Determination of Flow Rates in Capillary Liquid Chromatography Coupled to a Nanoelectrospray Source using Droplet Image Analysis Software

Liquid chromatography coupled to electrospray tandem mass spectrometry (LC-ESI-MS/MS) is widely used in proteomic and metabolomic workflows. Considerable analytical improvements have been observed when the components of LC systems are scaled down. Currently, nano-ESI is typically done at capillary LC flow rates ranging from 200 to 300 nL/min. At these flow rates, trouble shooting and leak detection of LC systems has become increasingly challenging. In this paper we present a novel proof-of-concept approach to measure flow rates at the tip of electrospray emitters when the ionization voltage is turned off. This was achieved by estimating the changes in the droplet volume over time using digital image analysis. The results are comparable with the traditional methods of measuring flow rates, with the potential advantages of being fully automatable and nondisruptive.