Must nitrogen is very important for successful fermentation. Irrigation can influence vine nitrogen availability, and therefore must nitrogen content and wine quality. The aim of this work was to assess the effect of vine water status on the nitrogen concentration of cv. Tempranillo grown under semiarid conditions. A rainfed and two regulated deficit irrigation (RDI) strategies, late (LDI) and early deficit irrigation (EDI) were applied during pre- and post-veraison respectively periods over two seasons in Badajoz (Spain). Twenty-eight amino acids, amines and ammonia were determined by high performance liquid chromatography (HPLC) in the must. Results showed an effect of the pre-veraison vine water status on the must amino acid concentration. Higher values of nitrogenous indices were reached in musts from LDI treatments, mainly in the dry year. Significant correlations between pre-veraison stem water potential and 19 amino acid concentrations were observed. Therefore, LDI was a more suitable strategy for increasing the content of nitrogenous substances in the must.