The Paraná pine is widely distributed in the native forests of southern Brazil and its seeds, the pinhões, have high starch content. The starches from unconventional sources, associated with the possibility of expanding their application as raw material in the production of packaging, polymeric composites and in the starch nanocrystals (SNCs) production have gain space and interest. The study was composed of pre-treatments on the starches (heat-moisture-treatment (HMT), annealing (ANN) and sonicatioin (SNT)) and subsequent obtaining of SNCs by acid hydrolysis (H2SO4 3.0 M) for 5 and 7 Days at 40 °C, followed by characterization of the SNCs by hydrolysis yield, chain distribution profile through gel permeation chromatography (GPC), relative crystallinity (RC), thermal stability (DSC). The yielding after hydrolysis ranged from 10.2 to 14.7%. The hydrolysis promoted the removal of the amylose chains that form the amorphous region of the starch granules and increased the RC of the SNCs by about 90%. The HMT and the SNT increased the yield of the SNCs (14.7 and 13.4, respectively), but they promoted losses in the RC of the same ones (35.9% and 34.1% respectively) comparing with native starch (42%), and with 7 days of hydrolysis the H2SO4 started the degradation of the crystalline region of the starch. After 7 days of hydrolysis, the pinhão starch SNCs showed two endothermic peaks. The first one was observed between 140.83 and 160.72 °C and the second between 159.69 and 179.62 °C. The ANN promoted better thermal stability of the SNCs. The Pinhão starch proved to be promising for the production of SNCs, due to the high yield and stability after H2SO4 hydrolysis, in addition, the HMT and SNT that showed themselves as efficient pre-treatments in the increase of the yield after 5 days of hydrolysis, compared to the native starch.