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Particularly in proteomics, nano liquid chromatography-mass spectrometry (nLC-MS) is frequently the tool of choice for the examination of very small samples. Although extremely powerful on its own, nLC-MS frequently depends on procedures for sample preparation that are done beforehand. It is possible to handle priceless samples effectively thanks to online upfront devices that are immediately connected to the nLC-MS analysis system. This paper examines upfront on-line interactions, specifically based on the use of solid phase extraction columns, for getting high sensitivity, high selectivity, and high robustness, with a focus on nLC-MS-based systems. Also covered are methods for obtaining quick sample preparation and little sample loss based on on-line reactions, such as on-line enzymatic digestion systems. The emphasis is on papers released between 2016 and mid-2020.