Neuropeptides are an important class of neuronal signalling molecules. In order to further characterise the peptides involved in the physiopathology of epilepsy, it is essential to monitor their concentration in the brain. Quantification of neuropeptides in dialysates is challenging due to their low extracellular concentrations, their low microdialysis efficiencies and the tendency of peptides to stick to surfaces. Therefore the use of very sensitive nano LC-MS/MS methods is required. In this study the feasibility of quantifying eight neuropeptides, possibly involved in epilepsy, is investigated, namely bradykinin, dynorphin A (1–13), galanin, ghrelin, neuromedin B, neuromedin N, neuropeptide Y and neurotensin. Samples were concentrated on a C18 precolumn (5 mm x 300 mm id, 5 mm) and back-flushed onto a nano C18 column (15 cm x 75 mm id) at 300 nl/min. Gradient elution was performed. The nano LC system was hyphenated to the nanosource of a Quattro Premier triple quadrupole MS using Picotip nanospray emitters (10 mm id). Detection was performed in ESI+ mode and quantification was executed in SRM mode. First, the compound-specific MS/MS parameters of the peptides are optimised. Second, all peptides were injected onto the nano LC-MS/MS system using a generic gradient. Optimisation of the LC method is necessary for ghrelin and dynorphin A (1–13) to decrease the peak width. Galanin and neuropeptide Y could not be measured at low concentrations. The limit of detection for bradykinin, neurotensin, neuromedin N and neuromedin B was found to be 7.0; 2.6; 1.5 and 23 pM respectively. Third, a screening was performed in basal hippocampal dialysates. Neuromedin N, neurotensin and ghrelin were detected. These preliminary results show that, after optimisation and validation, the quantification of the selected neuropeptides with nano LC-MS/MS is feasible.