The emergence and the fate of organic compounds in environmental, food, and biological samples have resulted in increased public concern because these compounds have the capacity to produce adverse health effects and have a negative influence on human life. This article presents an overview of the applicability of large-volume sample injection (LVSI) in column-switching high-performance liquid chromatography (HPLC) systems for the trace analysis of organic compounds (i.e., pesticides, herbicides, fungicides, pharmaceuticals, illegal food additives, toxins, and other organic contaminants) in different matrices (i.e., environmental, food and biological) as an alternative approach to preconcentrating and lowering the limits of detection of chromatographic methods.