Our results demonstrate two volume independent contributions to extra-column variance due to the injection valve. One of these arises from axial and mixing dispersion that occurs inside the injection valve groove, and is an exponential function of flow rate. We report for the first time a mathematical model that describes this relationship. The overall extra-column volume was calculated to be 35nL after optimization of the instrument and eliminating/reducing all possible sources of extra-column dispersion. The theoretical corrections for column performance were verified using a new nano-flow pump with integrated injection system having a smaller sample loop and different geometry. The chromatographic efficiency doubled with the use of the new injection valve. The extra-column variance results in a measured efficiency (N) that is approximately half of the actual column efficiency for a non retained analyte, 6% less for a compound with a retention factor of approximately 2, and approximately 1% less for a compound with a retention factor >3.5. Therefore, the extra-column band broadening should be characterized in order to determine the actual performance of capillary LC columns.