. Taking irreversible transesterification reaction of vinyl acetate and ethanol as an example, this technique has been used for the high-throughput evaluation of enzymes, fast optimizations, as well as real-time monitoring of reaction catalyzed by different enzymes. In addition, it is even softer than traditional electrospray ionization. The present method can also be used for the monitoring of other homogenous and heterogeneous reactions in liquid phases, which will show potentials in the catalysis industry.

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