Since its first introduction by Karlsson and Novotny in 1988 nano-LC has emerged as a complementary and/or competitive separation method to conventional HPLC, offering several advantages such as higher efficiency, ability to work with minute sample sizes and lower consumption of mobile phases, and better compatibility with MS, etc. Although its use was not so extended initially, in the last years new and interesting applications have appeared which deserve to be carefully considered. The aim of this review is therefore to provide an updated and critical survey of different nano-LC applications in analytical chemistry.