© 2022 The AuthorsNanoscale separation techniques (Capillary/nano-LC, CE, CEC and Chip based LC/CE/CEC) present a prospective trend in modern analytical chemistry due to allowing high speed analysis, low reagent consumption, high separation efficiency and green chemistry. These techniques are miniaturized techniques offering unique advantages over conventional ones in bioanalytical science and technology, and are gaining ground in a wide research area (e.g. omics, foods, cell structures). These systems are also promising tools for the analysis of very limited samples, especially in omics, and forensic sciences. Focusing on nanoscale separation-based techniques, this review presents recent research on the use of nano-columns. Special attention is given to recent achievements based on nanoscale separation techniques while their potentiality is discussed. Some recent and selected applications with or without mass spectrometry (e.g., low resolution/high resolution -MS) in the period of 2019-to present are also reported.