Nanoflow-HPLC/MS was proven a valuable tool for the analysis of highly complex proteome samples.Separations performed on low volume (sub-micro-liter) columns with flow rates in the nano-liter per minute range, result in excellent sensitivity when combined with nano-electro-spray ionization MS.However, use and maintenance of Nanoflow HPLC/MS systems are very challenging and require high user-proficiency.To obtain the best chromatographic performance, connections must be made carefully to minimize band broadening impairing separation performance.Good chromatographic performance can enhance the mass spectrometric results for instance by reducing ionization suppression effects between co-eluting peaks in the source; thus a greater number of peptides can be detected.